大网膜包裹构建血管化组织工程骨的实验研究
发布时间：2018-09-17 09:36
【摘要】：目的探讨利用大网膜包裹构建血管化组织工程骨修复兔桡骨大段骨缺损能力,评估大网膜包裹构建大段血管化组织工程骨的可行性及其对骨缺损的修复效果,为组织工程骨用于临床提供及补充实验依据。 方法采用全骨髓贴壁培养法结合换液传代分离培养、纯化兔骨髓间充质干细胞(rBMSCs),通过观察细胞形态、多向分化诱导及电镜下观察成骨诱导前后细胞的结构改变对其进行鉴定,MTT法测定第三代细胞的增殖情况,并绘制出生长曲线。取成骨诱导后生长状态良好第三代细胞兔骨髓间充质干细胞接种到自制的生物衍生骨支架材料上构建组织工程骨,将体外构建的组织工程骨(TEB)植入体内,分为大网膜包裹组织工程骨组和单纯组织工程骨组。术后不同时间点观察两组移植物的表面变化及周围组织反应,术后4周,8周,12周行X线摄片检查移植物修复骨缺损情况,行标本大体观察及组织学检测,观察成骨及血管化情况。 结果两组移植物植入兔桡骨缺损处后,术口生长良好,未发现明显的全身及局部的毒性和炎症反应,每只动物的手术切口均达到一期愈合。大网膜包裹组在术后12周时X线表明植入物与宿主骨连接处基本愈合,而单纯组术后12周与宿主骨愈合不甚满意。同时大网膜包裹组骨修复区的骨灰度值都高于单纯组织工程骨组,且差异有统计学意义(P.05)；术后4周,8周,12周通过分析骨组织学切片,表明不同时间点大网膜包裹组新生骨小梁面积都明显高于单纯组织工程骨组,且差异有统计学意义(P0.01)；同时从骨组织学分析材料的血管化面积,表明在同一时间点大网膜包裹组新生血管面积均高于单纯组织工程骨组(P0.05),差异有统计学意义。 结论利用具有丰富血管网的大网膜包裹组织工程骨植入兔桡骨缺损处,促进组织工程骨的血管化及成骨能力,提高了组织工程骨修复骨缺损的质量。
[Abstract]:Objective to evaluate the feasibility of vascularized tissue engineering bone encapsulated with greater omentum to repair the defect of rabbit radius, and to evaluate the feasibility of constructing vascularized tissue engineered bone with greater omentum and its effect on repairing bone defect. To provide clinical and supplementary experimental basis for tissue engineering bone. Methods the (rBMSCs), of rabbit bone marrow mesenchymal stem cells was purified by whole bone marrow adherent culture combined with the culture of fluid exchange, and the morphology of the cells was observed. The structural changes of osteoblasts before and after osteoblast induction were observed by multidirectional differentiation and electron microscopy. The proliferation of the third generation cells was determined by MTT assay and the growth curve was drawn. After osteogenic induction, bone marrow mesenchymal stem cells (BMSCs) of the third generation of rabbits in good growth state were inoculated into self-made scaffold of bioderived bone to construct tissue engineered bone, and the tissue engineered bone (TEB) was implanted in vivo. It was divided into two groups: omentum wrapped tissue-engineered bone group and simple tissue-engineered bone group. The surface changes and tissue responses of the two groups were observed at different time points after operation. X ray films were performed at 4 weeks and 8 weeks to 12 weeks after operation, and the bone defects were repaired by the grafts. The gross and histological changes of the grafts were observed. Osteogenesis and vascularization were observed. Results after the two groups of grafts were implanted into the radius defect of rabbits, the operation mouth grew well, no obvious systemic and local toxicity and inflammatory reaction were found, and the surgical incision of each animal was healed in one stage. In the omentum encapsulated group, X-ray showed that the junction between the implant and the host bone was basically healed at 12 weeks after operation, but the healing of the host bone in the simple group was not satisfactory at 12 weeks after operation. At the same time, the degree of bone ashes in the greater omentum encapsulated group was higher than that in the simple tissue engineered bone group (P.05), and the histological sections of bone were analyzed at 4 weeks and 8 weeks to 12 weeks after operation. The results showed that the trabecular area of the new bone in the greater omentum encapsulated group was significantly higher than that in the simple tissue engineered bone group at different time points (P0.01), and the vascularized area of the material was analyzed from the bone histology. The results showed that the area of neovascularization in omentum encapsulated group was higher than that in simple tissue engineered bone group at the same time point (P0.05), and the difference was statistically significant. Conclusion the tissue engineering bone encapsulated by greater omentum with abundant vascular network was implanted into the defect of rabbit radius, which promoted the vascularization and osteogenesis of tissue engineering bone, and improved the quality of tissue engineering bone in repairing bone defect.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
本文编号：2245465
[Abstract]:Objective to evaluate the feasibility of vascularized tissue engineering bone encapsulated with greater omentum to repair the defect of rabbit radius, and to evaluate the feasibility of constructing vascularized tissue engineered bone with greater omentum and its effect on repairing bone defect. To provide clinical and supplementary experimental basis for tissue engineering bone. Methods the (rBMSCs), of rabbit bone marrow mesenchymal stem cells was purified by whole bone marrow adherent culture combined with the culture of fluid exchange, and the morphology of the cells was observed. The structural changes of osteoblasts before and after osteoblast induction were observed by multidirectional differentiation and electron microscopy. The proliferation of the third generation cells was determined by MTT assay and the growth curve was drawn. After osteogenic induction, bone marrow mesenchymal stem cells (BMSCs) of the third generation of rabbits in good growth state were inoculated into self-made scaffold of bioderived bone to construct tissue engineered bone, and the tissue engineered bone (TEB) was implanted in vivo. It was divided into two groups: omentum wrapped tissue-engineered bone group and simple tissue-engineered bone group. The surface changes and tissue responses of the two groups were observed at different time points after operation. X ray films were performed at 4 weeks and 8 weeks to 12 weeks after operation, and the bone defects were repaired by the grafts. The gross and histological changes of the grafts were observed. Osteogenesis and vascularization were observed. Results after the two groups of grafts were implanted into the radius defect of rabbits, the operation mouth grew well, no obvious systemic and local toxicity and inflammatory reaction were found, and the surgical incision of each animal was healed in one stage. In the omentum encapsulated group, X-ray showed that the junction between the implant and the host bone was basically healed at 12 weeks after operation, but the healing of the host bone in the simple group was not satisfactory at 12 weeks after operation. At the same time, the degree of bone ashes in the greater omentum encapsulated group was higher than that in the simple tissue engineered bone group (P.05), and the histological sections of bone were analyzed at 4 weeks and 8 weeks to 12 weeks after operation. The results showed that the trabecular area of the new bone in the greater omentum encapsulated group was significantly higher than that in the simple tissue engineered bone group at different time points (P0.01), and the vascularized area of the material was analyzed from the bone histology. The results showed that the area of neovascularization in omentum encapsulated group was higher than that in simple tissue engineered bone group at the same time point (P0.05), and the difference was statistically significant. Conclusion the tissue engineering bone encapsulated by greater omentum with abundant vascular network was implanted into the defect of rabbit radius, which promoted the vascularization and osteogenesis of tissue engineering bone, and improved the quality of tissue engineering bone in repairing bone defect.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
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