RFP标记及bFGF/BMP-2信号诱导的牙组织工程的体内实验研究  

An experimental study of RFP marker and bFGF/BMP-2 signal induced tooth tissue engineering in vivo

作　　者：张悦 董红宾[2] 邬薇薇 韩祥祯 李晶[2] 单健良 何惠宇[2] 

机构地区：[1]新疆医科大学第五附属医院口腔科,新疆乌鲁木齐830011 [2]新疆医科大学第一附属医院口腔修复科,新疆乌鲁木齐830054 [3]乌鲁木齐军区总医院口腔科,新疆乌鲁木齐830054

出　　处：《口腔医学》2018年第2期97-103,共7页Stomatology

基　　金：国家自然科学基金(81660177)

摘　　要：目的 RFP标记的牙胚细胞和b FGF/BMP-2转染的骨髓间充质干细胞混合培养,复合3D打印聚乙烯醇(PVA)/双相陶瓷骨支架(DCCP)材料构建牙组织工程的体内孵育,探究其分化能力。方法取健康SD大鼠128只,随机分为4组:细胞团块组;PVA/DCCP支架材料组;细胞团块+PVA/DCCP支架材料组;空白对照组,不做任何干预,均在全麻下植入大鼠肾被膜下。术后按4个时间点(5、10、14、28 d各8只)分期取材,进行免疫组化染色和激光共聚焦切片观察。结果免疫组化:析因分析显示细胞团块+PVA/DCCP支架材料组在5、10 d DMP-1、BMP-4表达量最高且均高于其余各组,随着时间推移表达量逐渐有减小趋势,差异有统计学意义P<0.05;DSP在各组各时间点均为阴性表达,差异无统计学意义P>0.05。激光共聚焦显微镜显示:细胞团块+PVA/DCCP支架材料组各时间点切片组织中都可观测到荧光标记的牙胚细胞。结论 RFP标记的牙胚细胞和b FGF/BMP-2转染骨髓间充质干细胞混合培养,复合PVA/DCCP支架材料,构建牙组织工程成活并分化,其分化机制有待进一步研究。Objective Mixed culture of RFP markers of tooth germ cells and bFGF/BMP-2 transfecting the bone marrow stem cells （BMSCs） was conducted and polyvinyl alcohol （PVA）/biphasic ceramic bone scaffold （DCCP） stand was painted three- dimensionally to construct heterogeneous tooth tissue engineering （HTYE）. In order to investigate their ability to differentiate in vivo. Methods 128 healthy SD rats were randomly divided into four groups： the cell pellet group; PVA/DCCP scaffold group; cell clumps ＋ PVA/DCCP scaffold group; control group which was without any intervention. The HTYE was implanted into the renal subcapsular of rats under general anesthesia. Four time points after surgery were chosen to collect samples for testing, Immunohistochemical staining and confocal slice observation. At the 5th, 10th, 14th, 28th day, 8 samples were collected from each group. Results Immunohisto- chemistry： Factorial analysis showed that cell clumps ＋ PVA/DCCP scaffold group in 5, 10 days DMP-1, BMP-4 expression were the highest and were higher than the rest groups, the expression level gradually decreased over time, and the difference was statistically sig- nificant P〈0.05; DSP in each group at each time point was negative, the difference was not statistically significant P〉0.05. Confoeal microscopy showed： cell clumps ＋PVA/DCCP scaffolds group at each time point biopsy tissue could be observed in tooth germ cell flu- orescently marker. Conclusion RFP labeled tooth germ ceils and bFGF/BMP-2 transfecting BMSCs mixed culture, composite PVA/ DCCP scaffold successfully construct tooth tissue engineering and tooth differentiation, but its differentiation mechanism needs further study.

关 键 词：红色荧光蛋白 牙胚细胞 骨髓干细胞 PVA/DCCP 牙组织工程 

分 类 号：R318[医药卫生—生物医学工程] R781[医药卫生—基础医学]