Chandra Janaki, Nakamura Shin, Shindo Satoru, Leon Elizabeth, Castellon Maria, Pastore Maria Rita, Heidari Alireza, Witek Lukasz, Coelho Paulo G, Nakatsuka Toshiyuki, Kawai Toshihisa
Department of Oral Science and Translational Research, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL 33328, USA.
Department of Pathophysiology-Periodontal Science, Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8525, Japan.
Biomedicines. 2024 Aug 12;12(8):1835. doi: 10.3390/biomedicines12081835.
Surface pre-reacted glass-ionomer (S-PRG) is a new bioactive filler utilized for the restoration of decayed teeth by its ability to release six bioactive ions that prevent the adhesion of dental plaque to the tooth surface. Since ionic liquids are reported to facilitate transepithelial penetration, we reasoned that S-PRG applied to root caries could impact the osteoclasts (OCs) in the proximal alveolar bone. Therefore, this study aimed to investigate the effect of S-PRG eluate solution on RANKL-induced OC-genesis and mineral dissolution in vitro. Using RAW264.7 cells as OC precursor cells (OPCs), TRAP staining and pit formation assays were conducted to monitor OC-genesis and mineral dissolution, respectively, while OC-genesis-associated gene expression was measured using quantitative real-time PCR (qPCR). Expression of NFATc1, a master regulator of OC differentiation, and the phosphorylation of MAPK signaling molecules were measured using Western blotting. S-PRG eluate dilutions at 1/200 and 1/400 showed no cytotoxicity to RAW264.7 cells but did significantly suppress both OC-genesis and mineral dissolution. The same concentrations of S-PRG eluate downregulated the RANKL-mediated induction of OCSTAMP and CATK mRNAs, as well as the expression of NFATc1 protein and the phosphorylation of ERK, JNK, and p38. These results demonstrate that S-PRG eluate can downregulate RANKL-induced OC-genesis and mineral dissolution, suggesting that its application to root caries might prevent alveolar bone resorption.
表面预反应玻璃离子(S-PRG)是一种新型生物活性填料,因其能够释放六种生物活性离子,防止牙菌斑粘附于牙齿表面,故而被用于龋牙修复。鉴于离子液体据报道可促进经上皮渗透,我们推测应用于根龋的S-PRG可能会影响牙槽骨近端的破骨细胞(OCs)。因此,本研究旨在体外探究S-PRG洗脱液对RANKL诱导的破骨细胞生成和矿物质溶解的影响。使用RAW264.7细胞作为破骨细胞前体细胞(OPCs),分别进行抗酒石酸酸性磷酸酶(TRAP)染色和蚀坑形成试验,以监测破骨细胞生成和矿物质溶解,同时使用定量实时聚合酶链反应(qPCR)测量与破骨细胞生成相关的基因表达。使用蛋白质免疫印迹法测量破骨细胞分化的主要调节因子NFATc1的表达以及丝裂原活化蛋白激酶(MAPK)信号分子的磷酸化。1/200和1/400的S-PRG洗脱液稀释液对RAW264.7细胞无细胞毒性,但确实显著抑制了破骨细胞生成和矿物质溶解。相同浓度的S-PRG洗脱液下调了RANKL介导的破骨细胞刺激跨膜蛋白(OCSTAMP)和组织蛋白酶K(CATK)mRNA的诱导,以及NFATc1蛋白的表达和细胞外信号调节激酶(ERK)、应激活化蛋白激酶(JNK)和p38的磷酸化。这些结果表明,S-PRG洗脱液可下调RANKL诱导的破骨细胞生成和矿物质溶解,提示其应用于根龋可能会预防牙槽骨吸收。