Summary:. Hamstring tendons (HT) represent a widely used autograft for ACL reconstruction. Harvesting, processing and pretensioning procedures together with the time out of the joint could theoretically hamper tendon cells (TCs) viability. The authors hypothesize that HT cells are not impaired at the end of the surgical procedures and their tenogenic phenotype may be strongly improved by exposure to PEMF. Methods. Remnants of semitendinosus and gracilis tendons were collected at the end of the surgical procedures before skin closure from 15 healthy donors who underwent ACL reconstruction with autologous hamstring tendons. To isolate TCs, the tendon was minced and digested with 0.3 % type I collagenase and the nucleated cells were plated at a density 5x10E3 cells/cm2 and cultured in chamber slides in differentiation medium composed of DMEM + 5ng/ml basic fibroblast growth factor (b-FGF) for 7, 14, 21 days. The following cell cultures were set up:. -. TCs cultured with differentiation medium + exposure to PEMF 8 h/day (PEMF generator system IGEA, intensity of magnetic field = 1.5 mT, frequency = 75 Hz). -. TCs cultured with differentiation medium without exposure to PEMF. At day 0, day 7, day 14 and day 21, immunofluorescence analysis was performed to evaluate the expression of collagen type I, collagen type VI, scleraxis and PCNA (proliferative marker). Subsequently, tendon explant cultures were set up to verify, at day 21, explant viability and the expression of collagen type I, collagen type VI, beta-catenin and PCNA. Results. The TCs from the tendon fragments at the end of the ACL reconstruction were alive and they expressed markers of proliferation and tendon phenotype at the end of the culture periods. The TCs in the presence PEMF 8h/day showed greater production of collagen type I, collagen type VI and scleraxis than that of TCs cultured without PEMF (p<0,05): the expression of this markers increased from 7 to 21 days of culture. The expression of PCNA, in the presence of PEMF stimulus, was significantly lower (p<0,05) than that of TCs cultured without PEMF. A similar behavior was surprisingly observed in tendon explant cultures. Conclusions. Hamstring tendons used for ACL reconstruction are not simple autologous tenoconductive scaffold but are a biologic structure rich in progenitor cells that show tenogenic behavior. Their tenogenic phenotype may be strongly improved by exposure to PEMF. In a future clinical perspective, the postoperative use of PEMF could be used to enhance the ligamentization processes of autologous hamstring tendons, when used as autografts for ACL reconstructions

Introduction and Objective. Objectives: To determine the effectiveness of LIA compared to ACB in providing pain relief and reducing opiates usage in hamstring graft ACL reconstructions. Materials and Methods. In a consecutive series of hamstring graft ACL reconstructions, patients received three different regional and/or anaesthetic techniques for pain relief. Three groups were studied: group 1: general anaesthetic (GA)+ ACB (n=38); group 2: GA + ACB + LIA (n=31) and group 3: GA+LIA (n=36). ACB was given under ultrasound guidance. LIA involved infiltration at skin incision site, capsule, periosteum and in the hamstring harvest tunnel. Analgesic medications were similar between the three groups as per standard multimodal analgesia (MMA). Patients were similar in demographics distribution and surgical technique. The postoperative pain and total morphine requirements were evaluated and recorded. The postoperative pain was assessed using the visual analogue scores (VAS) at 0hrs, 2hrs, 4hrs, weight bearing (WB) and discharge (DC). Results. There was no statistically significant difference in opiates intake amongst the three groups. When comparing VAS scores; there were no statistical difference between the groups at any of the time intervals that VAS was measured. However, the GA+LIA group hospital's LOS (m=2.31hrs, SD=0.75) was almost half that of GA+ACB group (m=4.24hrs, SD=1.08); (conditions t(72)=8.88; p=0.000). There was no statistical significance in the incidence of adverse effects amongst the groups. Conclusions. The LIA technique provided equally good pain relief following hamstring graft ACL reconstructions when compared to ACB, while allowing for earlier rehabilitation, mobilisation and discharge

Flexion Deformity of knee is the most common deformity in post polio residual deformity. Wilson's release, supracondylar osteotomy etc have been described for its treatment. We present our result of fractional hamstring lengthening followed by gradual distraction using threaded rod in hollow tube to treat flexion deformity of knee. This retrospective study included 150 cases (80 males and 70 females) with the mean of 15 years (8-22yrs). The mean duration of deformity was 6 years (2 – 14yrs) with mean follow up 0f 3 years. The mean preoperative flexion deformity was 45degree (110 – 30 degree) with a mean pre operative further flexion of 110 degree (130 – 90) .20 cases were had a crawling gait and 10 cases were wheel chair bound. Flexion got corrected to 0 degree in 110 cases (P value <0.01). Post operative mean arc of motion was 80degree We had 10 cases who could not tolerate plaster and hence were put on traction . 20 cases had knee stiffness on removal of plaster which could not improve on physiotherapy. 10 cases had superficial infection cured with dressings. Our findings indicate that this method is very effective in the treatment of flexion deformity of knee with complication of knee stiffness in older cases

