Current bone grafts include allograft and autografts, both of which have limitations. Tissue engineering biotechnology has shown considerable promise in improving grafts. A competent graft material should ideally have osteoconductive and osteoinductive properties and comprise of bone forming cells and osteoinductive growth factors. In this study, we have evaluated the in vitro formation of bone and have used human demineralised bone matrix [DBM] and human insoluble collagenous matric [ICM] as scaffolds for mesenchymal stem cells [MSCs] and osteogenic protein [OP-1]. The objective was to determine whether combined addition of OP-1 and MSCs resulted in a superior bone graft substitute by improving the inherent osteoinductive property.

DBM and ICM were prepared and combined with rhOP [1.4 mg/0.25 mg of bone] and MSCs [1 x 105/ ml]. Statistically significant differences in MSC proliferation were seen between materials with and without OP-1 [P< 0.05}, n=8] in DBM on day 1, and both DBM and ICM on day 7 and 14. Enhanced osteogenic differentiation was observed in the presence of OP-1 when compared to DBM alone and on DBM and ICM with OP-1. In conclusion MSCs and OP-1 can be seeded together on DBM and ICM and Von Kossa staining and X-ray analysis confirmed in vitro de novo bone formation, with DBM + MSCs + OP-1 being more successful in this regard.

Conclusion: To date, no other study, to the author’s knowledge, has used MSCs and OP-1 together on a graft material; this funding, therefore, has very important clinical implications.

Limitations of allografts and autografts for bone repair have increased the demand for a synthetic bone graft substitute for load-bearing and non-load bearing osseous defects. Tissue engineering of bone has thus been implicated to circumvent and eliminate the limitations of existing therapies, with living cell-scaffold constructs ultimately “integrating” with the patients own tissue. Bone engineering requires cells, growth inducing factors and a scaffold for delivery of cells to the anatomic site, creation of 3-D space for tissue formation and mechanical support. In this study, we investigated whether addition of osteogenic Protein-1 (OP-1) enhanced the osseoinductive properties of hydroxyapatite (HA) loaded with mesenchymal stem cells (MSCs). The study was conducted over a fourteen day period and the two groups HA/MSC and HA/MSC loaded with OP-1 were analysed qualitatively by SEM and quantitatively by assessment of proliferation (Alamar blue assay and total cellular DNA) and differentiation marker alkaline phosphatase activity (ALP). HA/MS/OP-1 showed a statistically significant (p< 0.05) increase in cell proliferation (286.52 ± 58.2) compared to the unloaded samples (175.62 ± 23.51). ALP activity (release) was also significantly enhanced (p < 0.05) in the loaded samples at day 14 (12.63 ± 1.58) compared to the control (2.73 ± 1.07).

Conclusion: the osseoinductive potential of HA was markedly improved by the incorporation of MSC’s and OP-1. This type of graft could provide improved mechanical stability at an earlier time point, and may influence future clinical application of HA for load bearing sites.

Implantation of autologous chondrocytes and matrix autologous chondrocytes are techniques of cartilage repair used in the young adult knee which require harvesting of healthy cartilage and which may cause iatrogenic damage to the joint. This study explores alternative sources of autologous cells.

Chondrocytes obtained from autologous bone-marrow-derived cells and those from the damaged cartilage within the lesion itself are shown to be viable alternatives to harvest-derived cells. A sufficient number and quality of cells were obtained by the new techniques and may be suitable for autologous chondrocyte and matrix autologous chondrocyte implantation.

Introduction: Autologous chondrocyte implantation (ACI) is a technique described for treating symptomatic osteochondral defects in the knee. It is contra-indicated, however, in a joint rendered unstable by a ruptured anterior cruciate ligament (ACL). We present our early experience of combined ACL and ACI repair.

Methods: Patients underwent arthroscopic examination and cartilage harvesting of the knee. Chondrocytes were then cultured in plasma and a second operation was undertaken approximately four weeks later to repair the ruptured ACL with hamstring graft and to implant the chondrocytes via formal arthrotomy. Patients then underwent a graduated rehabilitation program and were reviewed at 6 and 12 months. Functional measurements were made using the Bentley functional scale and the modified Cincinnati rating system, with pain measured on a visual analogue scale. All patients also underwent formal clinical examination at each review.

