Autologous osteochondral grafting has demonstrated positive outcomes for treating articular cartilage defects by replacing the damaged region with a cylindrical graft consisting of bone with a layer of cartilage, taken from a non-loadbearing region of the knee. Despite positive clinical use, factors that cause graft subsidence or poor integration are relatively unknown. The aim of this study was to develop finite element (FE) models of osteochondral grafts within a tibiofemoral joint and to investigate parameters affecting osteochondral graft stability. Initial experimental tests on cadaveric femurs were performed to calibrate the bone properties and graft-bone frictional forces for use in corresponding FE models, generated from µCT scan data. The effects of cartilage defects and osteochondral graft repair were measured by examining contact pressure changes using in vitro tests on a single cadaveric human tibiofemoral joint. Six defects were created in the femoral condyles which were subsequently treated with osteochondral autografts or metal pins. Matching µCT scan-based FE models were created, and the contact patches were compared. Sensitivity to graft bone properties was investigated. The bone material properties and graft-bone frictional forces were successfully calibrated from the initial tests with good resulting levels of agreement (CCC=0.87). The tibiofemoral joint experiment provided a range of cases to model. These cases were well captured experimentally and represented accurately in the FE models. Graft properties relative to host bone had large effects on immediate graft stability despite limited changes to resultant cartilage contact pressure. Model confidence was built through extensive validation and sensitivity testing, and demonstrated that specimen-specific properties were required to accurately represent graft behaviour. The results indicate that graft bone properties affect the immediate stability, which is important for the selection of allografts and design of future synthetic grafts. Acknowledgements. Supported by the EPSRC-EP/P001076

Articular cartilage is often damaged, and its treatment is usually performed by surgical operation. Today, tissue engineering offers an alternative treatment option for injuries or diseases with increasing importance. Infrapatellar fat pad (IPFP) is a densely vascularized and innervated extra synovial tissue that fills the anterior knee compartment. Adipose-derived stem cells from infrapatellar fat pad (IPFP-ASCs) have multipotency means that they can differentiate into connective tissue cells and have age-independent differentiation capacity as compared to other stem cells. In this study, the osteochondral tissue construct was designed with different inner pattern due to original osteochondral tissue structure and fabrication of it was carried out by 3D printing. For this purpose, alginate (3% w/v) and carboxymethylcellulose (CMC) (9%w /v) were used as bioink. Also, IPFP-ASCs were isolated with enzymatic degradation. Osteogenic and chondrogenic differentiation of IPFP-ASCs were investigated with Alizarin Red and Alcian Blue staining, respectively. IPFP-ASCs-laden osteochondral graft differentiation will be induced by controlled release of growth factor BMP-2 and TGF-β. Before this step, nanocapsules formation with double emission technique with model protein BSA was carried out with different concentration of PCL (5%,10% and 20%). The morphology and structure of the nanocapsules were determined with scanning electron microscopy (SEM). Also, we successfully designed and printed alginate and CMC based scaffold with 20 layers. Chondrogenic and osteogenic differentiation of IPFP-ASCs with suitable culture conditions was obtained. The isolation of IPFP-ASCs, formation of the nanocapsules, and 3D printing of osteochondral graft were carried out successfully

Osteochondral (OC) grafting is one available method currently used to repair full thickness cartilage lesions with good results clinically when grafting occurs in patients with specific positive prognostic factors. However, there is poor understanding of the effect of individual patient and surgical factors. With limited tissue availability, development of Finite Element (FE) models taking into account these variations is essential. The aim of this study was to evaluate the effect of altering the material properties of OC grafts and their host environment through computer simulation. A generic FE model (ABAQUS CAE 2017) of a push-out test was developed as a press-fit bone cylinder (graft) sliding inside a bone ring (host tissue). Press-fit fixation was simulated using an interference fit. Overlap between host and graft (0.01mm–0.05mm) and coefficient of friction (0.3–0.7) were varied sequentially. Bone Young's moduli (YM) were varied individually between graft and host within the range of otherwise derived tissue moduli (46MPa, 82MPa, 123MPa). Increasing both overlap and frictional coefficient increased peak dislodging force independently (overlap: 490% & frictional coefficient: 176% across range tested). Increasing bone modulus also increased dislodging force, with host bone modulus (107%, 128%, and 140% increase across range, when Graft YM = 123MPa, 82 MPa, and 46MPa, respectively) having a greater influence than graft modulus (28%, 19% and 10% increase across range, when Host YM = 123 MPa, 82MPa and 46MPa, respectively). As anticipated increasing overlap and friction caused an increase in force necessary to dislodge the graft. Importantly, differentially changing the graft and host material properties changed the dislodging force indicating that difference between graft and host may be an important factor in the success or failure clinically of osteochondral grafting

