生物活性玻璃58S/丝素蛋白膜促进人牙髓干细胞增殖与分化

Bioactive glass 58S/silk fibroin membrane premotes proliferation and differentiation of human dental pulp stem cells

目的:研究生物活性玻璃58S/丝素蛋白(BG58S/SF)膜对人牙髓干细胞(h DPSCs)增殖、分化的影响。方法:制备纯SF膜(A组)和1 mg/m L(B组)、5 mg/m L(C组)BG58S/SF膜预孵育24 h后分别接种h DPSCs。于培养4、7、14、21 d时,CCK-8法检测h DPSCs的增殖能力;扫描电镜和荧光染色观察h DPSCs在膜材料表面的粘附和增殖状态;ALP活性试验评估h DPSCs的分化潜能;Real-Time PCR检测h DPSCs向成牙本质细胞分化特异性相关基因的表达水平。结果:h DPSCs在纯SF膜和BG58S/SF膜材料上粘附良好;从第7天起,与A组相比,B、C组细胞ALP活性显著增高,h DPSCs向成牙本质细胞分化特异性相关基因表达水平上调更加显著(P<0.05),且C组均高于B组。结论:BG58S/SF膜能促进h DPSCs粘附、增殖与分化。

AIM： To investigate the effect of bioactive glass 58S/silk fibroin（ BG58S/SF） membrane on the proliferation and differentiation of human dental pulp stem cells（ h DPSCs）. METHODS： h DPSCs were seeded on pure SF membrane and BG58 S / SF with BG58 S at 1 mg / m L and 5 mg / m L respectively after the materials were incubated in cell culture medium for 24 hours. Cell proliferation was assessed by CCK- 8 kit after 4,7,14 and 21 d culture respectively. The adhesion of h DPSCs on the material surface was evaluated by SEM and cytoskeleton staining; ALP activity was measured by ALP kit and the expression of odontoblastic differentiation- related genes was measured by RT- PCR. One- way ANOVA and paired test were used for statistical analysis. RESULTS： h DPSCs proliferated well on the surface of the membranes throughout the culture period. ALP activity was enhanced in all the 3 groups from day 7,BG58 S / SF membrane groups showed higher ALP activity than SF group（ P〈0. 05）. BG58 S / SF groups showed higher up- regulation of odontoblastic differentiation- associated genes than to SF group（ P〈0. 05）. ALP activity and orodontoblastic differentiation- associated gene expression level in 5 mg / m L BG58 S / SF group was higher than that in1 mg / m L group（ P〈0. 05）. CONCLUSION： BG58 S / SF membrane may promote the proliferation and odontoblastic differentiation of h DPSCs.