乏氧环境下骨骺干细胞株在软骨载体中的相容性、增殖和分化成软骨的实验研究  

Study on the compatibility and proliferation and chondrometaplasia of precartilaginous stem cells Transfected PTHrp gene in cartilage tissue engineering scaffold by HIF-1 under hypoxia

作　　者：赵守军[1] 熊文化[1] 郭宁峰[1] 

机构地区：[1]宁波市第二医院,宁波315010

出　　处：《现代实用医学》2012年第4期375-377,380,F0003,共5页Modern Practical Medicine

基　　金：宁波市社会发展科研项目(2009C50022)

摘　　要：目的观察乏氧环境中转染PTHrP基因的骨骺干细胞株在不同浓度HIF-1作用下与组织工程软骨载体材料的相容性,以及在软骨载体材料中骨骺干细胞株的增殖、分化成软骨作用。方法乏氧环境中,携带骨骺干细胞株的软骨载体置于含不同浓度HIF-1(0、20、40、80、100、120、140及160 pmol/L)的培养诱导液中培养,MTT法检测骨骺干细胞在载体上的黏附率及骨骺干细胞增殖情况;骨骺干细胞经转化生长因子1(TGF-1)诱导2周后免疫组化检测Ⅱ型胶原(collagenⅡ)、Aggrecan软骨蛋白聚糖表达情况;诱导后的骨骺干细胞与载体复合培养后4周取材电镜下行组织学观察软骨结节形成情况。结果细胞在载体上黏附率90.5%;乏氧环境中HIF-1可促进骨骺干细胞增殖,增殖效应与HIF-1剂量有一定的依赖性;经TGF-1诱导2周后collagenⅡ、Aggrecan免疫组化染色阳性;诱导后的骨骺干细胞在载体内生长增殖良好,4周时可见新生类软骨样组织形成。结论乏氧环境下转染PTHrP基因的骨骺干细胞与载体具有良好的相容性;乏氧环境下HIF-1能促进骨骺干细胞株的增殖、分化成软骨,其促进作用有一定的剂量依赖性。Objective To observe the compatibility and proliferation and chondrometaplasia of precartilaginous stem cells（PSCs） transfected PTHrp gene in cartilage tissue engineering scaffold by HIF-1αunder hypoxia.Methods Scaffold combined with induced PSCs was cultured in the medium by HIF-1α（0,20,40,80,100,120,140,160 pmol/L） under hypoxia. The number of PSCs attached to scaffold and the proliferation of cells were determined with MTT colorimetric microassay. PSCs were cultured in the medium of DMEM with TGF-1α for 14 days. Immunohis- tochemistry was used for detecting the expression of type II collagen and S-100 protein. Scaffold was combined with induced PSCs and cultured for 4 weeks and samples were prepared for histological examination.Results HIF-1α could promote the proliferation of PSCs under hypoxia .The effect of promotion was depended on the dose of HIF-1α to a certain extent .The cell adhesion rate was 90.5%. The result of type II collagen and S-100 protein was positive by immunohistochemistry in cells cultured in the medium of DMEM with TGF-1α. The induced PSCs were proliferated successfully and new cartilage phenotype was found by the end of 4 weeks.Conclusions PSCs transfected PTHrp gene were primely compatible in cartilage tissue engineering scaffold. HIF-1α tcould promote the proliferation and chondrometaplasia of PSCs under hypoxia. The effect of promotion was depended on the dose ofHIF-1α to a certain extent.

关 键 词：乏氧环境 骨骺干细胞 载体 HIF-1 软骨分化 

分 类 号：R714.246[医药卫生—妇产科学]