Results: 4 out of the 5 patients reported an improvement in pain as measured on visual analogue scale, with 1 patient reporting no difference. 4 patients had stable knees as determined by negative anterior draw, negative Lachman’s test and negative pivot shift test; one patient showed improvement, but remained pivot shift positive. Improvements in Bentley scores were noted in 3 patients. Cincinnati scores were markedly improved in 3 patients and slightly improved in the remaining 2 patients. The only operative complications were a traction neuropraxia to the saphenous nerve of one patient requiring no treatment and a manipulation under anaesthesia for poor mobilisation in another patient, which was successful in improving range of movement. A further patient required arthroscopic trimming of the cartilage graft which had overgrown; this was also successful.

Conclusion: Symptomatic cartilage defects and ACL deficiency may co-exist in many patients and represent a treatment challenge. Our early results suggest that a combined ACL and ACI repair is a viable option in this group of patients and should reduce the anaesthetic and operative risks of a two-stage repair. More patients and longer follow up will be required to fully assess this technique.

Introduction and aim: Several authors have suggested that hyaline repair tissue following autologous – chondrocyte implantation (ACI) gives better clinical results than either mixed hyaline and fibrocartilage or fibrocartilage alone. This data is based on the use of periosteum as a covering membrane in these previous studies. We have for some years been using a porcine collagen type 1/III membrane (ACI-C) instead of periosteum and have now the opportunity to analyze the clinical results when compared with the histology of the repaired defect. We have also analysed the influence on the result of age and sex of the patient, the etiology of the lesion, the duration of the knee symptoms, number of previous knee procedures, the site and size of defect and the preoperative functional scores.

Method: Until 2004, 234 patients underwent autologous chondrocyte implantation at our centre. The patients were assessed clinically by their modified Cincinnati scores prospectively from 1 to 4 years from surgery. Also at arthroscopy (1 to 3 years following ACI-C) they underwent biopsy of the implant where possible and the neo-cartilage was graded as hyaline (H), mixed fibrohyaline (F.H), fibrocartilagenous (F.C) and fibrous (F).

Results: The clinical results showed that older patients had poorer results (p< 0.001) and a high preoperative modified Cincinnati score predicted a good result (p< 0.001). Concerning the cause of the defect, the percentage of patients with excellent and good results were significantly low among those with previously failed ACIs and mosaicplaties (12.5%) compared with those following trauma, osteochondritis dessicans and chondromalacia patellae (67% to 77%). At 4 year follow-up, 75% of patients with hyaline neo-cartilage had excellent and good modified Cincinnati scores whereas those with mixed fibro-hyaline and fibro-cartilage had fewer excellent and good results (44.4% and 54.5% respectively). The other parameters such as gender, the site of defect, duration of knee symptoms and the number of previous procedures and the size of the defect did not significantly influence the outcome. In conclusion, patients most likely to benefit from autologous chondrocyte implantation using a collagen membrane (ACI – C) are younger patients with higher preoperative functional scores and those who develop hyaline neo-cartilage.

Study Purpose: The cause of intervertebral disc degeneration (IVDD) is multifactorial. One proposed mechanism is that IVDD originates in the nucleus pulposus (NP) and progresses radially to the annulus fibrosis (AF). Failure of current treatment modalities in preventing and treating IVDD and thereby low back pain have led to a growing interest in tissue-engineered solutions where a biological repair is induced. By preventing the abnormality at the NP it may be possible to halt further progression of IVDD. Injection of NP cells into an early degenerative IVD, where the AF is still intact, may retard the degenerative process and is presently under investigation. Using a three-dimensional scaffold that could be successfully introduced into the NP cavity through minimally invasive techniques would prevent the loss of chondrocytic phenotype of the cells and be an improvement over the existing technique by which cells are directly injected into the NP cavity.

Methods: (1) CaSO4 and CaCO3 alginates were injected into the NP cavity of a bovine tail. After 90 minutes the tail was dissected to reveal the gel. (2) NP cells released from pooled bovine NP tissue were dispersed into the CaSO4 and CaCO3 alginate gels (10x106 cells.mL-1) with and without Synvisc® and cultured for 21 days.

