Abstract
Enterococcus faecalis (E. faecalis), one of the main pathogens responsible for refractory periapical periodontitis and nosocomial infections, exhibits markedly higher pathogenicity in biofilms. Objectives Studies have shown that caseinolytic protease P (ClpP) is involved in biofilm formation. However, to date, few studies have investigated the role of ClpP in the survival of E. faecalis, and in enhancing biofilm formation. Therefore, we investigated the role of ClpP in the formation of E. faecalis biofilms.Methodology In our study, we used homologous recombination to construct clpP deleted and clpP complement strains of E. faecalis ATCC 29212. A viable colony counting method was used to analyze the growth patterns of E. faecalis. Crystal violet staining (CV) and confocal scanning laser microscopy (CLSM) were used to characterize biofilm mass formation and scanning electron microscopy (SEM) was used to observe the biofilm microstructure. Data was statistically analyzed via Student’s t-test or one-way analysis of variance (ANOVA).Results The results exhibited altered growth patterns for the clpP deletion strains and depleted polysaccharide matrix, resulting in reduced biofilm formation capacity compared to the standard strains. Moreover, ClpP was observed to increase biofilm formation in E. faecalis.Conclusion Our study shows that ClpP can increase biofilm formation in E. faecalis and emphasizes the importance of ClpP as a potential target against E. faecalis.
Highlights
Enterococcus faecalis (E. faecalis) is one of the primary etiologic pathogens of refractory periapical periodontitis, secondary root canal infections, and pulp infection
We investigated the role of caseinolytic protease P (ClpP) in the formation of E. faecalis biofilms
Upstream and downstream primers for the clpP gene were used in a PCR-based overlap extension method to obtain the ΔclpP fragment (1096 bp)
Summary
Enterococcus faecalis (E. faecalis) is one of the primary etiologic pathogens of refractory periapical periodontitis, secondary root canal infections, and pulp infection. It is the main pathogen of nosocomial infections, such as bacterial endocarditis and meningitis.. The ability of E. faecalis to form biofilms directly determines its survival, adaptability to the environment and pathogenicity. In the field of Endodontics, the medicaments used to remove bacteria from the root canal system primarily include calcium hydroxide and antibiotics. These agents are associated with poor efficiency and have been shown to cause adverse effects.. Investigating the mechanism associated with E. faecalis biofilm formation is essential to inform the development of effective treatment methods
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