Study on constructing cartilage tissue with adipose-derived stem cells-seeded acellular cartilaginous matrix / 中国矫形外科杂志

ABSTRACT

[Objective]To investigate the methods of preparing acellular cartilaginous matrix(ACM) and constructing the tissue engineered cartilage with adipose-derived stem cells(ADSCs)-seeded acellular cartilaginous matrix in vitro.[Method]The ADSCs were isolated from adult New Zealand albino rabbits by collagenase,cultured and amplified in vitro.Fresh cartilage isolated from adult New Zealand albino rabbits were freeze-dried for twelve hours and then treated with Triton X-100,Dnase and RNase so as to obtain the ACM.After sterilized with ultra-violet,the ADSCs were seeded in the acellular cartilaginous matrix at a final density of 2?107/L and cultured in chondrogenic differentiation medium for two weeks in order to construct tissue engineered cartilage.Histology,immunohistochemistry and transmission electron microscope(TEM) were applied to examine the fresh ACM and tissue engineered cartilage.[Result]The test of hematoxylin-eosin(HE) staining and TEM showed no cellular structure in the ACM with only recesses left by removed cells.The ACM had suitable interval porosity and aperture size.After integrated with ADSCs,cells migrated into the ACM and adhered to the surface of material and grew well.After cultured in chondrogenic differentiation medium for two weeks,immunohistochemical staining of type Ⅱ collagen showed part of the cells in the material had enhanced expression of type Ⅱ collagen.[Conclusion]ACM can be used as scaffold material in cartilage tissue engineering.If it was seeded with ADSCs and cultured in chondrogenic differentiation medium,ACM can be used to construct cartilage tissue successfully.