Results: (1) Injectable alginate suspensions formed solid viscoelastic gels, filling the exact shape of the NP cavity. (2) NP DNA and ECM synthesis was significantly greater in the CaCO3 alginate gel than in the CaSO4 alginate gel (p< 0.05). (3) Synvisc® significantly increased sulphated GAG (p< 0.01) and collagen (p< 0.05) production. These effects were supported histologically and immunohistologically where cells in the CaCO3 and Synvisc® gels stained more intensely for proteoglycan and collagen type II.

Conclusions: Both CaCO3 alginate gel and CaSO4 alginate gel are injectable and are capable of sustaining NP cells in-vitro. Cells remain viable, maintain their phenotype, proliferate and produce ECM during the culture period. The CaCO3 alginate gel provides a three-dimensional matrix more favourable to NP cellular activity than the CaSO4 alginate gel. Synvisc® behaves as a chondro-stimulant significantly enhancing NP cell metabolic activity. This study demonstrates a successful tissue-engineered approach for replacing the NP and, subject to further studies, may be used for retarding mild-to-moderate IVDD, alleviating lower back pain and restoring a functional NP through a minimally invasive technique.

Ninety knees were treated in the same centre between1995 and 1999 each had Low Contact Stress (LCS) knee prosthesis with patelloplasty; none of the patients had resurfacing of the patella. The average follow-up period was seven years (ranging from five to nine years). There were no exclusion criteria, almost all the patients included in this study were available for follow-up, the mean age was 75 years, and the majority of the patients were affected by osteoarthritis. All the operations were done by the same surgeon or under his supervision according to the protocol.

Evaluation was performed with using The Knee Society Clinical Rating System and new Patellar Scoring System. The final scoring was done by an independent Senior Physiotherapist. The Knee Society’s radiological evaluation system was used to assess the pre and post-operative alignment of the knee and the prosthesis. Image Tool (IT accessories UTHSCSA) software used to for measurements of patellar tracking and alignments on the digitalised radiographs.

Pre-operatively, the mean Knee Society score on a scale ranging from 0-200 points was 81.50 points (range 25 to 124 points); postoperatively this score improved to a mean of 150.76 points (range 46 to 195 points). Statistical analysis using paired-comparison t-tests showed the score differences were statistically significant at p-values < 0.0001.

None of the patients required resurfacing during the follow-up. There were no reported cases of significant anterior knee pain, dislocation, maltracking or fractured patella.

Purpose: To assess costs and health status outcomes following ACI and mosaicplasty used to treat chondral knee defects (1).

Methods: Patients received ACI or mosaicplasty at the Royal National Orthopaedic Hospital between 1997 and 2001, or, were on a waiting list for ACI. Resource use per patient was collected to two years post-operatively. A postal questionnaire collected sociodemographic characteristics, knee-related (Modified Cincinnati Knee Rating System) and general health status (EQ-5D).

Results: 53 ACI, 20 mosaicplasty and 22 patients waiting for ACI participated in this study. The average cost per patient was higher for ACI (£10,600: 95%CI £10,036-£11,214) than for mosaicplasty (£7,948: 95%CI £6,957-£9,243). Estimated average EQ-5D social tariff improvements for QALYs (quality adjusted life years) were 0.23 for ACI and 0.06 for mosaicplasty. Average costs per QALY were: £23,043 for ACI and £66,233 for mosaicplasty. The ICER (incremental cost effectiveness ratio) for providing ACI over mosaicplasty was £16,349. Post-operatively, ACI and mosaicplasty patients (combined) experienced better health status than patients waiting for ACI. ACI patients tended to have better health status outcomes than mosaicplasty patients, although this was not statistically significant.

Conclusions: Average costs were higher for ACI than for mosaicplasty. However, both the estimated cost per QALY and ICER fell beneath an implicit English funding threshold of £30,000 per QALY. To our knowledge this is the first study to compare the costs and utility of ACI with alternative ‘best’ treatments for people with chondral knee problems. Prospective studies are required to confirm these results.

Bovine and human articular chondrocytes were seeded in 2% alginate constructs and cultured for up to 19 days in a rotating-wall-vessel (RWV) and under static conditions. Culture within the RWV enhanced DNA levels for bovine chondrocyte-seeded constructs when compared with static conditions but did not produce enhancement for human cells. There was a significant enhancement of glycosaminoglycans and hydroxyproline synthesis for both bovine and human chondrocytes. In all cases, histological analysis revealed enhanced Safranin-O staining in the peripheral regions of the constructs compared with the central region. There was an overall increase in staining intensity after culture within the RWV compared with static conditions. Type-II collagen was produced by both bovine and human chondrocytes in the peripheral and central regions of the constructs and the staining intensity was enhanced by culture within the RWV. A capsule of flattened cells containing type-I collagen developed around the constructs maintained under static conditions when seeded with either bovine or human chondrocytes, but not when cultured within the RWV bioreactor.

