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目的:评价Nel样I型分子(Nell-1)基因修饰骨髓基质细胞(bone marrow stromal cells,bMSCs)复合β-磷酸三钙提升兔上颌窦底的效果。方法:抽取兔骨髓进行bMSCs培养,体外采用腺病毒载体携带Nell-1基因(AdNell-1)及绿色荧光蛋白EGFP基因(AdEGFP)转染bMSCs,GFP表达检测转染效率、Nell-1免疫细胞化学检测目的基因的表达。碱性磷酸酶(ALP)染色、半定量检测及钙结节茜素红染色检测细胞成骨分化。将基因修饰bMSCs与β-磷酸三钙颗粒复合用于兔上颌窦底提升,分别在术后2周和8周取材,HE染色,测量成骨面积,并采用SPSS11.0软件包对2组间数据进行t检验。结果:AdEGFP基因修饰组GFP表达效率可达60%～80%,Nell-1细胞化学染色显示,AdNell-1基因修饰组呈阳性表达。AdNell-1基因修饰组ALP染色及钙结节茜素红染色均高于AdEGFP基因修饰组,ALP半定量检测具有统计学差异(P<0.05)。体内实验研究中,AdNell-1基因修饰组新骨形成面积在8周时显著高于AdEGFP基因修饰组(P<0.05)。结论:采用AdNell-1基因转染兔bMSCs可促进其成骨分化,体内可促进上颌窦底提升的效果。
Objective: To evaluate the effect of Nel-like type I (Nell-1) gene-modified bone marrow stromal cells (β-TCP) combined with β-tricalcium on the maxillary sinus floor in rabbits. Methods: BMSCs were extracted from rabbit bone marrow and transfected into bMSCs by adenoviral vector carrying Nell-1 gene (AdNell-1) and green fluorescent protein EGFP gene (AdEGFP) in vitro. Transfection efficiency was detected by GFP expression. Nell-1 immunocytochemistry Detection of target gene expression. Alkaline phosphatase (ALP) staining, semi-quantitative detection and calcium nodular alizarin red staining of osteogenic differentiation. The genetically modified bMSCs and β-tricalcium phosphate particles were used to enhance the maxillary sinus floor in rabbits, and were harvested at 2 weeks and 8 weeks after operation respectively. HE staining was used to measure the osteogenic area. SPSS11.0 software package Data was t-tested. Results: The efficiency of GFP expression in AdEGFP gene modified group was 60% -80%. Nell-1 cells showed positive expression in AdNell-1 gene modified group. ALP staining and calcified nodules alizarin red staining of AdNell-1 gene modified group were higher than that of AdEGFP gene modified group, and there was a significant difference (P <0.05) in semi-quantitative detection of ALP. In vivo, the area of new bone formation in AdNell-1 gene modified group was significantly higher than that in AdEGFP gene modified group at 8 weeks (P <0.05). CONCLUSION: Transfection of rabbit bMSCs with AdNell-1 gene can promote osteogenic differentiation and promote the improvement of maxillary sinus floor in vivo.