重庆医学2016,Vol.45Issue(21):2890-2893,4.DOI:10.3969/j.issn.1671-8348.2016.21.003
Ⅱ型胶原糖胺聚糖复合支架对 hUCM SCs成软骨诱导的实验研究
Experimental research of type Ⅱ collagen composite glycosaminoglycan scaffold in hUCMSCs chondrogenic induction
摘要
Abstract
Objective To investigate the chondrogenic feasibility of the human umbilical cord derived mesenchymal stem cells (hUCMSCs)as cartilage tissue engineering seed cells ,type Ⅱ collagen composite glycosaminoglycan scaffold as the cellular carrier and cell‐scaffold complex .Methods The type Ⅱ collagen composite glycosaminoglycan scaffolds was prepared .The pore diameter , porosity and hydrophilia of scaffold materials were observed and measured by electronic microscope .The corresponding histological analysis on the scaffold materials was performed .hUCMSCs of P3 generation were cultured and identified .The hUCMSCs suspen‐sion was inoculated in the type Ⅱ collagen composite glycosaminoglycan scaffold for conducting culture without adding inducer .The samples were taken out after 3 weeks and performed the toluidine blue and safranin O staining ,type Ⅱ collagen immunohistochemi‐cal staining and SEM scanning .Results hUCMSCs of P3 generation highly expressed the mesenchymal cell marker CD29 and CD105 ,while hardly expressed endothelial cells of CD34 and hematopoietic cell markers .The type Ⅱ collagen composite glycosami‐noglycan scaffold presented white porous foam like ,the porosity was (91 .8 ± 2 .17)% ,the average pore diameter was 110‐230 μm , which was homogeneously distributed and had interpenetration .The scaffold showed good hydrophilicity with the water absorption expansion rate of (213 .71 ± 1 .31)% .The scaffold staining of toluidine blue ,safranin O and type Ⅱ collagen was positive .The car‐tilage‐like tissues were observed ,and gradually increased in the surface of cell‐scaffold complex along with culture ,which were posi‐tive in Toluidine blue ,safranin O and type Ⅱ collagen staining ,the electronic microscopic observation displayed that the cells were actively proliferated in the scaffold ,closely adhered with the materials ,the cartilage‐like cells and a large number of peripheral colla‐gen fibers with zigzag connection could be seen .Conclusion Compositing hUCMSCs and type Ⅱ collagen composite glycosamin‐oglycan scaffold could construct tissue‐engineering cartilage in vitro without induction ,which lays a certain experimental foundation for the repair of cartilage damage .关键词
人脐带间充质干细胞/种子细胞/糖胺聚糖/支架/组织工程软骨Key words
human umbilical cord mesenchymal stem cells/seed cells/glycosaminoglycan/scaffold/tissue-engineering carti-lage分类
基础医学引用本文复制引用
蔡第心,何鹏举,谭洪波,丁晶,余开富,张颖,周田华,杨军,徐永清..Ⅱ型胶原糖胺聚糖复合支架对 hUCM SCs成软骨诱导的实验研究[J].重庆医学,2016,45(21):2890-2893,4.基金项目
国家自然科学基金资助项目(81360274);第57批博士后科学基金面上资助项目(2015M572718);云南省应用基础研究自筹经费项目(2014F2106);成都军区昆明总医院院管项目(2012YG12)。 ()