Ali Mohamed R W, Mustafa Manal, Bårdsen Asgeir, Bletsa Athanasia
Faculty of Medicine, Department of Clinical Dentistry, University of Bergen, Bergen, Norway.
Oral Health Centre of Expertise in Western Norway, Bergen, Norway.
Eur J Oral Sci. 2019 Jun;127(3):261-268. doi: 10.1111/eos.12613. Epub 2019 Apr 8.
Tricalcium silicate cements (TSCs) are used in endodontic procedures to promote wound healing and hard tissue formation. The aim of this study was to evaluate and compare the effect of commonly used TSCs [mineral trioxide aggregate (MTA), Biodentine, and TotalFill] on cellular metabolism and osteogenic/angiogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) in vitro. We tested the null hypothesis of no difference between MTA, Biodentine, and TotalFill in stem cell responses. Cells were subjected to eluates of the tested materials for up to 14 d. Cell viability was evaluated using the 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Real-time PCR was used to determine the levels of expression of the osteogenic factors alkaline phosphatase (ALP), osteoprotegerin (OPG), osteocalcin (OC), and collagen 1A (COL1A1), and the angiogenic factors vascular endothelial growth factor A (VEGFA) and fibroblast growth factor 1 (FGF1). ELISAs were used to measure the levels of VEGFA and ALP in culture supernatants. Mineralization in vitro of hBMSCs was assessed using Alizarin Red staining. The hBMSCs tolerated exposure to TSCs well, with Biodentine showing the most favorable effect on cell viability. Expression of ALP, COL1A1, OPG, and VEGFA were differentially affected by the materials, with Biodentine and TotalFill inducing earlier changes at gene level. Increased mineralization was observed with time, after exposure to all TSCs tested, with MTA showing the greatest effect. The results revealed different responses of hBMSCs to TSCs in vitro.
硅酸三钙水门汀(TSCs)用于牙髓治疗程序,以促进伤口愈合和硬组织形成。本研究的目的是评估和比较常用的TSCs[矿物三氧化物凝聚体(MTA)、生物陶瓷和TotalFill]对人骨髓间充质干细胞(hBMSCs)体外细胞代谢和成骨/血管生成分化的影响。我们检验了MTA、生物陶瓷和TotalFill在干细胞反应方面无差异的零假设。将细胞暴露于测试材料的洗脱液中长达14天。使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法评估细胞活力。实时聚合酶链反应用于测定成骨因子碱性磷酸酶(ALP)、骨保护素(OPG)、骨钙素(OC)和胶原蛋白1A(COL1A1)以及血管生成因子血管内皮生长因子A(VEGFA)和成纤维细胞生长因子1(FGF1)的表达水平。酶联免疫吸附测定法用于测量培养上清液中VEGFA和ALP的水平。使用茜素红染色评估hBMSCs的体外矿化情况。hBMSCs对暴露于TSCs耐受性良好,生物陶瓷对细胞活力显示出最有利的影响。材料对ALP、COL1A1、OPG和VEGFA的表达有不同影响,生物陶瓷和TotalFill在基因水平上诱导更早的变化。在暴露于所有测试的TSCs后,随着时间的推移观察到矿化增加,MTA显示出最大的影响。结果揭示了hBMSCs在体外对TSCs的不同反应。