L-ascorbic acid-2-phosphate and Fibrin Substrate Enhances the Chondrogenic Differentiation of Human Adipose-derived Stem Cells

Abstract

Background: Adipose-derived stem cells (ASCs) have been developed as a substitute for chondrocytes in cartilage tissue engineering. This study aimed to evaluate the potency of ascorbic acid to induce the chondrogenic differentiation of human ASCs cultured on fibrin substrate. Methods: Human ASCs were isolated by an enzymatic process and characterized. The fibrin scaffold was fabricated from human blood as a natural scaffold. The ASCs were cultured in 10% fetal bovine serum (FBS) supplemented by ascorbic acid in various concentrations (1, 3, 6 mg/ml) and a control without an ascorbic acid supplementation. The chondrogenic differentiation of the ASCs was evaluated involving glycosaminoglycan analysis, and the mRNA expression of type 2 collagen, aggrecan and type 1 collagen. Results: Chondrogenic differentiation occurred when the ASCs were supplemented by 3 mg/ml of ascorbic acid and cultured on a fibrin scaffold confirmed by glycosaminoglycan (GAG) analysis, and the mRNA expression of type 2 collagen and aggrecan. The expression of type 1 collagen, which is an osteogenic marker, was low. Conclusions: ASCs with a supplementation of 3mg/ml of ascorbic acid cultured on a fibrin substrate promotes chondrogenic differentiation.