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目的探讨提取的牙骨质非胶原蛋白(cementum noncollagenous proteins,CNCPs)对乳牙牙髓干细胞(Stem cell from human exfoliated deciduous teeth,SHED)增殖、分化和功能的影响。方法拔除2岁龄牛健康牙,提取牙骨质基质的盐酸胍提取物。采用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析盐酸胍提取物的组分和分子量。采用MTT、ALP染色、茜素红染色和RT-PCR等方法观察牙骨质非胶原蛋白对SHED的影响。结果 CNCPs对SHED的增殖无促进作用,CNCPs(1ug/mL)组作用于SHED,在7d和14d时,同不含CNCPs组相比,能明显上调SHED的碱性磷酸酶活性(P<0.01),CNCPs(1ug/mL)可明显促进SHED表达DSPP、DMP1、BSP和Runx-2(P<0.05)及矿化。结论 CNCPs能够促进SHED向成牙本质细胞的分化及矿化。
Objective To investigate the effects of extracted cementum noncollagenous proteins (CNCPs) on the proliferation, differentiation and function of dental pulp stem cells derived from human exfoliated deciduous teeth (SHED). Methods Two-year-old cattle healthy teeth were extracted and the guanidine hydrochloride extract of cementum was extracted. The components and molecular weights of the guanidine hydrochloride extract were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The effect of cementless collagen on SHED was observed by MTT, ALP staining, alizarin red staining and RT-PCR. Results CNCPs had no effect on the proliferation of SHED. The CNCPs (1ug / mL) treated SHED at 7d and 14d significantly increased the alkaline phosphatase activity of SHED compared with those without CNCPs (P <0.01) , CNCPs (1ug / mL) significantly promoted SHED expression of DSPP, DMP1, BSP and Runx-2 (P <0.05) and mineralization. Conclusion CNCPs can promote the differentiation and mineralization of SHED into odontoblasts.