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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: Osteoblasts and local vascular formation of seed cells are important factors in the repair of bone tissue engineering. OBJECTIVE: To investigate the functional expression of osteoblasts induced by mesenchymal stem cells (MSCs) in vitro and the expression of vascular endothelial growth factor (VEGF) in vitro. To investigate the osteogenic differentiation of MSCs and its possible effect on the formation of vascular endothelial cells. Design: Cells as the research object, a single sample study, repeated observation and measurement. Unit: A University Department of Embryology. MATERIALS: The experiment was performed at Department of Embryology, Guangdong Medical College from March to December 2002. The object is rabbit bone marrow mesenchymal stem cells. Methods: Bone marrow mesenchymal stem cells (MSCs) were isolated and cultured. MSCs were induced to differentiate into osteoblasts with conditioned medium containing dexamethasone, β-glycerophosphate and vitamin C, and alkaline phosphatase (ALP) staining , Calcified nodule formation and VEGF expression. MAIN OUTCOME MEASURES: ① Cell morphology observation. ② cultured cells osteogenic differentiation identification. ③ induced cultured osteoblasts VEGF expression. RESULTS: After induction of MSCs, the cell ALP was strongly positive staining, forming mineralized nodules, and VEGF expression was enhanced, the gray value of 132.3 ± 4.6, compared with the gray value of VEGF expression of MSCs (148.5 ± 5.3), the difference There was significant (P <0.05). CONCLUSION: Rabbit MSCs can differentiate into osteoblasts after induction, and VEGF expression is enhanced, which may be involved in the formation of endothelial cells.