腺病毒介导BMP-2转染对骨髓间充质干细胞生物学功能的影响

发布时间：2018-01-08 02:05

本文关键词：腺病毒介导BMP-2转染对骨髓间充质干细胞生物学功能的影响　出处：《吉林大学》2011年硕士论文　论文类型：学位论文

【摘要】：由肿瘤、外伤、炎症等多种因素引起的颌骨缺损和吸收是口腔医学领域比较常见的临床难题。成骨细胞是参与骨形成的主要细胞,其前体细胞骨髓间充质干细胞在适当的条件下能分化成多种细胞,如成软骨细胞、成骨细胞、成脂肪细胞、成肌细胞等。通过对骨髓间充质干细胞向成骨细胞的分化和功能调控的研究,不仅对正确认识骨形成的机理具有重要的理论意义,而且对进一步治疗骨吸收相关疾病、促进骨再生具有实际的指导意义。骨形成是一个受多种细胞因子和生长因子调控的过程,如何利用生长因子更好的促进骨组织愈合,是目前的研究热点之一。研究表明,骨髓间充质干细胞复合骨源性生长因子能显著促进骨再生。BMP-2作为一种骨形成调控因子,在促进骨折愈合和骨缺损修复过程中作用显著。本实验拟通过转基因方法将外源性BMP-2导入骨髓间充质干细胞,使其高效、稳定的表达,以达到促进骨形成的目的。本研究通过建立体外骨髓间充质干细胞培养方法,用所构建的腺病毒载体携带BMP-2基因转染体外培养的骨髓间充质干细胞。通过MTT、碱性磷酸酶活性检测和茜素红染色等方法检测BMP-2对骨髓间充质干细胞增殖和成骨向分化的影响,结果表明BMP-2能促进骨髓间充质干细胞的增殖,并能促进其向成骨细胞的分化。因此,骨髓间充质干细胞这一培养体系的建立为体外研究骨髓间充质干细胞向成骨细胞分化和功能的调控提供了切实可行的方法;研究骨髓间充质干细胞转基因的可行性,为进一步利用转基因技术治疗骨吸收性疾病、促进骨形成奠定理论基础。
[Abstract]:By the tumor, trauma, inflammation caused by a variety of factors such as the jaw defect and absorption is difficult in clinical dentistry more common. Osteoblasts are important cells in bone formation, the precursor cells of bone marrow mesenchymal stem cells can differentiate into multiple cell types in appropriate conditions, such as cartilage, bone cells, adipocytes, myoblasts. The bone marrow mesenchymal stem cells on the differentiation and function of regulation of bone cells, the mechanism not only has important theoretical significance on the formation of a correct understanding of bone, but also for the further treatment of bone resorption related diseases, promote bone regeneration has practical significance.
Bone formation by a variety of cytokines and growth factors regulating the process, how to make use of growth factors to better promote bone healing, is one of the hot research at present. The study shows that bone marrow mesenchymal stem cells combined with bone derived growth factor can significantly promote bone regeneration.BMP-2 as a regulatory factor in bone formation. To promote fracture healing and bone defect repair process significantly. The experiment of transgenic methods of exogenous BMP-2 bone marrow mesenchymal stem cells, to make it efficient and stable expression, in order to achieve the purpose of promoting bone formation.
In this study, through the establishment of in vitro culture of bone marrow mesenchymal stem cell by adenovirus vector constructed by carrying BMP-2 gene transfection in vitro cultured bone marrow mesenchymal stem cells. By MTT method, and detected by alizarin red staining and alkaline phosphatase activity detection of BMP-2 of bone marrow mesenchymal stem cells proliferation and osteogenic differentiation the results show that BMP-2 can promote the effect of bone marrow mesenchymal stem cell proliferation, and can promote the osteogenic differentiation.
Therefore, bone marrow mesenchymal stem cells in this culture system was established for the study of bone marrow mesenchymal stem cells and provides a feasible way to regulate osteoblast differentiation and function; study the feasibility of bone marrow mesenchymal stem cells genetically modified, for the further use of transgenic technology for the treatment of bone resorptive diseases, promote bone lay to form a theoretical basis.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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