Uncemented total hip arthroplasty (THA) implants have become the standard for younger patients on account of increased implant survivorship and multiple other advantages. Nevertheless, uncemented THA remains controversial in elderly patients. The evidence base for this is limited, as previous studies have compared octogenarians to a younger control group. The aim of this prospective cohort study is to evaluate the outcome of octogenarian patients undergoing uncemented THA with a control group of similarly aged patients undergoing hybrid THA with a minimum 5 years follow up. Clinical outcomes including intra and postoperative complications, blood transfusion, revision rate and mortality were recorded. Radiological analysis of pre and postoperative radiograph assessed bone quality, implant fixation and any subsequent loosening. 143 patients, (mean age 86.2 yrs.) were enrolled in the study. 76 patients underwent uncemented THA and 67 underwent hybrid THA. The uncemented cohort had fewer intraoperative and postoperative complications. The uncemented cohort also had a lower transfusion rate (p=0.002). Mean hospital stay (p=0.27) was comparable between the 2 groups. Two patients underwent revision surgery in either cohort. Our study demonstrates uncemented THA is safe for the octogenarian patient and we recommend that age should be not be a barrier of choice of implant. However intraoperative assessment of bone quality should guide surgeon to the optimum decision regarding uncemented and hybrid implant.

Restoring native hip anatomy and biomechanics is important to create a well-functioning total hip arthroplasty (THA). Hip offset and leg length are regarded as the most important biomechanical characteristics. This study investigated their association with clinical outcomes including patient reported outcome measures (PROMs) and functional tests. This prospective cohort study was conducted in 77 patients undergoing primary THA (age=65±11 years). Hip offset and leg length were measured on anteroposterior radiographs of the hip pre- and postoperatively. Participants completed the Western Ontario & McMaster Universities Osteoarthritis Index (WOMAC) and performed functional tests (i.e. gait, single leg stance, sit-to-stand, block step-up) preoperatively, and 3 and 12 months postoperatively. A wearable motion sensor was used to derive biomechanical parameters. Associations between radiographic and functional outcomes were investigated with the Spearman's rho correlation coefficient. Subgroup comparisons were conducted for patients with more than 15% decreased or increased femoral offset after THA. Differences in postoperative offset and leg length had little impact on clinical outcomes. Femoral offset subgroups demonstrated no significantly different WOMAC function scores. In functional tests, patients with >15% decreased femoral offset after THA demonstrated more sagittal plane motion during block step-up (14.43° versus 10.66°; p=0.04) while patients with >15% increased femoral after THA demonstrated more asymmetry of frontal plane motion during block step-up (34.05% versus 14.18%; p=0.03). To create a well-functioning THA, there seems to be a reasonable safe zone regarding the reconstruction of offset and leg length.

SL-PLUS MIA stem (Smith & Nephew Orthopaedics AG) is a modified implant of Zweymuller type SL-PLUS standard stem (Smith & Nephew Orthopaedics AG). We constructed finite element (FE) models and analysed equivalent stresses in the femur. In addition, we measured bone mineral density (BMD) in the femur by dual-energy X-ray absorptiometry (DEXA) after THA. The purpose of this study was to investigate the equivalent stress and to compare the results of the FE analyses with changes in BMD after THA. Twenty-one patients (18 women and 3 men) who underwent primary cementless THA with SL-PLUS MIA or SL-PLUS formed the basis of this study. Eleven patients received SL-PLUS MIA and ten patients received SL-PLUS. Zones were defined according to Gruen's system (zones 1∼7). Computed-tomography (CT) images of the femur were taken before and at 1 week after THA. FE models of the femur and prosthesis were obtained from CT data by Mechanical Finder (Research Center of Computational Mechanics Inc., Tokyo, Japan), software that creates FE models showing individual bone shape and density distribution. Equivalent stresses were analysed in zones 1 to 7 and compared to the DEXA data. FE studies revealed that there was no significant difference in equivalent stress between SL-PLUS MIA and SL-PLUS. BMD was maintained after THA in zones 3, 4, and 5, whereas BMD decreased in zones 2, 6, and 7. In zone 1, BMD decreased in SL-PLUS MIA stem group by 14%, while BMD was maintained in SL-PLUS standard stem.

Patients with a hindfoot deformity impose a particular challenge when performing a total knee arthroplasty (TKA). This could be attributed to the lack of insights concerning the outcome towards the hindfoot alignment. Our objective was to perform a systematic review of the literature to investigate the influence of TKA on hindfoot alignment and vice-versa. In accordance with the Methodological Index For Non-Randomized Studies (MINORS) statement standards, we performed a systematic review. Electronic databases Pubmed, EMBASE, Web of Science, Google Scholar and Cochrane Library were searched to identify capable studies studying the influence between TKA and hindfoot malalignment. We indentified four prospective cohort studies, seven retrospective cohort studies and one case-control study. All twelve articles addressed the influence of TKA on hindfoot alignment. Seven out of nine studies which noticed an improvement of hindfoot alignment after TKA, found a significant improvement (p<0.05). Aditionally three of these studies reported a significant improvement only in valgus hindfeet (p<0.05). On the topic of hindfoot alignment influencing TKA, we identified two studies. These studies reported an impact of hindfoot alignment on the weightbearing and described that 87% of hindfeet remained in valgus alignment after TKA. Available data suggests that alignment in valgus hindfeet can improve after TKA, though long term results are not present. Contrary to last, improvement of hindfoot alignment is not expected in varus hindfeet. Furthermore hindfoot alignment deformity may cause a reduction of the long term survival of the knee prosthesis and therefore should be taken in to account.

Tourniquets have been used for many years during total knee arthroplasty (TKA). With a growing demand for TKA in recent years, tourniquet use has been surrounded by ongoing controversy due to many conflicting advantages and disadvantages of tourniquet use. Quantifying the case for or against tourniquet use in TKA, in terms of patient focused outcomes, is a priority. This meta-analysis analysed, the never before assessed, impact of tourniquet use during TKA on post-operative pain. We completed a systematic review and meta-analysis using PRISMA reporting guidelines to assess the impact of tourniquet use on patients post-TKA. Post-operative pain was the primary outcome. Secondary outcomes were post-operative range of motion (ROM) and length of stay (LOS). The initial search yielded 230 studies, of which 14 met the inclusion criteria. A post-operative increase in pain and reduction in ROM when using a tourniquet appeared significantly more likely when compared to no tourniquet use during TKA, yet with no overall difference in post-operative LOS. Subgroup meta-analysis demonstrated a trend that favoured the half-course tourniquet for reduced post-operative pain in patients when compared to full tourniquet use during TKA. This systematic review and meta-analysis concluded that the after-effects of tourniquet use in TKA patients and its impact on post-operative pain and ROM are indeed significant. We recommend further randomized controlled trials (RCTs) focusing on TKA patient outcomes of post-operative pain and ROM. Conflict of interest: The authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

Capturing objective data of the postoperative changes in the mobility of patients is expected to generate a better understanding of the effect of postoperative treatment. Until recently, the collection of gait-related data was limited to controlled clinical environments. The emergence of accurate wearable accelerometers with sufficient runtime, however, enables the long-term measurement and extraction of mobility parameters, such as “real-world walking speed”. An interim analysis of 1967 hours of actibelt data (3D accelerometer, 100 Hz) from 5 patients (planned total 20) with a femur fracture and 5 patients (planned total 20) with a humerus fracture from a geriatric population at two different sites of the university hospital of the Ludwigs-Maximilian-University in Munich was performed. Mobility data was captured during several days of stationary treatment starting directly after surgery and during a short follow-up visit six weeks after the surgery. Preliminary results show an increase of the mean walking speed between the two visits independent of the type of fracture. Patients with a humerus fracture tended to walk faster than patients with a femur fracture during both visits. The data also reveals an unexpected low level of mobility during the stationary stay. Mobile accelerometry can be used to evaluate different postoperative mobilisation strategies and even provide near-time feedback in geriatric trauma patients.

Revision total hip arthroplasty (THA) presents with increasing challenges, potentially compromising the integrity of a revision. The objective of this study was to assess radiologic outcomes of patients who underwent revision THA with a modular tapered stem (Reclaim, DePuy Synthes). This study retrospectively examined all revision Reclaim THAs between 2012 and 2016. Radiologic assessment compared x-rays at two time points: immediately after surgery and the most recent x-ray available. Leg length discrepancy, subsidence and line-to-line fit was assessed. Significant subsidence was considered ≥10mm. Adequate line-to-line fit was considered ≥30mm of bicortical contact. Descriptive statistics included clinical factors (i.e. age, Paprosky classification). P values <0.05 were considered significant. A total of 81 femoral revisions were completed. There were 42 females and 38 males with a mean age of 71 years (range, 46–89). Of these, 6 were revised (dislocation, fracture or infection), and 7 were lost to follow up. Average follow up time was 18 months (range, 1–46 months). Femoral revisions were classified as Paprosky 3a or 3b. Mean stem subsidence was 4.15mm (range, 0–25.6mm). Subsidence of the femoral stem was <10mm in 88% of patients. A total of 62% of patients had both subsidence <10mm and ≥30mm of bicortical contact. In patients with <10mm subsidence, 70% had ≥30mm of bicortical contact. There was a positive trend between cortical contact and stem stability (OR 2.3). The Reclaim modular femoral system has demonstrated radiographic stability. Inadequate initial fit is a potential determinant of subsidence.

Acetabular fractures in the elderly are associated with high levels of morbidity and mortality and are becoming more common. Treatment is complicated by osteoporosis and multiple comorbidities. We present the early results of the use of a coned hemi-pelvis component and total hip arthroplasty in the primary treatment of these injuries. We have prospectively monitored a series of seventeen patients (18 cases) with a mean follow-up of sixteen (4–36) months. They have been reviewed clinically and radiographically. The mean patient age was 78 (64–87), and they had a mean ASA score of 3.3 (3–5). There were (Letournel classification) three elementary fractures, and 15 associated fractures. Mean operative time was 94 (61–134) minutes. There were seven minor post-operative complications. One patient suffered a pre-operative bilateral sciatic nerve injury, partially resolved. Sixteen of 17 patients were allowed to mobilise full weight bearing day one post-operatively. Mean length of hospital stay was 12 (5–27) days. Mortality at 30 days was 0%, and at one year 8%. There have been no thromboembolic events, dislocations or deep infections and no cases of prosthesis migration. Early weight bearing is essential for a successful outcome in this cohort. The coned hemi-pelvis bypasses the fracture, creating an immediately stable construct that allows immediate weight bearing. This is the first description of an innovative use of this prosthesis in the treatment of a complex fracture that is traditionally associated with poor outcomes. Early results suggest this to be a safe technique with an acceptable early complication rate.

Acute multiligament knee injuries (MLKI) are rare, high energy traumatic injuries associated with an increased risk of lower limb complications. The objectives of this study were to investigate the adequacy of clinical assessment for neurovascular status, compartment syndrome, and deep vein thrombosis in the emergency department (ED) following acute MLKI. The authors conducted a retrospective case note review of 19 patients with MLKI presenting at the ED of a Major Trauma Centre during a 7.5-year period between June 2009 and December 2016. MLKIs were diagnosed by MRI or examination under anaesthesia and confirmed intraoperatively. Arterial assessment consisted of documented capillary refill time, dorsalis pedis and posterior tibial pulse assessment (through palpation or Doppler ultrasound), and ankle-brachial pressure index (ABPI) calculation. Neural assessment was adequate if there was documented assessment of both sensory and motor function of the superficial peroneal, deep peroneal and tibial nerves individually. Data was collected for 19 patients (17 male, 2 female). The mean age was 34 (range: 14–61). The most common injury mechanism was road traffic accident. Neurovascular assessment was suboptimal in all categories: only one patient received a satisfactory lower limb neurological assessment and no patients received complete vascular assessments. Neurovascular assessment of multiligament knee injuries was suboptimal. Reasons for this included poor documentation and lack of certain specific clinical assessments, such as ABPI calculation. We propose the introduction of an acute knee injury pro forma highlighting the components of a full lower limb neurovascular examination to rectify this problem.

Balloon kyphoplasty (BKP) is a minimally invasive surgical technique used to correct kyphosis and vertebral compression fractures. BKP uses cement to fill a void created by the inflation of a balloon in a vertebra, it can be used as an alternative to vertebroplasty to reduce cement extravasation. Issues such as poor inter digitisation of the cement and the trabecular bone can arise with the BKP method. This can be due to a compacted layer created during the procedure which can cause complications post-surgery. The primary aim of this study was to investigate alternative cement application methods which could improve the mechanical strength of the bone-cement interface. Three alternative methods were investigated, and cylindrical bone-cement specimens were created for all methods (BKP and three alternatives). An important part of this study was to replicate the compacted layer created by the inflation of the balloon tamp in BKP. Synthetic trabecular bone specimens (Sawbones®, Pacific Research Laboratories, Vashon Island, Washington, USA) were pre-loaded in compression and the resultant compacted layers were found to replicate the compacted layers found in surgery. Mechanical testing was carried out with an MTS Model 858 Bionix^® Servohydraulic load frame using static tensile and torsion loads. Static tests revealed that two of the three alternative methods were an improvement on BKP, with a high statistical significance in relation to the mechanical performance of the bone-cement interface (P < 0.001). This data illustrates the potential to improve the standard BKP technique, in terms of bone-cement interface performance.

There is an inherent risk of iatrogenic new neurological deficit (NND) arising at the spinal cord, cauda equina and nerve root during spinal surgery. Intraoperative neurophysiological monitoring (IONM) can be employed to preserve spinal cord function during spinal surgery. IONM techniques include somatosensory and motor evoked potentials, amongst others. A Canadian survey of 95 spinal surgeons showed that 62.1% used IONM and a similar survey in France of 117 spinal surgeons showed that only 36% used IONM. Unavailability was a common reason for its disuse. Current literature by the British Society of Clinical Neurophysiology has outlined the importance of IONM in preventing NND and the need for the implementation of guidelines for IONM. The lack of an established guideline has resulted in a varied approach in the use of IONM in England. There has been no previous attempt to ascertain the current use of IONM in England. Our study is aimed at assessing the variability of the use of IONM in England as well as identifying the rationale amongst surgeons that dictate their use of IONM. We are in the process of investigating the indications of use of IONM for cervical and lumbar spine procedures in 252 spinal surgeons from 33 hospitals with spinal services. Our survey will illustrate the current use of IONM in spinal surgery in England. It will highlight some of the reasons for the variability of use of IONM and identify factors that can contribute to a more standardised use of IONM in spinal surgery.

Lower limb fractures are commonly treated with cast immobilization, and as a main consequence of strict immobilization this typically leads to loss in muscle mass, decrease of bone density and decline in functional abilities. Body-worn sensors are increasingly used to assess outcome in clinical trials by providing objective mobility parameters in a real-world environment. The aim of this study is to investigate the usability aspects and potential changes in mobility parameters in partial-immobilization patients in real-world conditions. Six healthy young males (age 22.2 ± 1.2 years; weight 76.5 ± 6.7 kg, height 185.8 ± 6.1 cm. Mean ± standard deviation) wore a leg cylinder cast with walker boot to immobilize their dominant leg for two consecutive weeks. Subjects were asked to continuously wear a tri-axial accelerometer on the waist (actibelt) during waking hours for 6 weeks including 2 weeks before, during and after cast immobilisation. The total amount of days of continuous recording was 339 days with a total wearing time of 120 days. Software packages which allow to detect steps and to estimate real-world walking speed were used to analyse the accelerometry data. It was suspected that knee immobilization would affect strongly the wave form of the signal with an impact on the accuracy of the speed algorithm, whereas the step detection should be more robust. This effect was confirmed in a preliminary study performed to quantify the accuracy under immobilization conditions. On the other hand, step numbers are known to be sensitive to fluctuations in wearing time which was not uniform throughout the entire study. We concluded that in this setting step frequency is the most reliable parameter. Step frequency showed a systematic decrease in the values during the immobilization period which recovered to pre-immobilisation values after cast removal. This confirms the usability of accelerometry and sensitivity of its mobility parameters for clinical outcome assessment.

Acetabular fractures in the elderly are associated with high levels of morbidity and mortality. Conservative management is reserved for those unfit for extensive reconstruction, or those who achieve ‘secondary congruence' of a complex fracture. We present demographic data and the results of conservative management in patients over 65 years of age. The Fracture Outcome Research Database (FORD) at our unit was interrogated for all patients over 65 years, who had sustained an acetabular fracture between June 2008 and June 2016. 410 patients were identified. Following exclusions, thirty-two patients were included for analysis. They had a mean age of 80 (66–91), and a mean ASA equivalent score of 3.1 (2–4). Mean follow up was five (1–9) years. Twenty-five patients lived in their own home and seven in a nursing home. Thirty had low energy injures, two high energy. Twenty-four (75%) had anterior column posterior hemitransverse fractures, seven (22%) had associated both column and one (3%) had a T-type fracture. The mean length of inpatient stay was 43 days (4–140). Maximum post-operative mobility was limited to a hoist in eight (25%), a frame with or without assistance in 15 (47%), a stick in five (16%) and independence in four (13%). Thirty-day mortality was 6%- and one-year mortality 22%. The data demonstrates that conservative treatment in this cohort leads to long inpatient stays, poor mobility and significant levels of mortality. Complex reconstruction remains demanding on both surgeon and patient. Innovative ways of managing these patients are needed to improve outcomes.

Hindfoot disorders are complex 3D deformities. Current literature has assessed their influence on the full leg alignment, but the superposition of the hindfoot on plain radiographs resulted in different measurement errors. Therefore, the aim of this study is to assess the hindfoot alignment on Weight-Bearing CT (WBCT) and its influence on the radiographic Hip-Knee-Ankle (HKA) angle. A retrospective analysis was performed on a study population of 109 patients (mean age of 53 years ± 14,49) with a varus or valgus hindfoot deformity. The hindfoot angle (HA) was measured on the WBCT while the HKA angle, and the anatomical tibia axis angle towards the vertical (TA_X) were analysed on the Full Leg radiographs. The mean HA in the valgus hindfoot group was 9,19°±7.94, in the varus hindfoot group −7,29°±6.09. The mean TA_X was 3,32°±2.17 in the group with a valgus hindfoot and 1,89°±2.63 in the group with a varus hindfoot, which showed to be statistically different (p<0.05). The mean HKA Angle was −1,35°±2.73 in the valgus hindfoot group and 0,4°±2.89 in the varus hindfoot group, which showed to be statistically different (p<0.05). This study demonstrates a higher varus in both the HKA and TA_X in valgus hindfoot and a higher tibia valgus in varus hindfoot. This contradicts the previous assumption that a varus hindfoot is associated with a varus knee or vice versa. In clinical practice, these findings contribute to a better understanding of deformity corrections of both the hindfoot and the knee.

An adult acquired flatfoot deformity (AAFD) is a complex 3D deformity. Surgical correction consists of a medial calcaneal osteotomy (MCO) but shows limitations due to the current 2D assessment. Therefore, the aim is to determine the influence of an MCO on the longitudinal foot arch assessed by 2D and 3D weightbearing CT (WBCT). Seventeen patients with a mean age of 44,5 years (range 18–66 yrs) were retrospectively included. MCO was indicated in a stage II AAFD (N=15) and a post-traumatic valgus deformity (N=2). Pre- and post-operative imaging was obtained from a WBCT. The height of the longitudinal foot arch was measured as the distance from the navicular tuberositas to the floor (Navicular Height, NH) on 2D CT images (NH_2D) and computed on 3D CT data (NH_3D). Additionally, 3D assessment could compute the degree of exorotation (α) of the navicular bone towards the vertical axis. The mean pre-operative NH_2D and NH_3D were respectively 29.57mm ± 7.59 and 28.34mm ± 6.51. These showed to be statistically different from the mean post-operative NH_2D and NH_3D, respectively 31.62mm ± 6.69 and 31.67mm ± 6.47 (p < 0,001). A statistical difference was also found when comparing the mean degree of exorotation in pre- and post-operative, respectively: α_pre=14.08° ± 4,92 and the α_post=19,88° ± 3.50 (p < 0,001). This study demonstrates a significant correction of the longitudinal foot arch after a MCO. The novelty is attributed to the accurate degree of rotation assessment using WBCT. This information could be assistive to optimise a pre-operative planning.

