Failure of endodontic treatment is commonly associated with the presence of Enterococcus faecalis. Studies have highlighted that E. faecalis can form a calcified biofilm in tough environmental conditions, such as within root canals. The aims of this study were to investigate the effects of chemicals used in root-canal disinfection on the adherence of E. faecalis to collagen, as well as to estimate the force of adhesion between E. faecalis and collagen after such treatment. The number of adhering bacteria after chemical treatment was determined using confocal laser scanning microscopy–based adherence assay. It was found that the calcium hydroxide–treated group had a statistically significant (p=0.05) increase in the population of bacteria adhering. The adhesion force between bacteria and collagen of the treatment group with the highest number of bacteria adhering was determined by using optical tweezers (1064 nm) and Equipartitition theorem-based stiffness measurements. The presence of calcium hydroxide was found to significantly increase the bacterium-collagen adhesion force. These experiments highlighted the potential advantage of using optical tweezers to study bacteria-substrate interactions. The findings from the present study suggests that the presence of calcium hydroxide increased the adhesion force and adherence of E. faecalis to type-I collagen.
Failure of endodontic treatment is commonly due to Enterococcal infection. In this study influence of chemical
treatments of type-I collagen membrane by chemical agents commonly used in endodontic treatment on Enterococcus
faecalis cell adherence was evaluated. In order to determine the change in number of adhering bacteria after chemical
treatment, confocal laser scanning microscopy was used. For this, overnight culture of E faecalis in All Culture broth
was applied to chemically treated type-I collagen membrane. It was found that Ca(OH)2 treated groups had statistically
significant (p value=0.05) increase in population of bacteria adherence. The change in adhesion force between bacteria
and collagen was determined by using optical tweezers (1064 nm). For this experiment, Type-I collagen membrane was
soaked for 5 mins in a media that contained 50% all culture media and 50% saturated Ca(OH)2 . The membrane was
spread on the coverslip, on which diluted bacterial suspension was added. The force of laser tweezers on the bacteria was
estimated at different trap power levels using viscous drag method and trapping stiffness was calculated using
Equipartition theorem method. Presence of Ca(OH)2 was found to increase the cell-substrate adherence force from
0.38pN to >2.1pN. Together, these experiments show that it was highly probable that the increase in adherence to
collagen was due to a stronger adhesion in the presence of Ca (OH)2.
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