Streptococcus mutans biofilm에 대한 광역동 치료의 최적조건에 관한 연구

최서정,박호원,이주현,서현우,이시영 大韓小兒齒科學會 2015 大韓小兒齒科學會誌 Vol.42 No.2

할로겐 광중합기를 광원으로, 치면세균막 착색제인 erythrosine을 광감각제로 사용하여 S. mutans biofilm에 대한 광역동 치료를 시행할 때의 최적조건을 알아보기 위해서 erythrosine의 농도와 광조사 시간 및 광감각제의 접촉시간에 따른 광역동 치료의 효과를 비교하였다. erythrosine의 농도를 0, 10, 20, 40, 80 μM로 변화시킨 결과 농도가 증가할수록 항균효과가 증가하는 경향을 보였으며 40 μM과 80 μM의 두 군에서 통계적 유의성이 나타났다. 광 조사 시간을 0, 5, 15, 30, 60, 75초로 변화시킨 결과 조사 시간이 길어질수록 항균효과가 증가하는 경향을 보였으며 30초, 60초, 75초의 세 군에서 통계적유의성이 나타났다. erythrosine 처리시간을 0분, 1분, 2분30초, 5분으로 변화시킨 결과 erythrosine과의 접촉시간이 늘어날수록 항균효과가 증가하는 경향을 보였으며 2분30초 군과 5분 군에서 통계적 유의성을 보였다. 이상의 결과로 광감각제로사용한 erythrosine의 농도를 20-40 μM , 광원으로 사용한 할로겐 광중합기의 광 조사시간을 30초 이상, 광감각제인erythrosine을 2분30초 이상 처리하여 사용하는 것이 치아우식 예방을 위한 광역동 치료에 효과적임을 알 수 있다. The purpose of this study was to evaluate the effects of Photochemotherapy using a combination of erythrosine and standard halogen dental curing lights on the viability of Streptococcus mutans in the biofilm phase. To investigate the optimum treatment parameters, the researchers controlled the concentration of erythrosine, light irradiation time and the treatment time of erythrosine. The higher concentration of erythrosine (0, 10, 20, 40, 80 M) in the presence of light irradiation created greater effects in reducing the viability of S. mutans. The results showed a statistically significant difference among the antimicrobial effects in 20, 40, 80 M erythrosine. The higher irradiation time of light (0, 5, 15, 30, 60, 75s) in the presence of erythrosine showed greater effects in reducing the viability of S. mutans. There was statistically significant difference in 30, 60, 75 seconds. The higher treatment time of erythrosine (0, 1, 2.5, 5min) in the presence of erythrosine created greater effects on reduction of S. mutans viability. Statistically significant differences were found between 2.5 and 5 minutes of erythrosine treatment time. The results of this study showed that the photochemotherapy on S. mutans using erythrosine and the halogen dental curing lights conventionally used in dental clinics is effective in the condition of 20-40 M erythrosine concentration, irradiation time over 30 seconds, and erythrosine treatment time over 2.5 minutes.

활성탄 흡착에 의한 Erythrosine의 제거 특성

이종집(Jong Jib Lee),윤성욱(Sung Wook Yoon) 大韓環境工學會 2009 대한환경공학회지 Vol.31 No.7

Erythrosine은 식용색소와 염료로 사용되지만 독성 때문에 미국에서는 거의 사용되지 않는 물질이다. 본 연구에서는 입상활성탄에 의한 erythrosine의 흡착특성을 회분식 및 흡착칼럼실험을 통하여 조사하였다. erythrosine에 대한 활성탄의 흡착능은 pH조절에 의해 크게 개선되었으며 pH 11에서 초기농도 1,000 mg/L의 98%를 제거할 수 있었다. 회분식흡착실험을 통해 흡착등온선을 구한 결과 erythrosine의 평형흡착관계는 10~1,000 mg/L의 농도범위에서 Freundlich식이 잘 적용되었다. 고정층 실험을 통하여 얻은 파과곡선으로부터 초기유입농도, 유입속도, 충전층의 높이가 흡착탑 운전에 중요한 설계변수가 될 수 있음을 알았다. Erythrosine is used a food coloring, ink and dye, etc. but erithrosine is rarely used in United States due to its known hazards. The adsorption characteristics of erythrosine by granular activated carbon were investigated in the batch adsorber and the packed column. The adsorptivity of activated carbon for erythrosine were largely improved by pH control. When the pH was 11 in the sample, the erythrosine could be removed 98% of initial concentration. It was estabilished that the adsorption equilibrium of erythrosine on granular activated carbon was successfully fitted by Freundlich isotherm equation in the concentration range from 10mg/L to 1,000mg/L. The characteristics of breakthrough curve of activated carbon packed column depend on the design variables such as initial concentration, bed height, and flow rate.

