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机构地区:[1]上海第二医科大学第九人民医院口腔医学研究所,200011
出 处:《华西口腔医学杂志》2003年第5期347-349,共3页West China Journal of Stomatology
基 金:国家自然科学基金 (编号 30 0 0 0 1 91 );中国博士后科学基金 (编号中博基 1 999第 1 7号 );上海市博士后基金 (编号沪科 2 0 0 0第 487号 )资助项目
摘 要:目的 建立表达骨形成蛋白 2 (BMP_2 )的人牙周膜成纤维细胞 (HPDLFs) ,观察其体内成骨作用 ,为牙周缺损修复基因疗法的建立提供实验依据。方法 采用脂质体载体将人BMP_2噬菌粒表达载体pBK_B2转染至HPDLFs。利用免疫组化ABC法检测转染细胞内BMP_2蛋白的表达 ,并于无菌条件下将转染细胞注射于裸鼠股部肌肉内 ,2 1d后取材 ,苏木精—伊红染色观察肌肉组织内骨形成情况。结果 BMP_2基因转染后HPDLFs内有BMP_2蛋白的表达。在注射BMP_2基因转染细胞的肌肉组织内可见明显的新骨形成。结论 BMP_2基因成功转入HP DLFs中并得到有效表达 。Objective\ To establish human periodontal ligament fibroblasts (HPDLFs) that express bone morphogenetic protein 2 and observe their osteogenesis in vivo, so as to provide fundamental data for future study on gene therapy of periodontal defect treatment.Methods\ The phagemid expression vector for BMP_2 (pBK_B2) was transfected into HPDLFs by using Lipofect AMINE. The BMP_2 expression was determined by immunohistochemical ABC method. The BMP_2 gene transfected HPDLFs were injected into thigh muscle of nude mice under sterile condition. Specimens were collected in 21 days after injection and underwent normal procedures for fixation, decalcification, embedding, incising and staining with HE.Results\ The results demonstrated that BMP_2 protein expressed in HPDLFs after BMP_2 gene transfection. The HPDLFs transfected with BMP_2 gene could induce new bone formation at the injection site.Conclusion\ The results indicated that BMP_2 gene was transfected successfully and expressed efficiently in HPDLFs. The HPDLFs transfected with BMP_2 gene are capable of inducing new bone formation in vivo.
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