In this study, we evaluated the labrum tear using radial sequence 3D Multiple Echo Recombined Gradient Echo (MERGE) MRI without arthrography based on modified Czerny's classification, comparing with actual arthroscopic findings. A total of 61 hips including 27 hips of femoroacetabular impingement (FAI), 19 hips of borderline development dysplasia of the hip (BDDH) and 15 hips of early stage osteoarthritis (OA) were enrolled this retrospective study. MRI findings evaluated in each three regions of interest; anterior region, anterolateral region, and lateral region. The cases with severe degeneration that is not concordant with any original Czerny's classification is defined as stage4. We compared MRI findings with arthroscopic findings and calculated the sensitivity, specificity, and likelihood ratio in terms of the existence of labrum tear. MRI findings revealed labrum tear more frequently in anterolateral than lateral (p<0.001). Especially in FAI group, labrum tear was more frequently observed by MRI in anterolateral than lateral (p=0.006). In comparison with MRI findings and arthroscopic findings, the sensitivity was 97%, specificity was 79% and likelihood ratio was 4.59 as average of all regions in terms of the existence of labrum tear. In each region, sensitivity and specificity was 97% and 50% in anterior, 97% and 100% specificity in anterolateral, 94% and 81% in lateral, respectively. Thus, MERGE MRI revealed excellent sensitivity and specificity for diagnosis of labrum tear, especially in anterolateral region. The cases with severely degenerated labrum were classified as newly defined stage 4, which was recognized frequently in OA cases.

High ankle sprains (HAS) cause subtle lesions in the syndesmotic ligaments of the distal tibiofibular joint (DTFJ). Current intrinsic anatomical parameters of the DTFJ are determined based on 2D imaging and uncertainty remains whether they differ in a HAS patients. The aim of this study is therefore two-fold: radiographic parameters will be determined in 3D and compared in a healthy vs sprained group. Ten patients with a mean age of 42,56 (SD = 15,38) that sustained a HAS and twenty-five control subjects with a mean age of 47,44 (SD = 6,55) were retrospectively included. The slices obtained from CT analysis were segmented to have a 3D reconstruction. The following DTFJ anatomical parameters were computed using CAD software: incisura width, incisura depth, incisura length, incisura angle, and incisura-tibia ratio. The mean incisura depth in the sprained group was 3,93mm (SD = 0,80) compared to 4,76 mm (SD = 1,09) in the control group, which showed a significant difference (P < 0.05). The mean incisura length in the group of patients with HAS was 30,81 mm (SD = 3,17) compared to 36,10mm (SD = 5,27) in the control group which showed a significant difference (P < 0.05). The other DTFJ anatomical parameters showed no significant difference. This study shows a significant difference in both incisura depth and incisura length between HAS patients and control subjects. These parameters could be used to identify potential anatomical intrinsic risk factors in sustaining a HAS.

Drilling through bone is a complex action that requires precise motor skills of an orthopedic surgeon. In order to minimize plunging and soft tissue damage, the surgeon must halt drill progression precisely following penetration of the far cortex. The purpose of this study was to create a low-cost and easy-to-use drilling simulator to train orthopedic residents in reducing the drill plunging depth. This prospective observational study was performed in the division of orthopedic surgery of a single tertiary medical center. The participants included 13 residents and 7 orthopedic specialists. The simulator consisted of a synthetic femur bone model and ordinary modeling clay, and the training unit consisted of a disposable plastic tube (∼US$14), clamps (∼US$58) and a power drill + drill bit (standard hospital equipment). Plunging depths were measured by the simulator and compared between orthopedic specialists, the 6 “senior residents” (3+ years) and the 7 “junior residents” during a training session. Measurements were taken again 2 weeks following the training session. Initially, the plunging depths of the junior residents were significantly greater compared to those of the orthopedic specialists (7.00 mm vs 5.28 mm, respectively, p < 0.038). There was no similarly significant difference between the senior residents and the orthopedic experts ([6.33 mm vs. 5.28 mm, respectively; p = 0.18). The senior residents achieved plunging depths of 5.17 mm at the end of the training session and 4.7 mm 2 weeks later compared to 7.14 mm at the end of the training session and 6 mm 2 weeks later for the junior residents. This study demonstrated the capability of a low-cost drilling simulator as a training model for reducing the plunging depth during the drilling of bone and soft tissue among junior and senior residents.

Universities have an obligation to ensure that Intellectual Property (IP) outputs are properly captured and exploited according to various National and European guidelines. There are two main ways which University technology development can take on the road to commercialisation: 1. Licensing the technology to an existing company: A license is permission to do something the granting party (the licensor) has the right to otherwise prohibit. In the context of IP licensing, it is a grant, by the owner of the property, to another (the licensee) of the right to use the IP in question for commercial purposes; 2. Starting a new company: An important university objective is to explore and pursue opportunities for the exploitation of its intellectual property rights. For universities and its inventors, spin out companies often provide an effective means to achieve this objective. A spin out is created when the University creates a new company out of one of its existing departments, institutions or by an inventor. The decision of which path to take is critical and various elements can effect this decision such as the inventors own objectives, the market niche for the technology, the stage of technical development, the potential reward for each option and the types of support structures available. This talk will summarise the main points to consider when deciding on the most appropriate way to commercialise technologies developed in Universities.

Patents are among the most important assets that a technology company can have. Building a patent portfolio involves balancing competing demands. Technical development, cashflow management and attracting investment must all happen against a backdrop of unforgiving patent application deadlines. Getting it right can pave the way to commercial success; but getting it wrong can kill a business before it even starts. Of course, filing patent applications too late can be fatal. But there's also a risk of filing too soon; an applicant can be locked into a spiral of patent fees before it can afford the cost. Drawing on more than thirty years of experience in the IP field, Sean Cummings will help to navigate the patent maze. He will explain how the international patent system works and how to exploit the system to maximise options while minimising outlay. He will give tips for accelerating the process when granted patents are needed quickly and for slowing down the clock when cashflow is tight. And he will identify the ‘sweet spot' for assessing inventions and attracting investment in the early stages when patent costs are still modest.

Surgical training in the UK is under increasing pressure with a high demand for service provision. This raises concerns about the resultant negative impact this is having on training opportunities for surgical trainees in theatre due to a high demand for surgical procedures to be performed expediently by consultants. This is due to the assumption that trainee take significantly longer time to operate in theatre and thus result in a slow progress of theatre lists. Our study evaluated the differences in operative time between orthopaedic trainees and orthopaedic consultants, as well as provide realistic timings for each stage encompassed within the entire duration a patient is in theatre. From our trauma unit electronic theatre database, we retrospectively collected data for six Joint Committee of Surgical Training (JCST) mandatory procedures. Information collected included patients' ASA grading, total surgical time and grade of surgeons. A total of 956 procedures were reviewed: 71.8% hip procedures, 14.2% intramedullary nail fixations and 14.2% ankle fixations. 46.2% and 53.8% of the procedures were performed by consultants and trainees as first surgeon, respectively. On average, consultants were found to be 13 minutes quicker in performing the hip procedures and this difference was found to be statically significant (p < 0.05). However, trainees were found to be quicker in performing intramedullary femoral nailings and simple ankle fixations, but consultant were faster at performing intramedullary tibial nailings and complex ankle fixations. However, the differences were not found to be statistically significant (p > 0.05).

Although achondroplasia has been cited as the most common form of rhizomelic dwarfism, no report in the literature has given the data on the ratio of their upper and lower limb segments. We performed a paired study of 91achondroplasia patients with age and gender matched normal control group. Their upper and lower extremity radiographs were evaluated, and their radio-humeral and tibia-femoral ratios were compared. The ratios were compared using the Wilcoxon rank sum test. A p value of <0.05 was considered significant. The mean age of the patients was 15.8 years (95% confidence interval (CI), 13.1–18.6), and there were 45 males and 46 females. The radio-humeral ratio for the upper extremity of achondroplasia patients was 0.76 (95% CI, 0.75–0.76) which was significantly different from the normal control group of 0.79 (95% CI, 0.77–0.80) (p=0.001). The tibio-femoral ratio of the achondroplasia patients was 0.79 (95% CI, 0.78–0.79), which was not significantly different from the normal control group of 0.78 (95% CI, 0.77–0.79) (p=0.346). Rhizomelia is the predominant form of dwarfism in upper extremity, but not in lower extremity in achondroplasia. The functional deficits from these patients seem to come from the generalized limb shortness, and not specifically from rhizomelia.

The process of gaining informed consent can be a complex and much debated pursuit, especially within a paediatric setting. The role of the trainee surgeon and its explanation to children and their families prior to an operation has not been explored from the resident surgeons' point of view. Ten face-to-face interviews were conducted with orthopaedic surgery trainees at a tertiary level paediatric hospital in Toronto, Canada. These were transcribed and subsequently thematically coded by 3 reviewers. Three main themes were identified from the interviews. 1) Surgical trainees feel their level of participation and autonomy gradually increases dependent on their observed skills and level of training. 2) Trainees feel the consent process is adequate but acknowledge it is often purposely vague with regards to their intra-operative involvement as this is often unpredictable and it avoids patient/family anxiety. 3) Trainees believe families are aware of their participation however most likely underestimate their role during operations. Trainees in surgical specialties believe their level of autonomy is variable dependent on a number of factors and that this impacts on the ability to be more specific when gaining informed consent. This must be balanced with a family's right to an appropriate understanding of their child's operation and who is performing it. It may be that further patient education regarding trainees and their role in operations would help develop a more thorough and patient centred informed consent process.

Conventional marker based optical motion capture (mocap) methods for estimating the position and orientation (pose) of anatomical segments use assumptions that anatomical segments are rigid bodies and the position of tracking markers is invariant relative to bones. Soft tissue artefact (STA) is the error in pose estimation due to markers secured to soft tissue that moves relative to bones. STA is a major source of pose estimation error and is most prevalent when markers are placed over joints. Mocap and bi-plane videoradiography data were recorded synchronously while three individuals walked on a treadmill. For all three, pose of the thigh and shank, and movement of markers relative to the bones, were determined from the videoradiography data (DSX, C-Motion). Independently, pose of thighs and shanks was estimated using mocap data (Visual3D, C-Motion). Our measures of error in the mocap pose estimation were the relative thigh and shank translations. X-ray data from two subjects were used to generate a regression model for the antero/posterior movement of the lateral knee marker against internal/external hip rotation. The mocap translation errors of the third subject, attributed to STA of the knee marker, were 15.6mm and 32.0mm respectively. The pose of the third subject was then estimated using a probabilistic algorithm incorporating our regression model. Mocap translation errors were reduced to 10.6mm (thigh) and 4.4mm (shank). The results from these data suggest that errors in pose estimation due to STA may possibly be reduced via the application of algorithms based on probabilistic inference to mocap data.

A suitable wound closure is an indispensable requirement for an uncomplicated and expedient recovery after an abdominal surgery. The closure technique will have a great impact on the healing process of the wound. Surgical complications, such as wound dehiscence (sometimes associated with evisceration), infection, hernia, nerve injury and incisional pain are very common in the postoperative period of an abdominal surgery. Besides, although their development can be promoted by other risk factors like age, sex, lifestyle, diet, health condition, the closure method can also influence the emergence of these undesirable complications. For this reason, and having the wellbeing and quality of life of the patients in mind, particularly high-risk patients, a closure system consisting of anchors applied on either side of the wound that aims to reduce the tension caused on the surrounding tissues of a wound and, consequently, decrease the risk of herniation was evaluated in a pilot animal study and compared with the traditional suture approach.

Intra-articular injections of human mesenchymal stromal cells (MSCs) and platelet-rich plasma (PRP) have been intensively investigated as therapies for knee osteoarthritis (OA) with positive outcomes. In this work we evaluated weather a combination of the treatments (MSCs + PRP) would be beneficial compared to MSCs alone (MSCs) and standard corticosteroid injection (Control group). Forty seven patients (24 males and 23 females; 53.3 ± 10.7 years old) with radiographic symptomatic knee OA (Dejour grades II–IV) were randomized to receive intra-articular injections of MSCs (n = 16), MSCs + PRP (n = 14) or corticosteroid (n=17). MSCs were obtained after mononuclear cells separation from bone marrow aspiration collected from both posterior iliac crests using Sepax automated closed system and expanded in culture until reaching the number of 4 × 10⁷. PRP was obtained by double-centrifugation of whole blood according to a protocol developed in house. After 12 months follow-up, the MSCs and MSCs+PRP groups achieved higher percentages of expected improvement when comparing to the corticosteroid group for the KOOS-symptoms, pain, function and daily living, domains and global score. For the population older or equal to 60 years old the MSCs+PRP group showed significant superiority for the KOOS-ADL domain at 12 months. Cytokines quantification evidenced anti-inflammatory aspects of the treatments. This work evidences the safety and efficacy of intra-articular injection of MSCs for the treatment of early knee OA, with greater improvement with PRP addition particularly to the older population.

Originally the use of Additive Manufacturing (AM) in the medical field begun with simple 3D printed tissue and organ models. The technology has now evolved giving the ability to produce polymers, metal, ceramic personalized implants. What is the status and use of AM technology in cranio-maxillofacial surgeries? What are the recent advances in personalized implants? What are the technical, clinical and legal challenges in the use of AM this entails? These and other interesting questions, will be discussed in this presentation.

Osteoporosis is a worldwide spread, silent disease steadily increasing due to demographic shift; it results in bone loss and increased porosity that lead to an increase in bone fragility and to low-energy fractures. In such a contest, we worked on the development of 3D scaffolds engineered to mimic the features of human healthy bone. Healthy and osteoporotic bone microCT scans were obtained from tissues discarded during surgical interventions (Istituto Ortopedico Rizzoli-Italy). The obtained .STL file was used to 3D print a type I collagen solution to mimic bone matrix whereas mesoporous bioactive glass/nano-hydroxyapatite were embedded within the collagen fibers to mimic the inorganic phase of human bone. The rheological properties of the Type I collagen/mesoporous glass suspensions were investigated at different collagen concentration and temperatures. The possibility of incorporating growth factors (IGF and β-TGF) in the scaffold struts was investigated proposing several approaches and their retained activity was assessed. Different co-culture of osteoblasts and osteoclasts set-ups were explored in order to define the influence of both chemical and topographical stimuli on the osteoblast-osteoclast coupling.

Current treatments of cartilage defects, including chondrocyte implantation and several tissue engineering strategies, often result in a repair tissue that does not replicate the architecture and depth-dependent properties of the native tissue. As a result, these therapies often only delay the occurrence degenerative diseases, such as osteoarthritis. Additionally, when the damage is extended to the subchondral bone, the regeneration of both bone and cartilage is major challenge, due to the dissimilar composition of the two tissues and the inherent challenge in recreating their strong interface, thus favouring the integration in vivo of the neo-tissue. The recent progresses in the field of biofabrication are opening new avenues for the treatment of damaged articulating joints. In particular, bioprinting technologies allow coordinating the deposition of multiple cell types and materials, thus permitting to mimic the complex architecture of osteochondral structures. In this lecture, the latest development in the field of (stem) cell-laden hydrogels, also termed bioinks, to create zonal-biomimetic cartilage constructs will be discussed, together with the integration of multiple (bio)printing strategies (i.e. co-fabrication of hydrogels, reinforcing polymers and bioceramics), and the impact of these technologies towards the generation of fully biofabricated, high-performance engineered osteochondral grafts, with potential application as tissue engineering constructs for regenerative medicine in orthopaedics.

Additive Manufacturing techniques such as Selective laser melting (SLM) are increasingly used in the fabrication of hip, knee and other orthopaedic implants. This is due to the ability of these techniques to print geometrically complex parts with osteoconductive features, resulting in a decreased chance of aseptic loosening. To facilitate wider adoption of SLM, in-situ process monitoring is required. This paper examines the robustness of a novel monitoring systems ability to detect voids within the bulk of a component with varying part density. This work reports the results of a printing study carried out with Ti6Al4V parts using a production scale Renishaw system. This system is equipped with the recently developed in-situ monitoring system, called InfiniAM Spectral. InfiniAM measures the level of optical emissions emitted during the build process. The Spectral software creates a 3D representation of the part, in near real time, based on the level of emissions detected. In this work, Spectral 3D images are compared with those generated after printing using a micro CT scanner. The latter creates a virtual 3D representation of the part and has the ability to detect part defects and voids, as well as quantify part density, within the body of a component. In this work, parts were designed with voids of diameters in the range 200 to 600 μm. The sensitivity of the in-situ monitoring system was correlated with post process analysis of the void dimensions. Additionally, the detection of part density variation due to a variation of input energy, was also evaluated.

There is a growing requirement by governmental and other funders of research, that investigators pay attention to and integrate considerations of sex and gender in their health research studies. Doing so, the argument goes, will reduce data waste, lead to the generation of more complete and accurate evidence to apply to the delivery of health care, and hopefully improve outcomes for both male and female patients. Yet, it is not always clear what sex and gender mean and how best to apply these to the study of diverse health conditions and health service delivery. In this presentation sex, gender and other related factors will be considered in the context of fractures, fracture repair, and post-operative management. Examples of sex and gender bias, sex and gender differences, and the integration of sex and gender in research on fracture and fracture repair will be presented.

We need to shift our focus to integrating sex and gender into research proposals, so we can answer some of the most basic unanswered questions in the field of fracture management. Current evidence in guidelines indicate a near-to-linear increase from the 1990s for inclusion of sex and gender. However, these recommendations remain expressed in absolute terms, with little explanatory power, affecting uptake and implementation in clinical practice. This co-branded session, with members of the Orthopaedic Research Society – International section of fracture repair (ORS-ISFR), will provide participants with guiding principles and tools to assist researchers and grant reviewers understand what it means to include sex and gender in meaningful ways: from formulating research questions, recruitment strategies, to conducting sex-stratified analyses. In this presentation, we will consider diverse approaches, methods and, analyses to elevate sex and gender within trauma. A strong emphasis on the ways and means of including marginalized and vulnerable populations in research will be addressed.

The presentation of musculoskeletal disease differs in men and women, and recognition of the differences between men and women's burden of disease and response to treatment is critical to optimizing care. In this presentation, I will discuss the expanding evidence in the literature that examine the role of sex and gender in musculoskeletal disease, including how its examination increases the innovations and contributions, as well as expands the knowledge about musculoskeletal disease, conditions, and injury in a broad sense. We will discuss the role that structural anatomy differences, hormones, and genetics play in differential disease expression, to the historical biases in the subject populations of clinical and basic research projects. Participants will be provided with examples and opportunities to evaluate orthopaedic science through a sex and gender lens, and what impact this may play in setting the stage for both clinical practice and scientific investigation.

The aim of this study was to evaluate the relationship between the location of the insertion point of the AT into the posterior aspect of the calcaneus and the PF. Two hundred and two feet were evaluated from MRI scans. Ninety-seven women and one hundred and five men with a mean age of 40.15±18.58 were included in this study. Two independent investigators measured the horizontal distance from the most anterior point of the calcaneus to the most posterior part of the PF (A), including the horizontal length of the calcaneus (B). Moreover, distance between the most inferior point of the calcaneus and the most inferior part of the AT insertion into the calcaneus (C) and height of the posterior aspect of the calcaneus (D) were measured. Patients were divided into three groups based on age (I - patients younger than 18, II − 18–65, III - older than 65The all obtained mean values showed high sexual dimorphism between genders. However, when standardized ratios were compared, no statistically significant sexual differences were noted (p>0.05). Although previous studies have reported a correlation between the PF, age and gender, this correlation was not found in our study. Based on the obtained results, this study concludes that age and sex do not influence the morphology of the PF. However, aging strongly affects the location of the AT insertion point. Therefore, we believe this is the key factor which influence the relationship between the AT and PF.

Femoroplasty is the process of injecting cement (cement augmentation) into the proximal femur to prevent osteoporotic hip fractures. Femoroplasty increases the strength and energy to failure of the femur and can be performed in a minimally-invasively manner with lower hospitalization costs and reduced recovery. Our hypothesis was that efficient cement augmentation strategies can be identified via computational optimization. Therefore, using patient-specific planning we can minimize cement volume while increasing bone strength and reducing the risk of fracture. We proposed an in-silico methodology that was validated with in vitro experiments. A discrete particle model for cement infiltration was used to determine the optimum volume and filling pattern of the cement such that the best outcome was achieved. Several artificial bones were scanned before and after cement augmentation to applied previous in silico methodology. Then those femurs were mechanically tested (non-augmented and augmented). Therefore, in silico methodology was validated. Cement augmentation significantly increased the yield load. Predicted yield loads correlated well with the experiments. Results suggest that patient-specific planning of femoroplasty reduces the risk of hip fracture while minimizing the amount of cement required.