Erythrosine의 농도와 LED 조사 시간에 따른Enterococcus faecalis에 대한 세균치사 효과

이시영 ( Si Young Lee ),이민선 ( Min Sun Lee ),마득상 ( Deuk Sang Ma ) 대한예방치과·구강보건학회 2014 大韓口腔保健學會誌 Vol.38 No.4

Objectives: The purpose of this study was to provide photodynamic bactericidal effect against Enterococcusfaecalis by erythrosine concentrations and LED irradiation times. Methods: Erythrosine was used as a photosensitizer and green LED (3 Watt, 520-530 nm) was used aslight source. E. faecalis ATCC 1943 and E. faecalis ATCC 29212 were used in this study. Approximately105 CFU of bacteria were added in wells of a 96-well microtitration plate. For examining the effects ofconcentrations of erythrosine, 0, 0.625, 1.25, 2.5, 5, and 10 M of erythrosine were added in wellscontaining bacteria. The irradiation time with LED was 30 sec. In another set of experiment, the effect ofirradiation time for killing of bacteria was investigated by increasing irradiation time from 0 to 30 s with10 M of erythrosine final concentration. After irradiation, each sample was serially diluted with PBS and50 l of diluents was spread on duplicate blood agar plates. The plates were incubated for 72 h at 37oCunder aerobic conditions and the number of CFU was determined. The experiments were repeated fourtimes. The results were analyzed using one-way ANOVA, and Tukey’s multiple comparison at a significance evel of 0.05. Results: When the erythrosine concentrations were more than 2.5 M, E. faecalis ATCC 29212 wassignificantly decreased (P<0.05). The more erythrosine concentrations increased, the more E. faecalisATCC 1943 decreased statistically significantly (P<0.05). In another set of experiment, when LED irradiationtime was more than 20 s, E. faecalis ATCC 1943 decreased significantly (P<0.05), and if theirradiation 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.

The Effect of Erythrosine-mediated Photodynamic Therapy on Intraorally Formed Biofilm on Titanium Surface

Se Hwan Park,Si Young Lee,Beom-Seok Chang,Heung-Sik Um,Jae-Kwan Lee KOREAN ACADAMY OF ORAL BIOLOGY 2012 International Journal of Oral Biology Vol.37 No.3

The purpose of this study was to assess the efficacy of photodynamic therapy (PDT) using erythrosine and a halogen light source to treat a biofilm formed on a machined surface titanium disk in vivo. Ten volunteers carried an acrylic appliance containing six machined surface titanium disks on the upper jaw over a period of five days. After the five days of biofilm formation period, the disks were removed. PDT using 20 μM erythrosine and halogen light was then applied to the biofilms formed on the disks. Experimental samples were divided into a negative control group (no erythrosine and no irradiation), E0 group (erythrosine 60s + no irradiation), E30 group (erythrosine 60s + halogen light 30s), and E60 group (erythrosine 60s + halogen light 60s). Following PDT, the bacteria in the biofilm were found to be detached from each disk. Each suspension with detached bacteria were diluted and cultivated on a blood-agar plate for five days under anaerobic conditions. The cultivated bacterial counts in the E60 group were significantly lower than the control group (86.4%) or E0 group (76.7%). In the experimental groups also, the light exposure time and bacterial counts showed a negative correlation. In conclusion, PDT using erythrosine and halogen light has bactericidal effects on biofilms formed on a titanium disk in vivo. Notably, applying 20 μM erythrosine and 60 seconds of halogen light irradiation had a significantly potent effect.

