Culture expanded chondrocytes isolated from non-load bearing region of osteoarthritic (OA) joint has been used to construct tissue engineered cartilage for treatment purposes. The aim of the study was to compare the histological properties of the cartilage tissue and morphological properties of the chondrocytes isolated from less and severely affected OA knee. Human articular cartilage was obtained as redundant tissue from consented patients with late-stage OA undergoing total knee replacement surgery at Universiti Kebangsaan Malaysia Medical Centre (UKMMC). Articular cartilage was graded ccording to Dougados and Osteoarthritis Research Society International (OARSI) classification. Articular cartilage was classified into less affected (LA; Grade 0-1) and severely affected (SA; Grade 2-3). Cartilage tissue from less and severely affected region was stained with Safranin O staining. Isolated chondrocytes from each group were cultured until passage 4 (P4). Their growth patterns, cell areas, and circularity were compared. LA-cartilage tissue shows uniform spread of safranin O staining indicating intact extracellular matrix (ECM) component. However, SA-cartilage shows significant reduction and unstable staining due to its degraded ECM. LA-chondrocytes showed an aggregated growth compared to SA-chondrocyte that remains monolayer. Moreover, LA-chondrocytes have significantly higher cell area with wider spreading at passage 0 and 4 compared to SA-chondrocytes. It was also found that chondrocyte circularity increased with passage, and circularity of LAchondrocytes was significantly higher than that of the SA-chondrocytes at passage 3. This study demonstrated the considerable difference in the cellular properties for less and severely affected chondrocytes and implication of these differences in cell-based therapy needed to be explored.
Comparative Study on Cartilage Tissue Collected From Less- and Severely-Affected Region of Osteoarthritic Knee
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Kondrosit yang dikembang biak hasil isolasi dari kawasan kurang terjejas akibat osteo atritis (OA) dijadikan sumber untuk membentuk konstruk rawan bagi tujuan rawatan. Tujuan kajian ini adalah untuk membandingkan ciri-ciri rawan secara histologi dan mengenalpasti morfologi kondrosit yang telah diisolasi daripada OA lutut manusia yang kurang dan teruk terjejas. Tisu rawan artikular manusia dalam kajian ini diperolehi daripada pesakit OA yang memerlukan pembedahan penggantian lutut. Prosedur dilakukan di Pusat Perubatan Universiti Kebangsaan Malaysia (PPUKM). Tisu rawan artikular dikelaskan mengikut sistem Dougados dan Osteoarthritis Research Society International (OARSI) iaitu kurang terjejas [less-affected (LA); Gred 0-1] dan teruk terjejas [severely affected (SA): Gred 2-3]. Tisu rawan dari kedua-dua kumpulan diwarnakan dengan larutan Safranin O. Sel kondrosit yang diisolasi daripada setiap kumpulan dikultur sehingga ke subkultur ke-4 [passage 4 (P4)]. Perbandingan telah dilakukan untuk mengenalpasti kadar pertumbuhan, luas dan kebulatan kondrosit. Pewarnaan sekata Safranin O atas tisu rawan kurang terjejas (LA) menunjukkan komponen matriks ekstraselular (ECM) masih dalam keadaan baik manakala tisu rawan yang teruk terjejas menunjukkan pengurangan warna ketara dan tidak stabil disebabkan oleh penguraian ECM. Kondrosit-LA menunjukkan pembentukan sel agregasi berbanding dengan kondrosit-SA di mana tiada sel agregasi dapat diperhatikan. Kondrosit-LA mempunyai keluasan dan penyerebakkan sel yang lebih tinggi pada subkultur -0 dan -4 berbanding kondrosit-SA. Kebulatan sel kondrosit juga didapati meningkat seiring dengan peningkatan subkultur. Kadar kebulatan kondrosit-LA di subkultur-3 ketara lebih tinggi daripada kondrosit-SA. Keseluruhan kajian menunjukkan perbezaan ketara di antara ciri-ciri sel kondrosit-LA dan –SA dan implikasi perbezaan dalam sel terapi ini perlu diterokai dengan lebih mendalam di masa akan datang.
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