The complex structural arrangement of bone gives rise to anisotropic, rate-dependent failure behaviour, which varies significantly depending on tissue composition and architecture. This presents significant challenges in the development of orthopaedic surgical cutting instruments, which are required to generate sufficient forces to penetrate bone tissue, while minimizing the risk of thermal and mechanical damage to the surrounding environment. Currently, instrument designers rely heavily on empirical-based strategies to understand tool-bone interactions, with significant amounts of prototyping and validation experiments required throughout the design process. The aim of this study is to develop an experimentally-validated predictive computational model of orthopaedic cutting processes in three dimensions to understand the role of various cutting parameters on cutting forces and chip formation. An experimental model of orthogonal cutting was developed, whereby an adaptable cutting tool fixture driven by a servo-hydraulic uniaxial test machine was used to carry out high-rate cutting tests on Sawbone® trabecular bone analogues. A three-dimensional computational model was also developed using Abaqus/Explicit. The constitutive model describing material behaviour considers strain-rate and pressure-dependant yield behaviour using a Drucker-Prager elastic-plastic damage model, with Strain-hardening and rate-dependent model constants determined through dynamic uniaxial high-strain rate compression tests of material cubes. An excellent correlation between experimental and computational models was found, with the computational model accurately predicting tool cutting forces and chip development ahead of the tool during the cutting process. It was identifying that lower tool rake-angles resulted in the formation of larger discontinuous chips and higher cutting forces, while higher rake angles tended to lead to more continuous chip formation and lower cutting forces.

The treatment of scapholunate (SL) ligament injuries is addressed by surgical procedures to stabilize the carpal joint. Open techniques include bone-ligament-bone transfers, tenodesis, partial fusions and carpectomies. Innovative procedures using wrist arthroscopy, offer minimally invasive fixation without full exposure of carpal bones; however, the success of the technique and its impact on the reduction on the range of carpal movement is as yet not well known. In this work, the performance of Corella tenodesis technique to repair the SL ligament is evaluated for a wrist type II by numerical methods. Human wrist can be classified based on the lunate morphology: type I for lunate that articulates with radius, scaphoid, capitate and triquetrum, and type II which has an extra surface to articulate with the hamate. A finite element model was constructed from CT-scan images, the model includes cortical and trabecular bones, articular cartilage and ligaments. Three scenarios were simulated representing healthy wrist, SL ligament sectioning and the Corella technique. The performance of the technique was assessed by measure the SL gap in dorsal and volar side as well as the SL angle to be compared to cadaveric studies. In intact position, the SL gap and the SL angle predicted by the numerical model is 2.8 mm and 44.8º, these values are consistent to the standard values reported in cadaveric experiments (2.0 ± 0.8 mm for SL gap and 45.8 ± 9.7 for SL angle). Virtual surgeries may help to understand and evaluate the performance of the techniques at clinical application.

Formation of micro-cracks occurs in bone due to daily activities. Through a mechanism of self-repair, these micro-cracks are detected, and the damaged areas are restored, avoiding further propagation. The Scissors Model suggests that the osteocyte processes that cross the micro-cracks break as consequence of the cyclic displacements of the micro-crack faces, due to fatigue, and this triggers the remodelling processes. A fresh bovine tibia bone was cut in sections oriented 20° from the transversal direction. The cortical bone was sliced using a circular saw and shaped to the dimensions: 20 × 10 × 1 (mm) and the surfaces were polished. µCT images were obtained from all the samples (μCT 40, Scanco Medical, Brüttisellen, Switzerland). From the DICOM files, the geometries were reconstructed and meshed using tetrahedrons, in ICEM CFD. The Elasticity Modulus (E) was determined in Bonemat, by applying an empirical relationship Elasticity-Density from the literature. The parts were then imported into ANSYS APDL to simulate micro-crack propagation in bone. This model will be validated with further experimental work where the micro-crack will be initiated in the prepared samples and propagated due to fatigue loading, and the osteocyte processes will be visualized in the Scanning Electron Microscope (SEM). This investigation aims to study how cyclic loading in bones and failure of osteocyte processes can trigger target the mechanism of bone remodelling. The resulting model can later contribute for the investigation of treatments for bone diseases such as osteoporosis and the response of bone to the presence of orthopaedic implants.

In order to reduce the risk of dislocation larger femoral heads in total hip arthroplasty (THA) are being used by surgeons in recent years. The standard head size of 28 mm used in 73% of all hip procedures in 2003 was used in only 29% in 2016; whereas head sizes of 32 mm and 36 mm combined, were used in 70%. The increase of head size effectively reduces the thickness of the acetabular cup, altering the load transfer. Herein, this research work investigates the effect of increasing the femoral head size on the stresses of the periacetabular bone at two selected regions: A1 (superior) and A2 (anterior). Three Finite Element models were developed from CT scan data of a hemipelvis implanted with a cemented all-polyethylene acetabular cup with a 50 mm outer diameter and inner diameter to accommodate three head sizes: 28 mm, 32 mm and 36 mm. The peak reaction force at the hip joint during one leg stand for an overweight patient with a body weight of 100 Kg was simulated for head sizes investigated. We found that highest average von Mises stress was 5.7 MPa and occurred in the cortical bone of region A1 which is located within Zone 1 boundaries (Charnley &DeLee); whereas a lower stress of 4.0 MPa occurred at region A2. In the two regions the stresses were the same for the three head sizes. Periacetabular bone was found to be insensitive to the increase of femoral head diameter in cemented THA.

A pin-on-disc tribometer test with a rotating disc and a sector-wise loaded pin was used to determine friction coefficients for different material pairings. The four pin materials porcine cartilage, subchondral bone of the porcine cartilage, UHMWPE, vitamin E enhanced, crosslinked UHMWPE (VEPE) in combination with the three-disc materials zirconia toughened alumina ceramic (ZTA), CoCr, carbon-fibre-reinforced carbon (CrC) were tested. Stepwise loading was employed with the forces 10 N, 5 N, 2 N and 1 N. Test duration was 1 h. Diluted calf serum according ISO 14242-1 was used to determine the friction coefficients. The surface topography of all pins was examined using optical profilometry before and after the rotation tribometer tests. - No wear related modifications of the surface roughness parameters could be found. The coefficients of friction (COF) were lowest for the cartilage pins against all three-disc materials, with steady-state values between 0.01 and 0.02 for the highest applied load (10 N). Friction of subchondral bone yielded COF in the range 0.2 … 0.6, depending on the counterpart material. The two polyethylene materials behaved similar in this friction test with COF of about 0.1. The Ra roughness values of the different pins reflect the COF results: Ra of subchondral bone was one order of magnitude higher than Ra of the cartilage. This is in-line with the COF-values of bone being one order of magnitude higher than those of cartilage. These results will be discussed in view of the use of the disc materials as orthopaedic hemi-prostheses.

Immune response in periprosthetic joint infection (PJI) is diverse. Resident macrophage and/or wandering monocyte are superb guardians to sense microbial attacks, take invaders and alarm the danger. Neutrophils are refined but momentary fighters to kill microbes with projectile weapons as well as predation. The swift action is usually effective at the forefront to prevent expansion of infectious foci. However, such characteristics often evokes overshooting via self-defeating of pus, thus leading to crucial soft tissue damage in the acute phase. Intervention of monocyte/macrophages follow and act as wise organizers. In addition, stromal fibroblasts also act in front for host defence. They equip innate immune sensors (TLRs, NLRs), which can sense dangers and trigger off inflammatory response, but also is usually self-regulated. These sensors not only interact each other, but also have possible contribution to selective autophagy (xenophagy and lysophagy) in PJI. In this presentation, overview of pathology in PJI will be summarized with a special attention to innate immune sensors (TLRs and NLRs), and selective autophagy.

While stable long-term clinical results have been achieved in total joint arthroplasty, periprosthetic joint infection (PJI) has been actualized as difficult issue in this decade. For accurate diagnosis, it is important to establish standard criteria such as MSIS criteria, and it is prevailing now. As an issue involving PJI, however, the existence of viable, but non-culturable (VNC) bacteria must be noticed. It is difficult to identify the VNC state infection, because microbiologic culture result shows negative and other markers tend to be negative. Here, molecular diagnosis based on polymerase chain reaction (PCR) has certain role as potential diagnostic tools for such VNC infection. We have applied a real-time PCR system for the diagnosis of PJI, which is able to detect methicillin-resistant Staphylococcus (MRS) and distinguish gram-positive from gram-negative bacteria. The prominent advantage is that PCR is the singular way to identify MRS in such culture negative cases. Recent development of full-automatic PCR system may improve the time efficiency for routine application. In this presentation, we will show the overall sensitivity and specificity of our PCR system for diagnosing PJI and discuss the current problem and future prospect.

Patients with knee prosthetic joint infection (PJI) frequently receive one- or two-stage revision. To explore the feasibility of a randomised controlled trial (RCT) comparing methods, we analysed a UK registry, interviewed patients and surgeons, systematically reviewed literature, held a consensus meeting, and assessed progress of an RCT in hip PJI. In 2014, in England and Wales, knee PJI was treated with one- or two-stage procedures in 19% and 71% of patients respectively. Between 2007 and 2014, use of one-stage procedures doubled and, in major centres, up to 42% of treatments were one-stage. We conducted in-depth interviews with 16 patients with knee PJI and 11 surgeons performing one- or two-stage revision. Patients considered randomisation acceptable with appropriate counselling and, depending on infecting organisms and health status, surgeons would randomise treatments. In meta-analysis, two-year re-infection rates in 10 one-stage series (423 patients) and 108 two-stage series (5,129 patients) were 7.6% (95%CI 3.4,13.1) and 8.8% (7.2,10.6) respectively. In a series of patients with knee PJI, surgeons from 2 major centres considered 6/15 patients eligible for either treatment, with 4 more potentially eligible after treatment of soft tissue infection. In an ongoing RCT of surgical treatment of hip PJI, 116 patients have been randomised at 14 centres in 3 years. Randomising patients with PJI is feasible but, as knee PJI is uncommon, a multicentre RCT would be required. Based on WOMAC score outcome and appropriate assumptions on eligibility and acceptability, 170 patients would need to be randomised over 4 years at 14 major centres.

There has been a significant increase in the demand of polymeric scaffolds with promising affects in bone regeneration. However, inflammation is still a problem in transplantations to overcome with local antibiotic therapy. In this study, it is aimed to develop a functional POSS nanocage reinforced chitosan scaffold (CS/POSS) coated with drug loaded chitosan composite nanospheres to provide a controlled antibianyiotic delivery at the defect site. Gentamicin and vancomycin were selected as model antibiotic drugs. Drug loaded nanospheres were fabricated with electrospray method and characterized in terms of morphology, hydrodynamic size, surface charge, FT-IR, in vitro drug release, antimicrobial activity and cytotoxicity. CS/POSS scaffolds were fabricated via lyophilisation and characterized with mechanic, swelling test, SEM and micro CT analyses. Positively charged nanospheres with uniform morphology were obtained. High drug encapsulation efficiency (80–95%) and sustained release profile up to 25 days were achieved with a cumulative release of 80–90%. In addition, the release media of the nanospheres (in 6 hours, 24 hours and 25 days of incubation period) showed a strong antimicrobial activity against S.aureus and E.coli, and did not show any cytotoxic effect to 3T3 and SaOS-2 cell lines. CS/POSS scaffolds were obtained with high porosity (89%) and 223.3±55.2μm average pore size. POSS reinforcement increased the compression modulus from 755.7 to 846.1Pa for 10 % POSS addition. In vitro studies of nanosphere coated bilayer scaffolds have showed high cell viability. Besides ALP activity results showed that POSS incorporation significantly increased the ALP activity of Saos-2 cells cultured on the scaffold. In conclusion, these composites can be considered as a potential candidate in view of its enhanced physico-chemical properties as well as biological activities for infection preventive bone tissue engineering applications.

Chronic osteomyelitis (OM) is a progressive, inflammatory infection of bone caused predominantly by S. aureus and requires treatment through surgical debridement and systemic antibiotic administration. We have previously reported the fabrication of an antibiotic-eluting scaffold which is responsive to microbial activity for the treatment of OM. Herein, we ventured to assess the capacity of this antibiotic-eluting scaffold to treat infection in a rabbit model of chronic OM. Infections were established in the radii of New Zealand White rabbits using inoculations of 2×10⁶ CFUs S. aureus JAR 060131 over a period of 4 weeks. Following surgical debridement (6mm), rabbits underwent treatment for a period of 8 weeks until euthanasia. The treatment groups were; 1) empty, 2) antibiotic-eluting scaffold (collagen/hydroxyapatite scaffold loaded with vancomycin) and 3) commercially available antibiotic-eluting fleece (Septocoll E®, collagen fleece loaded with gentamicin). During the treatment period, all groups received systemic antibiotics (Cefazolin 25mg/kg) administered subcutaneously twice daily for 4 weeks. Inoculation of the radius resulted in the development of a sequestrum containing S. aureus, demonstrating the successful establishment of OM. After the 8-week treatment period, 4/5 rabbits in the empty group were still infected, indicating that systemic antibiotic administration following debridement was insufficient to treat the infection. Fewer rabbits in both the antibiotic-eluting scaffold group (2/4) and the antibiotic-eluting fleece group (1/3) were infected. This work demonstrates that the implantation of an antibiotic-eluting biomaterial into a defect following debridement enhances bacterial clearance in conditions of chronic OM.

Recently in traumatology various methods of impregnation biodegradable implants and allografts with antibiotics are widely used. Among them the soaking, shaking and ionophores are common used. We aimed to choose the optimal method of impregnation with the antibiotic of the head of the femur, taken from patients after arthroplasty. We studied 6 femoral heads after hip replacement. Head №1 the iohexol (Omnipaque) was injected through circular ligament and through the neck of the femur. Head №2 through the circular ligament, head №3 through the neck of the femur, head №4 through the circular ligament and through the neck of the femur, head №5 through 4 pre-drilled channels a brilliant green solution was injected. The head №6 was soaked in a brilliant green solution. Head №1 assessed by radiology. All the heads, treated with brilliant green, were cut in half to assess the degree of impregnation. On the X-ray image of head №1 the contrast agent has spread enough. In osteotomy, the impregnation with brilliant green head №2, №3, №4, №5 was seen in 3–4 mm around the needle passage place. Head №6 the bone was not impregnated. Despite the fact that the radiograph showed a sufficient spread of contrast agent, on the sections of the head, treated with brilliant green, showed the spread of liquid 3–4 mm around the needle passage place. This indicates that the impregnation of large bone is not effective.

Current strategies for bone repair have accepted limitations and the search for synthetic graft materials or for scaffolds that will support ex vivo bone tissue engineering continues. Bioprospecting has led to increased interest in potential applications for marine organisms and their by-products and biomimetic strategies have led to the investigation of naturally occurring porous structures as templates for bone growth. As a rich source of mineralising porous organisms, our seas and oceans could provide new directions for bone tissue engineering that may enhance in vivo and ex vivo bone formation.

Diatoms are unicellular microalgae whose cell walls are composed of remarkably uniform, hierarchical micro/nanopatterned, amorphous biosilica that cannot be replicated synthetically. Each species hosts its own unique morphology which is identically replicated generation-to-generation. There are currently estimated to be over 200,000 different diatom species, each with their own unique shape and morphology. This offers a huge array of surface topographies, particle sizes and shapes, each with the same silica precursor. Our research to date has shown that diatom-biosilica is non-cyctotoxic to J774.2 macrophages and hBMSC cells and does not invoke an immunological response or organ toxicity (kidney, spleen and liver) when tested in a murine model. Before testing diatom-biosilica in vivo in an animal fracture model, methods to incorporate the frustules into the defect are being investigated. Two methods have been developed 1) using a bioresorbable hydrogel and 2) 3-D printed polymer-biosilica scaffolds. Both methods have shown promising results with enhanced mechanical properties with the addition of the diatom-biosilica. Work is ongoing to further map and quantify the role of surface topography and chemical cues on cell fate through the systematic in vitro studies of different species of diatom-biosilica.

Human mesenchymal stem cells are considered the golden standard for clinical application in regenerative medicine for their multilineage differentiation potential, best candidates to treat diseases such as osteoarthritis and osteogenesis imperfecta. In the past few years several molecules have been described to induce the hMSCs differentiation into osteo cell progenitors, mainly discovered by screening of single metabolites bioactivity. However, hMSCs osteogenic differentiation potential is still poor, and the discovery of differentiation inducers with high efficiency is needed. Thanks to automated processes, High Throughput Screening (HTS) strategies shorten the metabolites bioactivity investigation timeline, allowing testing of many molecules simultaneously. In this work, reliable assays for natural products bioactivity investigation detection were developed using HTS methodologies and validated by testing 15 purified compounds derived by marine fungi and sponges. The HTS cytotoxicity investigation using HepG2 cells allowed to test in a single experiment, 15 metabolites in 4 concentrations ranging from 1 to 20µM. Low cytotoxicity was detected for metabolites concentrations from 1 to 10µM and so set as treatment concentrations to be tested in further assays. Anti-inflammatory bioactivity was tested on THP1 cells triggered by LPS. Five out of 15 metabolites showed to prevent the LPS induced THP1 inflammatory activation by lowering the TNF-α production. The metabolites pro-osteogenic potential was investigated using hMSCs: their differentiation was evaluated by calcium mineralization after 10 days differentiation. Pro-osteogenic molecules were not detected in this screening, but the method validation represents a powerful tool for future natural product and synthetic molecules libraries screenings.

A substantial body of evidence supports the use of extracorporeal shock wave therapy (ESWT) for fracture non-unions in human medicine. However, the success rate (i.e., radiographic union at six months after ESWT) is only approximately 75%. Detailed knowledge regarding the underlying mechanisms that induce bio-calcification after ESWT is limited. The aim of the present study was to analyse the biological response within mineralized tissue of a new invertebrate model organism, the zebra mussel Dreissena polymorpha, after exposure with extracorporeal shock waves (ESWs). Mussels were exposed to ESWs with positive energy density of 0.4 mJ/mm² or were sham exposed. Detection of newly calcified tissue was performed by concomitantly exposing the mussels to fluorescent markers. Two weeks later, the fluorescence signal intensity of the valves was measured. Mussels exposed to ESWs showed a statistically significantly higher mean fluorescence signal intensity within the shell zone than mussels that were sham exposed. Additional acoustic measurements revealed that the increased mean fluorescence signal intensity within the shell of those mussels that were exposed to ESWs was independent of the size and position of the focal point of the ESWs. These data demonstrate that induction of bio-calcification after ESWT may not be restricted to the region of direct energy transfer of ESWs into calcified tissue. The results of the present study are of relevance for better understanding of the molecular and cellular mechanisms that induce formation of new mineralized tissue after ESWT. Specifically, bio-calcification following ESWT may extend beyond the direct area of treatment.

Bone strength is influenced by bone quality besides its density. This study aimed to evaluate the effects of teriparatide on changes of bone strength as well as trabecular and cortical bone microstructures at femoral neck in female ovariectomized (OVX) rats. Eighteen female Wister rats were divided into three groups: the sham control, OVX and treatment (Tx) groups. All of them were sacrificed after 3-month intermittent teriparatide intervention in Tx group. All left femurs were removed and scanned using micro-CT and followed by mechanical test for each femoral neck. Regarding micro-CT, four trabecular parameters including bone volume fraction (BV/TV), trabecular thickness (TbTh), trabecular separation (TbSp), and trabecular number (TbN) and three cortical parameters including volumetric bone mineral density (vBMD), cortical cross-sectional area (CtAr) and cortical thickness (CtTh) were measured at femoral neck region. All data were analyzed and was presented as median ± SEM. The mean bone strength of femoral neck significantly decreased in OVX group when compared to the control group (p < 0.05) and was significantly restored in Tx group (p < 0.01). Regarding the trabecular parameters, the BV/TV and TbTh significantly decreased in OVX group while compare to Tx group. However, no significant difference was observed in TbSp and TbN between the groups. Regarding the cortical parameters, CtTh was significantly greater in Tx group than that in OVX group (p<0.01). As our findings, intermittent teriparatide can improve the deteriorated bone strength of femoral neck due to ovarian deficiency via changing both trabecular microarchitecture and cortical morphology.

