Department of Operative Dentistry and Endodontics, School of Stomatology, Wuhan University,Wuhan, China.
J Endod. 2010 Dec;36(12):1978-82. doi: 10.1016/j.joen.2010.08.038. Epub 2010 Oct 8.
The purpose of this study was to assess the effects of iRoot SP root canal sealer (Innovative BioCreamix Inc, Vancouver, Canada) on the expression of mineralization-related genes in human MG63 osteoblast-like cells.
Specimens (5 mm in diameter and 2 mm in height) of iRoot SP and AH Plus (Dentsply DeTrey, Konstanz, Germany) were extracted from a 5-mL culture medium. The MG63 cells were exposed to various dilutions (1/1, 1/2, and 1/4) of the extracts. The 3,(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide MTT assay was used for assessing the dental materials' nonspecific cytotoxicity. The expression of mineralization-related genes, including collagen type I (COL I), osteocalcin (OCN), bone sialoprotein (BSP) and osteopontin (OPN), was detected on days 1, 3, and 6 by a real-time polymerase chain reaction. An enzyme-linked immunosorbent assay experiment was used for evaluating COL I and BSP protein changes. The data were analyzed with one-way analysis of variance and Tukey tests.
In the MTT assay, the undiluted extracts of iRoot SP were noncytotoxic, whereas the undiluted extracts of AH Plus were rated as slightly cytotoxic. iRoot SP up-regulated COL I, OCN, and BSP messenger RNA expression after 3 and 6 days. In the enzyme-linked immunosorbent assay experiment, iRoot SP increased COL I and BSP protein levels compared with AH Plus and the control group on day 6.
In the presence of iRoot SP, MG63 cells can produce more mineralized matrix gene and protein expression. Based on these results, iRoot SP can be considered as a favorable material for cell-material interaction.
本研究旨在评估 iRoot SP 根管封闭剂(加拿大创新生物制药公司,温哥华)对人 MG63 成骨样细胞矿化相关基因表达的影响。
从 5 毫升培养基中提取 iRoot SP 和 AH Plus(登士柏迪特雷,康斯坦茨,德国)的标本(直径 5 毫米,高 2 毫米)。将 MG63 细胞暴露于不同稀释度(1/1、1/2 和 1/4)的提取物中。3,(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物(MTT)测定法用于评估牙科材料的非特异性细胞毒性。通过实时聚合酶链反应检测矿化相关基因,包括Ⅰ型胶原(COL I)、骨钙素(OCN)、骨涎蛋白(BSP)和骨桥蛋白(OPN)的表达,在第 1、3 和 6 天。酶联免疫吸附试验实验用于评估 COL I 和 BSP 蛋白变化。采用单因素方差分析和 Tukey 检验对数据进行分析。
在 MTT 测定中,iRoot SP 的未稀释提取物无细胞毒性,而 AH Plus 的未稀释提取物被评为轻微细胞毒性。iRoot SP 在第 3 天和第 6 天上调 COL I、OCN 和 BSP 信使 RNA 表达。在酶联免疫吸附试验中,与 AH Plus 和对照组相比,iRoot SP 在第 6 天增加了 COL I 和 BSP 蛋白水平。
在 iRoot SP 的存在下,MG63 细胞可以产生更多的矿化基质基因和蛋白表达。基于这些结果,iRoot SP 可以被认为是一种有利于细胞-材料相互作用的材料。