摘要
[目的]探讨软骨脱细胞基质支架材料的制备,及其体外复合脂肪源性干细胞构建软骨组织的技术方法。[方法]成年新西兰大白兔脂肪源性干细胞获得,培养,扩增。成年新西兰大白兔新鲜软骨,低温冻干12 h,后经曲拉通、DNA、RNA酶等处理制备成为软骨脱细胞基质支架材料,终浓度为2×107/L的脂肪干细胞种植于软骨脱细胞基质中于软骨细胞方向诱导培养基中培养2周,构建软骨组织。新鲜制备的软骨脱细胞基质及构建的软骨组织分别行组织学、免疫组织化学及透射电镜检测。[结果]实验制备的软骨脱细胞基质支架材料内无细胞结构存在,仅残留空白软骨陷窝。具有合适的孔隙率和孔径大小;复合脂肪源性干细胞后细胞向材料内部迁移,粘附,生长良好。部分载体内细胞Ⅱ型胶原免疫组化染色阳性。[结论]软骨脱细胞基质可作为支架材料应用于软骨组织工程,复合脂肪源性干细胞培养可成功构建软骨组织。
[ 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 × 10^7/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 ceils 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.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2009年第6期451-453,共3页
Orthopedic Journal of China
基金
贵州省科技厅和珠海市医学重点专科建设基金资助(编号:黔科合NY字[2006]3036珠卫[2008]80)
关键词
软骨组织
脱细胞基质
脂肪源性干细胞
cartilage tissue
acellular matrix
adipose-drived stem cells