Pulsed electromagnetic fields (PEMFs) have been considered a potential treatment modality for fracture healing. As bone fracture healing and osseointegration share the same biological events, the application of PEMF stimulation to facilitate the osseointegration process of orthopedic implants has been suggested. However, the mechanism of their action remains unclear. Mammalian target of rapamycin (mTOR) signaling may affect osteoblast proliferation and differentiation. This study aimed to assess the osteogenic differentiation of mesenchymal stem cells (MSCs) under PEMF stimulation and the potential involvement of mTOR signaling pathway in this process. PEMFs were generated by a novel miniaturized electromagnetic device (MED). Potential changes in the expression of mTOR pathway components, including receptors, ligands and nuclear target genes, and their correlation with osteogenic markers and transcription factors were analyzed. PEMF exposure increased cell proliferation, adhesion and osteogenic commitment of MSCs. Osteogenic-related genes were over-expressed following PEMF treatment. Our results confirm that PEMFs contribute to activation of the mTOR pathway via upregulation of the proteins AKT, MAPP kinase, and RRAGA, suggesting that activation of the mTOR pathway is required for PEMF-stimulated osteogenic differentiation. In summary, the findings of the present study revealed that MED-generated PEMFs stimulate osteogenic differentiation and the maturation of the adipose tissue-derived MSCs via activation of the mTOR pathways. Even though further research is required to determine an optimal stimulation timing and flux density both in-vitro and in-vivo, this study results may serve a source for an adjuvant therapy to improve orthopedic implant stability, longevity and enhance fracture healing.

The human amniotic membrane (hAM) contains cells of stem cell characteristics with low immunogenicity and anti-inflammatory properties and has for centuries been applied in the clinics especially for ophthalmology and wound care. It has recently been shown to be promising for novel applications such as tissue engineering and regenerative medicine. Towards these novel applications, we have demonstrated the potential of hAM in toto to differentiate towards bone, cartilage, Schwann like cells and recently also a producer of surfactant. We have further investigated the relevance of the location of origin for the therapeutic potential of the membrane. We show that placental and reflected hAM differs distinctly in morphology and functional activity. The placental region has significantly higher mitochondrial activity, however lower levels of reactive oxygen species, which suggests that placental and reflected regions may have different potential for tissue regeneration. We have further investigated the suitability of hAM to support therapeutic cells and have improved its maintenance in vitro towards xeno-free conditions.

The growth in the popularity of tissue engineering principles in the treatment of musculoskeletal disorders has been complemented greatly with research investment into tissue specific scaffolds. Biological scaffolds produced by means of decellularising native tissues have the advantage of providing the natural complex hierarchical matrix and, in doing so, replicating the specific biomechanical and biological functions of the tissue in question. Decellularisation treatments are multi-faceted, vary considerably between different processes and may involve many lengthy treatment steps. Some of these bio-processes may cause undesirable structural changes to the extracellular matrix of tissues and, by association, their mechanical properties. Thus, it is of paramount importance to ensure that the properties of the scaffolds are not affected to the extent of reducing their integration, biomechanical performance and longevity. This talk consists of a body of work detailing investigations into bio-process optimisation, sterilisation strategies and the regenerative and functional capacity of decellularised xenogeneic and allogeneic tendon, ligament and bone scaffolds. In addition, on-going work concerning advanced pre-clinical assessment, stratification of these products to particular patient populations and the importance of the manufacturing value chain in their translation will be discussed.

Mesenchymal stem cells (MSC) have a well recognised potential for tissue repair. This potential is two pronged: they can differentiate into the functional cell types of the damaged tissues and they can support tissue recovery by secreting trophic factors, depositing an extracellular matrix (ECM) and dampening inflammation. Three-dimensional microscopy recently shown that MSCs in the bone marrow create an intricate proteo-cellular scaffold with the ECM forming an interconnected cellular continuum whose structure is guided by the deposited ECM. This proteo-cellular scaffold controls bone marrow functions from hematopoiesis to osteogenesis. In the current study we aimed to optimise ECM production under in vitro conditions by immortalised MSCs with the view that the generated ECM can be utilised for tissue repair. With immunocytochemistry we determined the deposition of bone marrow-characteristic ECM proteins: collagen I, III, IV, V, VI, laminin and fibronectin. While primary MSCs produced slightly higher amount ECM proteins than immortalised MSCs, the relative abundancy of the ECM proteins was very similar. In order to isolate the ECM, we optimised a decellularisation method based on gentle lysis with sodium-deoxycholate and DNase digestion. Immunostaining for collagen I, III, VI and fibronectin and labelling the nuclei with Hoechst-33342 confirmed removal of all cells while retaining the ECM in its original architecture. Ideally, the decellularised ECM retains associated cytokines and chemokines, such as CXCL12, important for attracting MSCs. To test this, we seeded Molm-13 leukemia cells on decellularised ECM as MSC-produced CXCL12- and other cytokines protect leukemia cells against chemotherapeutics. We found that the decellularisation process however removed these factors and thus for therapeutic purposes, the decellularised ECM would need to be re-loaded with the essential chemo/cytokines. Overall, we developed a system for decellularised ECM production by immortalised MSCs and the results warrant further exploration of this avenue.

Macromolecular crowding (MMC) accelerates matrix deposition through excluded volume effect (EVE). Herein, we ventured to identify the optimal decellularisation protocol of MMC enhanced fibroblast cultures as a new cell formed platform model. Human dermal fibroblasts (hDF), human lung fibroblasts (hLF), and human mammary fibroblasts (hMF)seeded at 50,000 cells/cm² were cultured for 10 days without and with MMC (100 μg/mL carrageenan) and 100 μM L-ascorbic acid phosphate. Subsequently, the cultured cell layers were decellularised using various decellularisation protocols [i.e., ammonium hydroxide (NH₄OH), sodium deoxycholate (DOC), SDS-EDTA mixed buffer, and nonident P40 (NP40)]. SDS-PAGE, hydroxyproline assay, sGAG assay, SEM, histological staining (i.e., picrosirius red stain and H&E), immunocytochemistry (i.e., collagen I, III and fibronectin), PicoGreen® assay. SDS-PAGE with complementary density and hydroxyproline analysis for assessing collagen deposition, and sGAG assay for total sGAG content assessment demonstrated significantly increased (p< 0.001) in the presence of MMC. SEM, histological and immunocytochemistry displayed enhanced ECM deposition, integrity, and maintenance of the matrix composition in the presence of MMC. PicoGreen® assay revealed efficient decellularisation with significant removal of DNA (p <0.001) in all matrices. MMC can be used effectively to accelerate ECM deposition by fibroblast from various tissue sources, to facilitate production of cell-derived matrix-rich constructs feasible as robust platform models.

Inflammation has been associated with early degradative changes in articular cartilage and immune responses are key factor influencing normal tissue regeneration and repair. With synovitis a prominent feature in osteoarthritis (OA) and associated with the progressive degradation of articular cartilage, immune factors need to be factored into efforts to achieve efficient cartilage repair/regeneration. Recent efforts have focused on the use of autologous or allogeneic mesenchymal stem/stromal cells (MSCs) to modulate the inflammatory environment in the injured or osteoarthritic joint. Intraarticular injection of MSCS has modulated cartilage degradation in a variety of pre-clinical OA models. Results from early clinical trials have also shown effects on pain and function-associated outcome measures. Other cell types may also have some capacity for use as a therapy for OA. For example, primary allogeneic chondrocytes also seem to have some immune-privilege in the synovial joint and are immunomodulatory in a rat model. Although MSCs isolated from bone marrow that are induced to undergo chondrogenic differentiation do not retain these properties, MSCs isolated from the synovium or chondroprogenitors generated from cartilage itself may represent the future of cell therapy for OA.

Harnessing the potential of mesenchymal stem cell (MSC) mediated endochondral ossification for the repair of large bone defects represents a promising avenue of investigation as an alternative option to autologous bone transplantation. To date, it has been shown that undifferentiated MSCs are somewhat immune-privileged. In order to induce bone formation from MSCs by endochondral ossification it is usually necessary to first differentiate these cells chondrogenically. However, the status of differentiated cells is less clear than that of undifferentiated MSCs. Furthermore, the fate of implanted bone forming constructs in an allogeneic setting is not known. The potential to use allogeneic MSCs for large bone defect repair would offer opportunities to researchers to develop new therapies using more potent MSC sources and in a more readily available manner with regard to the patient. I will present our research investigating the interactions between chondrogenically primed MSCs and immune cell subsets, namely T cells and dendritic cells. Furthermore, I will discuss the ability of human paediatric MSCs to form bone in the in vivo allogeneic setting.

Industrialized countries experience a population aging. Elderly patients, due to the experienced immunity, have a constant pro-inflammatory milieu. Little is known on how adaptive immunity impacts the tissue homeostasis and regeneration. The standardized housing of lab animals is specific pathogen free (SPF). However, this housing condition hinders antigen exposure and thus an aging of the adaptive immune system. We hypothesized that exposure to antigens and a developing adaptive immunity will impact tissue homeostasis and regeneration in mice. Mice kept under SPF housing or non-SPF were examined towards their immune status via flow cytometry, bone structure via microCT and bone competence via biomechanical torsional testing. MSCs from these mice were analyzed regarding their differentiation potential and ECM production under various immune cell signaling. Bone regeneration was analyzed in vivo in a mouse osteotomy model. The memory and effector compartment of the adaptive immunity was significantly increased in mice under non-SPF housing. This housing led to an increased femoral cortical thickness and torsional stiffness (p<0,05), whereas the tissue mineral density was not affected. The differentiation potential of stem cells under the influence of an aged immune milieu was significantly reduced. Bone formation was highly affected by the immune status and availed of a naïve immune cell milieu. Adaptive immunity directly impacts bone tissue formation, by exhibiting a constant stress, leading to structural differences in bone tissue organization as well as mechanical competence. For experimental settings, it appears highly relevant if mouse models have had the chance to develop an experienced immune system.

Human synovium harbours macrophages and T-cells that secrete inflammatory cytokines, stimulating chondrocytes to release proteinases like aggrecanases and matrix metalloproteinases (MMPs) during the development of Osteoarthritis (OA). Inflammation of the synovium is a key feature of OA, linked to several clinical symptoms and the disease progression. As a prelude to testing in an OA mouse model, we have used the tetracycline system (Tet) to modify mouse mesenchymal stem cells (mMSCs) to over-express viral interleukin 10 (vIL10), an anti-inflammatory cytokine, to modulate the osteoarthritic environment and prevent disease development. MSCs isolated from the marrow of C57BL/6J mice expressed CD90.2, SCA-1, CD105, CD140a, and were negative for CD34, CD45 and CD11b by flow cytometry. Adenoviral transduction of MSCs carrying CMVIL10 and TetON as test, and untransduced, AdNull and TetOFF as negative controls was successful and tightly controlled vIL10 production was demonstrated by CMVIL10 and TetON MSCs using a vIL10 ELISA kit. Co-incubation of vIL10MSC CM with lipopolysaccharide activated bone-marrow derived murine macrophages (BMDMs) resulted in reduction of TNF-α, IL-6 levels and elevated production of IL-10 by ELISA and high iNOS release by Griess assay. Co-culture of active macrophages with TetON MSCs, resulted in polarisation of macrophage cell population from M1 to M2 phase, with decrease in pro-inflammatory MHC-II (M1 marker) and increase in regulatory CD206 (M2 marker) expression over time. The PCR profiler array on MSC CM treated BMDMs, also showed changes in gene expression of critical pro-inflammatory cytokines and receptors involved in the TLR4 pathway. The biscistronic TetON transduced MSCs proved to be most immuno-suppressive and therefore feasible as efficient anti-inflammatory therapy that can utilised in vivo.

Recently, we could illustrate how tightly the bone and the immune system are interconnected during normal homeostasis but even stronger during bone regeneration. Specifically, the patient´s individual ratio of CD8+ effector T cells (TEFF, already identified as potential unfavorable cells for successful healing) to CD4+ regulatory T cells (TREG, one counterpart to CD8+ TEFF in controlling intratissue inflammation) prior to injury/ surgery appears to determine the healing outcome after fracture. We hypothesized that concentrating CD4+ TREG could serve as innovative therapeutic strategy to improve bone healing. We used an adoptive CD4+ TREG transfer in our well-established mouse osteotomy model. Before treatment, we identified the pre-surgery ratio of CD8+ TEFF/ CD4+ TREG by flow cytometry to characterize the healing potential of individual animals. Thereafter, we performed an adoptive CD4+ TREG transfer to reshape inflammation for supporting osteotomy healing. Across all groups, healing outcome was analyzed after 21 days post-surgery by µCT. Whereas TREG were highly supportive in SPF mice, we observed a heterogeneous clustered healing outcome in the non-SPF mice: TREG responder (improved healing outcome; p = 0.038) and TREG non-responder (impaired healing outcome; p = 0.024). Interestingly, the pre-/peri-surgery ratio of CD8+ TEFF/ CD4+ TREG was higher in the TREG non-responder (p=0.057). Thus, the amount of adoptively transferred CD4+ TREG was not sufficient to improve the healing outcome due to initial unfavorable high CD8+ TEFF/CD4+ TREG ratio. These results clearly show the importance of determining the individual immune status of each patient in the clinic before applying an immunotherapeutic approach.

Research in orthopaedics is now moving away from permanent metallic implants, and looking towards the use of bioresorbable polymers (e.g. PLLA, PGA and related co-polymers) that, when implanted into the injured site, bioresorb as the tissue heals. However, reports of a delayed inflammatory reponse occurring in the late stages of polymer degradation has limited the wide scale use of these polymers. Few studies assess the long-term biocompatibility of these polymers and with an increasing market for bioresorbable materials it is anticipated that this will be a future issue. This work aims to develop a predictive tool that can be used to assess the delayed inflammatory response of poly(D,L-lactide-co-glycolide) (PDLGA) using in vitro tests. An elevated temperature accelerated test (47^oC) was developed and utilitised to induce predetermined amounts of degradation in PDLGA. This was used to mimic a range of clinically relevant in vivo implantation times up to 5–6 months. All pre-degradion work was performed under sterile conditions, in PBS solution. At predetermined time intervals, indicators of late stage inflammation will be assessed using an MTT cytotoxicity assay, an inflammation antibody array and an ELISA analysis for inflammatory factors, with mouse L929 fibroblasts, RAW264.7 and primary BMDM macrophages. It is hypothosised that at the later degradation time intervals signs of inflammatory factors will be observed. The methodologies developed in this work can be applied to the optimisation of polymer degradation profiles to minimise late-stage inflammatory repsonse and identification of beneficial additives in this regard.

Improvements in arthroplasty design and materials led to superior lifetime of the implants. Nevertheless, aseptic loosening due to particulate debris is still one of the most frequent late reasons for revision of hip and knee replacements. The complex process of inflammation and osteolysis due to wear particles is not understood in detail so far. A cellular and receptor mediated response to wear particles results in a release of pro-inflammatory cytokines and induces an inflammatory reaction causing periprosthetic osteolysis. The overall cellular response is influenced by particle volume as well as characteristics. But there is still a lack of data concerning all signalling pathways that are involved. To answer some open questions appropriate in vivo models are shown closing the loop between wear simulation, particle analysis, generation of sterile particles and biological evaluation. Beyond that, new aspects of particle effects and deposits in retrieved human tissue are given.

Total joint replacement is a successful clinical intervention. However, aseptic loosening due to wear related particulate debris is still one of the most frequent reasons for late revision of total joint replacement. This lecture gives an overview about the application of methods to study wear and friction in total joint replacements (e.g. hip, knee, shoulder). This involves complex joint simulation conditions as well as analytical assessments. Regarding joint simulation the focus will be on ligament stabilized joints. New approaches will be shown and discussed. Furthermore, analytical methods to study the release of wear products in term of solid particles and soluble complexes like metal ions will be presented.

Instability accounts for approximately 20% of all revision total knee arthroplasty (TKA), however diagnostic tests remain crude and subjective. The aim of this examination was to evaluate the feasibility of pressure mat (SB Mat, TekScan) analyses of functional tasks to differentiate instability in a clinical setting. Five patients (M = 4; age = 69.80±7.05 years; weight = 79.73±20.12 kg) with suspected TKA instability were examined compared to five healthy controls (M = 1; age = 46.80±7.85 years; weight = 71.54±16.17 kg). Peak pressure and time parameters were measured during normal gait and two-minute bilateral stance. Side-to-side pressure distribution was calculated over 10-second intervals during the second minute. Pressure distributions were expressed relative to bodyweight (%BW). T-tests compared loading parameters between groups (significance level = p<0.05). Analyses showed subtle differences in pressure distribution in unstable TKA patients versus healthy controls. Stance time during gait was indifferent. TKA patients tended to exhibit longer heel contact time (0.76 vs. 0.64 sec) and reduced weight acceptance (50.75% vs. 56.75%) on the operated versus non-operated limb. Side-to-side differences in toe-off forces were significantly more pronounced in TKA patients versus controls (9.25% vs. 3.75%; p=0.0088). Uneven loading was significantly greater – favouring the non-operated limb – in TKA patients during bilateral stance compared to controls (p<0.05). This feasibility work demonstrates subtle differences in limb loading and biomechanics during simple clinical tests in unstable TKA patients that might be undetectable to the naked eye. Pressure analyses may therefore be a useful diagnostic tool. These findings warrant further investigation.

Total knee arthroplasty is a well-established treatment for degenerative joint disease, on the other hand metal ion release of cobalt or chromium and particle formation can trigger intolerance reactions. Biotribological examinations can help to assess the metal ion release in different settings. The purpose of this study was the evaluation of inter-laboratory differences in the metal ion concentration analysis. Samples were generated in a 3+1 station knee wear simulator (EndoLab GmbH, Thansau, Germany) with a medium size Columbus Knee System with or without AS multilayer coating. The wear simulation was performed under highly demanding activity (HDA) profile and samples were taken after 0.5, 5.0, 5.5. and 8.0 million cycles. The samples were blinded and sent to three different laboratories and the content of chromium, cobalt, molybdenum, nickel, and zirconium was assessed by inductively coupled plasma mass spectrometry (ICP-MS). The AS multilayer coating clearly reduced the release of chromium, cobalt and molybdenum. Mean levels were: Chromium 9329.78µg/l ± 985.44 vs 503.75µg/l ± 54.19, cobalt 10419.00µg/l ± 15.517.53 vs 2.60µg/l ± 1.35, molybdenum 2496.33µg/l ± 102.62 vs 2.46µg/l ± 2.31. Interestingly we found especially for nickel and zirconium big inter-laboratory differences in the metal assessment. There were up to 10-fold higher values in comparison of one laboratory to another. The data demonstrate that results of metal ion assessment should be evaluated by interlaboratory comparison and should be critically interpreted.

Tear pattern and tendon involvement are risk factors for the development of a pseudoparalytic shoulder. However, some patients have similar tendon involvement but significantly different active forward flexion. In these cases, it remains unclear why some patients suffer from pseudoparalysis and others with the same tear pattern show good active range of motion. Moment arms (MA) and force vectors of the RC and the deltoid muscle play an important role in the muscular equilibrium to stabilize the glenohumeral joint. Biomechanical and clinical analyses were conducted calculating different MA-ratios of the RC and the deltoid muscle using computer rigid body simulation and a retrospective radiographic investigation of two cohorts with and without pseudoparalysis and massive RC tears. Idealized MAs were represented by two spheres concentric to the joints centre of rotation either spanning to the humeral head or deltoid origin of the acromion. Individual ratios of the RC /deltoid MAs on antero-posterior radiographs using the newly introduced Shoulder Abduction Moment (SAM) Index was compared between the pseudoparalytic and non-pseudoparalytic patients.

Decrease of RC activity and improved glenohumeral stability (+14%) was found in simulations for MA ratios with larger diameters of the humeral head which also were consequently beneficial for the (remaining) RC. Clinical investigation of the MA-ratio showed significant risk of having pseudoparalysis in patients with massive tears and a SAM Index <0.77 (OR=11). The SAM index, representing individual biomechanical characteristics of shoulder morphology has an impact on the presence or absence of pseudoparalysis in shoulders with massive RC tears.

