【作者】 李佳； 霍晓炎； 鲁晓琪； 李淑娟；

【Author】 LI Jia;HUO Xiao-yan;LU Xiao-qi;LI Shu-juan;Stomatology Hospital of Hebei Medical University;

【机构】 河北医科大学口腔医院；

【摘要】 目的 :将不同浓度的槲皮素作用于SD大鼠牙龈间充质干细胞(GMSCs),以探究其对GMSCs增殖能力的作用。方法 :选择4周龄的SD大鼠,提取牙龈组织,组织块法培养GMSCs,体外进行成骨及成脂分化潜能鉴定,取生长状况良好的第4代GMSCs接种于96孔板,实验分为6组,24小时后,弃去旧培养液,分别加入槲皮素浓度为0mM、20mM、40mM、60mM、80mM、100mM的培养液,于0d、1d、2d、3d时加入CCK8检测液,孵育2h后酶标仪测定450nm波长下的OD值。结果 :通过体外成骨诱导液诱导,茜素红染色中可观察到明显矿化结节生成,证明GMSCs可向成骨方向分化。通过体外成脂诱导液诱导,油红O染色中可观察到红染的脂滴生成,证明GMSCs可向脂肪样细胞方向分化。CCK8检测结果显示,不同浓度的槲皮素均可促进GMSCs的增殖,其中浓度为80mM时促进细胞增殖能力的作用较强,但这种促进能力随时间增长逐渐消失。结论 :SD大鼠分离培养的GMSCs具有成骨及成脂分化潜能,将槲皮素作用于GMSCs可在较短时间内促进细胞增殖,浓度为80mM时尤为明显,但促进作用会随时间消失。更多还原

【Abstract】 Objective: Different concentrations of quercetin were applied to SD rat gingival mesenchymal stem cells(GMSCs) to explore its effect on the proliferation of GMSCs. Methods: 4-week-old SD rats were selected to extract gingival tissue. GMSCs were cultured by tissue block method. The osteogenic and adipogenic differentiation potential of GMSCs were identified in vitro. The fourth generation GMSCs were inoculated into 96 well plate and divided into 6 groups. After 24 hours, the old culture medium was discarded, and the culture medium with quercetin concentration of 0 mM, 20 mM, 40 mM, 60 mM, 80 mM and 100 mM was added respectively. CCK8 was added at 0 d, 1 d, 2 d and 3 d. After 2 hours of incubation, OD value at 450 nm wavelength was determined by a microplate reader. Results: Mineralized nodule formation was observed in alizarin red staining by induction of osteogenic induction solution in vitro, which proved that GMSCs could differentiate into osteogenic direction. Lipid droplet formation was observed in oil red O staining induced by adipogenic induction solution in vitro, which proved that GMSCs could differentiate into adipoid cells. CCK8 detection results showed that different concentrations of quercetin could promote the proliferation of GMSCs, and the effect of 80 mM quercetin on the proliferation of GMSCs was stronger, but the promotion ability gradually disappeared with time. Conclusions: GMSCs isolated from SD rats have the potential of osteogenic and adipogenic differentiation. Quercetin can promote the proliferation of GMSCs in a short time, especially at the concentration of 80 mm, but it will disappear with time.更多还原