Use of a Cellulase from Trichoderma reesei as an Adjuvant for Enterococcus faecalis Biofilm Disruption in Combination with Antibiotics as an Alternative Treatment in Secondary Endodontic Infection

Abstract

Apical periodontitis is an inflammation leading to the injury and destruction of periradicular tissues. It is a sequence of events that starts from root canal infection, endodontic treatment, caries, or other dental interventions. Enterococcus faecalis is a ubiquitous oral pathogen that is challenging to eradicate because of biofilm formation during tooth infection. This study evaluated a hydrolase (CEL) from the fungus Trichoderma reesei combined with amoxicillin/clavulanic acid as a treatment against a clinical E. faecalis strain. Electron microscopy was used to visualize the structure modification of the extracellular polymeric substances. Biofilms were developed on human dental apices using standardized bioreactors to evaluate the antibiofilm activity of the treatment. Calcein and ethidium homodimer assays were used to evaluate the cytotoxic activity in human fibroblasts. In contrast, the human-derived monocytic cell line (THP-1) was used to evaluate the immunological response of CEL. In addition, the secretion of the pro-inflammatory cytokines IL-6 and TNF-α and the anti-inflammatory cytokine IL-10 were measured by ELISA. The results demonstrated that CEL did not induce the secretion of IL-6 and TNF-α when compared with lipopolysaccharide used as a positive control. Furthermore, the treatment combining CEL with amoxicillin/clavulanic acid showed excellent antibiofilm activity, with a 91.4% reduction in CFU on apical biofilms and a 97.6% reduction in the microcolonies. The results of this study could be used to develop a treatment to help eradicate persistent E. faecalis in apical periodontitis.

Silica nanoparticles containing nano-silver and chlorhexidine to suppress Porphyromonas gingivalis biofilm and modulate multispecies biofilms toward healthy tendency

“…Previous studies reported an acidic microenvironment containing peptides and heme-derivatives facilitates the selective expansion of periodontitis-promoting microorganisms, mainly P. gingivalis [ 43 , 44 ]. nMS-nAg-Chx would destroy the beneficial conditions for periodontitis-promoting bacterial growth for the preferential release in an acid microenvironment [ 31 , 45 , 46 ].…”

Study of CAZymes from >i<Thermothelomyces thermophilus>/i< M77 as antibiofilm agents: an oxidative and hydrolytic approach

“…Additional examples are provided for various biofilms, including B. cereus biofilm 141 , mono-and dual-species biofilms of P. aeruginosa 142 , S. aureus biofilms in a wound model 143,144 , Salmonella variants 145,146 , and Enterococcus faecalis 147 .…”

“…From all the cited works, it is evident that a high concentration (ranging from 1 to 30 mg/mL) of various cocktails of fungal cellulases from different strains, such as T. virididae (marketed as Cellulase R-10) 108,135,139,140,146 , T. reesei (marketed as Celluclast) 137,[147][148][149] , and A. niger [142][143][144] , has been used for biofilm eradication. These mixtures are primarily formulated for the hydrolysis of plant cellulose and typically consist of cellobiohydrolases, endoglucanases, and glucosidases in varying proportions.…”