Efeito do álcool perílico sobre periodontopatógenos e na modulação da resposta inflamatória de macrófagos

Abstract

Purpose: The study demonstrates an antibacterial activity and effect of perillyl alcohol (POH) on the modulation of the inflammatory response of macrophages in vitro. Main methods: The minimum inhibitory concentration (MIC) and minimum bactericidal (MBM) of POH on periodontal pathogens were determined by macrodilution and subculture, respectively. The cytotoxicity of POH on RAW 264.7 macrophages was assessed by Trypan Blue and MTS. The effect on cell proliferation was evaluated by Trypan Blue at periods of 24, 48 and 72 hours. The production of reactive oxygen species (ROS) was analyzed by flow cytometry and the M1/M2 profile by the gene expression of TNF and arginase1 by real-time PCR. Key findings: POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC = 250 μg / mL. Up to 100uM, no cytotoxicity was observed on macrophages. Cell proliferation was inhibited from 48 hours at 100μM (p <0.05) and 250μM (p <0.01). POH increased ROS production at both 10μM and 100μM (p <0.05) in unstimulated cells. PMA-induced production of ROS was not affected by POH, whereas 100 uM significantly reduced LPSinduced ROS. Expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Significance: POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100μM. In addition, POH did not affect ROS production by macrophages and favored the M1-phenotype, as expression of arginase-1 was inhibited in M2-polarized macrophages.