To investigate changes in quadriceps and hamstrings muscle groups during sustained isokinetic knee flexion and extension. 125 paediatric participants (45 males and 80 females, mean age 14.2 years) were divided into two groups: participants with a confirmed ACL tear (ACLi, n = 64), and puberty- and activity-level matched control participants with no prior history of knee injuries (CON, n = 61). Participants completed a series of 44 repetitions of isokinetic knee flexion and extension at 90 deg/ sec using a Biodex dynamometer (Biodex Medical Systems Inc, Shirley, New York). Surface EMG sensors (Delsys Incorporated, Natick, MA) simultaneously recorded the quadriceps and hamstring activations. Muscle function was assessed as the change in quadriceps activation and extension torque were calculated using the percent difference between the mean of the first five trials, and the mean of the last five trials. ACLi participants had significantly higher percent change in quadriceps activation for both healthy and injured legs, in comparison to CON dominant leg. As such, the healthy leg of the ACLi participants is activating significantly more than their health matched controls, while also demonstrating reduced muscular endurance (less torque in later repetitions). Therefore, we conclude that the non-injured limb of the ACLi participant is not performing as a healthy limb. Since return to activity clearance following ACLi implies return to sport against age- and activity matched opponents, clearing young athletes based on the non-injured contralateral limb may put them at greater risk of reinjury

To address the current challenge of anterior cruciate ligament (ACL) reconstruction, this study is the first to fabricate a braided collagen rope (BCR) which mimics native hamstring for ACL reconstruction. The study aims to evaluate the biological and biomechanical properties of BCR both in vivo and vitro. Rabbit ACL reconstruction model using collagen rope and autograft (hamstring tendon) was conducted. The histological and biomechanical evaluations were conducted at 6-, 12-, 18, 26-week post-operation. In vitro study included cell morphology analysis, cell function evaluation and RNA sequencing of the tenocytes cultured on BCR. A cadaver study was also conducted to verify the feasibility of BCR for ACL reconstruction. BCR displays satisfactory mechanical strength similar to hamstring graft for ACL reconstruction in rabbit. Histological assessment showed BCR restore ACL morphology at 26 weeks similar to native ACL. The superior dynamic ligamentization in BCR over autograft group was evidenced by assessment of cell and collagen morphology and orientation. The in vitro study showed that the natural collagen fibres within BCR enables to signal the morphology adaptation and orientation of human tenocytes in bioreactor. BCR enables to enhance cell proliferation and tenogenic expression of tenocytes as compared to hydrolysed collagen. We performed an RNA-Sequencing (RNA-seq) experiment where RNA was extracted from tenocyte seeded with BCR. Analysis of enriched pathways of the up-regulated genes revealed that the most enriched pathways were the Hypoxia-inducible factor 1-alpha (HIF1A) regulated networks, implicating the possible mechanism BCR induced ACL regeneration. The subsequent cadaver study was conducted to proof the feasibility of BCR for ACL reconstruction. This study demonstrated the proof-of-concept of bio-textile braided collagen rope for ACL reconstruction, and the mechanism by which BCR induces natural collagen fibres that positively regulate morphology and function of tenocytes

Anterior cruciate ligament reconstruction (ACLR) using a semitendinosus (ST) autograft, with or without gracilis (GR), results in donor muscle atrophy and varied tendon regeneration. The effects of harvesting these muscles on muscle moment arm and torque generating capacity have not been well described. This study aimed to determine between-limb differences (ACLR vs uninjured contralateral) in muscle moment arm and torque generating capacity across a full range of hip and knee motions. A secondary analysis of magnetic resonance imaging was undertaken from 8 individuals with unilateral history of ST-GR ACLR with complete ST tendon regeneration. All hamstring muscles and ST tendons were manually segmented. Muscle length (cm), peak cross-sectional area (CSA) (cm. 2. ), and volume (cm. 3. ) were measured in ACLR and uninjured contralateral limbs. OpenSim was used to simulate and evaluate the mechanical consequences of changes in normalised moment arm (m) and torque generating capacity (N.m) between ACLR and uninjured contralateral limbs. Compared to uninjured contralateral limbs, regenerated ST tendon re-insertion varied proximal (+) (mean = 0.66cm, maximum = 3.44cm, minimum = −2.17cm, range = 5.61cm) and posterior (+) (mean = 0.38cm maximum = 0.71cm, minimum = 0.02cm, range = 0.69cm) locations relative to native anatomical positions. Compared to uninjured contralateral limbs, change in ST tendon insertion point in ACLR limbs resulted in 2.5% loss in peak moment arm and a 3.4% loss in peak torque generating capacity. Accounting for changes to both max isometric force and ST moment arm, the ST had a 14.8% loss in peak torque generating capacity. There are significant deficits in ST muscle morphology and insertion points following ST-GR ACLR. The ST atrophy and insertion point migration following ACLR may affect force transmission and distribution within the hamstrings and contribute to persistent deficits in knee flexor and internal rotator strength

