D-methionine eradicates Porphyromonas gingivalis biofilms by suppressing the cyclic di-GMP level

Abstract

Objective To investigate the mechanisms by which D-methionine (D-Met) eradicates Porphyromonas gingivalis biofilms by suppressing cyclic dimeric GMP (c-di-GMP) levels. Methods Cell viability, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured to determine the effective concentrations of D-Met, which were subsequently used in the following experiments. During the P. gingivalis biofilm formation inhibition experiment and the mature biofilm disassembly experiment, biofilm biomass, exopolysaccharide (EPS), biofilm morphology, integrity of the cell membrane, and the level of c-di-GMP were determined. Results D-Met < 40 mmol/L was biocompatible. During the biofilm formation inhibition and mature biofilm disassembly experiments, D-Met ≥ 20 mmol/L decreased the biofilm biomass and the production of EPS. SEM analysis showed that the extracellular matrix and bacterial density were drastically reduced by D-Met ≥ 20 mmol/L. TEM detection showed that 35 mmol/L D-Met ruptured the cell membrane during biofilm formation and increased the permeability of the cell membrane in the disassembly phase of mature biofilms. C-di-GMP levels decreased with increasing concentrations of D-Met in a concentration-dependent manner. Conclusion D-Met ≥ 20 mmol/L could eradicate P. gingivalis biofilms by suppressing c-di-GMP levels.

Keywords

• biofilm，
• porphyromonas gingivalis，
• d-methionine，
• cyclic diguanosine monophosphate，
• periodontitis，
• peri-implantitis，
• exopolysaccharide，
• cell membrane，
• high performance liquid chromatography，