We examined osteochondral autografts, obtained at a mean of 19.5 months (3 to 48) following extracorporeal irradiation and re-implantation to replace bone defects after removal of tumours. The specimens were obtained from six patients (mean age 13.3 years (10 to 18)) and consisted of articular cartilage (five), subchondral bone (five), external callus (one) and tendon (one). The tumour cells in the grafts were eradicated by a single radiation dose of 60 Gy. In three cartilage specimens, viable chondrocytes were detected. The survival of chondrocytes was confirmed with S-100 protein staining. Three specimens from the subchondral region and a tendon displayed features of regeneration. Callus was seen at the junction between host and irradiated bone.

Background

Based on decellularisation and cleaning processes of trabecular bone and fibrocartilage, an osteochondral allograft has been developed.

Introduction

Articular hyaline cartilage has a unique structural composition that allows it to endure high load, distribute load to bone and enables low friction movement in joints. A novel acellular xenogenic graft is proposed as a biological cartilage replacement, for repair of osteochondral defects. Acellular porcine cartilage has been produced using repeated freeze thaw cycles and washing using hypotonic buffers and sodium dodecyl sulphate solution (SDS; Keir, 2008). DNA content of the acellular matrix was reduced by 93.3% compared to native cartilage as measured by nanodrop spectrophotometry of extracted DNA, with a corresponding reduction in glycosaminoglycan (GAG) content.

The clinical success of osteochondral autografts is heavily reliant on their mechanical stability, as grafts which protrude above or subside below the native cartilage can have a negative effect on the tribological properties of the joint [1]. Furthermore, high insertion forces have previously been shown to reduce chondrocyte viability [2]. Commercial grafting kits may include a dilation tool to increase the diameter of the recipient site prior to insertion. The aim of this study was to evaluate the influence of dilation on the primary stability of autografts. Six human cadaveric femurs were studied. For each femur, four 8.5 × 8mm autografts were harvested from the trochlear groove and implanted into the femoral condyles using a Smith & Nephew Osteochondral grafting kit. Two grafts were implanted into dilated recipient sites (n=12) and two were implanted with no dilation (n=12). Insertion force was measured by partially inserting the graft and applying a load at a rate of 1 mm/min, until the graft was flush with the surrounding cartilage. Push-in force was measured by applying the same load, until the graft had subsided 4mm below congruency. Significance was taken as (p<0.05). Average maximum insertion force of dilated grafts was significantly lower (p<0.001) than their non-dilated equivalent [28.2N & 176.7N respectively]. There was no significant difference between average maximum push-in force between the dilated and non-dilated groups [1062.8N & 1204.2N respectively]. This study demonstrated that significantly less force is required to insert dilated autografts, potentially minimising loss of chondrocyte viability. However, once inserted, the force required to displace the grafts below congruency remained similar, indicating a similar degree of graft stability between both groups