Following anterior cruciate ligament reconstruction (ACLR) using a semitendinosus (ST) autograft measures such as length, cross-sectional area, and volume may not fully describe the effects of tendon harvest on muscle morphology as these discrete measures cannot characterize three-dimensional muscle shape. This study aimed to determine between-limb ST shape similarity and regional morphology in individuals with a unilateral history of ACLR using a ST graft, and healthy controls. A secondary analysis of magnetic resonance imaging was undertaken from 18 individuals with unilateral history of ST ACLR and 18 healthy controls. ST muscles were manually segmented, and shape similarity were assessed between limbs and groups using Jaccard index (0-1) and Hausdorff distance (mm). ST length (cm), peak cross-sectional area (CSA) (cm. 2. ), and volume (cm. 3. ) was compared between surgically reconstructed and uninjured contralateral limbs, and between the left and right limbs of control participants with no history of injury. Cohen's d was reported as a measure of effect size. Compared to healthy controls, the ACLR group had significantly (p<0.001, d= −2.33) lower bilateral ST shape similarity. Furthermore, the deviation in muscle shape was significantly (p<0.001, d= 2.12) greater in the ACLR group. Within the ACLR group, maximum Hausdorff distance indicated ST from the ACLR limb deviated (23.1±8.68 mm) from the shape of the healthy contralateral ST, this was observed particularly within the distal region of the muscle. Compared to the uninjured contralateral limb and healthy controls, deficits in peak cross-sectional area and volume in ACLR group were largest in proximal (p<0.001, d= −2.52 to −1.28) and middle (p<0.001, d= −1.81 to −1.04) regions. Findings highlight morphological features in distal ST not identified by traditional discrete morphology measures. ST shape was most different in the distal region of the muscle, despite deficits in CSA and volume being most pronounced in proximal and middle regions. ST shape following ACLR may affect force transmission and distribution within the hamstrings and contribute to persistent deficits in knee flexor and internal rotator strength

Although remnant-preserved ACL reconstruction (ACLR) restores knee joint stability and dampens the problem of acute ACL rupture-induced knee pain, an increasing number of patients still develop post-traumatic osteoarthritis (PTOA) after 10 to 15 years of ACLR. We previously found that remnant-preserved ACLR with concomitant medial and lateral meniscus repair may not prevent cartilage degeneration and weaken muscle strength, while the clinical features of PTOA are not clear. We hypothesized that remnant-preserved ACLR with concomitant medial and lateral meniscus tears is related to early cartilage damage, worse function recovery, patient-reported outcomes (PROs) and delayed duration to return to sports. The aim is to evaluate the remnant-preserved ACLR with complicated meniscal injuries in predicting which patients are at higher risk of osteoarthritic changes, worse function and limited activities after ACLR for 12 months. Human ethical issue was approved by a committee from Xi'an Jiaotong University. 26 young and active patients (24 male, 2 female) with ACL injuries (Sherman type I and II) with concomitant medial and lateral meniscus within 2 months were included from January 2014 to March 2022. The average age of the ACLR+ meniscus repair was 26.77±1.52 (8 right, 5 left) and isolated ACLR control was 31.92±2.61 years old (7 left, 6 right). Remnant-preserved ACLR with a 5- to 6-strand hamstring tendon graft was operated on by the same sports medicine specialists. MRI CUBE-T. 2. scanning with 48 channels was conducted by a professional radiologist. The volume of the ACL graft was created through 3 dimensional MRI model (Mimics 19, Ann Arbor). Anterior Cruciate Ligament OsteoArthritis Score (ACLOAS) was applied to score visible cartilage damage. IKDC 2000 score and VAS were assessed by two blinded researchers. Results were presented as mean± SEM of each group. The cross-sectional area and 3D volume of the ACL graft were greater in the remnant-preserved ACLR+meniscus group compared with isolated ACLR (p=0.01). It showed that ACLR+ meniscus group had early signs of joint damage and delayed meniscus healing regarding ACLOAS compared to control group (p=0.045). MRI CUBE-T. 2. prediction of radiographic cartilage degeneration was not obvious in both groups post remnant-preserved ACLR over 12 months (p>0.05). However, higher VAS scores, lower IKDC scores, and long-last joint swelling were reported in the ACLR+ meniscus repair group at the end of 12 months follow-up. Although remnant-preserved ACLR+ meniscus was able to maintain the restore the knee function, it showed delayed timing (>12 months) to return to play at the pre-injury stage, while no difference between the timing of returning to the normal daily routine of their ACLR knee compared to control (p=0.30). The cost of ACLR+ meniscus (average 10,520.76$) was higher than the control group (6,452.92$, p=0.018). Remnants-preserved ACLR with concomitant injured medial and lateral meniscus repair shows a higher risk of cartilage damage, greater cost, worse functional performance, and longer time for young male patients to return to sports after 12-month follow-up compared to isolated ACLR. Further evidence and long-term follow-up are needed to better understand the association between these results and the risk of development of PTOA in this patient cohort

After anterior cruciate ligament (ACL) rupture, reconstructive surgery with a hamstring tendon autograft is often performed. Despite overall good results, ACL re-rupture occurs in up to 10% of the patient population, increasing to 30% of the cases for patients aged under 20 years. This can be related to tissue remodelling in the first months to years after surgery, which compromises the graft's mechanical strength. Resident graft fibroblasts secrete matrix metalloproteinases (MMPs), which break down the collagen I extracellular matrix. After necrosis of these fibroblasts, myofibroblasts repopulate the graft, and deposit more collagen III rather than collagen I. Eventually, the cellular and matrix properties converge towards those of the native ACL, but full restoration of the ACL properties is not achieved. It is unknown how inter-patient differences in tissue remodelling capacity contribute to ACL graft rupture risk. This research measured patient-specific tissue remodelling-related properties of human hamstring tendon-derived cells in an in vitro micro-tissue platform, in order to identify potential biological predictors for graft rupture. Human hamstring tendon-derived cells were obtained from remnant autograft tissue after ACL reconstructions. These cells were seeded in collagen I gels on a micro-tissue platform to assess inter-patient cellular differences in tissue remodelling capacity. Remodelling was induced by removing the outermost micro-posts, and micro-tissue compaction over time was assessed using transmitted light microscopy. Protein expression of tendon marker tenomodulin and myofibroblast marker α-smooth muscle actin (αSMA) were measured using Western blot. Expression and activity of remodelling marker MMP2 were determined using gelatin zymography. Cells were obtained from 12 patients (aged 12–51 years). Patient-specific variations in micro-tissue compaction speed or magnitude were observed. Up to 50-fold differences in αSMA expression were found between patients, although these did not correlate with faster or stronger compaction. Surprisingly, tenomodulin was only detected in samples obtained from two patients. Total MMP2 expression varied between patients, but no large differences in active fractions were found. No correlation of patient age with any of the remodelling-related factors was detected. Remodelling-related biological differences between patient tendon-derived cells could be assessed with the presented micro-tissue platform, and did not correlate with age. This demonstrates the need to compare this biological variation in vitro - especially cells with extreme properties - to clinical outcome. Sample size is currently increased, and patient outcome will be determined. Combined with results obtained from the in vitro platform, this could lead to a predictive tool to identify patients at risk for graft rupture

