论文部分内容阅读
目的:应用组织工程方法,探讨根尖乳头干细胞(SCAP)分别在炎性环境(肿瘤坏死因子-α)及正常环境下向牙髓牙本质复合分化的可能性。方法:分离大鼠根尖乳头组织,采用酶消化法结合组织块法获得SCAP,鉴定细胞来源,进行MTT生长曲线的测定,分正常组、炎症组(10 ng/m L TNF-α)及空白对照组行体外成骨能力茜素红及免疫组化实验,并将3个组分别移植到大鼠背部皮下观察8/12周后新生组织的形成情况。结果:MTT结果显示10 ng/m L TNF-α组显著高于正常SCAP组,差异有统计学意义(P<0.5);茜素红染色显示均有矿化结节生成,与炎性组(10 ng/m L TNF-α)相比,正常组SCAP染色显著,矿化结节形成的范围大且呈深橘色;免疫组化染色显示SCAP胞浆中DSP/BSP均有表达;正常、炎症组移植物可见牙髓牙本质样结构形成,且DSP/BSP均阳性表达,空白对照组未见矿化组织形成。结论:SCAP具有高增殖及成骨向分化能力,可进行牙髓牙本质复合体的再生,炎性因子TNF-α对SCAP的生长增殖有促进作用同时不同程度上对SCAP矿化及成牙成骨向分化有抑制作用。
OBJECTIVE: To explore the possibility of composite differentiation of apical papillary stem cells (SCAP) into dentin in inflammatory environment (tumor necrosis factor-α) and normal environment respectively by using tissue engineering methods. Methods: The apical papillary tissues of rats were isolated, and SCAP was obtained by enzymatic digestion and tissue block method. The cell sources were identified and MTT growth curve was determined. The normal and inflammation groups (10 ng / m L TNF-α) and blank Control group in vitro osteogenic capacity Alizarin red and immunohistochemistry experiments, and the three groups were transplanted into the back of the rat subcutaneously observed 8/12 weeks after the formation of new tissue. Results: The results of MTT showed that TNF-α of 10 ng / m L group was significantly higher than that of normal SCAP group (P <0.5). Alizarin red staining showed mineralized nodules were formed, 10 ng / m L TNF-α), SCAP staining of normal group was significant, the formation of mineralized nodules was large and dark orange; immunohistochemical staining showed that SCAP cytoplasm DSP / BSP were expressed; normal, Inflammatory group showed dental pulp dentin-like structure formation, and DSP / BSP were positive expression, no control group formed mineralized tissue. CONCLUSIONS: SCAP has high proliferative and osteogenic differentiation ability, which can regenerate dental pulp dentin complex. The inflammatory factor TNF-α can promote the growth and proliferation of SCAP. At the same time, SCAP mineralization and tooth formation Bone differentiation to inhibit.