牙本质胞外基质对体外血管形成的影响

Effects of dentin extracellular matrix components on angiogenesis

目的:探讨牙本质胞外基质成分(dentinextracellularmatrixcomponents ,DMCs)对体外血管形成的影响。方法:10 %EDTA提取健康人牙中的DMCs。血管内皮细胞 成纤维细胞共培养体系在含有DMCs (0 .0 0 0 1～5mg/ml)的培养液中培养11d ,PECAM 1和vonWillebrand因子显色血管后采用图像分析系统测量血管总数、长度及分支数。添加刺激因子VEGF组及抑制因子Suramin组分别作为阳性和阴性对照组,单纯培养液为空白对照组。各项指标与空白对照进行Student T统计分析。结果:低浓度DMCs(0 .0 0 0 1～0 .0 1mg/ml)可产生类似VEGF组刺激血管形成的效果,且最佳作用浓度为0 .0 1mg/ml。较高浓度DMCs (0 .1～5mg/ml)则对血管形成具有显著的抑制效应(P <0 .0 0 1)。结论:DMCs对血管形成具有计量依赖型影响效应。提示这一计量依赖型效应可能正是牙髓损伤后导致不同修复结果的原因。

Objective:To investigate whether preparations of dentine matrix components show angiogenic effects in vitro.Method: A total EDTA-soluble preparation of dentine matrix components was prepared by extraction of dissected sound human dentine with 10% EDTA. An endothelial cell - fibroblast co-culture system was used to investigate the angiogenic effects of the dentine matrix components (concentration range 0.0001～5.0 mg/ml), which were assessed after culture 11 days by development of branched, tubular capillary-like structures showing positive immunoreactivity for PECAM-1 and von Willebrand factor. Stimulatory effects of vascular endothelial growth factor (VEGF) (positive control) and inhibitory effects of suramin (negative control) were examined as well as media alone. Data were statistically compared with those from blank control using student-T method.Result: Stimulation of angiogenesis was observed in cultures with the VEGF (Positive control) and the dentine matrix components at concentration 0.001～ 0.01 mg/ml with optimal effects at 0.01 mg/ml as compared to the media alone. Higher concentrations of dentine matrix components (0.1～ 5 mg/ml ) inhibited angiogenesis significantly, as was suramin (negative control).Conclusion: Dentine matrix components can stimulate angiogenesis in a dose-dependent manner and release of such components after dental injury may contribute to variable angiogenesis and repair in the pulp.