As arthroplasty demand grows worldwide, the need for a novel cost-effective treatment option for articular cartilage (AC) defects tailored to individual patients has never been greater. 3D bioprinting can deposit patient cells and other biomaterials in user-defined patterns to build tissue constructs from the “bottom-up,” potentially offering a new treatment for AC defects. The aim of this research was to create bioinks that can be injected or 3D bioprinted to aid osteochondral defect repair using human cells.

Novel composite bioinks were created by mixing different ratios of methacrylated alginate (AlgMA) with methacrylated gelatin (GelMA). Chondrocytes or mesenchymal stem cells (MSCs) were then encapsulated in the bioinks and 3D bioprinted using a custom-built extrusion bioprinter. UV and double-ionic (BaCl2 and CaCl2) crosslinking was deployed following bioprinting to strengthen bioink stability in culture. Chondrocyte and MSC spheroids were also produced via 3D culture and then bioprinted to accelerate cell growth and development of ECM in bioprinted constructs.

Excellent viability of chondrocytes and MSCs was seen following bioprinting (>95%) and maintained in culture over 28 days, with accelerated cell growth seen with inclusion of MSC or chondrocyte spheroids in bioinks (p<0.05). Bioprinted 10mm diameter constructs maintained shape in culture over 28 days, whilst construct degradation rates and mechanical properties were improved with addition of AlgMA (p<0.05). Composite bioinks were also injected into in vitro osteochondral defects (OCDs) and crosslinked in situ, with maintained cell viability and repair of osteochondral defects seen over a 14-day period. In conclusion we developed novel composite AlgMA/GelMA bioinks that can be triple-crosslinked, facilitating dense chondrocyte and MSC growth in constructs following 3D bioprinting. The bioink can be injected or 3D bioprinted to successfully repair in vitro OCDs, offering hope for a new approach to treating AC defects.

As arthroplasty demand grows worldwide, the need for a novel cost-effective treatment option for articular cartilage (AC) defects tailored to individual patients has never been greater. 3D bioprinting can deposit patient cells and other biomaterials in user-defined patterns to build tissue constructs from the “bottom-up,” potentially offering a new treatment for AC defects. The aim of this research was to create bioinks that can be injected or 3D bioprinted to aid osteochondral defect repair using human cells.

Novel composite bioinks were created by mixing different ratios of methacrylated alginate (AlgMA) with methacrylated gelatin (GelMA). Chondrocytes or mesenchymal stem cells (MSCs) were then encapsulated in the bioinks and 3D bioprinted using a custom-built extrusion bioprinter. UV and double-ionic (BaCl2 and CaCl2) crosslinking was deployed following bioprinting to strengthen bioink stability in culture. Chondrocyte and MSC spheroids were also bioprinted to accelerate cell growth and development of ECM in bioprinted constructs.

Excellent viability of chondrocytes and MSCs was seen following bioprinting (>95%) and maintained in culture over 28 days, with accelerated cell growth seen with inclusion of MSC or chondrocyte spheroids in bioinks (p<0.05). Bioprinted 10mm diameter constructs maintained shape in culture over 28 days, whilst construct degradation rates and mechanical properties were improved with addition of AlgMA (p<0.05). Composite bioinks were also injected into in vitro osteochondral defects (OCDs) and crosslinked in situ, with maintained cell viability and repair of osteochondral defects seen over a 14-day period.

In conclusion we developed novel composite AlgMA/GelMA bioinks that can be triple-crosslinked, facilitating dense chondrocyte and MSC growth in constructs following 3D bioprinting. The bioink can be injected or 3D bioprinted to successfully repair in vitro OCDs, offering hope for a new approach to treating AC defects.

Patients with osteoarthritis (OA) of the knee commonly alter their movement to compensate for deficiencies. This study presents a new numerical procedure for classifying sit-to-walk (STW) movement strategies.

Ten control and twelve OA participants performed the STW task in a motion capture laboratory. A full body biomechanical model was used. Participants were instructed to sit in a comfortable self-selected position on a stool height adjusted to 100% of their knee height and then stand and pick up an object from a table in front of them. Three matrices were constructed defining the progression of the torso, feet and hands in the sagittal plane along with a fourth expressing the location of the hands relative to the knees. Hierarchical clustering (HC) was used to identify different strategies. Trials were also classified as to whether the left (L) and right (R) extremities used a matching strategy (bilateral) or not (asymmetrical). Fisher's exact test was used to compare this between groups.