In 2011, approximately 1.6 million total hip arthroplasties (THAs) were conducted in 27 of the 34 member countries in the Organization for Economic Cooperation and Development (OECD) However, approximately 10–15% of patients still require revision surgery every year. Therefore, new technologies are required to increase the life-spam of the prosthesis from the current 10–15 years to at least 20–30 years. Our strategy focuses on surface modification of the bearing materials with a hydrophilic coating to improve their wear behaviour. These coatings are biocompatible, with high swelling capacity and antifouling properties, mimicking the properties of natural cartilage, i.e. wear resistance with permanent hydrated layer that prevents prosthesis damage. Clear beneficial advantages of this coating have been demonstrated in different conditions and different materials, such as UHMWPE, PEEK, CrCo, Stainless steel, ZTA and Alumina. Using routine tribological experiments, the wear for UHMWPE substrate was decreased by 75% against alumina, ZTA and stainless steel. For PEEK-CFR substrate coated, the amount of material lost against ZTA and CrCo was at least 40% lower. Further experiments on hip simulator adding abrasive particles (1-micron sized aluminium particles) during 3 million cycles, on a total of 6 million, showed a wear decreased of around 55% compared to uncoated UHMWPE and XLPE. In conclusion, CIDETEC‘s coating technology is versatile and can be adapted to protect and improve the tribological properties of different types of surfaces used for prosthesis, even in abrasive conditions.

Total knee arthroplasty (TKA) is becoming more prevalent as the average age of the general population increases and is generally considered to be a very effective and successful surgery. However, functional recovery post-surgery can often be less than optimal. Neuromuscular electrical stimulation (NMES) is a beneficial therapy proven to improve haemodynamics and muscle strength and may be of great benefit in improving functional recovery in the acute phase post-TKA. The objective of the study was to assess functional recovery in the period immediately following TKA and hospital discharge in response to a home-based NMES programme. Twenty-six TKA patients were randomized into a NMES stimulation or placebo-controlled group. All participants were given a research muscle stimulator to use at home post-discharge for 90 minutes per day over a period of 5 weeks. In the stimulation group, application of stimulation resulted in an electrically activated contraction of the soleus muscle. Patients in the placebo-controlled group received sensory stimulation only. Outcome measures were physical activity levels, joint range of motion and lower limb swelling, which were measured pre-surgery and on a weekly basis post-discharge up until the sixth post-surgical week. 90 minutes per day NMES stimulation significantly increased the Activity Time (P = 0.029 week 1 post-discharge) and the number of Stepping Bouts (P < 0.05 weeks 1 to 4 post-discharge) in the early post-discharge phase. While there was a trend towards a greater knee flexion with use of NMES, this did not reach statistical significance (P = 0.722). No effect of NMES was observed on swelling (P > 0.05 for all measures). Compliance to the NMES therapy was measured by an on-board SIM card in the NMES device, with a 95% and 94% time compliance rate for the stimulation and placebo-controlled groups respectively. The results of this study suggest that NMES may be very useful in improving functional recovery through increasing physical activity levels in the early post-TKA discharge phase. The results of this study warrant further investigation into the use of an optimized NMES protocol whereby improvements in knee range of motion and swelling may also be observed.

The relevance of physical activity (PA) for general health and the value of assessing PA in the free-living environment especially for assessing orthopaedic conditions and outcome are discussed. Available methods for assessing PA such as self-reports, trackers, phone apps and clinical grade monitors are introduced. An overview of devices such as accelerometers for research quality assessments is given and aspects for choosing them such as wear location, usability or study population are reviewed. Basic principles to derive mobility parameters from the PA related sensor signals are presented. The symposium explains mobility parameters, their types, definitions, validity, analysis and those with particular relevance to assess orthopaedic conditions. The application of activity monitors is orthopaedic patient studies is demonstrated in various examples such as knee and hop osteoarthritis and total joint arthroplasty, in frail elderly subjects at fall risk or patients with shoulder pathologies.

Efficient, repeatable and reliable insertion of microneedles into skin is paramount to ensure efficacious drug and vaccine delivery, as well as effective microneedle-based biosensing. Through maintaining robust mechanical adhesion, this microneedle platform offers significant potential in therapeutic delivery and longitudinal wearable applications. Here, we have shown that an angled microneedle design, which is conducive to self-administration, has the potential to address key limitations in existing microneedle technology.

3D measurement of joint angles so far has only been possible using marker-based movement analysis, and therefore has not been applied in (larger scale) clinical practice (performance test) and even less so in the free field (activity monitoring). 3D joint angles could provide useful additional information in assessing the risk of anterior cruciate ligament injury using a vertical drop jump or in assessing knee range of motion after total knee arthroplasty. We developed a tool to measure dynamic 3D joint angles using 6 inertial sensors, attached to left and right shank, thigh and pelvis. The same sensors have been used for activity identification in a previous study. To validate the setup in a pilot study, we measured 3D knee and hip angles using the sensors and a Vicon movement lab simultaneously in 3 subjects. Subjects performed drop jumps, squats and ran on the spot. The mean error between Vicon and sensor measurement for the maximum joint angles was 3, 7 and 8 degrees for knee flexion, ad/abduction and rotation respectively, and 9, 7 and 10 degrees for hip flexion, ad/abduction and rotation respectively. No calibration movements were required. A major part of the inaccuracy was caused by soft tissue effects and can partly be resolved by improved sensor attachment. These pilot results show that it is feasible to measure 3D joint angles continuously using unobtrusive light-weight sensors. No movement lab is necessary and therefore the measurements can be done in a free field setting, e.g. at home or during training at a sport club. A more extensive validation study will be performed in the near future.

In least 12% of patients with symptomatic OA, the cause is joint injury that progressed over time to post-traumatic OA. Human adult articular cartilage has a limited innate ability to regenerate. Available treatment options are unable to restore native structure and function of hyaline cartilage. Agili-C (CartiHeal, Israel) is a first-in-class acellular scaffold consisted of two layers corresponding to cartilage and bone that is capable of attracting stem cells and guide a regenerative process in both tissues. Agili-C has been extensively tested in vitro in our laboratory using human normal cartilage and in vivo in preclinical and currently clinical studies. This scaffold consists of a natural crystalline aragonite, derived from corals, to which hyaluronic acid is added. It showed a great ability to induce regeneration of chondral and osteochondral lesions and attract chondrocytes and stems cells to fill the defect area. Cells remained viable over the course of the study (up to 2 months). Signs of the extracellular matrix formation were evident inside 3D structure of the scaffold. PG synthesis and gene expression of collagen type II and aggrecan were elevated by more than 2.5-fold in cartilage with the scaffold vs corresponding controls. Agili-C scaffold displays a potential in the treatment of focal chondral and osteochondral defects.

For many decades, we have viewed osteoarthritis (OA) as a homogeneous disease characterised by “wear and tear”. However, this view has been challenged recently and it is now clear that OA is a heterogeneous and low-grade inflammatory disease with multiple aetiologies and phenotypes. Each of these different phenotypes may be identified and targeted differently, opening up multiple pathways for therapeutic intervention. Combining imaging and carefully selected panels of biochemical markers can achieve enhanced patient stratification and lead to better-designed clinical trials. Analyses of observational studies and clinical trial datasets are underway to understand better the phenotypes responsible for why people develop OA and why, prognostically, they have differences in terms of disease progression. The aim of this presentation is to discuss the underlying mechanisms involved in common OA phenotypes, with a particular focus on low-grade inflammation and metabolic alterations. Aberrant cellular metabolism has been implicated in the pathogenesis of OA and this talk will summarise the current state of knowledge on the role of impaired metabolism in the cells of the osteoarthritic joint and highlight areas for future research, such as the potential to target metabolic pathways and mediators therapeutically.

While osteophytes are a hallmark feature of knee osteoarthritis (OA), there is limited information regarding their location. In particular, it is unknown whether osteophytes develop in patient-specific locations or if there are consistent osteophyte locations among OA knees. This lack of data mainly stems from the fact that osteophytes have been mostly assessed with scores quantifying their size or severity but not their location. Given the important role that bone could play in OA development and the option it offers for OA treatment, there is a need to better understand the osteophyte locations. This study aimed to develop a method to compare osteophyte locations among knees and determine the overlapping ratio. CT arthrogram of 11 medial-compartment OA tibias (Kellgren-Lawrence grade ≥ 3) were segmented to locate the osteophytes and a bone matching technique was used to report the osteophyte locations of the 11 knees on a single reference tibia. This newly proposed method was highly reproducible (intra-operator ICC = 0.89). When used to compare the 11 tibias, it showed that more than 60% of the overall subosteophytal area, defined as the reference bone area covered by at least one osteophyte from one knee, was common to less than two tibias. Moreover, less than 20% of the overall subosteophytal area was common to five or more tibias. The results of this study suggest that osteophyte locations are specific to each knee. Future work should determine the relationships with mechanical loading, as this could explain the high inter-patient variability.

As a part of the European Union BIOMED I study “Assessment of Bone Quality in Osteoporosis,” Sixty-nine second lumbar vertebral body specimens (L2) were obtained post mortem from 32 women and 37 men (age 24–92 years). Our initial remit was to study variations in density of the calcified tissues by quantitative backscattered electron imaging (BSE-SEM). To this end, the para-sagittal bone slices were embedded in PMMA and block surfaces micro-milled and carbon coated. Many samples were re-polished to remove the carbon coat and stained with iodine vapour to permit simultaneous BSE imaging of non-mineralised tissues - especially disc, annulus, cartilage and ligament - uncoated, at 50Pa chamber pressure. We have now studied most of these samples by 30-μm resolution high contrast resolution X-ray microtomography (XMT), typically 72 hours scanning time, thus giving exact correlation between high resolution BSE-SEM and XMT. The 3D XMT data sets were rendered using Drishti software to produce static and movie images for visualisation and edification. We have now selected a set of the female samples for reconstruction by 3D printing - taking as examples the youngest, post-menopausal, oldest, best, worst, and anterior and central compression fractures and anterior collapse with fusion to L3 - which will be attached to the poster display. The most porotic cases were also the most difficult to reconstruct. A surprising proportion of elderly samples showed excellent bone architecture, though with retention of fewer, but more massive, load-bearing trabeculae.

Osteoporosis is a worldwide disease with a high prevalence in elderly population; it results in bone loss and decreased bone strength that lead to low-energy fractures. Since antiresorptive treatments could lead to long-term adverse effects, the ERC BOOST project aims to propose a biomimetic 3D-printed scaffold reproducing the architecture and chemistry of healthy bone. In this study, the structural parameters of healthy bone were studied in order to reproduce them through 3D printing; furthermore, structural and mechanical differences between healthy and osteoporotic (OP) bones were assessed. Healthy and OP humeral heads discarded during surgical interventions (following ethical approval by Istituto Ortopedico Rizzoli-Italy) were tomographically analysed to obtain bone structural parameters. Successively, 8 mm diameter biopsies were harvested from the heads and underwent compression and nanoindentation tests to investigate macroscopic and microscopic mechanical properties, respectively. XRD measurements were performed on bone fragments. OP bone samples exhibited inferior mechanical properties to their less interconnected and more anisotropic structure, with thinner trabeculae and larger pores. On the other hand, nanoindentations performed on OP trabeculae showed increased Young Modulus compared to healthy samples probably due to their increased hydroxyapatite crystal size, as revealed by XRD. Osteoporosis causes the weakening of the trabecular structure that leads to a decrease of bone mechanical properties. However, OP trabeculae are stiffer due to increased dimensions of hydroxyapatite crystals.

The expression of the mechanosensor, integrin αvβ3, is reduced in osteoporotic bone cells compared to controls. MLO-Y4 osteocytes experience altered mechanotransduction under estrogen deficiency and it is unknown whether this is associated with defective αvβ3 expression or signalling. The objectives of this study are to (1) investigate αvβ3 expression and spatial organisation in osteocytes during estrogen deficiency, and (2) establish whether altered responses of osteocytes under estrogen deficiency correlate to defective αvβ3 expression and functionality. MLO-Y4 cells were cultured as follows: Ctrl (no added estradiol), E+ (10nM 17β-estradiol for 5 days), and Ew (10nM 17β-estradiol for 3 days and withdrawal for 2 days). Cells were cultured with/without 0.5µM IntegriSense750 (αvβ3 antagonist). Laminar oscillatory fluid flow of 1Pa at 0.5Hz was applied for 1hr. αvβ3 content was quantified using an ELISA. The location and quantity of αvβ3 and focal-adhesions was determined by immunocytochemistry. Estrogen withdrawal under static conditions led to lower cell and focal-adhesion area (p<0.05), compared to E+ cells. Fluid flow led to higher αvβ3 content (p<0.05) in all groups, compared to static counterparts, with αvβ3 blocking altering this response. Fluid flow on Ew cells had the highest αvβ3 levels (p<0.05), but αvβ3 did not localise at focal-adhesions sites. Cell morphologies were similar after treatment with the αvβ3 antagonist to the Ew group. These results suggest there are fewer functional focal-adhesion sites at which αvβ3 integrins localise to facilitate mechanotransduction. To further understand these results, we are analysing osteocyte mechanotransduction by quantifying PGE2 and gene expression (COX-2, RANKL, OPG, SOST).

I still remember as a green 16-year-old being completely seduced by Newman's portrait of a university – the ideal of a liberal education. I was completely charmed not only by Newman's seductive prose – but by the humanising ideals of the effects of an excellent education. The picture was compelling and inspirational to the daughter of a small farmer whose parents were forced to leave school at 12 years of age to go and earn a living. I was sitting in the “lap of luxury” in a boarding school for girls, whose excellent principal generated a huge respect for, and absolute belief in, the right to and the ability to gain from a rigorous and serious education – which for me at that time in the 1970s extended at least to the end of secondary schooling – a luxury no one in my family had access to in the previous generation. What are universities for? Many authors have considered this issue since Newman's time – in recent times for example Boyd (1979), Graham (2005), Collini (2012). They all, in different ways suggests the need not only to respond to societal / economic needs, but also the need for a more balanced, holistic conception of university activity. Leaders of universities in the 21^st century must try to articulate this, seek greater understanding of it. We must lobby government for greater recognition, understanding and support for the university's role not only for the present but also for the future. Contingency, vulnerability, adaptability, recognising the provisional nature of knowledge (and control); the caring versus the careless – all of this implies the need for diversity of disciplines, gender and experiences among university leadership in both the national and the international arena.

The key points of this talk are: (a) STEM skills are increasingly required by employers across a broad range of sectors. These skills help to foster systemic and critical thinking in a number of areas and are not confined to four subjects alone. Due to the increasing digitalisation of society and the world of work the demand for STEM skills will only intensify. (b) There is a need to increase the number of STEM-qualified people in Ireland and across Europe with employers highlighting a specific shortage of people with these skills. This is particularly apparent as concerns engineers, computer scientists and data analysts. To achieve this, it is necessary to raise awareness of the STEM-related careers that are available and to promote participation in STEM courses and studies, notably among women. (c) Parents, teachers, employers and education and training providers, both through their individual and joint actions, have a key role to play in fostering STEM skills acquisition.

The Electrospinning Company designs, develops and manufactures biomaterials for use in regenerative medical devices. Since 2012, Ann has led the growth of the company from start-up to supplier of innovative, clinical-grade product to an FDA-approved medical device, evolving the business model and adding capabilities in innovation, manufacturing, quality and alliance management. Ann will share some of the highs and lows of the journey from her perspective as a female leader of a diverse team.

Biomimicry is defined as the design and production of materials, structures, and systems that are modelled on biological entities and processes. Within the medical device sector, biomimicry uses an ecological standard to judge the “rightness” of biomaterial components and devices. After 3.8 billion years of evolution, nature has learned what works, what is appropriate, and what lasts. Biomimicry is a new way of viewing and valuing nature, and it introduces an era based not on what we can extract from the natural world, but on what we can learn from it. Original design manufacturing biomaterial projects that leverage the practice of biomimicry will be discussed. Both natural and synthetic polymer platforms will be reviewed for soft tissue and hard tissue applications. Given the complexity of musculoskeletal tissue structures, the key challenge is identifying the most appropriate materials and forms for recapitulating the native function in a tissue scaffold design. The general field of biomimicry will be reviewed along with specific examples in the regenerative medicine sector.

There are clinical situations in fracture repair, e.g. osteochondral fragments, where current implant hardware is insufficient. The proposition of an adhesive enabling fixation and healing has been considered but no successful candidate has emerged thus far. The many preclinical and few clinical attempts include fibrin glue, mussel adhesive and even “Kryptonite” (US bone void filler). The most promising recent attempts are based on phosphorylating amino acids, part of a common cellular adhesion mechanism linking mussels, caddis fly larvae, and mammals. Rapid high bond strength development in the wetted fatty environment of fractured bone, that is sustained during biological healing, is challenging to prove both safety and efficacy. Additionally, there are no “predicate” preclinical animal and human models which led the authors to develop novel evaluations for an adhesive candidate “OsStic^tm” based on calcium salts and amino acids. Adhesive formulations were evaluated in both soft (6/12 weeks) and hard tissue (3,7,10,14 & 42 days) safety studies in murine models. The feasibility of a novel adhesiveness test, initially proven in murine cadaver femoral bone, is being assessed in-vivo (3,7,10,14 & 42 days) in bilateral implantations with a standard tissue glue as the control. In parallel an ex-vivo human bone model using freshly harvested human donor bone is under development to underwrite the eventual clinical application of such an adhesive. This is part of a risk mitigation project bridging between laboratory biomaterial characterisation and a commercial biomaterial development where safety and effectiveness have to meet today´s new medical device requirements.

In 2009, a multidisciplinary team of orthopaedic surgeons, material scientists, and cell biologists created a consortium focused on developing novel biomaterials for cartilage regeneration. After years of hard work across scientific boundaries, the team discovered a solution that could benefit a large number of patients. However, the research team was faced with a question on how to proceed. Whether to continue the scientific path of unravelling the mysteries of cartilage regeneration or to focus on bringing the invention from bench to bedside? The latter would mean commercialisation of the invention, and for the scientists, taking a completely new career path. Taking this turn would mean risking the team members' scientific career, since running a start-up would inevitably mean lesser publications and other scientific merits in the forthcoming years. On the other hand, there was the potential to help a vast amount of patients. The team decided that the invention, a biodegradable weight-adaptive medical device for cartilage regeneration, was too promising to be left aside, so they made the choice to transform from academic researchers to entrepreneurs. Thus, Askel Healthcare Ltd was founded in March 2017. For a start-up operating in medical device sector, the company has a unique feature: the founding team is all-female. Not intentionally, but by a mere “side effect” of gathering the best talents to get the job done. The team continues to foster its strong scientific background, which is a true asset for a company focusing on tackling the big unmet medical need of cartilage regeneration.

Mesenchymal Stromal Cells (MSC) have been proposed as a potential therapy for a broad range of diseases including those affecting the musculoskeletal system. MSCs have received market authorization for treatment of graft versus host disease and fistulizing Crohn's disease. In addition, there are clinical trials underway for diseases affecting all organ systems. GMP manufactured cells are required for these clinical trials and suitable facilities with regulatory approval are thus crucial for the translational process. In this presentation I will review the process whereby such a facility has been constructed at NUI Galway and discuss challenges in operations and sustainability. Researchers at REMEDI and spin out company Orbsen Therapeutics are currently involved in 7 clinical trials using MSCs, 4 of which are EU wide consortia funded by the EU Commission. The presentation will also discuss issues such as source of MSCs, cell sorting, use of bioreactors and xeno-free processes.

Blood transfusion, organ and bone marrow transplantation and allogeneic tissue grafting create the potential for significant immunological challenges through the introduction of non-genetically identical major (HLA) and minor histocompatibility antigens (“allo-antigens”) into the body. Strategies to avoid the complications of immune responses against allo-antigens (transfusion reactions, rejection and graft versus host disease) include HLA matching, immunosuppressive therapies and immune tolerance promoting protocols. In the case of allogeneic mesenchymal stem/stromal cells (allo-MSC), it was initially believed that their combined properties of low HLA expression and inherent immune modulatory functions would render them invisible to the host immune system and, therefore, capable of being permanently accepted without further interventions. For clinical indications such as bone and tendon repair, in which permanent engraftment of allo-MSC or MSC-derived tissue constructs is particularly desirable, this model of “immune privilege” seemed almost too good to be true – and indeed, a decade of experimental research in this area has now convincingly demonstrated that allo-MSC typically elicit cellular (T-cell) and humoral (B-cell/antibody) immune responses in immunocompetent hosts – raising concern about their safety and long-term efficacy in human conditions. However, questions related to the immunogenicity of allo-MSC have evolved beyond a simple yes/no scenario to involve interesting observations and concepts about the potency, diversity, duration, functional characteristics and even potential clinical benfits of immunological responses to allo-MSC. In this presentation, I will summarise and critically evaluate current understanding of allo-MSC immunogenicity under experimental and clinical trial conditions with an emphasis on the implications for orthopaedic therapeutics.

