梯度弹性模量凝胶诱导牙髓干细胞向成牙本质细胞分化的研究
A study on the differentiation of dental pulp stem cells into odontoblasts by gel with gradient elastic modulus
苏小营 1李章一 2黄建永 3姚睿 1刘晓智4
作者信息
- 1. 天津市口腔医院儿童口腔科,天津300041
- 2. 天津市第五中心医院口腔科,天津300450
- 3. 北京大学力学与工程科学系,北京100871
- 4. 天津市第五中心医院中心实验室,天津300450
- 折叠
摘要
目的:探讨梯度弹性模量固体凝胶诱导牙髓干细胞(DPSCs)向成牙本质细胞分化的特点.方法:原代分离培养和鉴定小鼠DPSCs;利用低熔点琼脂糖与改良EaSe培养基分别配制成弹性模量＜25、25 ～ 50、50 ～75、75 ～ 100 kPa 4个梯度的固体凝胶,其所对应的琼脂糖浓度分别为4.8、19.2、76.8、307.2 g/L.将第4代DPSCs分别接种于上述凝胶表面,21 d后用茜素红染色检测其矿化结节的形成情况,RT-PCR和Western Blot检测其牙本质基质蛋白-1(DMPl)、涎蛋白(DSP)、磷蛋白(DPP)的mRNA和蛋白表达水平.结果:流式细胞术鉴定结果表明,所培养细胞高表达间充质干细胞标记蛋白CD29和CD90,几乎不表达造血干细胞标记蛋白CD34和CD45;DPSCs诱导分化结果显示,随着弹性模量增高,固体凝胶表面形成的矿化结节随之增多,同时成牙本质细胞标记基因DMPl、DSP、DPP的mRNA和蛋白质表达水平也逐级增加.结论:DPSCs具有顺培养基质表面硬度梯度逐级向成牙本质细胞分化的特点.
Abstract
AIM:To study the differentiation of dental pulp stem cells (DPSCs) into odontoblasts induced by gel with gradient elastic modulus.METHODS:Mouse DPSCs of passage 4 were seeded on the gel surface with 4 gradient elastic modulus respectively:less than 25 kPa,25 ～ 50 kPa,50 ～ 75 kPa and 75 ～ 100 kPa,which were prepared by a mixture of low melting point agarose and modified Eagle medium,and cultoured for 21 d.The mineralized nodules in the culture were detected using alizarin red staining,and the mRNA and protein expression of dentin matrix protein 1 (DMP1),dentin sialoprotein (DSP) and dentin phosphoprotein (DPP) was tested by reverse transcription polymerase chain reaction and western blot method respectively.RESULTS:The cultured cells expressed CD29 and CD90,almost did not express CD34 and CD45.With the increase of elastic modulus of the solid gel,the mineralized nodule in the culture increased,and mRNA and protein expression level of DMP1,DSP and DPP also increased.CONCLUSION:DPSCs may differentiate into odontoblasts when seeded on the gel surface with different elastic modulus in a gradient dependent form.
关键词
牙髓干细胞/牙本质/弹性模量/分化Key words
dental pulp stem cells/dentin/elastic modulus/differentiation引用本文复制引用
基金项目
天津市卫生和计划生育委员会科技基金(2014KY23)
天津滨海新区卫生和计划生育委员会科技基金(2015BWKY003)
出版年
2017
NETL
NSTL
维普
万方数据