Clustering of the torso matrix dichotomised the trials in two major clusters; subjects leaning forward (LF) or not. The feet and hands matrices revealed sliding the foot backward (FB) and moving an arm forward (AF) strategies respectively. Trials not belonging in the AF cluster were submitted to the last HC of the fourth matrix exposing three additional strategies, the arm pushing through chair (PC), arm pushing through knee (PK) and arm not used (NA). The control participants used the LF+FBR+PK combination most frequently whereas the OA participants used the AFR+PCL. OA patients used significantly more asymmetrical arm strategies, p=0.034.

The results demonstrated that control and OA participants favour different STW strategies. The OA patients asymmetrical arm behaviour possibly indicates compensating for weakness of the affected leg. These strategy definitions may be useful to assess post-operative outcomes and rehabilitation progress.

One of the most common bacteria in orthopaedic prosthetic infections is Staphylococcus Aureus. Infection causes implant failure due to biofilm production. Biofilms are produced by bacteria once they have adhered to a surface.

Nanotopography has major effects on cell behaviour. Our research focuses on bacterial adhesion on nanofabricated materials. We hypothesise that surface nanotopography impacts the differential ability of staphylococci species to adhere via altered metabolomics and may reduce orthopaedic implant infection rate.

Bacteria were grown and growth conditions optimised. Polystyrene and titanium (Ti) nanosurfaces were studied. The polystyrene surfaces had different nanopit arrays, while the Ti surfaces expressed different nanowire structures. Adhesion analysis was performed using fluorescence imaging, quantitative PCR and bacterial percentage coverage calculations. Further substitution with ‘heavy’ labelled glucose into growth medium allowed for bacterial metabolomic analysis and identification of any up-regulated metabolites and pathways.

Our data demonstrates reduced bacterial adhesion on specific nanopit polystyrene arrays, while nanowired titanium showed increased bacterial adhesion following qPCR (P<0.05) and percentage coverage calculations (P<0.001). Further metabolomic analysis identified significantly increased intensity counts of specific metabolites (Pyruvate, Aspartate, Alanine and Carbamoyl aspartate).

Our study shows that by altering nanotopography, bacterial adhesion and therefore biofilm formation can be affected. Specific nanopatterned surfaces may reduce implant infection associated morbidity and mortality. The identification of metabolic pathways involved in adhesion may allow for a targeted approach to biofilm eradication in S. aureus. This is of significant benefit to both the patient and the surgeon, and may well extend far beyond the realms of orthopaedics.

The most common bacteria in orthopaedic prosthetic infections are Staphylococcus, namely Staphylococcus Epidermidis (SE) and Staphylococcus Aureus (SA). Infection causes implant failure due to biofilm production. Biofilms are produced by bacteria once they have adhered to a surface.

Nanotopography has major effects on cell behaviour. Our research focuses on bacterial adhesion and biofilm formation on nanofabricated materials. Bacteria studied were clinically relevant from an orthopaedic perspective, SA and SE. We hypothesise that that nanosurfaces can modulate bacterial adherence and biofilm formation and may reduce orthopaedic implant infection rate.

Isolated bacteria were grown and growth conditions optimised. Bacterial concentrations were calculated by using qPCR. Statistical analysis allowed identification of optimal biofilm growth conditions. These were refined on standard, non-nanopatterned surfaces, and then control and nanopatterned polystyrene (nanopits) and titanium plates (nanowires). Adhesion analysis was performed using fluorescence imaging and quantitative PCR.

4 bacterial strains were isolated and cultured. Growth kinetics based on 24hr cultures allowed isolation of optimal media for biofilm conditions (Dulbecco's Modified Eagle Medium with additional supplements). Highest bacterial concentrations were found following 2hrs incubation with Lysozyme during qPCR. Bacterial concentration significantly increased between 30, 60 and 90 minutes incubation. Differences in percentage coverage on different polysyrene nanosurfaces (nanopits) were noted varying. This was confirmed by qPCR extractions that showed different bacterial concentrations on different nanopatterns. Titanium nanowire surfaces significantly increased bacterial adhesion (P<0.05).

Our study cultured and quantified bacterial biofilm and suggests that by altering nanotopography, bacterial adhesion and therefore biofilm formation can be affected. Specific nanopatterned surfaces may reduce implant infection associated morbidity and mortality. Clearly this is of significant benefit to the patient, the surgeon and the NHS, and may well extend far beyond the realms of orthopaedics.

In recent years there has been growing interest in enhanced recovery regimes in lower limb arthroplasty due to potential clinical benefits of early mobilisation along with cost-savings. Following adoption of this regime in a district general hospital, it was observed that traditional dressings were a potential barrier to its success with ongoing wound problems in patients otherwise fit for discharge. The aim of this audit was to assess current wound care practice, implement a potentially improved regime and re-evaluate practice.