Intra-synovial tendon injuries affect compressed tendon within a synovial environment (eg Rotator cuff tears of the shoulder) and frequently demonstrate ‘failed healing'. Current therapeutic methods for tendon tears (intra-synovial corticosteroid medication and surgical debridement) offer poor outcomes and new strategies for enhancing repair are needed. We have therefore evaluated two different approaches involving the use of mesenchymal stem cells and scaffolds. Bone marrow- and synovial-derived stem cells were capable of adhering to cut surfaces of tendon in vitro and modulating the release of extracellular matrix into the media. However, when administered in vivo into the digital flexor tendon sheath in naturally-occurring deep digital flexor tendon tears in horses and in an experimental model in sheep, neither cell type was capable of healing the tendon defect. Superparamagnetic iron oxide particle labelling of the implanted cells imaged using MRI and histologically revealed that cells only engraft into the synovium. In contrast a non-cellularised bilayered electrospun and woven polydioxanone scaffold, when used in the same experimental sheep model via a modified open approach and sutured over the created defect resulted in no local or systemic signs of excessive inflammation 3 months after implantation. All the tendon lesions healed with only a mild local inflammatory reaction and minimal-to-mild adhesion formation. Significant proliferative fibroblast infiltration was observed within and immediately adjacent to the implanted scaffold. The cellular infiltrate was accompanied by an extensive network of new blood vessel formation within the new tissue. In conclusion, the use of a scaffold to cover the defect appears to be a more successful strategy to repair intra-synovial tendon defects than intra-synovially injected mesenchymal stem cells. It remains to be tested whether the combination of the two techniques might offer an even better healing response.

This talk will initially give a brief overview of the motivations behind open access publishing and explain the practicalities of the different business models from an author's point of view. The talk will then discuss open access policy, particularly in Europe, and how the publishing landscape is constantly changing, with new initiatives and mandates being introduced all the time. Innovation in peer review such as transparent peer review and registered reports will be outlined and evaluated with examples from the BMC journals portfolio. The talk will then explain some of the funding options available to authors for open access publishing, and introduce the Springer Nature funding support service, which is available to anyone wishing to find out their options. Finally, the importance of data sharing will be discussed, as will the relatively new area of open access books.

You have a great research question or an idea for an innovation that will change your field. You have worked tirelessly to develop the project and are excited with the outcome. Now it is time to disseminate your findings to the world. This talk will give some insight into how to maximise the impact of your writing to reach the largest possible audience. It will discuss what makes a great paper, and provide pointers for navigating the editorial process, from your initial interactions with the editor to handling the sometimes-difficult process of peer review.

Congenital talipes equinovarus (CTEV), also known as club foot or talipes is a common congenital disorder. Parents are using the Internet more and more as a source of information about health care. Unfortunately, the quality of health care information on the Internet varies. This study looked at information available to parents using two instruments for judging the equality of information on the internet. The top five search engines were searched on Google. Three of these were also included in the top 50 sites in Ireland so these 3 sites were used. The phrases CTEV and club foot were searched from all 3 platforms. Websites were then scrutinized using the HON code and the DISCERN tool. 54 organic sites were found for the 3 search engines using the key word club foot. For the key word CTEV 55 matches were returned for the three search engines. 4 websites displayed the HON code. Using the discern tool CTEV websites had a mean score of 60 with a standard deviation of 17. While club foot had a mean score of 56.8 with a standard deviation of 13. Max score 80. Large volumes of information are available to parents on the Internet. Often parents find comfort in sharing experiences and feel empowered by learning about their children's illnesses. However, information provided on the interned can also be ambiguous and disingenuous. Practitioners should be aware of a number of key websites that parents can be directed towards.

The widespread dissemination of high-quality research facilitates keeping up to date with evidence-based practice, but the vast quantity can be overwhelming to physicians and surgeons. Information graphics, abbreviated to infographics, convey information using visualisations and images in an engaging manner. This format of presenting research format is preferable to 80% of clinicians when compared to text articles, and the long-term retention of information has been shown to be improve by a factor of 6.5 when methods were compared. Journal club was audited in our institution over 3 months. A multi-choice questionnaire was constructed weekly so as to test the attendees' recall of the research presented on a weekly basis. After five weeks, infographics were introduced, and the attendees' recall was assessed again on a weekly basis at the end of each journal club. The introduction of infographics to journal club saw improved test results from the journal club attendees. Not only was information retention improved, but the duration of journal club reduced following the intervention. Research can be disseminated efficiently using infographics in place of conventional journal club presentations. Satisfaction rates among clinicians, both with information retention and journal club duration, demonstrate the benefit of their use in teaching hospitals.

Tailoring your message to the people you are trying to engage will make it easier for them to listen, absorb and act on your message. Audiences can vary in a lot of ways such as gender, age, socio economic status, and ethnicity. Workshop attendees will learn how to identify specific audiences and deliver the topic of their research accordingly. This will help development of key messages to communicate research depending on what information would be interesting to the audience, the amount of science education the audience has, and how the audience will use the information afterwards.

Being able to communicate what your research entails quickly and effectively will ensure that you get the most important points across in a limited amount of time. Workshop attendees will develop a concise, compelling introduction to their research that can be communicated in a short message. This skill is beneficial in multiple scenarios, particularly when introducing yourself in an interview or a networking event. Additionally, this will help develop more effective ways of communicating through social media, press releases and conversations with people who do not have a scientific background.

Robust repair relies on blood flow. This vascularization is the major challenge faced by tissue engineering on the path to forming thick, implantable constructs. Without this vasculature, oxygen and nutrients cannot reach the cells located far from host blood vessels. To make viable constructs, tissue engineering takes advantage of the mechanical properties of synthetic materials, while combining them with extracellular matrix proteins to create a natural environment for the tissue- specific cells. Tropoelastin, the precursor of the elastin, is the extracellular matrix protein responsible for elasticity in diverse tissues, including robust blood vessels. We find that tropoelastin contributes a physical role in elasticity and also substantially to the biology of repairing tissue. The emerging model from a range of our in vivo studies is that tropoelastin encodes direct biological effects and has the versatility to promote repair. We have discovered that tropoelastin substantially improves healing by halving the time to repair bone in small animals and large animal preclinical models; tropoelastin elicits this response with early stage neo-angiogenesis, recruitment of endogenous cells with consistently accelerated repair. This potency is marked by the concerted appearance of blood vessels, tissue and phased cellular contributions that work together to accelerate repair.

CAR (CARSKNKDC) is a systemically administered wound-homing peptide that specifically recognizes angiogenic blood vessels and extravasates into sites of injury. CAR peptide requires heparan sulfate proteoglycans (HSPGs) for its cell penetrating activity. Syndecan-4 (SDC4) is a HSPG and binding to it triggers the wound re-epithelialization process. We have discovered that CAR peptide has the inherent ability to promote wound healing; wounds close and re-epithelialize significantly faster in CAR treated mice than in control groups (PBS and mutant peptide, i.e. mCAR injections). To delineate the molecular mechanism by which CAR accelerates wound healing, we focused on the requirement of HSPG binding for CAR peptide function. We demonstrate that CAR peptide endocytosis and its stimulation of keratinocyte cell migration are both dependent on SDC4. Finally, we show that the systemic administration of CAR peptide stimulates wound re-epithelialization only in WT mice, but not in SDC4 knockout (KO) mice. As SDC4 has very restricted expression in skin wounds, we propose that CAR peptide activates SDC4 function to promote re-epithelialization. CAR peptide may provide an entirely new way of enhancing wound healing, and perhaps tissue regeneration in general. This therapeutic approach is systemic, yet target organ- and cell- specific, and dependent on the naturally occurring SDC4 dependent migratory pathway that is crucial for tissue regeneration.

Collagen and hyaluronic acid are two major components of intervertebral disc (IVD). They give resistance and hydration to Nucleus Pulposus. In this study, we assessed the impact of Collagen (COLL) and Hyaluronic acid-Tyramine (THA) contents on the mechanical properties and the structure of composite hydrogels. For this purpose, a range of composites were obtained using a 4 mg/mL collagen concentration and different COLL/THA ratios from 8:1 to 1:5 (w/w). Composite gelling was performed by pH increase, triggering collagen fibrillogenesis and oxidative coupling of tyramine moieties in THA catalyzed by H₂O₂ and horseradish peroxidase (HRP). To modulate the THA gelling kinetic, different HRP concentrations (0.05; 0.1 and 0.5 U/mL) were used. Composites with a low THA content exhibited a fibrillar structure and possessed mechanical properties close to those of pure collagen hydrogels (200 Pa). From the ratio 1:1, the storage modulus increased to reach c.a 1200 Pa for the ratio 1:5. From the ratio 1:2, the fibrillar structure disappeared and sheets, characteristic of THA hydrogels, were observed. The HRP activity dramatically impacted the physical properties. A rapid THA gelling associated with a high THA content tended to destabilize collagen fibrils and promoted the formation of covalent bond between collagen and THA. On the opposite a slow gelling kinetic favored collagen fibril formation up to the COLL/THA ratio 1:2. Taken together, these results show that a slow gelling and an 8 mg/mL THA concentration are the appropriate conditions to obtain biomimetic biomaterials for the treatment of Nucleus Pulposus.

The enthesis is a specialised zonal tissue interface between tendon and bone, essential for adequate force transmission and composed by four distinct zones (tendon, fibrocartilage, mineralized fibrocartilage and bone). After injury, the native structure is often not re-established and a mechanically weaker fibrovascular scar is formed. Traditionally used monotherapies have failed to be effective, posing the need for multi-cargo localized delivery vehicles. We hypothesize that multilayer collagen-based scaffolds can serve as delivery vehicles for specific bioactive molecules with tenogenic, chondrogenic and osteogenic potential to enhance the functional regeneration of the enthesis. Three-layer scaffolds composed by a tendon-like layer of collagen type I, a cartilage-like layer of collagen type II and a bone-like layer of collagen type I and hydroxyapatite were fabricated by an iterative layering freeze-drying technique. The scaffolds were cross-linked with varying concentration of 4-arm polyethylene glycol (4s-PEG) and the biological and mechanical properties were assessed. Each layer was functionalized with platelet-derived growth factor, insulin growth factor, heparan sulfate or bone morphogenetic protein 7 and their tenogenic, chondrogenic and osteogenic potential on bone-marrow derived stem cells was investigated in vitro. Scaffolds cross-linked with 1 mM 4s-PEG showed 60% free amines reduction respect to non-cross-linked scaffolds, were stable in collagenase over 24 hours and had a compression modulus of 30 kPa. The bioactive molecules had a sustained release profile (approximately 50 ng/mL) over 5 days as a function of cross-linking. Preliminary in vitro studies confirmed the chondrogenic potential of heparin sulfate and insulin growth factor by the increase of proteoglycans.

Tissue engineering is a promising approach to regenerate damaged skeletal tissues. In particular, the use of injectable hydrogels alleviates common issues of poor cell viability and engraftment. However, uncontrolled cell fate, resulting from unphysiological environments and degradation rates, still remain a hurdle and impedes tissue healing. We thus aim at developing a new platform of injectable hyaluronic acid (HA) hydrogels with a large panel of properties (stiffness, degradation…) matching those of skeletal tissues. Hence, HA with different molecular weights were functionalized with silylated moieties. Upon injection, these hydrogels formed through a sol-gel chemistry within 5 to 20 minutes in physiological conditions, as demonstrated by rheological characterization. By varying the crosslinking density and concentration, we obtained hydrogels spanning a large range of elastic moduli (E = 0.1–20 kPa), similar to those of native ECMs, with tunable biodegradation rates (from 24 hours to > 50 days) and swelling ratios (500 to 5000% (w/w)). Cell viability was confirmed by Live/Dead assays and will be completed by in vivo subcutaneous implantations in mice to study the foreign body reaction and degradation rate. We further developed hybrid HA/biphasic calcium phosphate granules hydrogels and demonstrated a strong mechanical reinforcement (E = 0.1 MPa) and a faster relaxation behaviour (τ_1/2 < 400s), with similar degradation rates. Ongoing in vitro differentiation assays and in vivo implantations in a rabbit femur model will further assess their ability to drive bone regeneration. Collectively, these results suggest that this hydrogel platform offers promising outcomes for improved strategies in skeletal tissue engineering.

The rheological properties of synovial fluid (SF) are largely attributed to the presence of high molecular weight hyaluronic acid (HA). In normal SF, HA has been shown to be an anti-inflammatory molecule able to increase the viscosity and promote endogenous production of HA. The aim of the present report was to investigate the possible effect of HA concentration in rheological properties (elastic modulus, G´ and viscous modulus, G´´) of osteoarthritic equine SF. For this purpose, SF from intercarpal, metacarpophalangeal and distal interphalangeal joint was aspirated by aseptic arthrocentesis from 60 Warmblood horses. For determining HA concentrations in equine SF samples, a commercially available ELISA kit was used. Additionally, full rheological sample characterization was carried out with an AR-G2 rheometer (TA Instruments Ltd., UK) in order to measure the elastic G´ and viscous G´´ moduli, at horse's body (37.5 ºC) temperature. The ANOVA findings revealed statistically significant main effects of the factors Joint Type (p = 0.001), and main effects of covariates Age (p = 0.019) and HA (p < 0.001) on the mean values of logG” and logG' measurements. Interpreting the coefficients of the covariate HA, a positive correlation of HA was detected on the response logG” and logG' measurements. Collectively, these data illustrate the role of HA in equine pathological SF.

Phosphate metabolism is central to the bone formation pathway. Phosphate is shuttled through the cell membrane to the mitochondria, where it is polymerised to form adenosine triphosphate. Once exocytosed the ATP may then be cleaved to form pyro and orthophosphates, the balance of which can determine whether mineralisation occurs or not. We are developing a range of materials at the University of Birmingham that have been formulated so that they can influence this balance, with the potential either to drive or prevent mineralisation from occurring. This talk will describe how we have used this process to develop materials that can be used to stimulate bone formation around an implant or to prevent the formation of pathological bone. It will also talk about the steps that we have taken to move these therapies towards clinical trial.

The nature of the initial interaction between calcium phosphate (CaP) thin films and osteoblasts can be mediated by the outermost surface properties of that material. As such, the phase, crystallinity, stoichiometry, composition and morphology of the CaP surfaces are seen as key parameters that must be accurately controlled in order to influence their potential biofunctionality with respect to osteoblasts. Hydroxyapatite [HA – Ca₁₀(PO₄)₆(OH)₂] has been extensively studied due to the structural and chemical similarities demonstrated with the main inorganic constituent of bone tissue and teeth. However, it is well documented that biological hydroxyapatite, which forms the mineral phases of calcified tissues, differ from pure and synthetically produced HA. Biological apatite is comprised of a mixture of calcium phosphate phases and trace elements, e.g., strontium, zinc, magnesium and silicon. As such, when designing CaP biomaterials for clinical use (both bulk materials and coatings) one proposed route would be to introduce multiple ionic substitutions into HA in order to mimic the complex chemistry of human bone and thereby improve the biological performance of such materials, both in vitro and in vivo. This presentation will explore a novel approach to depositing substituted and co-substituted CaP systems onto a range of different substrates types, namely metal and polymers. In particular, this presentation will examine how the surface properties of bioinert polymers, such as Poly(etheretherketone) (PEEK) accurately controlled in order to provide an enhanced in vitro performance. The presentation will also look at how resorbable magnesium implants can also be manipulated to provide both enhanced bioactivity and to provide a route to control how they resorb in a physiological environment.

The enhancement of current bone cement properties is a challenging issue that has been the focus of much research. Developing bone composites with high level of cytocompatibility, mechanical and antibacterial properties is a challenging task. We overcome this challenge by designing a nanocomposite that contain two-dimensional (2D) nanosheets. To develop our novel bone cement nanocomposite, 2D nanosheets were synthesized, mixed in different ratios, and then added to the PMMA matrix. The results reveal that the incorporation of 2D nanosheets into the PMMA matrix leads to increase in the antibacterial properties of the bone cement composite against E. coli bacteria. In addition, the 2D nanosheets improve the compression strength of the bone cement nanocomposite significantly. We also show that nanosheets increased the bioactivity of the bone cements. Finally, MTT assay results indicate that PMMA as a control sample has the lowest cytocompatibility, however, our novel nanocomposites have the highest amount of cytocompatibility. Thus, the current study suggests that 2D nanosheets are potential filler components for the next generation of PMMA bone cement nanocomposites. The findings of this work reveal that the excellent performance of the proposed bone composite can result in a paradigm shift in design of state-of-the art bone cement composites.

Crosslinking has been already used for about 80 years to enhance the longevity of polyethylene cables. The polymer alteration has been achieved with peroxide, silane or irradiation. The medical devices industry discovered the benefit of this technology for its tribological applications like hip or knee bearings in the 2000s as crosslinking improves considerably the abrasion resistance of the material. The more current methods used are Gamma and Beta irradiation. On the basis of economical (rising prices of Cobalt), environmental (the radioactive source can not be turned off), technological (low dose rate) drawbacks for Gamma respectively low penetration for Beta irradiation we decided to investigate an alternative technology: the X-Ray irradiation, which provides a homogeneous crosslinking in a relatively short time. We analyzed the wear, mechanical, thermal, oxidative and network properties of two vitamin E doped UHMWPE: first crosslinked with E-Beam, second with X-Ray. There wasn't any significant difference between the X-Ray and the E-Beam crosslinked material.

Hydrogels are hydrated 3-dimensional (3D) polymer networks that can be chemically or physically crosslinked. Interest in the use of hydrogels for tissue engineering applications has been growing in the past few decades due to their excellent biocompatibility and biodegradability. One of the major drawbacks of the use of hydrogels in such applications is their lack of structural strength. To address this, in this work, we have combined two hydrogel types, namely gelatin and alginate. In this work, a 1 ml volume of gelatin alginate hydrogel was molded in each well of a 24 well-plate and crosslinked with different concentrations of calcium chloride (CaCl₂) (20, 40, 60, 80, and 100 mM) to investigate the influence of concentration on hydrogel properties and cell viability. The hydrogel was characterized using Fourier transform infrared (FTIR) spectrometry, environmental scanning electron microscopy (ESEM), and an Alamar blue assay to assess the chemical structure, the surface morphology, and the epithelial cell viability of the hydrogel, respectively. The FTIR analysis shows that network formation improved with increasing concentration; decreased ion-polymer interactions have been noted for concentrations ≤ 60 mM. This appears to be in agreement with ESEM images that show an evolution from a smooth, featureless surface to the appearance of surface pore structure for concentrations ≥ 80 mM. Perhaps as ion concentration increases and network formation improves, the effect is evidenced as surface porosity; low concentrations result in swelling and a smooth surface. In terms of cell viability, viability has been found to increase with increasing concentration. The cell viability is 90 % at 100 mM CaCl₂, in contrast to 50 % for a concentration of 20 mM after 9 days of incubation. It is possible that the reduced viability can be attributed to the high proportion of uncrosslinked polymer chains at low concentrations. Overall, these results provide useful information about the role of crosslinking concentration on hydrogel properties, knowledge that may be applied to 3D bioprinting.

Porcine and fish by-products in particular are rich sources for collagen, which is the main component of the extracellular matrix (ECM). Although there are studies investigating different collagen derived from various tissue sources for the purpose of creating biomaterials, the comparison of biophysical, biochemical and biological properties of type II collagen isolated from cartilaginous tissues has yet to be assessed. In addition, it has been shown from previous studies that sex steroid hormones affect the collagen content in male and female animals, herein, type II collagens from male and female porcine cartilage were assessed in order to investigate gender effects on the property of collagen scaffolds. Moreover, type II collagen has a supportive role in articular cartilage in the knee joint. Therefore, the aim is to assess the properties of type II collagen scaffolds as a function of species, tissue and gender for cartilage regeneration. Type II collagen was extracted from male and female porcine trachea, auricular, articular cartilage and cartilaginous fish through acid-pepsin digestion at 4°C. SDS-PAGE was conducted to confirm the purity of extracted collagen. Collagen sponges were created via freeze-drying. Scaffold structure and pore size were evaluated by scanning electron microscopy (SEM). Thermal stability was assessed by differential scanning calorimetry (DSC). Sponges were seeded with human adipose derived stem cells to assess chondro-inductive potential of collagen sponges after 7, 14 and 21 days of culture. In conclusion, collagen sponges support the proliferation and differentiation of human adipose derived stem cells to different extents.

