Molecular characterisation of chondrocytes in autologous chondrocyte implantation

Abstract

Autologous chondrocyte implantation (ACI) relies on the use of cultured cells. However, the biosynthetic profile of cultured chondrocytes is shown to be altered during in vitro expansion. The purpose of this study therefore, was to examine the cellular phenotype of chondrocytes cultured for ACI and to determine the apoptotic index of cells implanted into patients. Using electron microscopy, immunohistochemistry, RT-PCR and flow cytometry analyses, we have investigated protein and gene expression of several chondrocyte-specific, or associated markers in cultured cells used for implantation in patients. They included S-100, type I and II collagen, aggrecan, transforming growth factor beta, glucocorticoid receptor alpha and beta and vitamin D3 receptor. We have also examined the apoptotic index of chondrocytes. Our results demonstrated that cultured cells for ACI display the characteristics of chondrocytes. These cells are round in shape, contain numerous small surface processes of cytoplasmic membrane and have an accumulation of glycogen within the cytoplasm. They express S-100, aggrecan TGF-beta, glucocorticoid receptor alpha and vitamin D3 receptor as evidenced by either immunohistochemistry or RT-PCR, however, there is variation in the expression of type I, type II collagen glucocorticoid receptor beta between cases. Chondrocyte used for implantation has relatively low level of apoptosis (<11%). In conclusion, although there was variation in the level of expression of these genetic markers, our data indicate that cultured cells used for ACI were of chondrocytic lineage cells and have low level of apoptotic cells.