D-aspartic acid protects against gingival fibroblasts inflammation by suppressing pyroptosis

• Published in: Molecular biology reports Vol. 49; no. 7; pp. 5821 - 5829
• Main Authors: Du, Xuechun, Li, Baosheng, Cai, Qing, Qiao, Shuwei, Wang, Zixuan, Li, Zhen, Li, Yuyang, Meng, Weiyan
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.07.2022; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Aspartic acid; Biomedical and Life Sciences; Caspase-1; Cell membranes; Cell viability; Cytotoxicity; D-aspartic acid; Enzyme-linked immunosorbent assay; Fibroblasts; Flow cytometry; Gene expression; High mobility group proteins; Histology; HMGB1 protein; IL-1β; Interleukin 18; L-Lactate dehydrogenase; Lactic acid; Life Sciences; Membrane permeability; Morphology; Original Article; Permeability; Propidium iodide; Protein expression; Proteins; Pyroptosis; Toll-like receptors; Peri-implantitis; Pyroptosis; HMGB1; D-aspartic acid; Fibroblast
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-022-07335-y

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.