Comparative evaluation of the effects of three hydraulic calcium silicate cements on odontoblastic differentiation of human dental pulp stem cells: an in vitro study.

Journal of applied oral science : revista FOB Pub Date : 2022-11-04 eCollection Date: 2022-01-01 DOI:10.1590/1678-7757-2022-0203
Hadi Assadian, Arash Khojasteh, Zahra Ebrahimian, Fereshteh Ahmadinejad, Helia Sadat Haeri Boroojeni, Mahboubeh Bohlouli, Mohammad Hossein Nekoofar, Paul Mh Dummer, Hanieh Nokhbatolfoghahaei
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引用次数: 1

Abstract

Objective: The study aimed to compare the response of human dental pulp stem cells (hDPSCs) towards three hydraulic calcium silicate cements (HCSCs) by measuring cytotoxicity and expression of dentinogenic genes.

Methodology: Dental pulps of five impacted mandibular third molars were extirpated as a source for hDPSCs. Next to culturing, hDPSCs were subjected to fluorescence-activated cell sorting after the third passage to validate stemness of the cells. Human DPSCs were exposed to diluted supernatants of OrthoMTA (OMTA), Biodentine (BD) and Calcium-Enriched Mixture (CEM) at concentrations 10, 25, 50 and 100% at the first, third and fifth day of culture. Then, cells were exposed to 10% concentrations supernatant of HCSCs to determine DSPP and DMP1 gene expression, using a quantitative polymerase-chain reaction. Data were analyzed using one-way and three-way ANOVA, followed by Tukey post hoc statistical tests.

Results: Optimal cell proliferation was observed in all groups, regardless of concentration and time-point. HCSC supernatants were non-cytotoxic to hDPSCs at all three time-points, except for 100% Biodentine on day five. On day seven, OMTA group significantly upregulated the expression of DSPP and DMP1 genes. On day 14, expression of DMP1 and DSPP genes were significantly higher in BD and OMTA groups, respectively.

Conclusion: Biodentine significantly upregulated DMP1 gene expression over 14 days, whereas CEM was associated with only minimal expression of DSPP and DMP1 .

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三种水合硅酸钙胶合剂对人牙髓干细胞成牙体分化影响的体外比较研究。
目的:比较人牙髓干细胞(hDPSCs)对三种水化硅酸钙水泥(HCSCs)的细胞毒性和牙本质形成基因的表达。方法:取5颗阻生下颌第三磨牙的牙髓作为hdpsc的来源。接着进行培养,在第三代传代后对hdpsc进行荧光激活细胞分选,以验证细胞的干性。在培养的第1、3和5天,将人DPSCs暴露于浓度分别为10、25、50和100%的OrthoMTA (OMTA)、Biodentine (BD)和calcium - fulmixture (CEM)的稀释上清液中。然后,将细胞暴露于10%浓度的HCSCs上清液中,采用定量聚合酶链反应检测DSPP和DMP1基因的表达。数据分析采用单因素和三因素方差分析,随后采用Tukey事后统计检验。结果:无论浓度和时间点,各组细胞增殖均达到最佳。HCSC上清在所有三个时间点对hdpsc均无细胞毒性,第5天100%百妥定除外。第7天,OMTA组显著上调DSPP和DMP1基因的表达。第14天,BD组和OMTA组DMP1和DSPP基因的表达量分别显著升高。结论:Biodentine在14天内显著上调DMP1基因表达,而CEM仅与DSPP和DMP1的少量表达相关。
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