Yoshino Patrícia, Nishiyama Celso Kenji, Modena Karin Cristina da Silva, Santos Carlos Ferreira, Sipert Carla Renata
Endodontics Division, Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo (HRAC-USP), Bauru, SP, Brazil.
Braz Dent J. 2013;24(2):111-6. doi: 10.1590/0103-6440201302115.
The aim of this study was to compare the in vitro cytotoxicity of white mineral trioxide aggregate (MTA), MTA Fillapex® and Portland cement (PC) on human cultured periodontal ligament fibroblasts. Periodontal ligament fibroblast culture was established and the cells were used for cytotoxic tests after the fourth passage. Cell density was set at 1.25 X10 4 cells/well in 96-well plates. Endodontic material extracts were prepared by placing sealer/cement specimens (5x3mm) in 1mL of culture medium for 72 h. The extracts were then serially two-fold diluted and inserted into the cell-seeded wells for 24, 48 and 72 h. MTT assay was employed for analysis of cell viability. Cell supernatants were tested for nitric oxide using the Griess reagent system. MTA presented cytotoxic effect in undiluted extracts at 24 and 72 h. MTA Fillapex® presented the highest cytotoxic levels with important cell viability reduction for pure extracts and at ½ and ¼ dilutions. In this study, PC did not induce alterations in fibroblast viability. Nitric oxide was detected in extract-treated cell supernatants and also in the extracts only, suggesting presence of nitrite in the soluble content of the tested materials. In the present study, MTA Fillapex displayed the highest cytotoxic effect on periodontal ligament fibroblasts followed by white MTA and PC.
本研究的目的是比较白色三氧化矿物凝聚体（MTA）、MTA Fillapex®和波特兰水泥（PC）对人培养的牙周膜成纤维细胞的体外细胞毒性。建立了牙周膜成纤维细胞培养体系，细胞传代至第四代后用于细胞毒性试验。在96孔板中，细胞密度设定为1.25×10⁴个细胞/孔。通过将封闭剂/水泥标本（5×3mm）置于1mL培养基中72小时来制备牙髓材料提取物。然后将提取物进行连续两倍稀释，并加入接种有细胞的孔中培养24、48和72小时。采用MTT法分析细胞活力。使用格里斯试剂系统检测细胞上清液中的一氧化氮。MTA在未稀释提取物作用24和72小时时呈现细胞毒性作用。MTA Fillapex®呈现出最高的细胞毒性水平，纯提取物以及1/2和1/4稀释度时细胞活力显著降低。在本研究中，PC未诱导成纤维细胞活力发生改变。在提取物处理的细胞上清液以及仅提取物中均检测到一氧化氮，表明受试材料的可溶成分中存在亚硝酸盐。在本研究中，MTA Fillapex对牙周膜成纤维细胞的细胞毒性作用最高，其次是白色MTA和PC。
