兔骨髓间充质干细胞向成骨分化对血管内皮细胞形成的可能影响(英文)

Possible effects of osteoblastogenesis of marrow mesenchymal stem cells on the formation of vascular endothelial cells in rabbits

背景:种子细胞的成骨功能及局部血管形成是骨组织工程修复重要影响因素。目的:体外研究兔骨髓间充质干细胞(mesenchymalstemcells,MSCs)诱导培养分化为成骨细胞的功能表现及血管内皮生长因子(vascularen-dotheliagrowthfactor,VEGF)的表达。探讨MSCs向成骨分化及对血管内皮细胞形成的可能影响。设计:以细胞为研究对象,单一样本研究,重复观察测量。单位:一所大学组织胚胎学教研室。材料:本实验于2002-03/12在广东医学院组织胚胎学教研室完成。对象为兔骨髓间充质干细胞。方法:分离、培养新生兔骨髓间充质干细胞,用含地塞米松、β-甘油磷酸钠和维生素C的条件培养液诱导MSCs向成骨细胞分化,观察诱导细胞碱性磷酸酶(ALP)染色、钙化结节形成以及VEGF的表达。主要观察指标:①细胞形态学观察。②培养细胞的成骨分化鉴定。③诱导培养的成骨细胞VEGF表达。结果:MSCs诱导培养后,细胞ALP呈强阳性染色,形成矿化结节,且VEGF表达增强,其灰度值为132.3±4.6,与MSCs的VEGF表达灰度值(148.5±5.3)比较,差异有显著性意义(P<0.05)。结论:兔MSCs经诱导可分化为成骨细胞,且VEGF表达增强,可能参与内皮细胞的形成。

BACKGROUND:The osteoblastogenetic function of stem cell and local vasoformation is an important influencing factor in bone tissue engineering restoration.OBJECTIVE:To investigate the function of osteoblast induced,cultured and differentiated from mesenchymal stem cells(MSCs) and the expression of vascular endothelial growth factors(VEGF) in rabbits in vitro. DESIGN:A single sample study and repetitive observational measurement by using cells as subjectsSETTING:Department of histology and embryology in a universityMATERIALS:The study was conducted in the Department of Histology and Embryology of Guangdong Medical College between March and December of 2002.The subjects were rabbits'MSCs.METHODS:The MSCs of newborn rabbits were isolated and cultured.The cultured cells were induced to osteoblastogenesis with an osteo inductive medium containing dexamethasone, ascorbic acid and β glycerophosphate to observe the coloration of alkaline phosphates(ALP) and the formation of mineralized nodule in the cultured cells.MAIN OUTCOME MEASURES:① observation of cell morphology, ② identification of osteoblastogenesis from the cultured cells③ expression of VEGF in osteoblasts derived from the cultured cellsRESULTS:After MSCs osteo inductive culture,cells had strong ALP staining with the formation of mineralized node,and the VEGF expression enhanced with the grey scale of 132.3± 4.6,which was significantly different from that of the VEGF expression in MSCs(148.5± 5.3) (P< 0.05).CONCLUS