载血管内皮细胞生长因子微球促进牙髓再生和血管形成的实验研究

Vascular endothelial growth factor-loaded microspheres promote dental pulp regeneration and vascularization

目的 评价载血管内皮细胞生长因子（vascular endothelial growth factor,VEGF）微球促进牙髓再生和血管形成的作用,以期为应用组织工程技术探索牙髓治疗新方法奠定基础.方法 应用前期制备的载VEGF微球进行体外释放实验和人脐静脉内皮细胞（human umbilical vein endothelial cell,HUVEC）迁移实验明确微球释放VEGF的特点;将牙髓干细胞（dental pulp stem cell,DPSC）与载VEGF微球共培养,观察细胞与微球的相容性;DPSC与载VEGF微球经根尖孔注入根管腔,植入裸鼠背部皮下,9周后进行组织学和免疫组织化学观察.以DPSC、载VEGF微球单独注入根管,空白根管作为空白对照组.结果 DPSC可在载VEGF微球上生长和增殖;DPSC和载VEGF微球体内形成牙髓样组织,长约9.0 mm,充满根管的根尖1/3和根中1/3,可达冠1/3,伴血管分布和成牙本质细胞样细胞分化.结论 载VEGF微球和DPSC可在体内促进牙髓再生和血管形成.

 Objective To evaluate the effect of vascular endothelial growth factor（VEGF）-loaded microspheres on dental pulp tissue regeneration and vascularization in vivo. Methods In vitro release experiment and human umbilical vein endothelial cell migration experiment were conducted with VEGF loaded microspheres. The dental pulp stem cells （DPSC） were co-cultured with VEGF microspheres to observe the compatibility between the cells and the microspheres. DPSC and VEGF loaded microspheres were injected into the root lumen through the apical foramen,which were then transplanted subcutaneously into nude mice. Histological and immunohistochemical features were observed after nine weeks. Results DPSCs attached and spread on the surface of the microspheres. HE staining showed that the regenerated pulp-like tissue fulfilled the whole apex and middle third of the root. Differentiated odontoblast-like cells aligned with the existing tubular root dentin. Conclusions VEGF-loaded microspheres promoted the regeneration of pulp-like tissues and formation of blood vessels.