Three dimensional fabrication of ADSCs-chitosan/gelatin hydrogel engineered cartilage

Abstract

It was considered to be promising in treating articular cartilage defect by fabricating tissue engineered cartilage in vitro. To assess the advantages of chitosan / gelatin hydrogel scaffold seeded with ADSCs to construct tissue engineered cartilage dynamically in vitro in contrast to traditional static culture,chitosan / gelatin hybrid was chosen as cartilage biomimetic scaffold and its physical properties were subsequently tested. After that,adipose-derived stem cells( ADSCs)were inoculated into the chitosan / gelatin hydrogel scaffold at density of 1 × 107 cells / m L and cultured in a spinner flask and T-flask with chondroinductive media for two weeks,respectively. Chondrogenic differentiation ability of ADSCs within hydrogel scaffold was investigated with Toluidine Blue and Safranine O staining,the cell metabolism of glucose and lactic acid was analyzed through detecting kits,while the cell distribution,adhesion and extracellular matrix secretion within scaffold were observed by Dead / Live staining and scanning electron microscopy( SEM),respectively. The results showed that the average pore size,porosity,swelling rate and elasticity modulus of chitosan / gelatin hydrogel scaffold with good biocompatibility were 118. 25 ± 19. 51 μm,82. 60 ± 2. 34%,361. 28 ± 0. 47%,and 61. 2 ± 0. 16 k Pa,respectively. The ADSCs,in good conditions,could differentiate into chondrocytes,thus were suitable as seeding cells in tissue engineered cartilage construction. In addition,it was confirmed that the induced cells could express more proteoglycans in the dynamic spinner flask,where the cell distribution within the scaffold was more uniform and the scaffold could be filled mostly by extracellular matrix. The spinner flask with immobilized scaffold enhanced proliferation and chondrogenic differentiation of ADSCs within chitosan / gelatin hydrogel scaffolds,thus accelerated the dynamic fabrication of cell-hydrogel constructs,which could be an excellent method to construct tissue engineered cartilage in vitro.