Background: Autologous chondrocyte implantation (ACI) was introduced over 15 years ago as a treatment for full-thickness chondral defects in the knee. Current understanding of ACI graft morphology and maturation in humans is limited. The aims of this study were determine the incidence of hyaline-like repair following ACI, and to clarify the relationship between repair morphology and clinical outcome.

Methods: A retrospective review of 194 ACI graft biopsies from 180 patients, and their clinical outcome was conducted. 154 Biopsies were performed 1 year after implantation and 40 biopsies were performed at 2 years. Three techniques of ACI implantation were used; Collagen covered ACI (ACI-C), periosteum covered ACI (ACI-P) and Matrix-Induced ACI (MACI).

Results: At 1 year, hyaline repair tissue was found in 48 (53%) ACI-C grafts, 7 (44%) ACI-P grafts, and 12 (36%) MACI grafts. The frequency of hyaline tissue found in biopsies performed at 2 years (84%) was significantly higher than those performed at 1 year (48.6%), p=0.0001, suggesting that grafts continue to remodel after the first year post implantation.

Clinical outcomes during the first two postoperative years did not vary according to repair morphology type, though hyaline repair was associated with better clinical outcomes beyond 2 years; At 1 year, good to excellent clinical scores were observed in 29 (78.4%) patients with hyaline-like repair, 23 (76.7%) patients with fibrohyaline repair, and 54 (74.0%) patients with fibrocartilage repair. By years 3 and 4 post-implantation, clinical scores further improved in patients with hyaline-like repair yet declined in those with fibrocartilage and fibrohyaline. The difference was significant at 3 years though not at 4 due to the small number of cases.

Conclusions: Achieving hyaline-like repair is critical to the longevity of cartilage repair. The finding of hyaline-like cartilage or fibrohyaline cartilage in 31 of 37 biopsies (84%) performed after 2 years is therefore encouraging and supports further use of the ACI technique.

Background: Autologous Chondrocyte Implantation (ACI) is widely used as a treatment for symptomatic chondral and osteochondral defects of the knee. Variations of the original periosteum cover technique include the use of porcine-derived type I/III collagen as a cover (ACI-C), and the use of a collagen bilayer seeded with chondrocytes (MACI).

Aim: To determine whether differences in clinical, arthroscopic and histological outcomes at 1 year exist between ACI-C and MACI techniques.

Methods: We have performed a prospective randomised comparison of ACI-C versus MACI for the treatment of symptomatic chondral defects of the knee on 91 patients of whom 44 received ACI-C and 47 received MACI grafts.

Results: Both treatments resulted in improvements of clinical scores after 1 year. Mean modified Cincinnati knee scores increased by 17.5 in the ACI-C group and 19.6 in the MACI group (p> 0.05). Arthroscopic assessments performed after 1 year demonstrated good to excellent ICRS graft repair scores in 79% of ACI-C grafts and 67% of MACI grafts. Hyaline-like or hyaline-like cartilage with fibrocartilage was found in the biopsies of 43% of ACI-C grafts and 36% of MACI grafts after 1 year. The rate of graft hypertrophy was 9% in the ACI-C group and 6% in the MACI group. The frequency of re-operation was 9% in each group.

Conclusions: We conclude that clinical, arthroscopic and histological outcomes are comparable for both ACI-C and MACI techniques. While the MACI technique is technically attractive, further long-term studies are required before widespread adoption of this new technique.

We prospectively studied the clinical, arthroscopic and histological results of collagen-covered autologous chondrocyte implantation (ACI-C) in patients with symptomatic osteochondritis dissecans of the knee. The study included 37 patients who were evaluated at a mean follow-up of 4.08 years.

Clinical results showed a mean improvement in the modified Cincinnati score from 46.1 to 68.4. Excellent and good clinical results were seen in 82.1% of those with juvenile-onset osteochondritis dissecans but in only 44.4% of those with adult-onset disease.

