CGRP体外促成骨、成神经效果及钛板表面CGRP缓释系统的建立

Effect of CGRP on Mock-microenvironment around Implants and the Building-up of CGRP Sustained Release System on Titanium Plate Surface

目的:探索降钙素基因相关肽(calcitonin gene-related peptide,CGRP)对人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)的影响及最佳作用浓度,并在钛板表面建立CGRP缓释系统,检测其促神经生长作用。方法:测定0~10-6 mol/L不同浓度CGRP对hBMSCs增殖和分化的作用,继而根据最佳细胞活性结果组CGRP浓度,测定其对hBMSCs碱性磷酸酶活性、成骨相关基因及蛋白表达的作用。通过聚多巴胺螯合CGRP黏附于钛板表面,ELISA法测定在28 d内钛板的CGRP释放浓度,并测定其对人神经母细胞瘤细胞(SH-SY5Y)的作用。结果:10-7 mol/L浓度CGRP在体外促进了hBMSCs和SH-SY5Y的成骨和成神经向分化。聚多巴胺螯合CGRP复合改性的钛板具有CGRP缓释效果,且具有促神经细胞增殖分化的作用。结论:聚多巴胺螯合CGRP复合改性钛板表面可为CGRP在种植体的临床应用,以促进种植体骨结合和骨感知提供新策略和途径。

Objective:To explore the effect of calcitonin gene related peptide(CGRP)on human bone marrow mesenchymal stem cells(hBMSCs)and its optimal concentration,and to build up a CGRP sustained-release system on the surface of titanium plate to detect its nerve growth promoting effect.Methods:The proliferation and differentiation of hBMSCs induced by different concentrations of CGRP(0-10-6 mol/L)were measured.According to the concentration of CGRP induced the best cell activity of hBMSCs,the effects of CGRP on the activity of alkaline phosphatase and the expression of osteogenic related genes and proteins were determined.The surface of titanium plate was modified by dopamine chelating CGRP compound.The sustained release effect of CGRP was checked by Elisa in 28 days,and its effect on human neuroblastoma cells(SH-SY5Y)was also measured.Results:CGRP with a concentration of 10-7 mol/L had the effect of promoting osteogenic differentiation of hBMSCs and neuronal differentiation of SH-SY5Y in vitro.Dopamine chelating CGRP composite modified titanium plate had a sustained release effect and promoted the proliferation and differentiation of nerve cells.Conclusion:Dopamine chelating CGRP composite modified titanium plate surface might build up a way to apply CGRP in the microenvironments round implants,forming a new strategy to promote the osseointegration and osseoperception of dental implants.