Introduction and Objective. The rupture of the anterior cruciate ligament is a common sports injury and surgical reconstruction is often required to restore full function of the knee. Hamstring tendons are usually used as autografts. In addition to knee pain and stiffness, infections are feared complications after surgery. Incubation of the autograft in a vancomycin solution until implantation reduced the infection rate by about ten-fold. Recent studies showed no negative effect of vancomycin on the biomechanical properties of porcine tendons. A negative effect of high vancomycin concentrations on chondrocytes and osteoblast is reported, but the effect on tendon and tenocytes is not known. Materials and Methods. Rat Achilles tendons or isolated tenocytes were incubated with an increasing concentration of vancomycin (0 – 10 mg). Tendons were incubated for 0 – 40 minutes, while tenoyctes were incubated for 20 minutes followed by culturing for up to 7 days. Cell viability was assessed with PrestoBlue Assay and live/dead stain. The potential effect of vancomycin on the expression of tendon specific genes and extracellular matrix (ECM) genes was quantified. Possible structural changes of the tendon are analyzed. Results. Incubation of the tendons or tenocytes with 5 mg vancomycin for 20 minutes (clinical use) had no negative effects on the cell viability in the tendons or the isolated tenocytes, while incubation with the toxic control (ethanol) significantly reduced cell viability. Even twice the concentration and a longer incubation time had no negative effect on the cells in the tendons or the isolated cells. Vancyomycin did not affect the expression of Col1a1, Col3a1, and the tenocyte markers mohawk, scleraxis and tenomodulin. Conclusions. The results showed that clinical practice of wrapping the autograft in vancomycin did not impair the tenocyte viability. The expression of collagens and tenocyte markers was also not affected, neither in the incubated tendons nor in the isolated cells. This indicates that vancomycin had no effect on cell phenotype and the formation of the extracellular matrix, which, in addition to cell viability, is important for the performance of the autograft

Introduction and Objective. Anterior cruciate ligament reconstruction (ACLR) with tendon autografts is the “gold standard” technique for surgical treatment of ACL injuries. Common tendon graft choices include patellar tendon (PT), semitendinosus/gracilis “hamstring” tendon (HT), or quadriceps tendon (QT). Healing of the graft after ACLR may be affected by graft type since the tissue is subjected to mechanical stresses during post-operative rehabilitation that play important roles in graft integration, remodeling and maturation. Abnormal mechanical loading can result in high inflammatory and degradative processes and altered extracellular matrix (ECM) synthesis and remodeling, potentially modifying tissue structure, composition, and function. Because of the importance of load and ligamentization for tendon autografts, this study was designed to compare the differential inflammatory and degradative metabolic responses to loading by three tendon types commonly used for autograft ACL reconstruction. Materials and Methods. With IRB approval (IRB # 2009879) and informed patient consent, portions of 9 QT, 7 PT and 6 HT were recovered at the time of standard of care ACLR surgeries. Tissues were minced and digested in 0.2 mg/ml collagenase solution for two hours and were then cultured in 10% FBS at 5% CO. 2. , 37°C, and 95% humidity. Once confluent, cells were plated in Collagen Type I-coated BioFlex® plates (1 × 10. 5. cells/well) and cultured for 2 days prior to the application of strain. Then, media was changed to supplemented DMEM with 2% FBS for the application of strain. Fibroblasts were subjected to continuous mechanical stimulation (2-s strain and 10-s relaxation at a 0.5 Hz frequency) at three different elongation strains (mechanical stress deprivation-0%, physiologic strain-4%, and supraphysiological strain-10%). 9. for 6 days using the Flexcell FX-4000T strain system. Media was tested for inflammatory biomarkers (PGE2, IL-8, Gro-α, and MCP-1) and degradation biomarkers (GAG content, MMP-1, MMP-2, MMP-3, TIMP-1, and TIMP-2). Significant (p<0.05) difference between graft sources were assessed with Kruskal-Wallis test and post-hoc analysis. Results are reported as median± interquartile range (IQR). Results. Differences in Inflammation-Related Biomarker Production (Figure 1): The production of PGE2 was significantly lower by HT fibroblasts compared to both QT and PT fibroblasts at all timepoints and strain levels. The production of Gro-α was significantly lower by HT fibroblasts compared to QT at all time points and strain levels, and significantly lower than PT on day 3 at 0% strain, and all strain levels on day 6. The production of IL-8 by PT fibroblasts was significantly lower than QT and HT fibroblast on day 3 at 10% strain. Differences in Degradation-Related Biomarker Production (Figure 2): The production of GAG by HT fibroblasts was significantly higher compared to both QT and PT fibroblasts on day 6 at 0% strain. The production of MMP-1 by the QT fibroblasts was significantly higher compared to HT fibroblasts on day 3 of culture at all strain levels, and in the 0% and 10% strain levels on day 6 of culture. The production of MMP-1 by the QT fibroblasts was significantly higher compared to PT fibroblasts at in the 0% and 4% strain groups on day 3 of culture. The production of TIMP-1 by the HT fibroblasts was significantly lower compared to PT fibroblasts on day 3 of culture. Conclusions. The results of this study identify potentially clinically relevant difference in the metabolic responses of tendon graft fibroblasts to strain, suggesting a lower inflammatory response by hamstring tendon fibroblasts and higher degradative response by quadriceps tendon fibroblasts. These responses may influence ACL autograft healing as well as inflammatory mediators of pain in the knee after reconstruction, which may have implications regarding graft choice and design of postoperative rehabilitation protocols for optimizing outcomes for patients undergoing ACL reconstruction. For any figures or tables, please contact the authors directly