Abstract. Objectives. Osteochondral grafting (OCG) is one treatment strategy for osteoarthritis with good clinical results. Decellularised tissues provide a promising alternative to standard autografts or allografts. This study aimed to compare the stability of traditional OCG and decellularised scaffolds upon initial implantation. Methods. Host cubes (N=16) were extracted from porcine femoral condyles around an artificial defect hole. Grafts (N=11) were harvested from the trochlear groove; porcine decellularized osteochondral scaffold (N=5) were prepared. Each host was secured in fixtures and submerged in PBS at 37 ºC. Each graft or scaffold was press fit into one of the hosts, then pushed in for 5 mm, using an indenter (Instron3365) and pushed out in the opposite direction for 10 mm. Parameters analysed were the force required to initiate movement (Dislodging Force) and the maximum force (Max Force). Results. The Dislodging Force of grafts (mean ± std. dev) was 133±15 N for the push in test and 109±11 N for the push out test. This was significantly higher than values for scaffolds: IN 24±1 N and OUT 26±5 N. The Max Force were also larger in the grafts than the scaffolds: IN 152±16 N vs. 41±4 N and OUT 118±14 N vs. 33±3 N. Conclusions. The force required to dislodge a graft or scaffold from a host environment was similar for the push in test and the push out test, suggesting it is a good measure of initial stability. Upon initial implantation, the decellularised scaffolds were easier to dislodge than the OCG. Previously, the decellularisation process was found to soften bone, relative stiffness may thus be an important consideration in graft fixation. A greater press fit may be necessary for decellularised scaffolds in order to achieve the same level of graft stability as natural OCG when used in vivo. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Osteoarthritis (OA) affects over 8.75 million people in the UK creating the need for early stage interventions. Osteochondral (OC) grafting has been used to repair full thickness lesions but the efficacy of this therapy is questionable. As a first step in developing a testing framework able to predict the potential and suitability of OC grafts for repair, here, we present experimental data to be used in informing boundary conditions, input parameters and testing sequences for developing and verifying an FE model of the interaction of OC grafts and surrounding host tissue. Ten OC cylindrical grafts (height: 10mm; diameter: n=5–6.5mm; n=5–8.5mm) were harvested from adult porcine femurs (60–70kg). Unconfined compression tests were conducted using an Instron3365 with a 500N load cell and a BioBath filled with PBS at 37ºC. The OC grafts (prior to separation of cartilage and bone) and cartilage underwent four 5% strain (of cartilage layer) steps with displacement rate of 0.005mm/sec, each followed by a 45-minute relaxation. Final strain was 20%. Bone underwent a single displacement of 20% strain of bone at same displacement rate. Young's moduli ranged from 6.2–42.0MPa, 0.7–3.9MPa, 46.8–123.7MPa for OC graft, cartilage and bone, respectively. The coefficient of variance between OC Grafts, cartilage and bone was 70.6%, 71.8%, and 25.2%, respectively. Dispersion between samples was high. This may be due to intrinsic tissue variability but also due to the testing protocol: for cartilage in particular, the load was at the low end of the load cell capacity and the sample aspect ratio was poor for compressive testing. This work provides insight in understanding the effect of individual patient's and/or individual grafts used during osteochondral grafting. The results compel the need to accurately model these tissues when developing specimen-specific FE models for OC grafting

Joint surface restoration of deep osteochondral defects represents a significant unmet clinical need. Moreover, untreated lesions lead to a high rate of osteoarthritis. The current strategies to repair deep osteochondral defects such as osteochondral grafting or sandwich strategies combining bone autografts with ACI/MACI fail to generate long-lasting osteochondral interfaces. Herein, we investigated the capacity of juvenile Osteochondral Grafts (OCGs) to repair osteochondral defects in skeletally mature animals. With this regenerative model in view, we set up a new biological, bilayered, and scaffold-free Tissue Engineered (TE) construct for the repair of the osteochondral unit of the knee. Skeletally immature (5 weeks old) and mature (11 weeks old) Lewis rats were used. Cylindrical OCGs were excised from the intercondylar groove of the knee of skeletally immature rats and transplanted into osteochondral defects created in skeletally mature rats. To create bilayered TE constructs, micromasses of human periosteum-derived progenitor cells (hPDCs) and human articular chondrocytes (hACs) were produced in vitro using chemically defined medium formulations. These constructs were subsequently implanted orthotopically in vivo in nude rats. At 4 and 16 weeks after surgery, the knees were collected and processed for subsequent 3D imaging analysis and histological evaluation. Micro-computed tomography (µCT), H&E and Safranin O staining were used to evaluate the degree of tissue repair. Our results showed that the osteochondral unit of the knee in 5 weeks old rats exhibit an immature phenotype, displaying active subchondral bone formation through endochondral ossification, the absence of a tidemark, and articular chondrocytes oriented parallel to the articular surface. When transplanted into skeletally mature animals, the immature OCGs resumed their maturation process, i.e., formed new subchondral bone, partially established the tidemark, and maintained their Safranin O-positive hyaline cartilage at 16 weeks after transplantation. The bilayered TE constructs (hPDCs + hACs) could partially recapitulate the cascade of events as seen with the immature OCGs, i.e., the regeneration of the subchondral bone and the formation of the typical joint surface architecture, ranging from non-mineralized hyaline cartilage in the superficial layers to a progressively mineralized matrix at the interface with a new subchondral bone plate. Cell-based TE constructs displaying a hierarchically organized structure comprising of different tissue forming units seem an attractive new strategy to treat osteochondral defects of the knee