Arthroscopy at one year revealed International Cartilage Repair Society grades of 1 or 2 in 21 of 24 patients (87.5%). Of 23 biopsies, 11 (47.8%) showed either a hyaline-like or a mixture of hyaline-like and fibrocartilage, 12 (52.2%) showed fibrocartilage.

The age at the time of ACI-C determined the clinical outcome for juvenile-onset disease (p = 0.05), whereas the size of the defect was the major determinant of outcome in adult-onset disease (p = 0.01).

We investigated the prognostic indicators for collagen-covered autologous chondrocyte implantation (ACI-C) performed for symptomatic osteochondral defects of the knee.

We analysed prospectively 199 patients for up to four years after surgery using the modified Cincinnati score. Arthroscopic assessment and biopsy of the neocartilage was also performed whenever possible. The favourable factors for ACI-C include younger patients with higher pre-operative modified Cincinnati scores, a less than two-year history of symptoms, a single defect, a defect on the trochlea or lateral femoral condyle and patients with fewer than two previous procedures on the index knee. Revision ACI-C in patients with previous ACI and mosaicplasties which had failed produced significantly inferior clinical results. Gender (p = 0.20) and the size of the defect (p = 0.97) did not significantly influence the outcome.

Introduction and Aims: ACI (autologous chondrocyte implantation) using a periosteum cover was developed by Peterson et al. Recently, the technique has been developed using a Type I/Type III collagen membrane (Chondro-Gide). A second technique MACI (matrix-induced autologous chondrocyte implantation) has evolved using a membrane with chondrocytes seeded onto its surface. Aim is to review the one and two-year results of the first 159 patients at a single regional centre.

Method: The two-stage procedure was performed with a standardised, progressive rehabilitation program. Patients were assessed clinically at three, six, nine, 12 and 24 months (pain score, Modified Cincinnati, Bentley), and arthroscopically at 12 and 24 months.

Results: 159 patients have been assessed at one year and 101 patients at two years. Of those patients reviewed at one year, 110 patients had the ACI repair with Chondro-Gide, 31 patients had the ACI repair with periosteum and 18 patients had the MACI repair. Sixty-nine percent had good or excellent results at one year and 60% at two years.

These figures represent the early results of this study performed at this unit.

Conclusion: We propose that the ACI technique is valuable for selected patients with Chondral and osteochondral defects of the knee even with large and multiple defects in the articular cartilage.

Introduction and Aims: Uni-compartmental knee arthroplasty (UKA) is appropriate for one in three osteoarthritic knees requiring replacement. An accelerated protocol enables patients undergoing UKA to be discharged within 24 hours of surgery. Before such an approach is universally accepted it must be safe, effective and economically viable. A study was performed to compare the new accelerated protocol with current standard care in a state healthcare system.

Method: A single blind RCT design was used. Patients eligible for UKA were screened for NSAID tolerance, social circumstances and geographical location before allocation to either an accelerated recovery group (Group A) or a standard non-accelerated group (Group S). The accelerated protocol included dedicated pain management and discharge support. Primary outcome was the Oxford Knee Assessment at six months post-operation, compared using independent t tests. Pain, range of movement and incidence of complications were also recorded by assessors blind to group allocation. Cost effectiveness was calculated in quality life adjusted years (QLAY) using the Euroqual instrument. The study power was sufficient to avoid type 2 errors. The study was supported by a NHS Regional R& D grant.

Results: Forty-one patients (21 group A, 20 group S) were included. Groups had comparable age and patient profiles. Average discharge time was 37 hours (1.5 days) for group A and 114 hours (4.3 days) for group S. Pain on hospital discharge was similar for both groups. No significant difference was found between groups for pain or range of movement at any time, although patients in group S regained pre-operative flexion faster than group A. One major complication occurred in each group; one infection (group S) and one manipulation for poor movement (group A). The cost per QLAY for the new protocol was 59% of the standard care.

Conclusion: The new protocol allows for safe accelerated discharge from hospital after UKA. The approach is cost-effective and should help to increase the throughput of patients who require knee replacement.

The purpose of this study was to examine the effects of hyaluronic acid supplementation on chondrocyte metabolism in vitro. The clinical benefits of intra-articular hyaluronic acid injections are thought to occur through improved joint lubrication. Recent findings have shown that exogenous hyaluronic acid is incorporated into articular cartilage where it may have a direct biological effect on chondrocytes through CD44 receptors.

