Hypoxia effects on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes / 中国组织工程研究

ABSTRACT

BACKGROUND:Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes. OBJECTIVE:To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes. METHODS:Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×1010/L. The cells were cultured in normoxia (20%O 2 ) and hypoxic (5%O 2 ) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II col agen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cellcomplex was measured. RESULTS AND CONCLUSION:In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix;alcian blue staining showed massive glycosaminoglycan formation;immunohistochemistry staining detected strongly positive expression of col agen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes. .