rhPTH(1-34)间断刺激对人根尖牙乳头干细胞增殖及成牙成骨分化的影响  

Effect of recombined human parathyroid hormone 1-34 on proliferation and odonto/osteogenic differentiation of human stem cells from apical papilla

作　　者：庄颖 庞希瑶[1,4] 蔡琴 曹灵 于金华[1,2] ZHUANG Ying;PANG Xiyao;CAI Qin;CAO Ling;YU Jinhua(Jiangsu Key Laboratory of Oral Diseases,Nanjing Medical University,Nanjing 210029,China;Department of Operative Dentistry and Endodontics,Affiliated Hospital of Stomatology,Nanjing Medical University,Nanjing 210029,China;Department of Stomatology,People＇s Hospital of Binhai,Binhai 224500,China;Department of Stomatology,Nanjing First Hospital,Nanjing 210006,China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学附属口腔医院牙体牙髓科,江苏南京210029 [3]江苏省滨海县人民医院口腔科,江苏滨海224500 [4]南京市第一医院口腔科,江苏南京210006

出　　处：《口腔生物医学》2018年第3期125-128,共4页Oral Biomedicine

基　　金：国家自然科学基金(81371144);江苏高校优势学科建设工程资助项目(2014-37)

摘　　要：目的:探讨重组人甲状旁腺素(1-34)[rh PTH(1-34)]对人根尖牙乳头干细胞(SCAPs)增殖和成牙成骨分化潜能的影响。方法:酶消化法原代培养SCAPs,分别用完全培养基、1×10^(-12)、1×10^(-11)、1×10-10、1×10-9、1×10^(-8)mol/L rh PTH(1-34)条件培养液间断性刺激SCAPs,碱性磷酸酶(ALP)检测法筛选最佳诱导浓度,CCK8法检测rh PTH(1-34)对SCAPs增殖的影响,实时定量RT-PCR检测rh PTH(1-34)作用于SCAPs后其成牙/成骨分化指标的变化。结果:1×10-8mol/L rh PTH(1-34)间断性刺激能显著提高SCAPs的ALP活性;CCK8结果显示rh PTH(1-34)间断性作用于SCAPs后,其增殖能力无明显改变(P>0.05);rh PTH(1-34)间断性刺激可增强SCAPs中骨钙素(OCN)、Osterix、Runt相关转录因子2(RUNX2)、牙本质涎蛋白(DSP)等基因的表达(P<0.05)。结论:rh PTH(1-34)间断刺激对人SCAPs的增殖无明显影响,但可促进其成牙/成骨向分化。Objective： To investigate the effect of recombined human parathyroid hormone rhPTH （1-34） on the proliferation and odonto/osteogenic differentiation of human stem cells from apical papilla （SCAPs） in vitro. Methods： Primary SCAPs were isolated by the method of enzyme digestion. SCAPs were intermittently stimulated by complete medium, 1×10^-12 , 1×10^-11 , 1×10^-10 , 1×10^-9 and 1×10^-8 mol/L rhPTH （1-34）. Then SCAPs were cultured in α-MEM supplemented with different concentrations of rhPTH（1-34） and the optimal concentration of rhPTH（1-34） was determined by the alkaline phosphatase （ALP） activity. CCK8 assay was used to detect the proliferation ability of SCAPs. RT-PCR was performed to investigate the odonto/osteogenic potential of SCAPs by detecting the expression of osteocalcin （OCN）, osterix （OSX）, runt-related transcription factor 2 （RUNX2） and dentin sialoprotein （DSP）. Results： ALP activity was significantly upregulated by 1×10^-8 mol/L rhPTH（1-34）, while CCK8 assay showed that there was no significant difference （ P 〉0.05） in the proliferation ability between the control group and the rhPTH（1-34） intermittently treated group. RT-PCR analysis revealed that the odonto/osteogenic markers （OCN, OSX, RUNX2, DSP） were significantly upregulated in rhPTH（1-34） intermittently treated group as compared with control group. Conclusions： The intermittently treated rhPTH （1-34） has no effect on the proliferation of SCAPs, but can significantly promote the capacity of their odonto/osteogenic differentiation.

关 键 词：重组人甲状旁腺素(1—34) 根尖牙乳头干细胞 增殖 分化 

分 类 号：R780.2[医药卫生—口腔医学]