碱性成纤维细胞生长因子和骨形成蛋白2在牙源性细胞分化中的表达及意义

The expression and modulatory effect of fibroblast growth factor 2 and bone morphogenetic protein 2 on the differentiation of dental derived cells

目的研究人牙髓细胞(DPCs)和牙周韧带细胞(PDLCs)矿化过程中碱性成纤维细胞生长因子(FGF2)和骨形成蛋白2(BMP2)的表达变化,以及FGF2和BMP2在牙髓牙周损伤修复动物模型中的分布,探讨FGF和BMP信号通路在DPCs和PDLCs向成牙本质/成骨样细胞分化及牙髓牙周组织损伤修复中的调控作用。方法酶消化法分离培养DPCs和PDLCs,取诱导前及矿化诱导14d的DPCs和PDLCs,实时荧光定量反转录聚合酶链反应检测FGF2和BMP2的表达变化并进行统计学分析。建立大鼠牙髓牙周联合损伤动物模型,HE染色观察4周后牙髓牙本质复合体及牙周组织修复情况,免疫荧光检测FGF2和BMP2在新生牙髓及牙周组织的表达和分布。结果 DPCs和PDLCs经矿化诱导,FGF2mRNA表达下调,BMP2mRNA表达上调,与对照组间差异有统计学意义(P<0.05)。免疫荧光示FGF2在新生的牙周韧带组织强表达,在新生牙髓及正常牙髓牙周组织弱表达,BMP2蛋白在损伤处的新生牙髓及牙周韧带组织细胞浆强表达,FGF2和BMP2在正常牙髓及牙周韧带组织均无表达。结论 FGF2和BMP2在DPCs和PDLCs体外成牙本质/成骨分化中可能具备独立的生物学功能,在体内牙齿损伤修复过程中可能存在协同调控作用。

Objective The purpose of this study was to investigate the expression and distribution of fibroblast growth factor 2（FGF2） and bone morphogenetic protein 2（BMP2） in human dental pulp cells（DPCs） and periodontal ligament cells（PDLCs） undergoing odontogenic/osteogenic differentiation and rat dental pulp and periodontal ligament with injury.Methods DPCs and PDLCs were isolated by enzymatic digestion and preceded to odontogenic/osteogenic induction for 14 d,and the cells without induction served as controls.The expressions of FGF2 and BMP2 were investigated by quantitative real-time RT-PCR.Lewis rat model with dental pulp and periodontal ligament injury was created and the distribution of FGF2 and BMP2 was determined by immunofluorescent staining.Results The mRNA level of BMP2 was significantly increased albeit FGF2 significantly decreased compared with the control group as DPCs and PDLCs undergoing odontogenic/osteogenic induction（P 0.05）.Immunofluorescent staining also revealed the FGF2 merely showed tense expression in the regenerated peridontal ligament tissue,whereas BMP2 signal was activated both in the newly formed dental pulp and periodontal ligament tissue compared with the normal ones.Conclusions FGF2 and BMP2 signaling may play individual modulatory role in the odontogenic/osteogenic differentiation of dental derived cells with in vitro culture,albeit FGF2 and BMP2 may cooperatively regulate the in vivo dental tissue regeneration after injury.