SF/COL/PLCL静电纺丝三维纳米纤维支架与人脐带血间充质干细胞的细胞相容性研究  

Cytocompatibility of electrospun SF/COL/PLCL blend three-dimensional nanofiber scaffold with human umbilical cord blood mesenchymal stem cells

作　　者：韩雨芝 王家奎[1] 何许霞 孙琪[1] 钟梅[1] 孙可墨 殷丽华[1] 余占海[1] 

机构地区：[1]兰州大学口腔医学院,甘肃兰州730000

出　　处：《中国生物制品学杂志》2014年第7期895-901,共7页Chinese Journal of Biologicals

基　　金：国家自然科学基金青年科学基金项目(81102712);甘肃省陇原青年创新人才扶持计划(B类)

摘　　要：目的研究SF/COL/PLCL静电纺丝三维纳米纤维支架与人脐带血(human umbilical cord blood,hUCB)间充质干细胞(mesenchymal stem cells,MSCs)的细胞相容性。方法分离、培养hUCBMSCs,并进行传代,取第3代hUCBMSCs,茜素红染色和Von Kossa染色检测其体外诱导成骨分化的能力;流式细胞术检测其表面相关抗原的表达。制备SF/COL/PLCL静电纺丝三维纳米纤维支架,扫描电镜观察其形貌表征,并检测其力学性能。将hUCBMSCs接种于静电纺丝三维纳米纤维支架上,观察细胞在支架上的生长及增殖情况。结果 hUCBMSCs具有成骨诱导分化能力,其表达CD44、CD29、CD90和CD105,不表达CD45和CD34。SF/COL/PLCL复合纳米纤维的纤维形貌良好,随着PLCL含量的增加,纤维的直径和力学性能均逐渐增加。hUCBMSCs能够在三维纳米纤维支架上很好地黏附,并相互连接向周围扩展,三维纳米纤维支架能很好地促进细胞黏附和增殖,与常规培养的细胞相比,差异有统计学意义(P<0.05或P<0.01),当SF/COL与PLCL的质量比为30∶70时,最有利于细胞的生长。结论 hUCBMSCs能够在SF/COL/PLCL静电纺丝纳米纤维支架上生长、增殖,这种支架材料具有良好的力学性能及细胞相容性,有望成为一种新型组织工程支架材料。Objective To investigate the cytocompatibility of electrospun SF / COL / PLCL blend three-dimensional nanofiber scaffold with human umbilical cord blood mesenchymal stem cells（hUCBMSCs）. Methods The hUCBMSCs were isolated,cultured and subcultured,of which the passage 3 were determined for the ability in induction of osteoplastic differentiation in vitro by alizarin red staining and Von Kossa staining,and for the expression of relevant antigen on surface by flow cytometry. SF / COL / PLCL blend three-dimensional nanofiber scaffold was prepared,observed for morphology by microscopy,and determined for mechanical properties,onto which hUCBMSCs were inoculated and observed for growth and propagation. Results The hUCBMSCs showed ability in induction of osteoplastic differentiation,in which CD44,CD29,CD90 and CD105 were expressed while CD45 and CD34 were not. SF / COL / PLCL complex nanofiber showed good morphology,of which the diameter and mechanical properties increased gradually with the increasing PLCL content. The hUCBMSCs showed good adhesion to the nanofiber scaffold,which were connected each other and expanded. The adhesion and propagation of hUCBMSCs on the nanofiber scaffold showed significant difference with those in routine culture（P〈0. 05）. A mass ratio of SF / COL to PLCL of 30 ∶ 70 was beneficial to the growth of hUCBMSCs. Conclusion The hUCBMSCs grew and were propagated on SF / COL / PLCL blend three-dimensional nanofibrous scaffold. With good mechanical properties and cytocompatibility,the nanofiber might be used as a novel scaffold material for tissue engineering.

关 键 词：脐带血 间充质干细胞 静电纺丝 丝素蛋白 胶原蛋白 聚乳酸 聚已内酯 纳米纤维 细胞相容性 

分 类 号：R608[医药卫生—外科学]