Introduction. Regular, repeated stretching increases joint range of movement (RoM), however the physiology underlying this is not well understood. The traditional view is that increased flexibility after stretching is due to an increase in muscle length or stiffness whereas recent research suggests that increased flexibility is due to modification of tolerance to stretching discomfort/pain. If the pain tolerance theory is correct the same degree of micro-damage to muscle fibres should be demonstrable at the end of RoM before and after a period of stretch training. We hypothesise that increased RoM following a 3 weeks hamstrings static stretching exercise programme may partly be due to adaptive changes in the muscle/tendon tissue. Materials and Methods. Knee angle and torque were recorded in healthy male subjects (n=18) during a maximum knee extension to sensation of pain. Muscle soreness (pain, creatine kinase activity, isometric active torque, RoM) was assessed before knee extension, and 24 and 48 hours after maximum stretch. An exercise group (n=10) was given a daily home hamstring stretching programme and reassessed after 3 weeks and compared to a control group (n=8). At reassessment each subject's hamstring muscles were stretched to the same maximum knee extension joint angle as determined on the first testing occasion. After 24 hours, a reassessment of maximum knee extension angle was made. Results. At the start of the study RoM was 71.3 ± 10.0 degrees and there was no significant difference between groups. After 3 weeks stretching RoM increased significantly (p=0.01) by 9 degrees; the control group showed no change. Stiffness did not differ for either group. Pain score and RoM were the most sensitive markers of muscle damage and were significantly changed 24 and 48 hours after the initial stretch to end of range, (p<0.005) and (p=0.004) respectively. Discussion. The results show that a 3 week stretching programme causes muscle adaptation resulting in an increase in the extensibility of the hamstring muscle/tendon unit but no change in stiffness. The lack of evidence of muscle damage suggests that participants in the stretching group are likely to have undergone a physical change/adaptation rather than simply an increase in pain threshold

Anterior cruciate ligament injury is the most common and economically costly sport injuries, frequently requiring expensive surgery and rehabilitation. Post-operative knee septic arthritis represents a serious complication with an incidence rate between 0.14% and 1.7%. A common practice to avoid septic arthritis is the “vancomycin wrap”, consisting in the soaking of the graft for 10–15 minutes within a sterile gauze swab previously saturated with 5 mg/mL vancomycin. Even though several studies have been conducted to investigate vancomycin toxicity on different musculoskeletal tissues or cells, little is known about the effect of such antimicrobial on tendon-derived cells. The aim of this study was to determine the in vitro toxicity of different concentrations of vancomycin at different time points on human primary tenocytes (hTCs). hTCs were isolated from hamstring grafts of patients undergoing anterior cruciate ligament reconstruction. After expansion, cells were treated with different concentrations of vancomycin (2.5, 5, 10, 25, 50 and 100 mg/mL) for 10, 15, 30 and 60 minutes. In vitro toxicity was evaluated measuring: metabolic activity through the reduction of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT Assay); cytotoxicity (Live/Dead assay); and cell apoptosis (Annexin V apoptosis kit). The metabolic activity of hTCs was affected by vancomycin treatment starting from 10 mg/mL at all time points (p < 0.05) and dropped down at 100 mg/mL at all time points (0.05 < p < 0.001). Cells viability resulted to be unaffected only by 2.5 mg/mL vancomycin at all time points. Vancomycin resulted to be cytotoxic starting from 10 mg/mL after 15 minutes of treatment and at all higher concentrations under study at all time points. Cells died when treated with vancomycin concentrations higher than 5 mg/mL but not through apoptosis, as confirmed by negative staining for Annexin V. In our experimental conditions, vancomycin resulted to be toxic on hTCs at concentrations higher than 5 mg/mL. The use of this antibiotic on tendons to prevent infections could be useful and safe for resident cells if used at a concentration of 2.5 mg/mL up to 1 hour of treatment

