转染携带BMP2腺病毒的鼠肌母细胞异位成骨能力的实验研究

发布时间：2019-01-26 21:19

【摘要】： 研究背景及目的有相关调查表明5%到10%的骨折病人发生延迟愈合或愈合不理想。这些不愈合的骨折通常发生于距骨颈和舟状骨折、感染、暴力损伤和骨质疏松的病人。对于此类的骨折,目前治疗措施通常有自体骨移植、带血管的骨移植、添加骨蛋白的异体骨移植等。通常愈后较差,且供骨区常发生感染和骨缺损等并发症。本课题立足于寻求一种表达BMP2能力高、取材方便且对机体创伤小的细胞载体,并将其用于临床,以寻求一种可替代、且行之有效的微创基因疗法来促进上述骨不连或延迟愈合的恢复。方法建立相应几组的小鼠模型,80只C_3H小鼠分为4组:按分组要求将移植物注入小鼠腓肠肌肌腹,转染Ad-BMP2的肌母细胞组(A组)、转染Ad-LacZ的肌母细胞组(B组),注入空白肌母细胞组(C组),注入等剂量Ad-BMP2组(D组),术后每周(连续3次)观察各组小鼠注射区的组织学变化情况及各组小鼠模型处的放射学表现。结果 (1)间接免疫荧光:Ad-BMP2转染的肌母细胞中BMP-2阳性细胞多见,未转染的细胞,未观察到阳性细胞,仅见微弱的背景染色。(2)ALP活性:空白肌母细胞和转染Ad-LacZ的肌母细胞的ALP活性含量极低,Ad-BMP2转染组中ALP活性含量较高。(3)4组动物模型骨折愈合的影像学表现:只有A组小鼠模型处有明确的异位成骨灶出现。结论 (1)构建的Ad-BMP2腺病毒能高效转染小鼠肌母细胞,且转染后的肌母细胞能在机体持续分泌有效量的BMP2。(2)本实验采用的肌母细胞孵育方法可快速培养、扩增所需载体细胞。(3)转染Ad-BMP2的鼠肌母细胞具有异位成骨能力。
[Abstract]:Background and objective: 5% to 10% of fracture patients have delayed healing or poor healing. These nonunion fractures usually occur in patients with talus and scaphoid fractures, infections, violent injuries, and osteoporosis. For such fractures, autografts, vascularized bone grafts and allogeneic bone grafts with bone protein are commonly used. Usually, the recovery is poor, and the donor area often has complications such as infection and bone defect. The purpose of this study is to seek a cell vector with high ability to express BMP2, easy to draw materials and less trauma to the body, and to use it in clinical practice to find an alternative. And effective minimally invasive gene therapy to promote the above-mentioned nonunion or delayed healing recovery. Methods 80 C3H mice were divided into 4 groups: the grafts were injected into the gastrocnemius muscle abdomen of the mice according to the request of grouping, and the myoblast cells transfected with Ad-BMP2 (group A). Ad-LacZ transfected myoblast group (group B), blank myoblast group (group C) and equal dose of Ad-BMP2 group (group D). The histological changes in the injection area of each group and the radiologic manifestations in the model of each group were observed every week (3 times in succession). Results (1) indirect immunofluorescence: BMP-2 positive cells were more common in Ad-BMP2 transfected myoblasts, but no BMP-2 positive cells were observed in untransfected cells. (2) ALP activity: the ALP activity of blank myoblasts and Ad-LacZ transfected myoblasts was very low. (3) the imaging manifestations of fracture healing in the four groups: only a clear ectopic osteogenic focus appeared in the model of group A mice. Conclusion (1) the constructed Ad-BMP2 adenovirus can efficiently transfect the murine myoblasts, and the transfected myoblasts can continuously secrete the effective BMP2. (2) the myoblast incubating method used in this experiment can be cultured rapidly. (3) murine myoblast transfected with Ad-BMP2 had ectopic osteogenic ability.
【学位授予单位】：同济大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R683;R346

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