镁黄长石浸提液影响人脂肪干细胞增殖和成骨分化的实验研究

Experimental Study on the Effect of Extracts of Akermanite on Proliferation and Osteo-Differentiation of Human Adipose-Derived Stem Cells

目的通过探讨不同浓度镁黄长石浸提液对人脂肪干细胞(Human adipose-derived stem cells,hADSCs)增殖和成骨分化的影响,初步阐明镁黄长石体外促进hADSCs成骨分化的机制。方法依照ISO/EN 10993-5标准,制备镁黄长石浸提液,将获得的hADSCs培养于不同浓度的浸提液中(1/2、1/4、1/8、1/16、1/32),利用MTT法检测细胞的增殖情况;在成骨诱导条件下,通过碱性磷酸酶染色、活性检测和茜素红染色以及钙离子定量检测,观察不同浓度浸提液对hADSCs成骨特性的影响。结果 1/2、1/4、1/8浓度的浸提液可以浓度依赖性地抑制hADSCs的体外增殖;1/4、1/8、1/16浓度的浸提液可以促进hADSCs的体外成骨分化;碱性磷酸酶染色和活性检测、茜素红染色和钙离子定量检测显示,1/4浓度的浸提液对hADSCs的体外成骨分化促进作用最强,此时的钙、镁和硅离子浓度分别为:2.36 mM、1.11 mM和1.03 mM。结论镁黄长石浸提液中的离子在适当浓度时,可以抑制hADSCs的体外增殖,同时促进hADSCs的体外成骨分化。

Objective To elucidate the underlying mechanism of the osteo-differentiation of akermanite on human adipose-derived stem cells（hADSCs） via observing the effect of different diluted rate extracts on the proliferation and osteo-differentiation of hDASCs.Methods The original extract was prepared according to the ISO/EN 10993-5 and hDASCs were cultured in the different diluted rate extracts（1/2,1/4,1/8,1/16,1/32）.The MTT assay was carried out to quantitatively evaluate the proliferation of the cells.Under the condition of osteo-induction,osteogenic differentiation of hADSCs cultured in the different extracts was detected by ALP expression and calcium deposition.Results The 1/2,1/4,1/8 extracts could concentration-dependently suppress the proliferation of hASCs,while the 1/4,1/8,1/16 extracts could promote the osteogenic differentiation of hADSCs.Among them,the 1/4 extract with the concentration of Ca2＋,Mg2＋ and Si4＋ were 2.36 mM,1.11 mM,1.03 mM,has the optimum one.Conclusion The ions in the extract of akermanite can suppress the proliferation of hADSCs while promoting the osteogenic differentiation of hADSCs in a suitable concentration.