Bone-patellar tendon-bone autografts, hamstring autografts or allografts are widely used grafts for ACL revision surgeries. Also use of quadriceps autograft for both primary and revision ACL surgeries is in an increasing popularity due to its biomechanical superior properties and less donor site morbidity. However, although several fixation techniques and devices for quadriceps tendon graft fixation on femoral side have been reported, literature lacks about biomechanical studies comparing properties of these different fixation techniques and devices. We aimed to investigate whether there is a difference between the fixation techniques of quadriceps tendon graft by using different fixation techniques and devices on the femoral side in terms of stiffness and amount of slippage in the tunnel. Full-thickness central parts of quadriceps tendons from paired knees of twenty five calf knees were fixed through a 10mm x 25mm tunnel in twenty five paired sheep femurs. Quadriceps tendon patellar side with soft tissue ending fixed with four different fixation devices (adjustable suspensory system (group 1), absorbable interference screw (group 2), titanium interference screw (group 3) and adjustable suspensory system + absorbable interference screw (group 4)) and quadriceps tendon with a patellar bone plug fixed with a titanium interference screw (group 5) were tested in a servohydraulic materials testing machine. 10 samples were included in each group. After applying a preload of 10 N, a cyclic force was applied for 20 cycles from 10N to 110N at a 1 hertz frequency. Amount of slippage in the tunnel was calculated as the difference measured in millimeters between length at 10 N after 20 cycles and starting length at 10 N (Graph 1). To determine the stiffness, a single load-to-failure cycle was performed at a strain rate of 20 mm/min as the last step (Figure 1). Rupture of the graft was not seen in any of the samples. Median values of amount of slippage in the tunnel were 6,41mm, 5,99mm, 3,01mm, 4,83mm, and 3,94mm respectively. Median values of maximum load at failure were 464N, 160N, 350N, 350N and 389N respectively. Amount of slippage in the tunnel was highest in the group 1 and was lowest in the group 3 (p<0.001). Group 1 was found to be most resistant group against load-to-failure test and group 2 was the weakest (p<0.001). However inter-group analyses between group 3 and 5 revealed that, although group 3 had the least slippage in the tunnel, group 5 was better in terms of stiffness, but there was no statistically significant difference (p=0,124 and 0,119 respectively). There was a significant difference between group 2 and 3 in both amount of slippage in the tunnel and stiffness (p=0,001 and 0.028 respectively)(Table 1). Our study revealed that, although quadriceps graft with a bone plug fixed with metal interference screws is widely presumed to be a stable fixation technique, there was no significant difference in terms of stiffness when compared with quadriceps graft with soft tissue ending fixed with a metal interference screw. Although adjustable suspensory device group was the best in the terms of resistance against load-to-failure, it was the worst in terms of amount of slippage from the tunnel. Thus, if a suspensory device is to be used, it must be kept in mind that a strong 20 cycles of intra-operative tension force must be applied to prevent further slippage of the graft in the tunnel which can result in failure of reconstruction. For any figures or tables, please contact the authors directly

Alarmins- also referred to as damage associated molecular patterns (DAMPS)- are endogenous molecules mobilized in response to tissue damage known to activate the innate immune system and regulate tissue repair and remodelling. The molecular mechanisms that regulate inflammatory and remodelling pathways in tendinopathy are largely unknown therefore identifying early immune effectors is essential to understanding the pathology. S100A8 and S100A9 are low molecular weight calcium binding proteins primarily released by activated phagocytes in an inflammatory setting and also secreted as a heterodimeric complex that exhibits cytokine like functions. Based on our previous investigations we sought evidence of S100A8/A9 expression in human tendinopathy and thereafter, to explore mechanisms whereby S100 proteins may regulate inflammatory mediators and matrix regulation in human tenocytes. Torn supraspinatus tendon (established pathology) and matched intact subscapularis tendon (representing ‘early pathology’) biopsies were collected from patients undergoing arthroscopic shoulder surgery. Control samples of subscapularis tendon were collected from patients undergoing arthroscopic stabilisation surgery. S100A8/A9 expression was analysed at transcript and protein level using quantitative RT-PCR and immunohistochemistry, respectively. Primary human tenocytes were cultured from hamstring tendon tissue obtained during hamstring tendon ACL reconstruction. The in vitro effect of recombinant human S100 A8/A9 on primary human tenocytes was measured using quantitative RT-PCR and ELISA. Immunohistochemistry of tendinopathic tissues demonstrated the presence of S100 A8/A9 in diseased tissues compared to control tissue. In addition, early pathological diseased tissue indicated greater S100A9 expression compared with established diseased pathology. These findings were reflected by data obtained at transcript level from diseased tissues. Recombinant human S100A8, A9 and A8/A9 complex led to significant increase in expression of inflammatory mediators, including IL-6 in vitro. Further analysis via quantitative RT-PCR demonstrated recombinant S100A8, A9 and A8/A9 complex treatment on tenocytes, in vitro, had no direct effect on the expression of genes involved in matrix remodelling. The presence of S100A8 and S100A9 in early tendinopathic lesions suggests expression is upregulated in response to cellular damage. S100A8 and S100A9 are endogenous ligands of Toll-like receptors (TLRs) and receptor for advanced glycation end products (RAGE). These receptors have known regulatory effects on immune mediated cytokine production. We propose S100A8 and S100A9 as active alarmins in the early stages of tendinopathy and thus targeting of its downstream signalling may offer novel therapeutic approaches in the management of human tendon disorders

