软骨脱细胞基质复合脂肪源性干细胞构建软骨组织的实验研究

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软骨脱细胞基质复合脂肪源性干细胞构建软骨组织

的实验研究

软骨脱细胞基质复合脂肪源性干细胞构建软骨组织的实验研究（作者:___________单位: ___________邮编: ___________）作者：

王兆杰，安荣泽，于立志，张强，齐新文，杨晋【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2107/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

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合脂肪源性干细胞培养可成功构建软骨组织。

【关键词】软骨组织; 脱细胞基质; 脂肪源性干细胞Abstract：

[Objective]To investigate the methods of preparing acellular cartilaginous matrix(ACM) and constructing the tissue engineered cartilage with adiposederived 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 freezedried for twelve hours and then treated with Triton X100,Dnase and RNase so as to obtain the ACM.After sterilized with ultraviolet,the ADSCs were seeded in the acellular cartilaginous matrix at a final density of 2107/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 hematoxylineosin (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

---------------------------------------------------------------最新资料推荐------------------------------------------------------ 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 coll agen.[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. Key words：

cartilage tissue; acellular matrix; adiposedrived stem cells 关节软骨因损伤或退行性病变引起缺损后，自身修复能力十分有限。

近年来，软骨组织工程的迅速发展，为解决这个难题提供了新的途径[1～ 3]。

而寻找一种合适的软骨细胞载体则是当前软骨组织工程研究的焦点之一。

脱细胞基质是近几年发展起来的天然支架材料，因其去除了组织中的抗原性成分，故具有良好生物相容性，在组织工程中应用广泛[4]。

2019 年 Zuk PA等[5]从人和大鼠的脂肪组织中发现了具有

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