【摘要】 背景:血运重建机制是组织工程化脂肪组织成功构建的决定性因素。目的:观察脂肪干细胞与外源性血管内皮生长因子和纤维蛋白胶复合物在体内构建血管化组织工程脂肪的可行性。方法:从健康成年人吸脂术后的脂肪组织中分离脂肪干细胞并行原代及传代培养,将第3代经BrdU标记的脂肪干细胞向脂肪细胞定向诱导2周后,制成5×1010L-1细胞悬液。由0.5mL细胞悬液、100μL的血管内皮生长因子工作液或DMEM培养基与0.5mL纤维蛋白胶组成实验组和对照组移植物,分别植入裸鼠背部皮下。结果与结论:术后8周取材时:①实验组可见血管增生并长入材料,呈轻度纤维包裹;对照组有少量血管长入材料中,也有轻度纤维包裹现象。实验组新生组织湿质量大于对照组(P<0.01)。②苏木精-伊红染色均可见移植物中有新生脂肪组织形成和不同程度的微血管长入;实验组微血管数多于对照组(P<0.01)。③新生组织免疫荧光染色示,两组新生脂肪细胞的胞核及部分微血管内皮细胞的胞核呈现绿色荧光。结果说明脂肪干细胞与外源性血管内皮生长因子和纤维蛋白胶复合物在体内可构建血管化组织工程脂肪,脂肪干细胞与外源性血管内皮生长因子共同参与新生脂肪组织的血管化过程。更多还原

【Abstract】 BACKGROUND: Revascularization mechanism is the decisive factor for the successful construction of tissue engineering adipose tissue. OBJECTIVE: To observe the feasibility of construction of vasuclarized tissue engineering adipose via adipose-derived stem cells and exogenous vascular endothelial growth factor (VEGF) carrier complex in vivo. METHODS: Adipose derived stem cells (ADSCs) were isolated from the subcutaneous adipose tissue of healthy adult after liposuction, and primary culture and subculture of ADSCs were conducted. After being induced towards adipocytes for 2 weeks, 5× 1010/L passage 3 cell suspension labeled by BrdU was prepared. The experiment was divided into two groups: group A (experimental group) complex in which 0.5 mL cell suspension, 100 μL VEGF and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice; group B (control group) complex in which 0.5 mL cell suspension, 100 μL DMEM and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice. RESULTS AND CONCLUSION: Eight weeks after operation, neogenetic vessels grew into the scaffolds and mild fiber encapsulation was observed in group A, while few vessels grew into the scaffolds and also mild fiber encapsulation was observed in group B. The wet weight of cambium in group A was higher than that in group B (P < 0.01). Hematoxylin-eosin staining showed the formation of neogenetic adipose tissues and the growth of micrangium in the implant. The number of micro vessels in group A was greater than that in group B (P < 0.01). The immunofluorescence staining of cambium showed that the cell nucleus of regenerated adipocytes and partial capillary endothel ium in groups A and B presented green fluorescence. It is feasible to prepare vasuclarized tissue engineering adipose via ADSCs and exogenous VEGF carrier complex in vivo. ADSCs and exogenous VEGF participate in neovascularization of neogenetic adipose tissue.更多还原