Si Xiaohui, Liu Zheng
Research Institute of Stomatology, Ninth People's Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2003 Oct;21(5):347-9.
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.
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.
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.
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.
构建表达骨形态发生蛋白2的人牙周膜成纤维细胞（HPDLFs）并观察其在体内的成骨情况，为牙周缺损基因治疗的进一步研究提供基础数据。
利用脂质体转染法将骨形态发生蛋白2（BMP-2）的噬菌粒表达载体（pBK-B2）转染至人牙周膜成纤维细胞。采用免疫组织化学ABC法检测BMP-2的表达。将转染BMP-2基因的人牙周膜成纤维细胞在无菌条件下注射到裸鼠大腿肌肉内。注射后21天取材，按常规进行固定、脱钙、包埋、切片及苏木精-伊红（HE）染色。
结果显示，BMP-2基因转染后人牙周膜成纤维细胞表达BMP-2蛋白。转染BMP-2基因的人牙周膜成纤维细胞可在注射部位诱导新骨形成。
结果表明，BMP-2基因成功转染并在人牙周膜成纤维细胞中高效表达。转染BMP-2基因的人牙周膜成纤维细胞在体内能够诱导新骨形成。