Tendon pathologies represent an unresolved clinical challenge where the patients suffer from pain and impaired mobility. One of the most frequently ruptured tendons is the Achilles tendon and primarily seen in recreational and professional athletes. A study from Sweden reported a significant increase in the incidence of Achilles tendon ruptures of 17% in men and 22% in women due to the demographic changes and the higher sportive activity of older adults (Huttunen TT Am J Sports Med 2014). The re-rupture rate is between 2–10%, and the patients suffer from an impairment over a long time accompanied with incapability to work. The healing process results in the formation of a mechanically insufficient scar tissue. A detailed knowledge on the cellular and molecular processes underlying human Achilles tendon healing is necessary to develop new treatment strategies and judge therapeutic success. The analysis of human Achilles tendon samples at different time points post rupture and the comparison to intact and degenerated tendon tissue provides important information on the healing process.

Muscle and tendon have an adaptive, symbiotic biomechanical relationship that is drastically altered following acute tendon injury. Such injuries, like Achilles tendon rupture (ATR), do not only lead to impairments in the resultant tendinous tissue, but also to irrecoverable atrophy in the connected muscle in series. As a result, a new relationship between muscle and tendon is established after ATR, leading to lasting functional deficits in the lower limb. It remains unclear how these develop, particularly since this imbalance may be influenced by the dependent relationship of the two tissues to each other. A further confounding factor is that tendon and muscle tissues adapt on different time scales in response to mechanical loading, such as those introduced during rehabilitation. Thus, it is warranted to perform assessments not only of the overall muscle-tendon unit, but also its constituent tissues. This presentation will discuss findings from both short-term and long-term follow-ups of ATR patients, with a focus on the recovery of gait and changes in the muscle-tendon unit tissues following ATR repair. Both the influence of the rehabilitation process and suggestions for future research directions will be additionally presented.

Unresolved inflammatory processes in tendon healing have been related to the progression of tendinopathies. Thus, the management of tendon injuries may rely on cell-based strategies to identify and modulate tendon inflammatory cues. Pulsed electromagnetic field (PEMF) has been approved by FDA for orthopedics therapies and has been related to a reduction in pain and to improve healing. However, the influence of PEMF in tendon healing remains largely unknown. Human tendon resident cells (hTDCs) were cultured in an inflammatory environment induced by exogenous supplementation of IL-1β and their response assessed after exposure to different PEMF treatments. This study demonstrates that IL-1β induced up-regulation of pro-inflammatory factors (IL-6 and TNFα) and extracellular matrix components (MMP−1, −2, −3) whereas reduces the expression of TIMP-1, suggesting IL-1β as a candidate inflammation model to study hTDCs response to inflammation cues. Moreover, in both homeostatic and inflammatory environments, hTDCs respond differently to PEMF treatment suggesting that cells are sensitive to magnetic field parameters such as strength (1.5 – 5mT), frequency (5–17Hz) and duration (10–50% duty cycle, dc). Among the conditions studied, PEMF treatment with 4mT/5Hz/50%dc suppresses the inflammatory response of hTDCs to the IL-1β stimulation, as evidenced by the decreases amount of IL-6, TNFα and downregulation of MMP-1, −2, −3 and COX-2, IL-8, IL-6, TNFα genes. These results demonstrate the potential of PEMF, in particular 4mT/5Hz/50%dc PEMF in treating tendon inflammation suppressing the inflammatory stimulation induced by IL-1β, which may be beneficial for tendon healing strategies.

During in vitro sub-culturing, tenocytes lose their phenotype which ultimately affects their functioning. As spindle-shaped fibroblasts, tenocytes have a unique thin elongated phenotype and they possess more spread-out shape through phenomena named dedifferentiation1. Given the link between cell shape and cell function, in this study, we first aimed to dedifferentiate tenocytes through in vitro sub-culturing in order to have a model system for dedifferentiation. For this, we isolated human flexor tendon cells from healthy female flexor digitorum longus and seeded at 5000 cells/cm² cell density, passaged every two days for six passages. In order to assess cell phenotype, we fixed with 4% paraformaldehyde and stained with phalloidin and DAPI to visualize the actin cytoskeleton and DNA respectively. We noted that in each passage, cells lost their spindle-shaped phenotype and became more pancake-shaped. At passage 1 and 2, the main cell phenotype is spindle-shaped. However, as the cells are further passaged, the phenotype of the cell population becomes more heterogeneous and at passage 5 and 6, they already display a more spread-out shape. Based on these results, we further hypothesized that they can be re-differentiated through matrix-mediated mechano-transduction and regain their morphology and function. For this aim, we generated decellularized tendon from porcine Achilles tendon and setup a mechanical loading system where we can provide mechanical loadings at physiological levels. This system will provide a new approach on in vitro tenocyte culturing.

Cellular therapies play an important role in tendon tissue engineering with tenocytes being described as the most prominent cell population if available in large numbers. In vitro expansion of tenocytes in standard culture leads to phenotypic drift and cellular senescence. Maintenance of tenogenic phenotype in vitro can be achieved by recapitulating different aspects of the tendon microenvironment. One approach used to modulate in vitro microenvironment and enhance extracellular matrix (ECM) deposition is macromolecular crowding (MMC). In addition, as tendon has been described to be a relatively avascular and hypoxic tissue and low oxygen tension can stimulate collagen synthesis and cross-linking through the activation of hypoxia-inducible factor 1-alpha (HIF1-α), we venture to assess the synergistic effect of MMC and low oxygen tension on human tenocyte phenotype maintenance. SDS-PAGE and immunocytochemistry analysis demonstrated that human tenocytes treated with MMC at 2 % oxygen tension showed increased synthesis and deposition of collagen type I. Moreover, immunocytochemistry for the tendon-specific ECM proteins collagen type III, V, VI and fibronectin illustrated enhanced deposition when cells were treated with MMC at 2 % oxygen tension. In addition, western blot analysis revealed increased expression of tendon-specific protein Scleraxis, while a detailed gene analysis illustrated upregulation of tendon-specific genes and downregulation of trans-differentiation genes again when cells cultured with MMC under hypoxic conditions. Collectively, results suggest that the synergistic effect of MMC and low oxygen tension can accelerate the formation of ECM-rich substitutes, which stimulates tenogenic phenotype maintenance.

This investigation of elite male collegiate basketball players aims to determine 1) the change in 3D dynamic functional variables across a single season and 2) correlate cross-season changes in functional variables with changes in clinical and quantitative ultrasound measures. Eleven male college basketball players (mean age 19, range 18–21 years) from a single team underwent baseline patellar tendon shear wave (SW) elastography and dynamic function at the start of the season (Visit1) and at a late-season time point (Visit2). Players reported their VISA-P scores every two weeks across their 24-week season. Each athlete performed a box-ground-box jump five times while 3D lower extremity kinematic and kinetic variables were collected. Functional measures included for landing (LAND) and take-off (TOFF) phases: knee valgus angle, valgus torque, and peak limb force. Knee valgus angular impulse and ground contact time were also measured. Paired t-tests and Pearson correlation coefficients (r) compared Visit1 and Visit2 variables and assessed the strength of linear dependency, respectively. The mean change in VISA-P score was 15.18 (+/-8.55). No functional variables were different across the season. Clinical, quantitative ultrasound and functional variables were moderately correlated with take-off valgus moment, landing force, take-off force and contact time. Other correlations were low (< 0.4). Our analyses have shown moderate correlations between important clinical, quantitative imaging and function measurements. These correlations reflect the changes that occur between relevant time points and which relate internal structure and external function.

Ceramics such as hydroxyapatite are routinely used in fracture repair. However, their effects could be significantly improved as its bioavailability is incredibly poor (issues including low solubility, anionic charge, tendency to agglomerate). Nanoscale hydroxyapatite are gaining much interest, demonstrating increased effectiveness when compared to their micro-sized counterpart. In this study, we have developed a bioactive cargo–polymer-based system that allowed for the sustained, localised non-viral delivery of hydroxyapatite nanoparticles using an amphipathic peptide as a capping agent. The nanoparticles were delivered from a polycaprolactone nanofibre reinforced novel Alg-co-PNIPAAm thermoresponsive hydrogel. The bioactive cargo–polymer-based system was characterised in terms of its physiochemical properties, in vitro properties and in vivo performance using a subcutaneous mouse model. From this study, we have demonstrated that osteogenesis and bone regeneration were significantly increased when our novel capping agent was used to limit the particle size distribution and optimised the physiochemical characteristics of nanoscale hydroxyapatite (i.e. reducing risk of agglomeration and increasing its bioavailability). Additionally, the dual functionality of the thermoresponsive hydrogel as a scaffold for bone regeneration and as a vehicle for the sustained, local delivery of hydroxyapatite nanoparticles over an extended period was successfully demonstrated.

All types of regenerative materials, including metal implants, porous scaffolds and cell-laden hydrogels, interact with the living tissue and cells. Such interaction is key to the settlement and regenerative outcomes of the biomaterials. Notably, the immune reactions from the host body crucially mediate the tissue-biomaterials interactions. Macrophages (as well as monocytes and neutrophils), traditionally best known as defenders, accumulate at the tissue-biomaterials interface and secrete abundant cytokines to create a microenvironment that benefits or inhibits regeneration. Because the phenotype of these cells is highly plastic in response to varying stimuli, it may be feasible to manipulate their activity at the interface and harness their power to mediate bone regeneration. Towards this goal, our team have been working on macrophage-driven bone regeneration in two aspects. First, targeting the abundant, glucan/mannan-recognising receptors on macrophages, we have devised a series of glucomannan polymers that can stimulate macrophages to secrete pro-osteogenic cytokines, and applied them as coating polymer of mesenchymal stem cells-laden hydrogels. Second, targeting the toll-like receptors (TLRs) on macrophages, we have screened TLR-activating polysaccharides and picked up zymosan (beta-glucan) to be modified onto titanium and glass implants. We evaluated both the efficacy of integration and safety of immune stimulation in both in vitro and in vivo models. Our future exploration lies in further elaborating the different roles and mechanisms of macrophages of various types and origins in the regenerative process.

Intra-articular infusions of adipose tissue-derived stem cells (ASCs) are a promising tool for bone regenerative medicine, thanks to their multilineage differentiating ability. One major limitation of ASCs is represented by the necessity to be isolated and expanded through in vitro culture, thus a strong interest was generated by the adipose stromal vascular fraction (SVF), the non-cultured fraction of ASCs. Besides the easiness of retrieval, handling and good availability, SVF is a heterogeneous population able to differentiate in vitro into osteoblasts, chondrocytes and adipocytes, according to the different stimuli received. We investigated and compared the bone regenerative potential of SVF and ASCs, through their ability to grow on SmartBone^®, a composite xenohybrid bone scaffold. SVF plated on SmartBone^® showed better osteoinductive capabilities than ASCs. Collagen I, osteocalcin and TGF↕ markedly stained the new tissue on SmartBone^®; microCT analysis indicated a progressive increase in mineralised tissue apposition by quantification of newly formed trabeculae (3391 ± 270,5 vs 1825 ± 133,4, p± 0,001); an increased secretion of soluble factors stimulating osteoblasts, as VEGF (153,5 to 1278,1 pg/ml) and endothelin 1 (0,43 to 1,47 pg/ml), was detected over time. In conclusion, the usage of SVF, whose handling doesn't require manipulation in an in vitro culture, could definitively represent a benefit for a larger use in clinical applications. Our data strongly support an innovative idea for a bone regenerative medicine based on resorbable scaffold seeded with SVF, which will improve the precision of stem cells implant and the quality of new bone formation.

Mesenchymal stromal cells (MSCs) are widely used in clinical trials for the treatment of many bone defects. Apatite-wollastonite glass ceramic (A-W) is an osteoconductive biomaterial shown to be compatible with MSCs. This is the first study comparing the osteogenic potential of two MSC populations, heterogeneous plastic adherence MSCs (PA-MSCs) and CD271-enriched MSCs (CD271-MSCs), when cultured on A-W 3D scaffold. The paired MSC populations were assessed for their attachment, growth kinetics and ALP activity using confocal or scanning electron microscopy and the quantifications of DNA contents and p-nitrophenyl (pNP) production. While the PA-MSCs and CD271-MSCs had similar expansion and tri-lineage differentiation capacity during standard 2D culture, they showed different proliferation kinetics when seeded on the A-W scaffolds. PA-MSCs displayed a well-spread attachment with more elongated morphology compared to CD271-MSCs, signifying a different level of interaction between the cell populations and the scaffold surface. PA-MSCs also fully integrated into the scaffold surface and showed a stronger propensity for osteogenic differentiation on the A-W scaffold as indicated by higher ALP activity than CD271-MSCs. Furthermore, A-W scaffold seeded uncultured bone marrow mononuclear cells (BM-MNCs) demonstrated a higher proliferation rate and greater ALP activity compared to freshly isolated CD271-enriched BM-MNCs. Our findings suggest that enrichment of CD271-positive population is not beneficial for osteogenesis when the cells are seeded on A-W scaffold. Furthermore, unselected heterogeneous MSCs or BM-MNCs are more promising for A-W scaffold-based bone regeneration, providing novel insight with potential clinical implications in regenerative medicine for bone defects using an innovative tissue engineering approach.

The human amniotic membrane (hAM) may be helpful as a support for bone regeneration. To assess its potential for bone repair, a wide heterogeneity of preservation methods of hAM has been studied. The objectives of this study were: i) to assess bone regeneration potential of fresh versus cryopreserved hAM, and ii) to characterize hAM depending on four preservation methods. hAM was used either fresh (F-hAM), cryopreserved (C-hAM), lyophilized (L-hAM) or decellularized and lyophilized (DL-hAM). First, critical calvarial bone defects were performed in mice. Defects remained empty or were covered by F-hAM or C-hAM. Then, the cytotoxicity of the four preservation methods of hAM was assessed in vitro on human bone marrow mesenchymal stem cells (hBMSCs), and, their biocompatibility was evaluated in vivo in a rat subcutaneous model. X-Rays analysis showed that no calvarial defect was regenerated ad integrum. Bone regeneration was slightly enhanced by C-hAM. In vitro, the decellularization and the lyophilization process did not confer any cytotoxicity of the tissue compared to other preservation methods. In vivo, L-hAM and DL-hAM were easier to handle. Histological analysis of explanted samples from the rat indicated a slight to moderate inflammatory reaction with hAM. One month after surgery, a complete resorption of F-hAM and C-hAM implants occured, whereas L-hAM and DL-hAM were still observed. C-hAM has a limited potential for GBR. L-hAM and DL-hAM are biocompatible without cytotoxic effects. These preservation methods should be suitable in the field of bone regeneration.

Segmental bone transport (SBT) with an external fixator has become a standard method for treatment of large bone defect. However, a long time-application of devices can be very troublesome and complications such as nonunion is sometimes seen at docking site. Although there have been several studies on SBT with large animal models, they were unsuitable for conducting drug application to improve SBT. The purpose of this study was to establish a bone transport model in mice. Six-month-old C57BL/6J mice were divided randomly into bone transport group (group BT) and an immobile control group (group EF). In each group, a 2-mm bone defect was created in the right femur. Group BT was reconstructed by SBT with external fixator (MouseExFix segment transport, RISystem, Switzerland) and group EF was fixed simply with unilateral external fixator (MouseExFix simple). In group BT, a bone segment was transported by 0.2 mm per day. Radiological and histological studies were conducted at 3 and 8 weeks after the surgery. In group BT, radiological data showed regenerative new bone consolidation at 8 weeks after the surgery, whereas high rate of nonunion was observed at the docking site. Histological data showed intramembranous and endochondral ossification. Group EF showed no bone union. In this study, experimental group showed good regenerative new bone formation and was similar ossification pattern to previous large animal models. Thus, the utilization of this bone defect mice model allows to design future studies with standardized mechanical conditions for analyzing mechanisms of bone regeneration induced by SBT.

The discussion will focus on new approaches to reduce bacterial adhesion on the surface of polymethylmethacrylate (PMMA) in contact with bone, comparing the clinical and engineering point of view. One possibility is to encourage and speed up direct interaction with the bone, for example by adding a bioactive phase in the cement (hydroxyapatite, glass and bioactive glass ceramic). A widespread strategy is also the addition of different types of antibiotics (gentamicin, tobramycin vancomycin, etc.), although they are known to have some drawbacks: not complete release, resistant strain development. Another strategy could be represented by the PMMA-based composite cements loaded with a completely inorganic filler consisting of a bioactive glass doped with ions whose bioactivity mechanism is well-known and encompasses a chemical and biological interaction with tissues promoting osteoinduction. Bioactive glasses can be doped with antibacterial ions (silver, copper, etc.) preserving their biocompatibility and bioactivity and, at the same time, acquiring antibacterial properties. Thus, it is possible to produce composite cements that combine the properties of the polymer matrix with those of the inorganic filler, overcoming the main problems associated with the use of antibiotics. An additional possibility is the addition of essential oils, vegetable oils with remarkable antibacterial properties.

Post-surgical infections are still one of the most frequent adverse events in the prosthetic surgery. PMMA-based cements are widely employed in orthopaedic surgery as filler or prosthetic fixing device. The main problems associated with this material are poor bone integration and infection development. Aiming to avoid bacterial adhesion and to extend the longevity of implants, different solutions were proposed, both in terms of operative procedures and new materials development. Regarding the materials advancement, innovative PMMA-based composite bone cements, contemporaneously bioactive and antibacterial (without the use of antibiotics), were developed. The composites are based on a PMMA matrix containing a bioactive glass, doped with antibacterial ions (Ag+ or Cu++); so, the same filler shows at the same time the ability of promoting bone ingrowth and an antibacterial effect. Composite cements were characterized in terms of morphology and composition, curing parameters and mechanical properties; in vitro tests were performed to verify the material ability to release antibacterial ions and to promote the precipitation of hydroxyapatite. Moreover, cytotoxicity and antimicrobial properties were verified. The cements characteristics were tested using different commercial matrix and different viscosities; therefore, the proposed formulations represent an innovative solution for a new family of antibiotic-free, bioactive and antibacterial cements.

Infection is one of the most serious complications of orthopedic surgery, particularly in implant-related procedures. Minimum inhibitory concentration (MIC) for identified bacteria is an important factor for successful antibiotic treatment. We investigated the MIC of antibiotics in Staphylococcus species from orthopedic infections, comparing with isolates from respiratory medicine. Staphylococcus species isolated in our laboratory from January 2013 to July 2016 were retrospectively reviewed. The MIC of vancomycin (VCM), arbekacin (ABK), teicoplanin (TEIC), linezolid (LZD), and rifampicin (RFP) was reviewed. Differences in the MIC of each antibiotic in orthopedic and respiratory samples were determined. A total of 259 isolates were evaluated (89 orthopedic, 170 respiratory). Staphylococcus aureus was the most commonly identified species (58%). In comparison with orthopedic samples, the number of isolates with a VCM MIC <0.5 μg/ml in methicillin sensitive staphylococcus aureus (MSSA) was significantly higher in respiratory isolates, while a MIC of 2 μg/ml was significantly lower (P = 0.0078). The proportion of isolates with a VCM MIC of 2 μg/ml in methicillin-resistant coagulase-negative staphylococci (MRCNS) was significantly higher in orthopedic isolates than that seen in respiratory isolates of methicillin-resistant staphylococcus aureus (MRSA; P < 0.001). When comparing MRCNS and other orthopedic Staphylococci, the rate of RFP MIC >2 μg/ml in MRCNS isolates was significantly higher (P = 0.0058). The MIC of VCM in Staphylococcus species from orthopedic infection was higher than that of respiratory samples, particularly in MRCNS from implant-related samples. MRCNS showed a significantly higher rate of resistance for RFP versus other orthopedic isolates.

Biphasic calcium phosphates (BCP) are the most frequently used materials because of their mineral analogy with bio-mineral part of bones. Their chemical synthesis can be modulated by doping, in order to respond to the biological needs. We present here the biological responses induced by copper ions in solution, to characterize its cytotoxicity and antibacterial activity. We also investigate the antibacterial property of Cu-doped BCP (Ca10 Cu0.1 (PO4)6 (OH)1.8 O0.2) on a strain of clinical interest: S. aureus, compared to undoped BCP. The sol-gel route has been used to prepare the BCP ceramics. Human BMC (Bone Marrow Cells) were obtained from metaphysal cancellous bone collected during hip arthroplasty and used for cytotoxicity evaluations. A strain of Staphylococcus aureus isolated from an osteoarticular infection after total knee arthroplasty was used to evaluate antibacterial activities. Results indicate that 3 ppm of copper ions leads to the death of all cultured bacteria in 24 hours and 25 ppm caused the death of all cells in 15 days. Regarding BCP, the undoped bioceramics increased the bacterial growth compared to a control without bioceramic. After 16 hours of contact, the copper ions released by the Cu-doped BCP induced a significant decrease of the bacterial concentration, indeed no viable bacteria were found. These materials seem to be a promising alternative for the preparation of multifunctional bone substitutes.