Dual mobility (DM) bearing implants reduce the incidence of dislocation following total hip arthroplasty (THA) and as such they are used for the treatment of hip instability in both primary and revision cases. The aim of this study was to compare lower limb muscle activity of patients who underwent a total hip arthroplasty (THA) with a dual mobility (DM) or a common cup (CC) bearing compared to healthy controls (CON) during a sit to stand task. A total of 21 patients (12 DM, 9 CC) and 12 CON were recruited from the local Hospital. The patients who volunteered for the study were randomly assigned to either a DM or a CC cementless THA after receiving informed consent. All surgeries were performed by the same surgeon using the direct anterior approach. Participants underwent electromyography (EMG) and motion analysis while completing a sit-to-stand task. Portable wireless surface EMG probes were placed on the vastus lateralis, rectus femoris, biceps femoris, semitendinosus (ST), gluteus medius and tensor fasciae latae muscles of the affected limb in the surgical groups and the dominant limb in the CON group. Motion capture was used to record lower limb kinematics and kinetics. Muscle strength was recorded using a hand-held dynamometer during maximal voluntary isometric contraction (MVIC) testing. Peak linear envelope (peakLE) and total muscle activity (iEMG) were extrapolated and normalized to the MVIC and time cycle for the sit to stand task. Using iEMG, quadriceps-hamstrings muscle co-activation index was calculated for the task. Nonparametric Kruskal Wallace ANOVA tests and Wilcoxon rank sum tests were used to identify where significant (p < 0.05) differences occurred. The DM group had greater iEMG of the ST muscle compared to the CC (p=0.045) and the CON (p=0.015) groups. The CC group had lower iEMG for hamstring muscles compared to the DM (p=0.041) group. The DM group showed lower quadriceps-hamstrings co-activation index compared to the CON group and it approached significance (p=0.054). The CC group had greater anterior pelvis tilt compared to both DM (p=0.043) and the CON (p=0.047) groups. The DM also had larger knee varus angles and less knee internal rotation compared to both groups, however this never reached significance. No significant differences in muscle strength existed between the groups. Higher ST muscle activity in the DM group is explained by the reduction in internal rotation at the knee joint as the ST muscle was more active to resist the varus forces during the sit-to-stand task. Reduced quadriceps activity in the CC group is explained by increased pelvic anterior tilt as this would shorten the moment arm and muscle length in the quadriceps, ultimately reducing quadriceps muscle activity. The reduced co-activation between quadriceps and hamstrings activity in the DM group compared to the CC and CON groups is related to better hip function and stability. Combining lower co-activation and larger range of motion for the DM group without impingement, this implant seems to offer better prevention against THA subluxation and less wear of the implant

Introduction. Patients with knee osteoarthritis (OA) often tell us that they put extra load on the joints of the opposite leg as they walk. Multiple joint OA is common and has previously been related to gait changes due to hip OA (Shakoor et al 2002). The aim of this study was to determine whether patients with medial compartment knee OA have abnormal biomechanics of the unaffected knee and both hips during normal level gait. Methods. Twenty patients (11 male, 9 female), with severe medial compartment knee OA and no other joint pain were recruited. The control group comprised 20 adults without musculoskeletal pain. Patients were reviewed, x-rays were examined and WOMAC and Oxford knee scores were completed. A 12 camera Vicon (Vicon, Oxford) system was used to collect kinematic data (100Hz) on level walking and the ground reaction force was recorded using three AMTI force plates (1000Hz). Surface electrodes were placed over medial and lateral quadriceps and hamstrings bilaterally to record EMG data (1000Hz). Kinematics and kinetics were calculated using the Vicon ‘plug-in-gait’ model. A co-contraction index was calculated for the EMG signals on each side of the knee, representing the magnitude of the combined readings relative to their maximum contraction during the gait cycle. Statistical comparisons were performed using t-tests with Bonferroni's correction for two variables and ANOVA for more than two variables (SPSS v16). Results. The mean age of the patients was 69 (SD 8.8). Mean gait speed was 0.95m/s (study group) and 1.44m/s (control group). Peak adduction moments for the OA group [OA Knee; Unaffected Knee; Ipsilateral Hip; Contralateral Hip; in Nm/Kg(±95% CI)] were: 0.55(0.06); 0.47(0.06); 0.73(0.09); 0.73(0.08). Control values for peak moments were 0.64 (0.06) for the knee and 0.81(0.07) at the hip. Mid-stance adduction moments for the OA group (listed as before) were: 0.44(0.08); 0.33(0.06); 0.64(0.06); 0.61(0.08). Control values for mid-stance moments were 0.14(0.03) and 0.40(0.04). [OA group vs. Controls: p=NS for peak moments at all 4 joints; p<0.01 for mid-stance moments at all joints]. Co-contraction indices for hamstrings and quads, [OA knee medial; and lateral; unaffected knee medial; and lateral; control medial; and lateral; 0<X. Discussion. Although the affected subjects all had only single joint OA, abnormal moments were present in the hips and knees of both legs during normal level gait, despite the reduced gait speed of the OA cohort. Abnormal hamstring and quadriceps co-contraction occurs bilaterally in patient with single joint OA. Increased trunk sway is a recognised compensation in knee OA and may be the cause of the abnormal hip and contra-lateral knee loading found in this study. Further investigation is warranted and may lead to improvements in the long term outcome for these patients. Acknowledgement. The study was funded by the North Wales NHS Trust