A prospective clinical audit was performed over a three month period involving 100 patients undergoing hip or knee arthroplasty. Fifty patients with traditional dressings were evaluated prior to change in practice to a modern dressing (Aquacel™ Surgical). Fifty patients were then evaluated with the new dressing to complete the audit cycle. Clinical outcome measures included wear time, number of changes, blister rate and length of stay. Statistical comparisons were performed using Mann Whitney or Fisher's Exact test (statistical significance, p<0.05).

Wear time for the traditional dressing (2 days) was significantly shorter than the modern dressing (7 days), p<0.001, and required more changes (0 vs. 3 days), p<0.001. 20% of patients developed blisters with the traditional dressing compared with 4% with the modern dressing (p=0.028). Length of stay was the same for the modern dressing (4 days) compared with the traditional dressing (4 days). However, in the modern group 75% of patients were discharged by day 4 whereas in the traditional group this took until day 6.

This audit highlights the problems associated with traditional dressings with frequent early dressing changes, blistering and delayed discharge. These adverse outcomes can be minimised with a modern dressing specifically designed for the demands of lower limb arthroplasty. Units planning to implement enhanced recovery regimes should consider adopting this dressing to avoid compromising patient discharge.

This study aimed to overcome the subjective nature of routine assessment of knee laxity and develop a repeatable, objective method using a hand-held force application device (FAD).

Eighteen clinicians (physiotherapists, consultants, trainees) volunteered to measure the coronal angular deviation of the right knee of a healthy volunteer using a validated non-invasive infrared measuring system. Effort was taken to ensure the knee flexion angle (∼2°) and hand positions were constant during testing. Three varus and valgus stress tests were conducted, in which maximum angular deviation was determined and subsequently averaged, in the following order of conditions: manual stress without the FAD up to a perceived end-point (before); with the FAD to apply a moment of 18 Nm; and again without the FAD (after). A repeated measures ANOVA was used to analyse the results.

All three groups of clinicians produced measurements of valgus laxity with consistent mean values and standard deviations (<1°) for each condition. For varus mean values were consistent but standard deviations were larger.

Valgus deviations varied significantly between conditions (p < 0.01), with deviations achieved using the FAD greater than both before (p < 0.01) and after (p < 0.05) indicating that the perceived endpoints were less than that achieved at 18 Nm. However varus perceived endpoints were no different to that achieved at 18 Nm, suggesting that clinicians usually apply a greater valgus moment than varus. Furthermore, the non-significant increase in valgus deviation between before and after (p = 0.123) is suggestive of a training trend, especially for trainees.

Our standardised knee laxity assessment may have a role in improving the balancing techniques of TKA and the diagnosis of collateral ligament injuries. Also, by quantifying the technique of senior clinicians, and with use of the FAD, the perceptive skills of more junior trainees may be enhanced.

Knee alignment is a fundamental measurement in the assessment, monitoring and surgical management of patients with OA. In spite of extensive research into the consequences of malalignment, there is a lack of data regarding the potential variation between supine and standing (functional) conditions. The purpose of this study was to explore this relationship in asymptomatic, osteoarthritic and prosthetic knees. Our hypothesis was that the change in alignment of these three groups would be different.

Infrared position capture was used to assess knee alignment for 30 asymptomatic controls and 31 patients with OA, before and after TKA. Coronal and sagittal mechanical femorotibial (MFT) angles in extension (negative values varus/hyperextension) were measured supine and in bi-pedal stance and changes analysed using a paired t-test. To quantify this change in 3D, vector plots of ankle centre displacement relative to the knee centre were produced.

Alignment in both planes changed significantly from supine to standing for all three groups, most frequently towards relative varus and extension. In the coronal plane, the mean±SD(°) of the supine/standing MFT angles was 0.1±2.5/−1.1±3.7 for asymptomatic (p=0.001), −2.5±5.7/−3.6±6.0 for osteoarthritic (p=0.009) and −0.7±1.4/ −2.5±2.0 for prosthetic knees (p<0.001). In the sagittal plane, the mean±SD(°) of the supine/standing MFT angles was −1.7±3.3/−5.5±4.9 for asymptomatic (p<0.001), 7.7±7.1/1.8±7.7 for osteoarthritic (p<0.001) and 6.8±5.1/1.4±7.6 for prosthetic knees (p<0.001). The vector plots showed that the trend of relative varus and extension in stance was similar in overall magnitude and direction between the groups.

The similarities between each group did not support our hypothesis. The consistent kinematic pattern for different knee types suggests that soft tissue restraints rather than underlying joint deformity may be more influential in dynamic control of alignment from lying to standing. This potential change should be considered when positioning TKA components on supine limbs as post-operative functional alignment may be different.