With an ever-increasing aging population, total hip and knee arthroplasty is projected to increase by 137% and 601%, respectively, between the period; 2005–2030. Prosthetic Join Infection (PJI) occurs in approximately 2% of total joint replacements (TJRs) in the U.S. PJI is primarily caused by adherence of bacteria to the surface of the prosthesis, ultimately forming an irreversibly attached community of sessile bacteria, known as a biofilm, highly tolerant to antibiotic treatment. Often the only resolution if the ensuing chronic infection is surgical removal of the implant – at high cost for the patient (increased morbidity), and for healthcare resources. Strategies to prevent bacterial adherence have significant potential for medical impact. Laser surface treatment using an automated continuous wave (CW) fiber laser system has shown promise in producing anti-adherent and bactericidal surfaces. Work presented here aims to investigate the effect of this approach on orthopaedic metals as a proof of concept, specifically Ti-6Al-4V (kindly supplied by Stryker Orthopaedics, Limerick). A coupon was surface treated using a laser (MLS-4030; Micro Lasersystems BV, Driel). Samples were incubated in Müller Hinton Broth (MHB) inoculated with methicillin resistant Staphylococcus aureus (MRSA; ATCC 43300) for 24h before Live/Dead staining (BacLight™ solution; Molecular Probes) and inspection by fluorescence microscopy (GXM-L3201 LED; GX Optical). Images were analysed using ImageJ software (NIH) and a significant reduction (p > 0.05, n=24) in total biofilm coverage and Live/Dead ratio was observed between the laser treated and as received surfaces. This data demonstrates the anti-adherent, and indeed bactericidal, effect of Laser-surface treatment.

Periprosthetic joint infections (PJIs) are uncommon but are devastating complications of total knee replacement (TKR). We analysed the risk factors of revision for PJI following primary TKR and their association with PJI at different post-operative periods. Primary TKRs and subsequent revision surgeries performed for PJI from 2003–2014 were identified from the National Joint Registry (NJR). Multilevel piece-wise exponential non-proportional hazards models were used to estimate the effect of the investigated factors at different post-operative periods. Patient, perioperative and healthcare system characteristics were investigated and data from the Hospital Episode Statistics for England were linked to obtain information on specific comorbidities. The index TKRs consisted of 679,010 primaries with 3,659 subsequently revised for PJI, 7% within 3 months, 6% between 3–6months, 17% between 6–12months, 27% between 1–2years and 43% ≥2 years from the index procedure. Risk factors for revision for PJI included male sex, high BMI, high ASA grade and young age. Patients with chronic pulmonary disease, diabetes and liver disease had higher risk of revision for PJI, as had patients who had a primary TKR for an indication of trauma or inflammatory arthropathy. Surgical procedure, fixation method, constraint and bearing type influenced the risk of revision for PJI. Their effects were period-specific. No or small associations were found with the operating surgeon grade, surgical volume and hospital surgical volume. These findings from the world's largest joint replacement registry show a more complex picture than the meta-analyses published to date with specific time-dependent effects for the identified risk factors.

Cells with stem/progenitor characteristics can be isolated from articular cartilage and may have utility in cartilage repair and regeneration therapies. Unlike other adult cell types with differentiation capabilities, clonal chondroprogenitors differentiate into cartilage that resembles stable cartilage rather than endochondral cartilage. We have isolated a large series of chondroprogenitor clones from normal human articular cartilage from individuals of one to forty-five years of age and characterized them with known and novel markers. The clones were isolated separately from different zones of the articular cartilage. As first reported by others, the cloneable cells were mainly found in the upper zones. However, there are clones with chondroprogenitor status in the deeper zones, albeit at far lower frequency. These deep zone clones have different characteristics to those from the upper zones. We have used selected clones to re-engineer stable cartilage with use of the right environmental conditions (growth factors, oxygen level etc).

One of the core tenets of our philosophy for tissue regeneration include the use of “raw materials,” where biomaterials themselves serve as both building blocks and bioactive signals. In recent years, a few groups around the world have gravitated toward cartilage matrix as a potentially chondroinductive material for cartilage regeneration. The major challenge to date in cartilage injury has been creating a biomaterial-only strategy that is capable of regenerating true hyaline-like cartilage without the addition of growth factors or exogenous cells. In the past few years, we have focused our efforts on establishing chondroinductivity in vitro, and in developing new materials synthesis strategies to provide ease of application for orthopedic surgeons in the operating room. By leveraging nanotechnology, we have developed a paste-like material constructed from cartilage matrix with encouraging mechanical performance post-crosslinking, and which avoids contraction after extended time. Looking to the future, we are working on next-generation approaches to chondroinductive materials. We have encouraging preliminary data which suggest the possibility of a chondroinductive response to a novel peptide sequence in vitro, which may be enhanced by simultaneous inclusion of adhesion peptides. Initial in vivo data in regeneration of rabbit femoral condyle cartilage defects may suggest promising regenerative capabilities with hydrogels based on these peptides. If indeed chondroinductive materials exist, and if they can be delivered easily, are safe, and can be provided at reasonable cost and with a reasonable regulatory strategy, chondroinductive materials may hold the potential to revolutionize cartilage regeneration.

Our musculoskeletal system has a limited capacity for repair. This has led to increased interest in the development of tissue engineering strategies for the regeneration of musculoskeletal tissues such as bone, ligament, tendon, meniscus and articular cartilage. This talk will review our attempts to use biomaterials and mesenchymal stem cells (MSCs) to bioprint functional articular cartilage and bone grafts for use in bone and joint regeneration. It will begin by describing how 3D bioprinting can be used to engineer biological implants mimicking the shape of specific bones, and how these bioprinted tissues mature into functional bone organs upon implantation into the body. Next, it will be demonstrated that different musculoskeletal injuries can be regenerated using 3D bioprinted implants, including large bone defects and osteochondral defects. The talk will conclude by describing how we can integrate biomaterials and MSCs into 3D bioprinting systems to engineer scaled-up tissues that could potentially be used regenerate entire diseased joints.

Symptomatic articular cartilage defects are one of the most common knee injuries, arising from acute trauma, overuse, ligamentous instability, malalignment, meniscectomy, osteochondritis dissecans. Surgical treatment options include bone marrow–stimulating techniques such as abrasion arthroplasty and microfracture, osteochondral mosaicplasty, corrective osteotomy, cartilage resurfacing techniques and tissue engineering techniques using combinations of autologous cells (chondrocytes and mesenchymal stem cells), bioscaffolds, and growth factors. Matrix induced autologous chondrocyte implantation (MACI) is considered the most surgically simple form of autologous chondrocyte implantation. Our group has involved in the development of MACI since 2000 and has led to the FDA approval of MACI as the first tissue engineering product for cartilage repair in 2016. In this article, we have documented the characterisation of autologous chondrocytes, the surgical procedure of MACI and the long term clinical assessment (15 years) of patients with treatment of MACI. We have also reported the retrospective survey in patients with MACI in Australia. Our results suggest that MACI has gained good to excellent long term clinical outcome and probably can delay total knee replacement. However, restoration of hyaline-like cartilage by MACI may be interrupted by the osteoarthritic condition of the joint in patients with progressed osteoarthritis. In addition, because articular cartilage and subchondral bone are considered a single functional unit that is essential for joint function, many cartilage repair technologies including MACI and microfractures have failed short to address the functional barrier structure of osteochondral unit. Further studies are required to develop tissue engineering osteochondral construct that is able to fulfil the function of articular cartilage-subchondral bone units.

Dynamic loading is necessary for the preservation of native cartilage, but mechanical disuse is one major risk factor for osteoarthritis (OA). As post-transcriptional regulators, microRNAs (miRs) represent promising molecules to quickly adjust the cellular transcriptome in a stimulus-dependent manner. Several miR clusters were related to skeletal development, joint homeostasis and OA pathophysiology but whether miRs are associated with mechanosensitivity and regulated by mechanotransduction is so far unknown. We aimed to investigate the influence of mechanical loading on miR expression and to identify mechanosensitive miR clusters characteristic for non-beneficial loading regimes which may serve as future tools for improved diagnosis or intervention during OA development. Loading regimes leading to an anabolic or catabolic chondrocyte response were established based on an increase or decrease of proteoglycan synthesis after loading of human engineered cartilage. miR microarray profiling at termination of loading revealed only small changes of miR expression (7 significantly upregulated miRs) by an anabolic loading protocol while catabolic stimulation produced a significant regulation of 80 miRs with a clear separation of control and compressed samples by hierarchical clustering. Overall regulation of 8/14 miR was confirmed by qRT-PCR with mean amplitudes of up to 2.5-fold for catabolic loading. Cross-testing revealed that 2 miRs were upregulated by both loading conditions and 6 were specifically elevated by the catabolic loading regime. Conclusively, this study defines the first mechanosensitive miR cluster associated with non-beneficial compressive cyclic loading of human engineered cartilage which can now be tested for its diagnostic potential in healthy versus OA-affected human cartilage.

The aim of this study was to evaluate the trochlear bone and cartilaginous regeneration of rabbits using a composite based on platelet rich plasma (PRP), chitosan and hydroxyapatite. The study was approved by the ethics committee of the Federal University of Campina Grande under number 72/2017. Surgical holes measuring four millimetres in diameter were performed in rabbit trochleae, one surgical hole in each animal remained empty and another one was filled with the composite. Clinical-orthopaedic and radiographic evaluations were carried out for 60 days, after which the animals were euthanized for histomorphometric evaluations. Clinical-evaluations exhibited lameness of two members of the treatment (T) group and one member of control (C) group. The radiographic evaluation of T group exhibited absence of subchondral bone reaction (33%); nonetheless, presence of moderate subchondral bone reaction was more frequently reported in group C with 67%. Microscopic evaluation revealed the presence of tissue neoformation, composed of dense connective tissue. Microscopic findings were similar in both groups, with a difference in the amount of neoformed tissue, which was confirmed after the morphometric analysis, revealing a significant difference in the quantity of newly formed tissue at the bone / cartilage / implant interface in the T group. The results indicate that the composite based on chitosan, hydroxyapatite and PRP enhanced bone and cartilage healing.

Calcium phosphate ceramics and bioactive glasses are frequently used in orthopedic surgery to stimulate the regeneration of bone tissue due to their superior compatibility to bone tissue. Nevertheless, the brittleness and lack of self-healing behavior of bioceramics are still considered as serious drawbacks. Therefore, these bioceramics have been combined with organic biomaterials for several decades. Since the 1990s, the emergence of nanotechnology has accelerated the progress with respect to the development of organic-inorganic nanocomposites of improved functionality compared to conventional composite biomaterials. This presentation focuses on the development of injectable (nano)composites with self-healing and/or load-bearing capacity. To this end, the affinity between polymeric and inorganic components was tuned by modifying non-covalent interactions between both composite components. Specifically, we exploited reversible interactions between hydrogel matrices and inorganic nanoparticles (traditional nanocomposites), hydrogel nanoparticles and inorganic nanoparticles (colloidal nanocomposites), as well as fibers and bioceramic matrices (fiber-reinforced cement composites). The resulting composite biomaterials were mechanically strong and self-healing, which may open up new avenues of research on the applicability of self-healing and load-bearing composite biomaterials for regenerative medicine.

The development of functional biomaterials scaffolds for bone tissue engineering applications includes the control of specific biological and mechanical parameters that are involved in the growth of bone tissue in a way that mimics the physiological process of healing bone defects. Here, we report on the development of composite scaffolds made from biodegradable natural and synthetic biomaterials with characteristic architectural features, functionalized with the osteoinductive growth factor bone morphogenetic protein BMP-2, and evaluating their osteogenic response in static and dynamic cell culture systems. The results show that scaffold designing with advanced technologies combined with appropriate biochemical and mechanical stimulating factors, results to an enhanced proliferative and osteogenic/chondrogenic differentiation response of cells cultured on the developed scaffolds, and thus controlling the new tissue formation and reconstruction.

Orbital floor (OF) fractures are commonly treated by implanting either bioinert titanium or polyethylene implants, or by autologous grafts. A personalized implant made of biodegradable and osteopromotive poly(trimethylene carbonate) loaded with hydroxyapatite (PTMC-HA) could be a suitable alternative for patients where a permanent implant could be detrimental. A workflow was developed from the implant production using stereolithography (SLA) based on patient CT scan to the implantation and assessment its performance (i.e. implant stability, orbit position, bone formation) compared to personalised titanium implants in a repair OF defect sheep model. Implants fabrication was done using SLA of photo-crosslinkable PTMC mixed with HA [1–3]. Preclinical study: (sheep n=12, ethic number 34_2016) was conducted by first scanning the OF bone of each sheep in order to design and to fabricate patient specific implants (PSI) made of PTMC-HA. The fabricated PSI was implanted after creating OF defect. Bone formation and defect healing was compared to manually shaped titanium mesh using time-laps X-ray analyses, histology (Giemsa-Eosin staining) and sequential fluorochrome staining over 3-months. Additionally, the osteoinductive property of the biomaterials was assessed by intramuscular implantation (IM). In this study, we showed that the composite PTMC-HA allowed for ectopic bone formation after IM implantation, without requiring any biotherapeutics. In addition, we could repair OF defect on sheep using SLA-fabricated PTMC-HA with a good shape fidelity (compared to the virtual implant) and a better bone integration compared to the titanium mesh. This study opens the field of patient-specific implants made of degradable and osteoinductive scaffolds fabricated using additive manufacturing to replace advantageously autologous bone and titanium implants.

The current gold standard bone substitute is still autologous bone, despite the fact that its harvest demands for a second operation site, causes additional pain, discomfort, potential destruction of the grafting site, and is limited in supply. Since newly developed clinical approaches like transplantation of cells are invasive and costly, and osteoinduction by bone morphogenetic proteins is expensive and is associated with mild to severe side effects, the optimization of osteoconduction appears as promising option to realize bone substitute-based bone tissue engineering. In the 90ties of the last century, the holy grail of pore size for scaffolds in bone tissue engineering was set between 0.3 and 0.5 mm. More recent, papers from others and us indicated that the optimal microarchitecture for bone tissue engineering scaffolds in terms of pore size, constrictions, rod thickness, or rod distance is still unknown. Additive manufacturing appears as an ideal tool to study those diverse microarchitecture options since it can generate scaffolds where size and location of pores and connections between pores can be tested. For the production of our test scaffolds, we applied laser sintering of titanium and lithography-based additive manufacturing of ceramics. Histomorphometry of calvarial defects in rabbits revealed that bone formation was significantly increased by scaffolds with pore diameters in the range of 0.7–1.2 mm. Scaffolds with pores of 1.5 and 1.7 mm perform significantly worse. Therefore, pore diameters in osteoconductive bone substitutes should be 1.0–1.2 mm and thus much bigger than previously suggested. In essence, osteoconductive microarchitectures of degradable bone substitutes can be realized by lithography based additive manufacturing and this methodology appears as a promising tool for the production of personalized bone tissue engineering scaffolds to be used in cranio-maxillofacial surgery, dentistry, and orthopedics.

Despite poly(vinyl alcohol) (PVA) hydrogel-based drug delivery systems have been extensively studied in the last years, so far there is no research investigating hydrogels in microspherical shape. In the present study, hydrogels for drug delivery systems were obtained from different formulations of poly(vinyl alcohol), poly(acrylic acid), ciprofloxacin and hydroxyapatite (Hap) aqueous solutions and shaped into spheres through dripping the solution into liquid nitrogen at extremely low temperatures. Hydrogels were then strengthened by freeze-thaw cycles. Characterisation of the samples produced aimed to evaluate the thermal (DSC), chemical (EDS), morphology (SEM), drug release properties of the hydrogel and to investigate the influence of each compound on PVA and their biocompatibility. Samples were able to maintain a spherical shape after the freeze-thawing cycles, also, cross-section of these samples revealed different internal structures depending on the components incorporated into the PVA, EDS revealed quantities of Ca and P into these hydrogels due to the HAp and the incorporation of drug, poly(acrylic acid) and hydroxyapatite increased both the melting point and the glass transition temperature of PVA. Ciprofloxacin release exhibited a burst release for approximately two hours, then stabilising the drug release to a maximum of 96.82%. PAA has acted as a release retardant and the burst release was significantly delayed. PAA chains helped encapsulating the drug and reinforced the three-dimensional structure of the hydrogel, hampering ciprofloxacin to be delivered, the total of drug release was 92.11%. Cells mortality rate (MTT) shows that PVA substrates is non-toxic for NRK cells after 24 hours of exposure.

Orthopaedic infection with bacteria leads to high societal cost and is detrimental to the life quality. Particularly, deep bone infection leading to osteomyelitis results in an inflammatory response whereby localized bone destruction occurs. Current treatments like antibiotic-containing polymethymethacrylate (PMMA) still has the high risk of bacterial resistance. Taking advantages of silver which has antibacterial and anti-inflammatory effect and bioactive collagen, we fabricated a silver nanoparticle (AgNP)-coated collagen membrane by sonication and sputtering. SEM showed good deposition of AgNPs on collagen membrane by both coating methods. The optimal coating concentration was finalized by assessing optimal antibacterial effect against cytotoxicity and finally collagen membrane coated with 1mg/mL AgNPs solution was selected. We also found that the coated collagen membrane demonstrating short-term cytotoxicity within 24 hours with damage to the cell membrane, which was evidenced by MTS and LDH release test, but had no significant influence (p > 0.05) thereafter. The amount of released AgNPs from coated collagen membrane had negligible cytotoxicity (p > 0.05). Confocal laser scanning microscope displayed similar cell morphology in both coated and uncoated collagen membrane. ELISA and qPCR presented the decreased secretion and expression (p < 0.001) of IL-6 and TNF-alpha. Upregulated expression (p < 0.001) of osteogenesis markers (RUNX2, ALP and OPN) could be found and this might be attributed to the modified collagen fibre surface coated by AgNPs. Collectively, the osteogenesis induced by AgNPs demonstrates a promising application in orthopaedic surgery for its use both as an antimicrobial agent, and to enhance bone regeneration.

There is a growing socio-economic need (i.e. “ageing society”) for effective and reproducible strategies to repair musculoskeletal tissue. In particular, acute tendon injury and chronic tendinopathies remain clinically challenging and novel treatment modalities are urgently needed. Tendons resemble a connective tissue rich in highly organized collagen fibers, displaying a remarkably high tensile strength. However, partly due to the low number of cells and their more or less avascular nature tendons heal relatively slowly. Ultimately, tendon regeneration encompasses the full restoration of the biological, biochemical and biomechanical properties, which are often impaired by endogenous healing cascades. Usually, a connective scar tissue forms at the injury site and the replaced tissue does not function adequately at high strain levels, increasing the chance of re-rupture. Despite significant advancements in tissue regeneration and engineering strategies, the clinical impact for the regeneration of tendon remains limited. For the development of novel methods to repair tendons we need to pin down the molecular and cellular mechanisms amenable to modulate endogenous (or exogenous) cell behaviour towards functional tissue regeneration. By comparing the gene expression profile of Achilles tendon tissue harvested from young-mature and old mice we demonstrate profound changes in the expression of ECM-related proteins and a previously unknown role of Secreted protein acidic and rich in cysteine (Sparc; also known as BM-40 or osteonectin) in tendons. Sparc levels in tendons are critical for proper collagen fibril maturation and its age-related decrease, together with a change in ECM properties potentially drives adipogenic differentiation of tendon stem and progenitor cells (TDSPCs) and consequently lipid accretion in tendons. Generally, the fate of stem/ progenitor cells is largely determined by stimuli from the stem cell niche. In tendons, we describe a novel cellular barrier, most likely preventing the leakage of blood-borne products into the tendon proper. We propose that this “blood-tendon barrier” is part of the stem cell niche in tendons controlling TDSCP fate, preventing erroneous differentiation. By investigating the developmental programs driving tendon tissue formation and on the other hand the mechanisms contributing to the senescence of tendons, ultimately resulting in decreased quality of tendons in the elderly, novel targets for clinical intervention potentially can be discovered.