Photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentration and LED irradiation times

Si Young Lee; Min-Sun Lee; Deuk-Sang Ma

doi:10.11149/jkaoh.2014.38.4.227

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Lee, Lee, and Ma: Photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentration and LED irradiation times

Original Article

J Korean Acad Oral Health 2014;38(4):227-231.

Published online: 12 December 2014

DOI: https://doi.org/10.11149/jkaoh.2014.38.4.227

Photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentration and LED irradiation times

Si Young Lee^1,³, Min-Sun Lee^2,³, Deuk-Sang Ma^2,³

¹Departments of Oral Microbiology, College of Dentistry, Gangneung-Wonju National University, Gangneung, Korea

²Departments of Preventive and Public Health Dentistry, College of Dentistry, Gangneung-Wonju National University, Gangneung, Korea

³Research Institute of Oral Science, Gangneung-Wonju National University, Gangneung, Korea

Corresponding Author: Deuk-Sang Ma Department of Preventive and Public Health Dentistry, College of Dentistry, Gangneung-Wonju National University, 7 Jukheon-gil, Gangneung 210-702, Korea Tel: +82-33-640-3194 Fax: +82-33-640-3103 E-mail: mads@gwnu.ac.kr

Received 6 October 2014 Revised 2 December 2014 Accepted 15 December 2014

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objectives

The purpose of this study was to provide photodynamic bactericidal effect against Enterococcus faecalis by erythrosine concentrations and LED irradiation times.

Methods

Erythrosine was used as a photosensitizer and green LED (3 Watt, 520-530 nm) was used as light source. E. faecalis ATCC 1943 and E. faecalis ATCC 29212 were used in this study. Approximately 10⁵ CFU of bacteria were added in wells of a 96-well microtitration plate. For examining the effects of concentrations of erythrosine, 0, 0.625, 1.25, 2.5, 5, and 10 M of erythrosine were added in wells containing bacteria. The irradiation time with LED was 30 sec. In another set of experiment, the effect of irradiation time for killing of bacteria was investigated by increasing irradiation time from 0 to 30 s with 10 M of erythrosine final concentration. After irradiation, each sample was serially diluted with PBS and 50 l of diluents was spread on duplicate blood agar plates. The plates were incubated for 72 h at 37°C under aerobic conditions and the number of CFU was determined. The experiments were repeated four times. The results were analyzed using one-way ANOVA, and Tukey’s multiple comparison at a significance level of 0.05.

Results

When the erythrosine concentrations were more than 2.5 M, E. faecalis ATCC 29212 was significantly decreased (P<0.05). The more erythrosine concentrations increased, the more E. faecalis ATCC 1943 decreased statistically significantly (P<0.05). In another set of experiment, when LED irradiation time was more than 20 s, E. faecalis ATCC 1943 decreased significantly (P<0.05), and if the irradiation times was more than 5 s, E. faecalis ATCC 29212 decreased significantly (P<0.05).

Conclusions

PDT using erythrosine and green LED was found to be an effective method in killing E. faecalis.

Keywords: Enterococcus faecalis, Erythrosine, LED, Photodynamic therapy

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Table 1.

CFU/Well (×10⁴) of E. faecalis by erythrosine concentration after PDT

Contents	LED irradiation time (seconds)	Erythrosine concentration (μxF06D;M)	CFU/Well (×10⁴) (Mean±SD)	P-value
E. faecalis ATCC 1943	30	10	2.39±0.82^a ^a	<0.001
		5	3.82±0.24^a
		2.5	11.70±0.66^b
		1.25	16.51±3.98^c
		0.625	17.30±0.99^c
		0	23.82±1.81^d
E. faecalis ATCC 29212	30	10	0.74±0.25^a	<0.001
		5	3.25±2.22^a
		2.5	7.27±0.66^b
		1.25	9.32±0.66^bc
		0.625	8.81±0.86^bc
		0	10.71±1.18^cd

^a ,b,c,dThe same letter indicates no significant difference by one-way ANOVA and Tukey's multiple comparison at =0.05.

Table 2.

CFU/Well (×10⁴) of E. faecalis by LED irradiation time after PDT

Contents	LED irradiation	CFU/Well (×10⁴)	P-value
Contents	time (seconds)	(Mean±SD)	P-value
E. faecalis	0	24.44±5.91^a	<0.001
ATCC 1943	1	26.56±7.22^a
	5	22.21±2.24^a
	10	21.84±3.30^a
	15	12.20±3.31^b
	20	9.12±1.29^b
	30	4.78±0.53^b
E. faecalis	0	7.26±1.20^a	<0.001
ATCC 29212	1	7.40±1.47^a
	5	6.36±1.15^a
	10	4.04±0.46^b
	15	2.93±0.27^bc
	20	2.54±0.38^bc
	30	0.89±0.81^c

^a ,b,cThe same letter indicates no significant difference by one-way ANOVA and Tukey’s multiple comparison at =0.05.

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