MIRB标记人脱落乳牙牙髓干细胞移植修复大鼠牙槽骨缺损的实验研究

Experimental study on transplantation of MIRB labeled human deciduous dental pulp stem cells to repair periodontal bone defects in rats

目的:探讨人脱落乳牙牙髓干细胞(stem cells from human exfoliated deciduous teeth,SHED)在牙槽骨缺损修复中的作用机制,利用Molday ION Rhodamine B (MIRB)标记示踪SHED在牙槽骨缺损修复中的归转。方法:利用MIRB标记SHED,检测细胞的标记效率及标记后细胞的存活、增殖及成骨分化能力。通过核磁影像、免疫组织化学-荧光共染色双模态示踪及H-E染色,分析MIRB标记的SHED移植到大鼠牙槽骨缺损模型后,在体内的存活、分化及促进损伤的牙槽骨组织愈合情况。采用SPSS 24.0软件包对数据进行统计学分析。结果:MIRB标记SHED不影响其生长及成骨分化的最佳标记浓度为25μg/mL,对SHED的标记效率达到100%。通过核磁成像示踪体内移植MIRB标记的SHED,可在体内存活8周以上,MIRB标记的SHED能够在体内分化成骨细胞且明显促进牙槽骨的缺损修复。结论:MIRB标记的SHED可以进行体内示踪,并观察到标记的SHED促进缺损牙槽骨修复。

PURPOSE:To trace the fate of transplanted stem cells from human exfoliated deciduous teeth(SHED) in the repair of periodontal bone defects,Molday ION rhodamine B(MIRB) was used to label SHED and explore the mechanism of SHED in the repair of periodontal bone defects.METHODS:In vitro cultured SHED were labeled by MIRB.The labeling efficiency,cell survival,proliferation and osteogenic differentiation of MIRB-labelled SHED were detected.The labeled cells were transplanted into the rat model with periodontal bone defect.The survival,differentiation and improvement of host periodontal bone healing of MIRB labeled SHED in vivo were analyzed by immunohistochemistry and fluorescence co-staining,nuclear magnetic imaging dual-mode tracking and H-E staining.The data were statistically analyzed with SPSS 24.0 software package.RESULTS:MIRB labeled SHED did not affect its growth and osteogenic differentiation.The optimal labeling concentration was 25 μg/mL,the labeling efficiency of SHED reached 100%.The transplantation of MIRB labeled SHED in vivo can survive for more than 8 weeks.It was found that MIRB labeled SHED could differentiate into osteoblasts in vivo and significantly promote the repair of alveolar bone defects.CONCLUSIONS:MIRB labeled SHED can be traced in vivo,and the effect of labeled SHED on the repair of defective alveolar bone was observed.