碱性成纤维细胞生长因子基因转染对人牙周韧带细胞的作用  

Effect of basic fibroblast growth factor gene transfection on the proliferation of human periodontal ligament cells

作　　者：李立恒[1] 高燕飞[2,3] 

机构地区：[1]河北北方学院附属第一医院,河北省张家口市075000 [2]河北北方学院 [3]张家口教育学院,河北省张家口市075000

出　　处：《中国组织工程研究》2012年第33期6147-6151,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:主要来源于因正畸或阻生拔除的健康牙培养而成的人牙周韧带细胞已成为牙周组织工程理想的种子细胞来源之一。目的:了解采用重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染对体外培养的人牙周韧带细胞增殖及细胞周期的影响。方法:体外培养人牙周韧带细胞,经重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染,分为3组:对照组、空载病毒组、碱性成纤维细胞生长因子转染组。用RT-PCR,Western blot检测人牙周韧带细胞在转染前后碱性成纤维细胞生长因子基因和蛋白的表达。应用细胞生长曲线、四甲基偶氮唑蓝比色法观察细胞生长的优化作用;采用流式细胞术测定细胞周期分布的变化。结果与结论:对照组、空载病毒组未检测到碱性成纤维细胞生长因子mRNA和蛋白表达;碱性成纤维细胞生长因子转染组碱性成纤维细胞生长因子mRNA和蛋白水平均有表达,细胞的生长速度明显增快,细胞周期G0/G1期减少,S期细胞数增多。各组间比较差异有显著性意义(P<0.05)。BACKGROUND： Human periodontal ligament ceils are one of the ideal seed cells in periodontal tissue engineering OBJECTIVE： To investigate the effect of basic fibroblast growth factor gene transfection with recombinant adeno-associated virus as a vector on the proliferation and cell cycle of cultured human periodontal ligament cells in vitro METHODS： Human periodontal ligament cells cultured in vitro were transfected with basic fibroblast growth factor gene mediated by recombinant adeno-associated virus, and then the transfected ceils were divided into three groups： control group, recombinant adeno-associated virus group and basic fibroblast growth factor gene transfection group. Reverse transcription-PCR and western blot were used to assess the gene and protein expression of basic fibroblast growth factor before and after transfection. Cellular proliferation was determined by the cell growth curve and MTT assay. The cell cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION： There was no basic fibroblast growth factor expression in the control and recombinant adeno-associated virus groups. After transfection, the expression of basic fibroblast growth factor mRNA and protein was found in the basic fibroblast growth factor gene transfection group, the cell growth was accelerated, the cell cycle of G0/G1 phase was decreased and cell number in S phase was increased. There was a significant difference among the groups （P 〈 0.05）.

关 键 词：碱性成纤维细胞生长因子 基因转染 人牙周韧带细胞 细胞周期 重组腺相关病毒 

分 类 号：R318[医药卫生—生物医学工程]