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Session: Poster session I
Session starts: Thursday 24 January, 15:00

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Meike W.A. Kleuskens (TU/e)
René van Donkelaar (TU/e)
Keita Ito (TU/e)

Abstract:
Current knee osteoarthritis (OA) treatments are sub-optimal or not long-lasting, and tissue engineering might provide a promising alternative. Within the RegMed XB/InSciTe Mimicart project, regenerative osteochondral implants for focal defects are being developed. The ability of the implants to regenerate cartilage and bone and integrate with the adjacent tissues needs to be explored and maximized. Aim of the present study is to characterize human osteochondral tissue at various stages of OA, and to develop an approach to culture OA tissue for 28 days while maintaining the biochemical and histological composition of the explants. A double chamber culture platform was used to culture Ø10mm human osteochondral explants. In this culture platform the cartilage and bone tissue of an osteochondral plug can be cultured in two separated media compartments, allowing for the supplementation of tissue specific medium components. Previous research has shown that Ø8mm porcine osteochondral explants can be cultured up to 56 days in this system, while maintaining cartilage tissue content, structure and mechanical properties.1 Osteochondral explants with either a smooth or fibrillated cartilage surface, representing different OA stages, were harvested from human tibia plateaus obtained from total knee replacement surgeries at the Maxima Medical Centre, Eindhoven. The native and cultured explants were evaluated for their viability using an MTT assay and an LDH assay, biochemical tissue content and cell gene expression. Moreover, samples were histologically stained with safranin-O/fast green and picrosirius red for analysis of the proteoglycan and collagen distribution and to determine the grade of OA using the Mankin scoring system. OA cartilage of various stages, including tissue with smooth and fibrillated surfaces, was evaluated and statistical differences were found in biochemical and histological analyses at day 0 (average Mankin score of 3.4 vs. 5.1, average proteoglycan content of 8.4±1.7%dw vs. 13.5±3.2%dw, average collagen content of 39.9±3.8%dw vs. 29.3±4.6%dw, for tissue with a smooth or fibrillated surface respectively). Preliminary results of the cultures reveal that chondrocyte viability was maintained over 28 days, but bone tissue was less viable. Osteochondral explant culture is ongoing to explore whether the biochemical and histological composition of the cartilage is preserved over time.