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目的探讨杨梅素对乳牙牙髓干细胞(SHED)增殖和成骨分化能力的影响。方法收集7~10岁儿童因滞留而拔除的乳牙,分离、培养SHED。加入含有1μmol/L(A1组)、5μmol/L(B1组)、10μmol/L(C1组)、15μmol/L(D1组)、20μmol/L(E1组)、50μmol/L(F1组)和100μmol/L(G1组)杨梅素的DMEM培养基200μl;对照1组加入不含杨梅素的DMEM培养基200μl;分别培养24、48和72h,观察杨梅素对SHED增殖能力的影响。加入含有1μmol/L(A2组)、5μmol/L(B2组)、10μmol/L(C2组)、15μmol/L(D2组)、20μmol/L(E2组)杨梅素的成骨诱导液200μl;对照2组加入不含杨梅素的成骨诱导液200μl;培养3、5和7d,检测SHED碱性磷酸酶(ALP)活性。结果 A1组和B1组SHED增殖能力与对照1组比较无统计学差异(P>0.05),C1组、D1组、E1组促进SHED的增殖(P<0.05),F1组和G1组抑制SHED的增殖(P<0.05)。各组ALP活性在5d时达到最高,7d时活性有所下降。与对照2组相比,C2组、D2组、E2组能促进ALP活性(P<0.05),且随着杨梅素浓度的增加,ALP活性增加(P<0.05)。结论适宜浓度的杨梅素可以促进SHED的增殖能力和早期成骨能力,以20μmol/L的杨梅素作用最为显著。
Objective To investigate the effect of myricetin on the proliferation and osteogenic differentiation of dental pulp stem cells (SHED) of deciduous teeth. Methods The deciduous deciduous teeth from children aged 7 to 10 were collected, and SHED was isolated and cultured. The cells were treated with 1μmol / L (group A1), 5μmol / L (group B1), 10μmol / L group C1, 15μmol / L group D1, 20μmol / L group E1 and 50μmol / 200μmol / L DMEM medium with 100μmol / L (G1 group); 200μl DMEM medium containing no myricetin in control group 1; cultured for 24, 48 and 72 hours respectively; the effect of myricetin on the proliferation of SHED was observed. 200μl of osteogenic induction solution containing 1μmol / L (A2), 5μmol / L (B2), 10μmol / L (C2), 15μmol / L (D2) and 20μmol / L Two control groups were added with 200μl osteosarcoma without myricetin induction. After incubated for 3, 5 and 7 days, alkaline phosphatase (ALP) activity of SHED was detected. Results There was no significant difference in SHED proliferation between group A1 and group B1 (P> 0.05), while group C1, group D1 and group E1 promoted SHED proliferation (P <0.05) Proliferation (P <0.05). The activity of ALP in each group reached the highest at 5d, and decreased at 7d. Compared with control group 2, group C2, group D2 and group E2 could promote ALP activity (P <0.05), and ALP activity increased with the increase of myricetin concentration (P <0.05). Conclusion Myricetin at a suitable concentration can promote SHED proliferation and early osteogenesis, with 20μmol / L myricetin being the most effective.