Huang Fu-Mei, Lee Shiuan-Shinn, Yang Shun-Fa, Chang Yu-Chao
Oral Medicine Center, Chung Shan Medical University Hospital, Chung Shan Medical University, Taichung, Taiwan.
J Endod. 2009 Mar;35(3):363-6. doi: 10.1016/j.joen.2008.11.020.
Histologic investigations have demonstrated that root canal sealers can induce mild to severe inflammatory alternations. A recently identified tumor necrosis factor family molecule, receptor activator of nuclear factor-kappaB ligand (RANKL), plays a critical role in the development of osteoclasts that result in bone resorption. The aim of this study was to investigate the effects of root canal sealers AH26, Canals, and N2 on the expression of RANKL in human osteoblast cell line U2OS cells. Freshly mixed materials were filled in glass rings (4 mm height and 10 mm in diameter) and eluted in 10 mL of culture medium for 1 day. Subsequently, various dilutions (final dilution: 1/2, 1/4, and 1/8) of these extraction media were prepared for this study. Cytotoxicity was measured by the propidium iodide fluorescence assay. The expression of RANKL was measured by reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. The results showed that AH26, Canals, and N2 were cytotoxic to U2OS cells in a concentration-dependent manner (P < .05). The exposure of U2OS cells to root canal sealers resulted in the up-regulation of RANKL mRNA and protein expression (P < .05). The expression of RANKL was up-regulated by root canal sealers in the following order: N2 > AH26 > Canals. Taken together, the activation of RANKL might play an important role in the pathogenesis of periapical bone destruction induced by root canal sealers.
组织学研究表明,根管封闭剂可引发轻度至重度的炎症改变。一种最近被鉴定出的肿瘤坏死因子家族分子,核因子-κB受体活化因子配体(RANKL),在导致骨吸收的破骨细胞发育过程中起关键作用。本研究的目的是探讨根管封闭剂AH26、Canals和N2对人成骨细胞系U2OS细胞中RANKL表达的影响。将新鲜混合的材料填充到玻璃环(高4毫米,直径10毫米)中,并在10毫升培养基中洗脱1天。随后,为本研究制备这些提取培养基的各种稀释液(最终稀释度:1/2、1/4和1/8)。通过碘化丙啶荧光测定法测量细胞毒性。通过逆转录聚合酶链反应和酶联免疫吸附测定法测量RANKL的表达。结果表明,AH26、Canals和N2对U2OS细胞具有浓度依赖性细胞毒性(P <.05)。U2OS细胞暴露于根管封闭剂导致RANKL mRNA和蛋白表达上调(P <.05)。根管封闭剂使RANKL表达上调的顺序为:N2>AH26>Canals。综上所述,RANKL的激活可能在根管封闭剂诱导的根尖周骨破坏的发病机制中起重要作用。