Background

In vitro expanded chondrocytes being tools of cell therapy in ACI and MACI have been evaluated for their phenotype and gene expression. In this study we report a 3D polymer scaffold platform which yields chondrocytes with enriched hyaluronic acid (HA) content and miRNA 140 which are considered hallmarks of healthy chondrocytes and are essential for maintenance of the homeostasis of cartilage tissue.

Materials And Methods

Osteo-arthritis (OA) affected knee cartilage, removed before TKA were used in this study (n=7). Small bits of the tissue, observed to be healthy by the orthopedician were harvested, (weighing 0.1 ~0.3 grams) enzyme digested and the isolated chondrocytes were cultured in monolayer (2D) and in a Thermoreversible gelation polymer (3D-TGP). 2D specimens were cultured until they degenerated and 3D group till 42nd day. No growth factors or fetal calf serum or eternally added hyaluronic acid were used during the entire culture duration. Cells were harvested and subjected to evaluation of HA and miRNA 14-0 expression by RT PCR and Immunohistochemistry.

Results

In 2D, the cells grew with fibroblast morphology whereas in 3D they started forming clusters and grew as tissues. miRNA 140 expression in 3D was 30-fold higher than 2D and the HA expression evaluated by staining for CD44 antibody was more intense (IHC score = +++) compared to the 2D (IHC score = +).

Conclusion

To our knowledge this is the first of its kind report in the literature reporting HA and miRNA enrichment in cultured human chondrocytes taken from damaged and inflamed OA affected cartilage, thus making the 3D platform a suitable for producing ideal type of cells for ACI and MACI. Further, encapsulation of cells with the scaffold during transplantation is worth further research as it may yield a better in vivo environment for enhanced engraftment and hence better clinical results.