镁黄长石浸提液诱导多潜能干细胞的成骨分化

Osteogenic differentiation of pluripotent stem cells induced by akermanite extracts

背景:镁黄长石属于硅酸盐生物活性陶瓷,在生物体内具有降解作用,降解溶出的离子产物能够诱导细胞成骨分化,是组织工程骨支架材料的良好选择。目的:通过研究不同浓度的镁黄长石浸提液对诱导性多潜能干细胞增殖和成骨分化的影响,确定镁黄长石浸提液促进诱导性多潜能干细胞成骨分化的最佳浓度。方法:将诱导性多潜能干细胞培养于不同浓度的镁黄长石浸提液中,用MTT法检测细胞的增殖情况;在第7,14,21天时,分别收集培养上清液,检测其中碱性磷酸酶、骨钙素、Ⅰ型胶原蛋白含量。结果与结论:镁黄长石浸提液促进诱导性多潜能干细胞增殖的作用具有时间依赖性,第3天时,诱导性多潜能干细胞增殖明显,第5天与第3天相比差异无显著性意义,至第7天时则明显减弱。同时,1/4浓度浸提液组促进诱导性多潜能干细胞增殖作用最强。作为细胞成骨分化标志的碱性磷酸酶和骨钙素含量随时间增加而增加,1/4浸提液浓度组含量最高。作为细胞成骨分化早中期标志的Ⅰ型胶原蛋白在第7天时各组均未检出,第14,21天时,同样是1/4浓度浸提液组含量最高,这些说明镁黄长石浸提液中的Ca、Mg、Si离子参与了诱导性多潜能干细胞的成骨分化,其中1/4浓度浸提液(Ca离子2.37 mmol/L、Mg离子1.12 mmol/L、Si离子1.05 mmol/L)促进诱导性多潜能干细胞体外成骨分化的效果最好。

BACKGROUND：Akermanite belongs to silicate bioactive ceramics, with degradation in vivo, and its ionic products can induce osteogenic differentiation, which is a good choice for bone tissue engineering scaffold. OBJECTIVE：By studying the different concentrations of akermanite extracts on the proliferation and osteogenic differentiation of pluripotent stem cel s, to determine the optimum concentration of akermanite extract to inducing the osteogenic differentiation of pluripotent stem cel s. METHODS：MTT assay was used to detect the proliferation of pluripotent stem cel s cultured in different concentrations of akermanite extracts （1/2, 1/4, 1/8, 1/16, 1/32）. After cultured with the extracts for 7, 14, 21 days, the culture supernatants were col ected to detect the levels of alkaline phosphatase, osteocalcin, type I col agen. RESULTS AND CONCLUSION：The MTT assay showed that the proliferation of pluripotent stem cel s was increased in a concentration-dependent manner after induction with akermanite extracts. The pluripotent stem cel s proliferated obviously at 3 days after induction, and then weaken at 7 days, but there was no difference at 3 and 5 days after induction. At 7 days after induction, the 1/4 extract had the best effect on promoting osteogenic differentiation of pluripotent stem cel s;the levels of alkaline phosphatase and osteocalcin were increased with time, especial y after induction with the 1/4 extract;but there was no expression of type I col agen. At 14 and 21 days after induction, the levels of alkaline phosphatase, osteocalcin, type I col agen were highest in the 1/4 extract group. These findings indicate that the 1/4 akermanite extract （Ca 2.37 mmol/L, Mg 1.12 mmol/L, Si 1.05 mmol/L） is the optimum to promote the osteogenic differentiation of pluripotent stem cel s.