Xuechun Du1,2, Baosheng Li1, Qing Cai1, Shuwei Qiao1,2, Zixuan Wang1,2, Zhen Li1,2, Yuyang Li1,3, Weiyan Meng4. 1. Department of Dental Implantology, Hospital of Stomatology, Jilin University, 130021, Changchun, Jilin, China. 2. Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Changchun, Jilin, China. 3. Department of Cariology and Endodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, Jiangsu, China. 4. Department of Dental Implantology, Hospital of Stomatology, Jilin University, 130021, Changchun, Jilin, China. mengwy@jlu.edu.cn.
Abstract
BACKGROUND: Peri-implantitis is the main cause of dental implant failure, which is associated with pyroptosis. The roles of D-aspartic acid (D-Asp) on pyroptosis and the mechanism of the protective effect of D-Asp on human gingival fibroblasts (HGFs) remain unknown. This study investigated the effects of D-Asp on the pyroptosis of HGFs induced by high mobility group box 1 protein (HMGB1). METHODS: The cytotoxic effects of D-Asp on HGFs was detected by Cell Counting Kit-8 assay, the membrane permeability was investigated by propidium iodide/ Hoechst 33,342 double staining, flow cytometry analysis, and lactate dehydrogenase releasing, The gene and protein expression levels were detected by real-time quantitative PCR, enzyme-linked immunosorbent assay, and Western blot, respectively. RESULTS: Cell viability analysis showed that D-Asp ≤ 30 mM had no cytotoxicity to HGFs. HMGB1 drastically raised the membrane permeability of HGFs, while 1/10/30 mM D-Asp suppressed the permeability and remained the integrity of the membrane. HMGB1 promoted the mRNA expression of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18, and the protein expression of IL-1β, IL-18, caspase-1, GSDMD, and NLRP3. CONCLUSIONS: With the pretreatment of HGFs with D-Asp of 1/10/30 mM for 24 h, the cell membrane permeability was reduced and the expression of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18 was significantly decreased compared with the HMGB1 group, indicating the competitive antagonism of D-Asp against HMGB1 on the binding with toll-like receptors. Hence, this study may provide a novel insight into preventing pyroptosis and propose a new strategy for the treatment of peri-implantitis.
BACKGROUND: Peri-implantitis is the main cause of dental implant failure, which is associated with pyroptosis. The roles of D-aspartic acid (D-Asp) on pyroptosis and the mechanism of the protective effect of D-Asp on human gingival fibroblasts (HGFs) remain unknown. This study investigated the effects of D-Asp on the pyroptosis of HGFs induced by high mobility group box 1 protein (HMGB1). METHODS: The cytotoxic effects of D-Asp on HGFs was detected by Cell Counting Kit-8 assay, the membrane permeability was investigated by propidium iodide/ Hoechst 33,342 double staining, flow cytometry analysis, and lactate dehydrogenase releasing, The gene and protein expression levels were detected by real-time quantitative PCR, enzyme-linked immunosorbent assay, and Western blot, respectively. RESULTS: Cell viability analysis showed that D-Asp ≤ 30 mM had no cytotoxicity to HGFs. HMGB1 drastically raised the membrane permeability of HGFs, while 1/10/30 mM D-Asp suppressed the permeability and remained the integrity of the membrane. HMGB1 promoted the mRNA expression of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18, and the protein expression of IL-1β, IL-18, caspase-1, GSDMD, and NLRP3. CONCLUSIONS: With the pretreatment of HGFs with D-Asp of 1/10/30 mM for 24 h, the cell membrane permeability was reduced and the expression of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18 was significantly decreased compared with the HMGB1 group, indicating the competitive antagonism of D-Asp against HMGB1 on the binding with toll-like receptors. Hence, this study may provide a novel insight into preventing pyroptosis and propose a new strategy for the treatment of peri-implantitis.
Authors: Panos N Papapanou; Mariano Sanz; Nurcan Buduneli; Thomas Dietrich; Magda Feres; Daniel H Fine; Thomas F Flemmig; Raul Garcia; William V Giannobile; Filippo Graziani; Henry Greenwell; David Herrera; Richard T Kao; Moritz Kebschull; Denis F Kinane; Keith L Kirkwood; Thomas Kocher; Kenneth S Kornman; Purnima S Kumar; Bruno G Loos; Eli Machtei; Huanxin Meng; Andrea Mombelli; Ian Needleman; Steven Offenbacher; Gregory J Seymour; Ricardo Teles; Maurizio S Tonetti Journal: J Periodontol Date: 2018-06 Impact factor: 6.993