Effect of Four Essential Oils on Cells Release Membrane and Biofilm Formation of Clinical Bacterial Isolated From Oral Infection

Abstract

Background: Bacterial biofilms forming are current resistant bacterial form to the treatment of oral diseases that colonizes in the gingival and sub-gingival regions of the mouth. The present study aims to screen the anti-biofilm potential and evaluate the effect of four essential oils on cells release membrane. Methods: Seven type isolate bacteria obtained during previous work were screen to select those who had ability to form biofilm using Congo Red Agar method, tube method and crystal violet method. The inhibitory parameter of biofilm forming was determine using microtiter plate method. The effect of essential oil on cell membrane release of each selected bacterial was put in evidence by measuring cellular material that absorb at 260 nm and 280 nm after 0 min, 30 min and 60 min of exposure and confirm by measuring DNA, RNA and proteins release by treated cells on extracellular medium using Nanodrop 1000 spectrophotometer. Results: The crystal violet method shows twelve (12) strong, five (05) moderate and five (05) weak biofilm forming bacteria. The anti-biofilm activity against the oral bacteria who shown that most of essentials oils have activity on different biofilm formation and the MICs ranged from 0.31 mg/mL to 1.25 mg/mL. Concentration of intracellular material released in extracellular medium ranged from 186,56 ± 2,35 ng/µL to 766,6 ± 2,84 ng/µL for DNA, 158,06 ± 1,87 ng/µL to 628,53 ± 2,05 ng/µL for RNA and 695,9 ± 2,11ng/µL to 1125,23 ± 2,15 ng/µL for proteins. Conclusion: This study demonstrates that the selected EOs have a significant anti-biofilm activity, acting on the cell surface and causing the disruption of the bacterial membrane. These EOs are interesting alternative to conventional antimicrobials for the control of oral microorganisms.

Keywords: Anti-biofilm activity, Biofilm, oral diseases