iRoot BP Plus 对脱落乳牙牙髓干细胞和人牙髓干细胞生物学行为的影响

doi:10.19439/j.sjos.2019.03.006

摘要/Abstract

摘要： 目的评估iRoot BP Plus和ProRoot MTA作为盖髓剂对脱落乳牙牙髓干细胞（stem cells from human exfoliated deciduous teeth,SHED）和人牙髓干细胞（human dental pulp stem cells,DPSC）生物学行为的影响。方法选择处于生理性牙根吸收期的乳牙和无龋的正畸减数前磨牙或第三磨牙,体外提取、分离和培养SHED和DPSC,制备iRoot BP Plus和ProRoot MTA标准膜片并提取浸提液,通过CCK-8法检测1、3、5、7 d时iRoot BP Plus和MTA浸提液对SHED和DPSC增殖能力的影响;Transwell小室和划痕修复实验观察iRoot BP Plus和MTA浸提液对SHED和DPSC迁移能力的影响;SHED和DPSC分别接种于iRoot BP Plus和MTA样品表面进行培养,分别在1、3、5 d使用鬼笔环肽(phalloidin)和DAPI（4',6-diamidino-2-phenylindole）进行免疫荧光染色,观察细胞骨架变化;SHED和DPSC分别在成骨诱导培养基、含MTA浸提液的成骨诱导培养基和含iRoot BP Plus浸提液的成骨诱导培养基中进行矿化诱导,7 d和14 d时进行碱性磷酸酶（alkaline phosphatase,ALP）染色以及ALP定量分析,21 d时进行茜素红染色和半定量分析钙盐沉积量,采用SPSS 19.0软件包对数据进行统计学分析。结果 iRoot BP Plus和MTA均能促进SHED和DPSC的细胞增殖;细胞迁移与黏附实验中,iRoot BP Plus和MTA均能促进SHED和DPSC的迁移与黏附,其中,iRoot BP Plus的作用更显著（P=0.000）;经矿化诱导后,iRoot BP Plus组的SHED和DPSC的ALP活性强于MTA组,茜素红染色和半定量分析钙盐沉积量结果显示,iRoot BP Plus和MTA均能促进细胞矿化,且iRoot BP Plus促进细胞矿化的能力显著强于MTA（P=0.000）。结论 iRoot BP Plus和MTA均具有较好的生物相容性,均能促进SHED和DPSC的细胞增殖,能够促进细胞黏附、迁移,具有较好的成骨分化能力;且iRoot BP Plus促进SHED和DPSC黏附、迁移和成骨分化的能力相对MTA更好,iRoot BP Plus和MTA均可作为乳牙和年轻恒牙牙髓治疗的盖髓剂。

关键词: SHED, DPSC, iRoot BP Plus, MTA, 增殖, 迁移, 黏附, 成骨诱导

Abstract: PURPOSE: To evaluate the effect of iRoot BP Plus as pulp-capping agents on the biological behaviors of stem cells from human exfoliated deciduous teeth (SHED) and human dental pulp stem cells (DPSC). Methods: iRoot BP Plus and ProRoot MTA sample disks were prepared and the extractive solution was extracted from iRoot BP Plus and ProRoot MTA sample disks. The influence of iRoot BP Plus and MTA extracts on the SHED and DPSC proliferation capacity was detected by CCK-8 method at 1, 3, 5 and 7 days.The influence of iRoot BP Plus and MTA extract on the SHED and DPSC migration capacity was observed by Transwell chamber and scratch repair experiments. SHED and DPSC were respectively inoculated on the sample disks of iRoot BP Plus and MTA for culture. Phalloidin and DAPI (4',6-diamidino-2-phenylindole) were used for immunofluorescence staining on 1, 3 and 5 days respectively to observe cytoskeleton changes. SHED and DPSC underwent mineralization induction respectively in osteoblastic induction medium, the osteoblastic induction medium containing MTA extract and the osteoblastic induction medium containing iRoot BP Plus extract. Alkaline phosphatase (ALP) staining and quantitative analysis of ALP were performed at 7 and 14 days, Alizarin Red staining and semi-quantitative analysis of calcium salt deposition were performed at 21 days. SPSS 19.0 software package was used for statistical analysis of the data. Results: iRoot BP Plus and MTA extracts could promote cell proliferation of SHED and DPSC. In cell migration and adhesion experiments, iRoot BP Plus and MTA both promoted migration and adhesion of SHED and DPSC, and iRoot BP Plus played a more significant role (P=0.000). After mineralization induction, the ALP activity of SHED and DPSC in iRoot BP Plus group was significantly greater than that of MTA. Alizarin red staining and semi-quantitative analysis of calcium salt deposition showed that both iRoot BP Plus and MTA could promote cell mineralization. Moreover, the ability of iRoot BP Plus to promote cell mineralization was significantly stronger than that of MTA (P=0.000). Conclusions: iRoot BP Plus and MTA has good biocompatibility and good osteogenetic differentiation ability, it can promote SHED and DPSC cell proliferation, adhesion, migration, and BP Plus has better affect of promoting iRoot SHED and DPSC adhesion, migration and distribution of differentiation than MTA, therefore iRoot BP Plus and MTA may be used as pulp capping agent both for deciduous and permanent teeth.

Key words: SHED, DPSC, iRoot BP Plus, MTA, Proliferation, Migration, Adhesion, Osteoblast induction

中图分类号:

R788.2

王静, 方滕姣子, 刘鹤. iRoot BP Plus 对脱落乳牙牙髓干细胞和人牙髓干细胞生物学行为的影响[J]. 上海口腔医学, 2019, 28(3): 251-258.

WANG Jing, FANGTENG Jiao-zi, LIU He. Effect of iRoot BP Plus on biological behavior of deciduous tooth pulp stem cells and human pulp stem cells[J]. Shanghai Journal of Stomatology, 2019, 28(3): 251-258.

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