Effect of a novel bioceramic root canal sealer on the angiogenesis-enhancing potential of assorted human odontogenic stem cells compared with principal tricalcium silicate-based cements

Abstract Objective: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). Methodology: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers’ instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6–8 h. Statistical analyses included Kruskal–Wallis, Mann–Whitney U, and Friedman and Wilcoxon signed rank tests. Results: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). Conclusion: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.

摘要
目的：本研究评估了硅酸三钙基矿物三氧化物凝聚体（ProRoot MTA）、Biodentine及新型生物陶瓷根管封闭剂（Well-Root ST）对人牙髓干细胞（human dental pulp stem cells, hDPSCs）、人牙周膜干细胞（human periodontal ligament stem cells, hPLSCs）与人牙胚干细胞（human tooth germ stem cells, hTGSCs）的促血管生成潜能。
方法：将ProRoot MTA、Biodentine或Well-Root ST试件（按制造商说明书制备）置于达尔伯克改良伊格尔培养基中孵育24 h制备条件培养基。将细胞接种于上述条件培养基中培养，并于第1、3、7、10及14天采用3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺苯基)-2H-四唑鎓（MTS）法检测细胞活力。于第1、7及14天采用夹心酶联免疫吸附试验（sandwich enzyme-linked immunosorbent assay, ELISA）检测血管生成生长因子[血小板衍生生长因子（platelet-derived growth factor, PDGF）、碱性成纤维细胞生长因子（basic fibroblast growth factor, FGF-2）及血管内皮生长因子（vascular endothelial growth factor, VEGF）]的表达水平。于6~8 h采用人脐静脉内皮细胞（human umbilical vein endothelial cell, HUVEC）迁移实验评估受试材料的血管活性。统计学分析采用Kruskal-Wallis检验、Mann-Whitney U检验、Friedman检验及Wilcoxon符号秩检验。
结果：所有硅酸三钙基材料均无细胞毒性，且均可诱导相似水平的血管生成生长因子（PDGF、FGF-2及VEGF）释放（p>0.05）。第14天时，所有硅酸三钙基材料处理组的hDPSCs活力最高（p<0.05）。HUVEC管腔形成实验显示，所有受试材料均可显著提升管腔形成能力（p<0.05）。
结论：硅酸三钙基材料可对各类干细胞产生促血管生成刺激作用，并可显著增强HUVEC的管腔形成能力。