Surgical reconstruction of articular surfaces by transplantation of osteochondral autografts has shown considerable promise in the treatment of focal articular lesions. During mosaicplasty, each cylindrical osteochondral graft is centred over the recipient hole and delivered by impacting the articular surface. Impact loading of articular cartilage has been associated with structural damage, loss of the viability of chondrocytes and subsequent degeneration of the articular cartilage. We have examined the relationship between single-impact loading and chondrocyte death for the specific confined-compression boundary conditions of mosaicplasty and the effect of repetitive impact loading which occurs during implantation of the graft on the resulting viability of the chondrocytes. Fresh bovine and porcine femoral condyles were used in this experiment. The percentage of chondrocyte death was found to vary logarithmically with single-impact energy and was predicted more strongly by the mean force of the impact rather than by the number of impacts required during placement of the graft. The significance of these results in regard to the surgical technique and design features of instruments for osteochondral transplantation is discussed

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

We evaluated the histological changes before and after fixation in ten knees of ten patients with osteochondritis dissecans who had undergone fixation of the unstable lesions. There were seven males and three females with a mean age of 15 years (11 to 22). The procedure was performed either using bio-absorbable pins only or in combination with an autologous osteochondral plug. A needle biopsy was done at the time of fixation and at the time of a second-look arthroscopy at a mean of 7.8 months (6 to 9) after surgery.

The biopsy specimens at the second-look arthroscopy showed significant improvement in the histological grading score compared with the pre-fixation scores (p < 0.01). In the specimens at the second-look arthroscopy, the extracellular matrix was stained more densely than at the time of fixation, especially in the middle to deep layers of the articular cartilage.

Our findings show that articular cartilage regenerates after fixation of an unstable lesion in osteochondritis dissecans.

The treatment of osteochondral lesions and osteoarthritis remains an ongoing clinical challenge in orthopaedics. This review examines the current research in the fields of cartilage regeneration, osteochondral defect treatment, and biological joint resurfacing, and reports on the results of clinical and pre-clinical studies. We also report on novel treatment strategies and discuss their potential promise or pitfalls. Current focus involves the use of a scaffold providing mechanical support with the addition of chondrocytes or mesenchymal stem cells (MSCs), or the use of cell homing to differentiate the organism’s own endogenous cell sources into cartilage. This method is usually performed with scaffolds that have been coated with a chemotactic agent or with structures that support the sustained release of growth factors or other chondroinductive agents. We also discuss unique methods and designs for cell homing and scaffold production, and improvements in biological joint resurfacing. There have been a number of exciting new studies and techniques developed that aim to repair or restore osteochondral lesions and to treat larger defects or the entire articular surface. The concept of a biological total joint replacement appears to have much potential.

Cite this article: Bone Joint Res 2013;2:193–9.

We developed a new porous scaffold made from a synthetic polymer, poly(DL-lactide-co-glycolide) (PLG), and evaluated its use in the repair of cartilage. Osteochondral defects made on the femoral trochlear of rabbits were treated by transplantation of the PLG scaffold, examined histologically and compared with an untreated control group.

Fibrous tissue was initially organised in an arcade array with poor cellularity at the articular surface of the scaffold. The tissue regenerated to cartilage at the articular surface. In the subchondral area, new bone formed and the scaffold was absorbed. The histological scores were significantly higher in the defects treated by the scaffold than in the control group (p < 0.05).

Our findings suggest that in an animal model the new porous PLG scaffold is effective for repairing full-thickness osteochondral defects without cultured cells and growth factors.

In this study a combination of autologous chondrocyte implantation (ACI) and the osteochondral autograft transfer system (OATS) was used and evaluated as a treatment option for the repair of large areas of degenerative articular cartilage. We present the results at three years post-operatively. Osteochondral cores were used to restore the contour of articular cartilage in 13 patients with large lesions of the lateral femoral condyle (n = 5), medial femoral condyle (n = 7) and patella (n = 1). Autologous cultured chondrocytes were injected underneath a periosteal patch covering the cores. After one year, the patients had a significant improvement in their symptoms and after three years this level of improvement was maintained in ten of the 13 patients. Arthroscopic examination revealed that the osteochondral cores became well integrated with the surrounding cartilage. We conclude that the hybrid ACI/OATS technique provides a promising surgical approach for the treatment of patients with large degenerative osteochondral defects.