CHONDROGENIC DIFFERENTIATION OF HUMAN WHARTON’S JELLY MESENCHYMAL STEM CELLS ON SILK FIBROIN-POLYVINYL ALCOHOL SCAFFOLD INDUCED BY INDIRUBIN

• Main Author: Eileen Wiguna, Nathalie
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/47037
• Institution: Institut Teknologi Bandung

Articular cartilage is frequently damaged because of trauma and degenerative joint disease. Its limited self-healing capacity makes damaged articular cartilage difficult to repair. Cartilage tissue engineering involving the combination of cells, biomaterial scaffold, and biofactors is a promising method for the repair of injured articular cartilage. In this work, human Wharton’s jelly mesenchymal stem cells (hWJ MSC) were grown on silk fibroin-polyvinyl alcohol (SF/PVA) scaffold with supplementation of indirubin. Silk fibroin (SF) and PVA are biocompatible and biodegradable material that are suitable for cartilage tissue engineering. Indiruin is a bioactive agent which is potential to suppress chondrocyte hypertrophy. This study was done to to analyze the comparison of chondrogenic differentiation of hWJ MSC on SF-PVA scaffold and SF scaffold. The other objective was to evaluate the potential of indirubin in chondrogenic differentiation of hWJ MSC and chondrocyte hypertrophy process. hWJ MSC were characterized using flow cytometry and multipotency assay. Cells were then seeded on scaffold which fabricated by salt leaching method. Scaffold morphology was analyzed using a scanning electron microscope (SEM). Cytotoxicity of indirubin and scaffold biocompatibility were done with MTT assay. Chondrogenic differentiation of hWJ MSC was evaluated through collagen type II visualization by immunocytochemistry (ICC) and GAG quantification assay with Alcian Blue staining. Chondrocyte hypertrophy process was evaluated through collagen type X visualization by ICC and calcium deposit quantification with Alizarin Red S staining. The result showed that hWJ MSC fulfill three criteria of MSC. Based on SEM analysis, SF/PVA scaffold formed 400 – 500 ?m pores. MTT assay showed that 0.05 and 0.1 ?M indirubin were non toxic and SF-PVA scaffold was biocompatible. On day 21, cells on SF scaffold showed more collagen II than cells on SF/PVA scaffold. There was no difference among various indirubin concentrations. Cells seeded on SF scaffold also produce more GAG than cells on SF/PVA scaffold at day 21 and there was no significant difference at the various indirubin concentrations. On day 21, collagen type X was more observed on SF scaffold groups than SF/PVA scaffold groups. There was no differences among various indirubin concentrations. Calcium deposit was more observed on SF/PVA scaffold groups. Calcium deposition was not significally different among various indirubin concentration. These results indicate that SF scaffold was better in supporting chondrogenic differentiation compared to SF/PVA scaffold. The addition of indirubin did not contribute in chondrogenic differentiation and in inhibition of chondrocyte hypertrophy.