Wang Jing, Fangteng Jiao-Zi, Liu He
Department of Pediatric Dentistry, Peking University Stomatological Hospital. Beijing 100081, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2019 Jun;28(3):251-258.
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).
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.
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).
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.
评估iRoot BP Plus作为盖髓剂对人乳牙脱落干细胞(SHED)和人牙髓干细胞(DPSC)生物学行为的影响。
制备iRoot BP Plus和ProRoot MTA样本盘,并从iRoot BP Plus和ProRoot MTA样本盘中提取提取液。采用CCK-8法在第1、3、5和7天检测iRoot BP Plus和MTA提取物对SHED和DPSC增殖能力的影响。通过Transwell小室和划痕修复实验观察iRoot BP Plus和MTA提取物对SHED和DPSC迁移能力的影响。将SHED和DPSC分别接种在iRoot BP Plus和MTA的样本盘上进行培养。分别在第1、3和5天使用鬼笔环肽和DAPI(4',6-二脒基-2-苯基吲哚)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提取物的成骨诱导培养基和含iRoot BP Plus提取物的成骨诱导培养基中进行矿化诱导。在第7和14天进行碱性磷酸酶(ALP)染色和ALP定量分析,在第21天进行茜素红染色和钙盐沉积半定量分析。使用SPSS 19.0软件包对数据进行统计分析。
iRoot BP Plus和MTA提取物均可促进SHED和DPSC的细胞增殖。在细胞迁移和黏附实验中,iRoot BP Plus和MTA均促进了SHED和DPSC的迁移和黏附,且iRoot BP Plus的作用更显著(P = 0.000)。矿化诱导后,iRoot BP Plus组中SHED和DPSC的ALP活性显著高于MTA组。茜素红染色和钙盐沉积半定量分析表明,iRoot BP Plus和MTA均能促进细胞矿化。此外,iRoot BP Plus促进细胞矿化的能力显著强于MTA(P = 0.000)。
iRoot BP Plus和MTA具有良好的生物相容性和良好的成骨分化能力,可促进SHED和DPSC细胞增殖、黏附、迁移,且iRoot BP Plus在促进SHED和DPSC黏附、迁移及分化分布方面比MTA具有更好的效果,因此iRoot BP Plus和MTA均可作为乳牙和恒牙的盖髓剂。