Bovine articular chondrocytes were isolated and seeded into alginate constructs. These were cultured in medium containing hyaluronic acid at varying concentrations. Samples were assayed for biochemical and histological changes.

There was a dose-dependent response to the exposure of hyaluronic acid to bovine articular chondrocytes in vitro. Low concentrations of hyaluronic acid (0.1 mg/mL and 1 mg/mL) significantly increase DNA, sulphated glycosaminoglycan and hydroxyproline synthesis. Immunohistology confirmed the maintenance of cell phenotype with increased matrix deposition of chondroitin-6-sulphate and collagen type II. These findings confirm a stimulatory effect of hyaluronic acid on chondrocyte metabolism.

Autologous chondrocyte implantation (ACI) is used widely as a treatment for symptomatic chondral and osteochondral defects of the knee. Variations of the original periosteum-cover technique include the use of porcine-derived type I/type III collagen as a cover (ACI-C) and matrix-induced autologous chondrocyte implantation (MACI) using a collagen bilayer seeded with chondrocytes. We have performed a prospective, randomised comparison of ACI-C and MACI for the treatment of symptomatic chondral defects of the knee in 91 patients, of whom 44 received ACI-C and 47 MACI grafts.

Both treatments resulted in improvement of the clinical score after one year. The mean modified Cincinnati knee score increased by 17.6 in the ACI-C group and 19.6 in the MACI group (p = 0.32). Arthroscopic assessments performed after one year showed a good to excellent International Cartilage Repair Society score in 79.2% of ACI-C and 66.6% of MACI grafts. Hyaline-like cartilage or hyaline-like cartilage with fibrocartilage was found in the biopsies of 43.9% of the ACI-C and 36.4% of the MACI grafts after one year. The rate of hypertrophy of the graft was 9% (4 of 44) in the ACI-C group and 6% (3 of 47) in the MACI group. The frequency of re-operation was 9% in each group.

We conclude that the clinical, arthroscopic and histological outcomes are comparable for both ACI-C and MACI. While MACI is technically attractive, further long-term studies are required before the technique is widely adopted.

Autologous chondrocyte implantation (ACI) is a technique used for the treatment of symptomatic osteochondral defects of the knee. A variation of the original periosteum membrane technique is the matrix-induced autologous chondrocyte implantation (MACI) technique. The MACI membrane consists of a porcine type-I/III collagen bilayer seeded with chondrocytes. Osteochondral defects deeper than 8 to 10 mm usually require bone grafting either before or at the time of transplantation of cartilage. We have used a variation of Peterson’s ACI-periosteum sandwich technique using two MACI membranes with bone graft which avoids periosteal harvesting. The procedure is suture-free and requires less operating time and surgical exposure. We performed this MACI-sandwich technique on eight patients, five of whom were assessed at six months and one year post-operatively using the modified Cincinnati knee, the Stanmore functional rating and the visual analogue pain scores.

All patients improved within six months with further improvement at one year. The clinical outcome was good or excellent in four after six months and one year. No significant graft-associated complications were observed. Our early results of the MACI-sandwich technique are encouraging although larger medium-term studies are required before there is widespread adoption of the technique.

An increasing number of patients are treated by autologous chondrocyte implantation (ACI). This study tests the hypothesis that culture within a defined chondrogenic medium containing TGF-β enhances the reexpression of a chondrocytic phenotype and the subsequent production of cartilaginous extracellular matrix by human chondrocytes used in ACI. Chondrocytes surplus to clinical requirements for ACI from 24 patients were pelleted and cultured in either DMEM (Dulbecco’s modified eagles medium)/ITS+Premix/TGF-β1 or DMEM/10%FCS (fetal calf serum) and were subsequently analysed biochemically and morphologically.

Pellets cultured in DMEM/ITS+/TGF-β1 stained positively for type-II collagen, while those maintained in DMEM/10%FCS expressed type-I collagen. The pellets cultured in DMEM/ITS+/TGF-β1 were larger and contained significantly greater amounts of DNA and glycosaminoglycans. This study suggests that the use of a defined medium containing TGF-β is necessary to induce the re-expression of a differentiated chondrocytic phenotype and the subsequent stimulation of glycosaminoglycan and type-II collagen production by human monolayer expanded chondrocytes.