血小板衍化生长因子A基因转染成骨细胞生长特性的影响

Influence of platelet-derived growth factor A gene transfection on osteoblasts growth

背景:采用基因转染技术加强或改造种子细胞的功能是当今组织工程研究中的热点问题,用血小板衍化生长因子A转染成骨细胞用于牙周组织再生少见报道。目的:观察血小板衍化生长因子A基因转染成骨细胞后对其生长特性的影响。方法:构建血小板衍化生长因子A真核表达质粒,分离培养3T3细胞并进行不同方式的转染,RT-PCR检测血小板衍化生长因子A基因在成骨细胞中的表达;MTT法检测细胞的增殖活性;体外细胞划痕实验观察细胞的迁移情况。结果与结论:RT-PCR检测结果显示,血小板衍化生长因子A真核表达质粒转染的细胞表达量明显增加、生长、迁移速度明显增快(P<0.05)。说明脂质体介导血小板衍化生长因子A基因成功转染成骨细胞,并能促进细胞的增殖和迁移,为血小板衍化生长因子A基因在牙周组织中的治疗奠定基础。

BACKGROUND： Today＇s hot issue in the tissue engineering is to strengthen or reconstruct the function of seeding cells by gene transfection technology. But osteoblasts transfected by platelet-derived growth factor A （PDGF-A） for periodonta tissue regeneration is rarely reported. OBJECTIVE： To observe the influence of PDGF-A gene transfection on osteoblasts growth. METHODS： The eukaryotic expression plasmid of PDGF-A was constructed. 3T3 cells were isolated and transfected with various methods. The gene expression of PDGF-A in osteoblasts was detected by reverse transcription-PCR method, and cells proliferation activity was detected with MTT method, as well as the migration of cells was observed by the experiment of cells scratch test in vitro. RESULTS AND CONCLUSION： The results of reverse transcriptien-PCR showed that the expression of osteoblasts transfected by eukaryotic expression plasmid carrying PDGF-A was significantly increased, and growth rate and migration speed was accelerated obviously （P 〈 0.05）. These findings suggest that the limposome mediator PDGF-A gene can transfect osteoblasts successfully to promote the growth, proliferation and migration of osteoblasts, laying a foundation for PDGF-A gene treatment in periodontal tissue regeneration.