脂肪干细胞外泌体对人牙髓干细胞增殖与成骨分化的影响

Effects of adipose-derived stem cell derived exosomes on proliferation and osteogenic differentiation of human dental pulp stem cells

目的观察人脂肪间充质干细胞来源外泌体对人牙髓干细胞增殖与成骨分化的影响。方法制备人脂肪来源的间充质干细胞和外泌体。将人牙髓干细胞随机分为3组,正常组进行常规培养,对照组进行成骨诱导分化,实验组在成骨诱导分化时添加5μg·mL^(-1)外泌体。用CCK-8法检测细胞增殖活性,用酶联免疫吸附实验法检测碱性磷酸酶活性,用Western blot法检测骨钙素和骨桥蛋白的表达情况。结果实验组、对照组和正常组的细胞相对增殖活性(光密度值)分别为0.54±0.02,0.34±0.01和0.32±0.01,碱性磷酸酶相对活性(光密度值)分别为10.37±1.26,4.89±0.94和0.71±0.03,骨钙素蛋白相对表达量分别为0.59±0.10,0.23±0.05和0,骨桥蛋白相对表达量分别为0.63±0.12,0.61±0.04和0,实验组的上述指标与对照组和正常组比较,差异均有统计学意义(均P<0.05)。结论人脂肪间充质干细胞来源的外泌体可促进人牙髓干细胞的增殖和成骨分化。

Objective To observe the effect of exosomes derived from human adipose-derived mesenchymal stem cells on the proliferation and osteogenic differentiation of human dental pulp stem cells.Methods Human adipose-derived mesenchymal stem cells and exosomes were prepared.Human dental pulp stem cells were randomly divided into three groups.Normal group was cultured routinely.Control group was osteogenic differentiation.Experimental group was supplemented with 5μg·mL^(-1)exosomes during osteogenic differentiation.Cell Counting Kit-8 was used to detect cell proliferation activity.Enzyme-linked immunosorbent assay was used to detect alkaline phosphatase activity.Western blot was used to detect the expressions of osteocalcin and osteopontin.Results The relative proliferation activities(optical density value)of the experimental,control and normal groups were0.54±0.02,0.34±0.01 and 0.32±0.01,the relative activities of alkaline phosphatase(optical density value)were 10.37±1.26,4.89±0.94 and 0.71±0.03,the relative expressions of osteocalcin were 0.59±0.10,0.23±0.05 and 0,the relative expressions of osteopontin were 0.63±0.12,0.61±0.04 and 0.Compared with the control group and normal group,there were statistically significant differences in the above indexes of the experimental group(all P<0.05).Conclusion The exosomes derived from human adipose mesenchymal stem cells can promote the proliferation and osteogenic differentiation of human dental pulp stem cells.