ROLE OF L-ASCORBIC ACID 2 PHOSPHATE AND PLATELET RICH PLASMA IN CARTILAGE ENGINEERING USING HUMAN ADIPOSE-DERIVED STEM CELL AND SILK FIBROIN SCAFFOLD

• Main Author: Musdalifah Amsar, Rizka
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/37042
• Institution: Institut Teknologi Bandung

The cartilage has the avascular characteristic so when it damaged, it difficult to repair due to limited regeneration capability. An alternative solution to repair cartilage’s damage is tissue engineering. Tissue engineering use cells, biomaterial and regulatory signals to regenerate cartilage. The aim of this study is to evaluate the cell penetration in silk fibroin scaffold 12% with pore size 500 ?m, the role of L-ascorbic acid 2 phosphate (LAA) 50 ?g/mL and Platelet Rich Plasma (PRP) 10% on the gene expression of N-cadherin, ?-catenin, cyclin D and Collagen type 2 as a marker of chondrogenic differentiation. Mesenchymal stem cells (MSC) were isolated from adipose and FACS analysis was performed to examine the expression of the MSC marker (CD 105+, CD 73+, and CD45-). Multipotent capability was analyzed by Alizarin Red, Oil Red O, Alcian Blue staining. HADSC were treated with PRP 10% and LAA 50 ?g/mL for a period of 21 days. Cell penetration and collagen type 2 content were analyzed with immunohistochemistry. The gene expression of N-cadherin, ?-catenin, cyclin D and collagen type 2 were analyzed with RT-qPCR. The results revealed that cell shows mesenchymal characterization and it called hADSC. Cell penetration in silk fibroin scaffold was occurred. In 21 days of culture, LAA and PRP upregulated the gene expression of N-cadherin and collagen type 2, however downregulated ?-catenin and cyclin D. Immunohistochemistry confirmed the presence of collagen type 2 protein. The finding of this study suggested that cell penetration occurred in silk fibroin scaffold, LAA and PRP treatment increased the gene expression of N-cadherin, involved in chondrogenesis in vitro by regulation of ?-catenin gene expression and enhanced collagen type 2 secretion.