Range of motion (ROM) is a well recognised outcome measure following TKA and combines both knee flexion and extension. In contrast to achieved flexion, fewer studies have recognised the importance of fixed flexion deformity (FFD). A post-operative FFD can adversely affect pain and functional knee scores and so its detection is important. The aim of this study was to investigate the accuracy of standard clinical ROM measurement techniques following TKA and determine their reliability for recognising FFD.

Thirty patients six weeks post-TKA had knee ROM measurements performed with an infrared (IR) tracking system of ±1 accuracy. The patients were also assessed independently by experienced arthroplasty practitioners using a standardised goniometric measurement technique. For goniometric clinically-measured flexion (Clin_flex) and extension (Clin_ext) linear models were generated using IR-measured flexion and extension (IR_flex and IR_ext), BMI and gender as covariables. Data for extension was categorised as FFD present or absent based on Ritter's criteria and agreement was assessed using Kappa.

For both models neither BMI nor gender were significant variables. Models were Clin_flex = 0.54 + 0.66∗IR_flex (r²_adj=0.521) and Clin_ext = 0.23 + 0.50∗IR_ext (r²_adj=0.247), showing that for every 10° increase in flexion, clinical measurement only increased by 7° and for every 10° increase in FFD angle, clinical measurement only increased by 5°. In identifying FFD there was moderate agreement between the two measurements (κ=0.44) with disagreement for nine patients all being patients with FFD that were not identified clinically.

For both flexion and extension there was a greater tendency for the goniometric clinical measurements to underestimate the actual angle. In the context of TKA outcome for maximum flexion this may be preferable to overestimating. In contrast, underestimating the degree of flexion when the knee is in the extended position is not desirable as it will potentially underreport the frequency and magnitude of FFD.

Assessment of coronal knee laxity via manual stress testing is commonly performed during joint examination. While it is generally accepted that the knee should be flexed slightly to assess its collateral restraints, the importance of the exact degree of flexion at time of testing has not been documented. The aim of this study therefore was to assess the effect of differing degrees of knee flexion on the magnitude of coronal deflection observed during collateral stress testing.

Using non-invasive infrared technology, the real-time coronal and sagittal mechanical femorotibial (MFT) angles of three asymptomatic volunteers were measured. A single examiner, blinded to the real-time display of coronal but not sagittal alignment, held the knee in maximum extension and performed manual varus and valgus stress manoeuvres to a perceived end-point. This sequence was repeated at 5° increments up to 30° of flexion. This provided unstressed, varus and valgus coronal alignment measurements as well as overall envelope of laxity (valgus angle – varus angle) which were subsequently regressed against knee flexion.

Regression analysis indicated that all regression coefficients were significantly different to zero (p < 0.001). With increasing knee flexion, valgus MFT angles became more valgus and varus MFT angles became more. The overall laxity of the knee in the coronal plane increased approximately fourfold with 30° of knee flexion.

The results demonstrated that small changes in knee flexion could result in significant changes in coronal knee laxity, an observation which has important clinical relevance and applications. For example the assessment of medial collateral ligament (MCL) injuries can be based on the perceived amount of joint opening with no reference made to knee flexion at time of assessment. Therefore, close attention should be paid to the flexion angle of the knee during stress testing in order to achieve a reliable and reproducible assessment.

The assessment of knee laxity by application of varus and valgus stress is a subjective clinical manoeuvre often used for soft tissue balancing in arthroplasty or for diagnosis of collateral ligament injuries. Quantitative adjuncts such as stress radiographs have enabled a more objective measurement of angular deviation but may be limited by variations in examination technique. The aim of this study was to quantify clinical knee laxity assessment by measurement of applied forces and resultant angulations.

A novel system for measuring the manually-applied forces and moments was developed. Both hardware and software components underwent laboratory validation prior to volunteer testing. Two clinicians performed multiple blinded examinations on two volunteers and the corresponding angular deviations were measured using a validated non-invasive system with a repeatability of ±1° for coronal alignment. The distance between the kinematically-determined knee and ankle centres was used as the moment arm.

Comparison of single measurements of laxity showed a wide intra- and inter-observer variation (up to 3°). However, when the median value of repeated measurements was used there was good repeatability for both a single surgeon on different days and between the two clinicians with angular measurements agreeing within 1°. In spite of this agreement, the magnitudes of the tangential forces and moments applied varied between clinicians and did not correlate with the corresponding angular deviations.

It was not possible to standardise clinical examination using the current system. Orientation of the applied force with respect to the leg was not quantified and during force measurement it became apparent that the assumed tangential direction of application was not true. This may explain the lack of correlation between the force and angulation data. However, for quantitative measurement of coronal knee laxity using non-invasive laxity measurements, the use of a repeated measures protocol may be accurate enough for clinical application.