Introduction. Numerous risk factors have been identified for patellar tendinopathy (PT), often in small population studies. The aim was to use an online questionnaire internationally to generate a large database and identify significant risk factors. Materials and Methods. Subjects were recruited from England, Spain and Italy with the questionnaire available in all three languages, with the questionnaire previously having been validated by Morton et al. (2014) as to be suitable for self-administration. The questionnaire can be viewed at: . http://patellartendinopathyquestionnaire.blogspot.co.uk/. (English), . http://tendinopatiarotuliana.blogspot.co.uk/. (Spanish) and . http://tendinopatiarotulea.blogspot.co.uk/. (Italian). All data was anonymised and password protected. 825 data sets were collected with 23.4% having clinically diagnosed PT. Results. Eight risk factors were included in the analysis based on a purposeful selection procedure: gender, hours of training, hamstring flexibility, previous patellar tendon rupture, previous knee injury, current/previous back pain, family history and age. To be female was found to be positively associated with PT, suggesting being female is protective (odds ratio (OR) = 0.70, 95% CI: 0.49–1.00, p=0.05). As hours of training increased the association with PT became stronger so that training >20 hours a week had a very significant OR of 8.94 (95%CI: 4.68–17.08, p<0.05), the most significant OR calculated. There was an association between a previous knee injury and PT (OR=2.10, 95% CI: 1.45–3.04, p<0.05) and having self-reported flexible hamstrings suggested some protection from PT (OR=0.61, 95% CI0.38–0.97, p=0.04). There was a trend towards association for back pain (OR=1.45, 95% CI: 0.99–2.14, p=0.06) and a family history of tendon problems (OR=1.51, 95% CI: 0.96–2.37, p=0.08). Discussion. Risk factors have been identified that are potentially modifiable in order to inform prevention and rehabilitation programmes; future research is required to establish causal relationships. Certain risk factors require investigation as they are not currently recognised in the literature

The objective was to seek evidence of hypoxia in early human tendinopathy and thereafter, to explore mechanisms whereby tissue hypoxia may regulate apoptosis, inflammatory mediators and matrix regulation in human tenocytes. Fifteen torn supraspinatus tendon (established pathology) and matched intact subscapularis tendon (representing ‘early pathology’) biopsies were collected from patients undergoing arthroscopic shoulder surgery. Control samples of subscapularis tendon were collected from 10 patients undergoing arthroscopic stabilisation surgery. Markers of hypoxia were quantified by immunohistochemical methods. Human tendon-derived primary cells were derived from hamstring tendon tissue obtained during hamstring tendon ACL reconstruction. The impact of hypoxia upon tenocyte biology ex vivo was measured using quantitative RT-PCR, multiplex cytokine assays, apoptotic proteomic profiling, immunohistochemistry and annexin V FACS staining. Increased expression of HIF 1a, Bcl-2 and clusterin (hypoxic and apoptotic markers) was detected in subscapularis tendon samples compared to both matched torn samples and non matched control samples (p<0.01). Hypoxic tenocytes exhibited increased production of proinflammatory cytokines (p<0.001), altered matrix regulation (p<0.01) with increased production of Collagen type III operating through a MAPK dependent pathway. Finally, hypoxia increased expression of several mediators of apoptosis and thereby promoted tenocyte apoptosis. Hypoxia promotes expression of proinflammatory cytokines, key apoptotic mediators and drives matrix component synthesis towards a collagen type III profile by human tenocytes. We propose hypoxic cell injury as a critical pathophysiological mechanism in early tendinopathy offering novel therapeutic opportunities in the management of tendon disorders

Summary Statement. ACL reconstruction using a quadriceps tendon autograft was quantitatively evaluated using a robotic testing system. Biomechanical results on joint stability and graft function support its use as an alternative to the hamstrings. Introduction. Recently, a number of surgeons have chosen the quadriceps tendon (QT) autograft as an alternative autograft over the hamstrings tendon for ACL reconstruction because its bone-to-bone healing on one side, large size, and preservation of lateral and rotatory knee function could lead to fewer post-operative complications. However, there have been little or no biomechanical studies that quantitatively evaluate knee function after reconstruction using a QT autograft. Therefore, the objective of this study was to assess the function of a reconstructed knee with a QT autograft and compare the results with a quadrupled semitendinosus and gracilis (QSTG) tendon autograft on the same knee. Methods. Ten human cadaveric knees (57.4 ± 4.2 years of age) were tested using a robotic/UFS testing system in 4 knee states: intact, ACL-deficient, and after ACL reconstruction with both QT and QSTG autografts. Reconstructions were performed in randomised order using posterolateral femoral tunnel placement. The knee kinematics in each state were measured at 5 flexion angles (full extension, 15°, 30°, 60°, and 90°) under 3 externally applied loading conditions: (1) 134 N anterior tibial load (ATL), (2) 134 N ATL with 200 N axial compression, and combined rotatory (CR) load of 10 Nm valgus and 5 Nm internal tibial torque (at 15° and 30°). Based on the established procedure, knee kinematics and in-situ forces were obtained using the principle of superposition. A repeated measures ANOVA was used to compare anterior tibial translation (ATT) and in-situ forces between the knee states at each flexion angle, with a Bonferroni post-hoc analysis. Results. Under the ATL, the ATT was found to be restored to within 1.1 mm of the intact knee for both reconstructions (P > 0.05). The in-situ forces in the grafts were also not significantly different from those in the intact ACL except in deep flexion (P < 0.05 at 90° for both grafts). With added axial compression, both reconstructions could still restore the ATT to within 2.4 mm of the intact joint at all flexion angles, and the in-situ forces in both grafts were within 25 N of the intact ACL at 15°, 30°, and 60° (P > 0.05). Under the CR load, knee kinematics and in-situ forces in the grafts were not significantly different from the intact ACL at any tested angle (P > 0.05). Further, no significant differences could be detected between the reconstructions under any experimental condition (P > 0.05). Discussion/Conclusion. ACL reconstruction with a QT autograft was found to restore knee function close to levels of the intact knee and similar to those reconstructed with a QSTG autograft. These results support clinical findings suggesting the QT autograft as a viable alternative for ACL reconstruction