CAJ | 학술논문

背景：转化生长因子β3与肝素联合作用后对人乳牙牙髓干细胞的增殖和矿化能力的作用目前仍有待研究。目的：探讨转化生长因子β3对人乳牙牙髓干细胞向成牙本质样细胞分化的作用。方法：采用酶消化法将人乳牙牙髓分离培养，获得人乳牙牙髓干细胞；对体外培养的第3代人乳牙牙髓干细胞单独加入25μg/L重组转化生长因子β3、10 U/mL 肝素，或者二者联合进行培养；通过Q-PCR和Western-blotting 方法，分别检测乳牙牙髓干细胞中牙本质涎磷蛋白基因及牙本质涎蛋白表达的情况；通过碱性磷酸酶试剂盒检测乳牙牙髓干细胞碱性磷酸酶活性的改变。结果与结论：乳牙牙髓干细胞在诱导体系中生长状态良好。25μg/L转化生长因子β3+10 U/mL肝素联合作用组的碱性磷酸酶活性明显增强，与转化生长因子β3单独作用组、肝素单独作用组以及对照组相比差异有显著性意义(P <0.01)；转化生长因子β3+肝素联合作用组的茜素红染色呈强阳性，Q-PCR和Western-blotting结果均显示，转化生长因子β3+肝素联合作用组的牙本质涎磷蛋白 mRNA 和蛋白表达均明显升高。结果可见在转化生长因子β3与肝素联合作用的诱导下，可促进乳牙牙髓干细胞分化为成牙本质样细胞。
배경：전화생장인자β3여간소연합작용후대인유아아수간세포적증식화광화능력적작용목전잉유대연구。목적：탐토전화생장인자β3대인유아아수간세포향성아본질양세포분화적작용。방법：채용매소화법장인유아아수분리배양，획득인유아아수간세포；대체외배양적제3대인유아아수간세포단독가입25μg/L중조전화생장인자β3、10 U/mL 간소，혹자이자연합진행배양；통과Q-PCR화Western-blotting 방법，분별검측유아아수간세포중아본질연린단백기인급아본질연단백표체적정황；통과감성린산매시제합검측유아아수간세포감성린산매활성적개변。결과여결론：유아아수간세포재유도체계중생장상태량호。25μg/L전화생장인자β3+10 U/mL간소연합작용조적감성린산매활성명현증강，여전화생장인자β3단독작용조、간소단독작용조이급대조조상비차이유현저성의의(P <0.01)；전화생장인자β3+간소연합작용조적천소홍염색정강양성，Q-PCR화Western-blotting결과균현시，전화생장인자β3+간소연합작용조적아본질연린단백 mRNA 화단백표체균명현승고。결과가견재전화생장인자β3여간소연합작용적유도하，가촉진유아아수간세포분화위성아본질양세포。
BACKGROUND:Studies have reported that the superfamily of transforming growth factors exert a role in the mineralization of various stem cells, but the combination effects of transforming growth factorβ3 and heparin on proliferation and mineralization ability of stem cells from human exfoliated deciduous teeth remains to be studied. OBJECTIVE:To explore the effect of transforming growth factorβ3 on odontoblast-like differentiation of stem cells from human exfoliated deciduous teeth. METHODS:Human deciduous teeth were col ected using enzyme digestion. The 3rd cells were incubated with 25μg/L recombinant human transforming growth factorβ3, 10 U/mL heparin or their combination. The dentin sialophosphoprotein mRNA and dentinsialoprotein expressions were detected by Q-PCR and western blot assay. Alkaline phosphatase activity was determined using alkaline phosphatase kit. RESULTS AND CONCLUSION:Stem cells from human exfoliated deciduous teeth grew wel after induction. The activity of alkaline phosphatase in the combination group was significantly higher than that in the transforming growth factorβ3, heparin and control groups (P<0.01). After combination induction, the cells were strongly positive for alizarin red staining. Results fromα-PCR and western blot assay showed that the expressions of dentin sialophosphoprotein were both remarkably increased at mRNA and protein levels. In summary, stem cells from human exfoliated deciduous teeth can differentiate into odontoblast-like cells under the induction of transforming growth factorβ3 plus heparin.