활성탄에 의한 에리스로신 염료수용액의 흡착특성에 관한 연구

이종집 ( Jong Jib Lee ) 한국공업화학회 2011 공업화학 Vol.22 No.2

입상활성탄에 대한 erythrosine의 흡착특성을 초기농도, 접촉시간 및 흡착온도를 변수로 하여 회분식실험을 통하여 조사하였다. Erythrosine에 대한 흡착동력학적 연구는 298 K에서 초기농도가 100, 250, 500 mg/L인 에리스로신 수용액에 대해 수행하였다. 흡착공정은 유사이차속도식에 잘 맞았으며 유사이차속도상수(k2)는 에리스로신의 초기농도가 높을수록 감소하였다. 에리스로신의 평형흡착관계는 298~318 K의 온도범위에서 Freundlich 등온식이 잘 적용되었다. 흡착 자유에너지변화(△G˚), 엔탈피변화(△H˚), 엔트로피변화(△S˚)를 계산하여 본 결과, 표준자유에너지 변화량이 -3.72~-9.62 kJ/mol로 자발적인 공정임을 알았다. 엔탈피변화량이 양의 값을 나타내어 활성탄에 대한 에리스로신 염료의 흡착이 흡열반응임을 알 수 있었다. Adsorption characteristics of erythrosine dye onto the activated carbon has been investigated in a batch system with respect to initial concentration, contact time and temperature. Kinetic studies of the adsorption of erythrosine were carried out at 298 K, using aqueous solutions with 100, 250 and 500 mg/L concentration of erythrosine. The adsorption process followed a pseuo second order model, and the adsorption rate constant (k2) decreased with increasing the initial concentration of erythrosine. The equilibrium process can be well discribed by Freundlich isotherm in the temperature range from 298 to 318K. Free energy of adsorption (△G˚), enthalpy (△H˚), and entropy (△S˚) change were calculated to predict the nature the adsorption. The estimated values for △Go were -3.72~-9.62 kJ/mol over the activated carbon at 250 mg/L, indicated toward a spontaneous process. The positve value for △H˚ indicates that the adsorption of erythrosine dye on activated carbon is an endothermic process.

Streptococcus mutans와 Streptococcus sobrinus에 대한 erythrosine의 LED 광역학치료 효과

이시영 ( Si Young Lee ),장범석 ( Beom Seok Chang ),엄홍식 ( Heung Sik Um ),마득상 ( Deuk Sang Ma ) 대한예방치과·구강보건학회 2012 大韓口腔保健學會誌 Vol.36 No.1

연구목적: 내성의 가능성이 낮은 광역학치료는 기존의 물리적 치료나 약물치료를 대신하거나 보조적 수단으로 활용할 수 있다. Erythrosine은 치과에서 흔히 사용하는 제제로 광역학치료에 있어 광감작제로 사용가능성이 이미 보고되었으나, 본 연구에서는 새로 개발된 4가지 파장의 LED 광원을 이용하여, 우식원인균을 대상으로 단축된 조사시간에서 가장 적합한 파장의 광원을 찾아내고 그 효과를 비교연구하고자 하였다. 연구방법: 광감작제는 erythrosine을 사용하였고, 광원으로는 청색(465 nm), 녹색(525 nm), 황색(590 nm), 적색(630 nm) 3 W power LED를 사용하였다. 연구대상 우식원인균은 Streptococcus mutans와 Streptococcus sobrinus을 사용하였다. 96 well plate에 세균 5×105 CFU/mL을 배양하고(350 μL/well), 5 μMerythrosine을 넣고 각 파장의 광원으로 직경 6 mm로 10 mm 거리에서 10초간 조사하였다. 조사후 세균은 PBS로 희석하여 blood agar plate에 도말하고 37oC에서 72시간 배양한 후 CFU 단위로 측정하였다. 효과는 광감각제와 광원을 사용하지 않은 대조군에 대한 상대적인 생존율로 비교검토하였다. 연구결과: S. mutans에 대한 생존률은 광원의 종류에 따라 통계적으로 유의한 차이가 없었으나(P>0.05), 녹색과 청색에서 살균효과가 우수한 경향이 있었다. S. sobrinus에 대한 생존률은 녹색과 청색 광원이 적색과 황색과 비교하여 통계적으로 유의한 차이가 있었다(P<0.05). 결론: 새로 고안한 3 W power LED는 erythrosine을 이용한 광역학치료에서 10초라는 단시간 동안 녹색(525nm)과 청색(465 nm) 파장에서 S. mutans와 S. sobrinus에 대하여 우수한 살균효과를 나타내었다.

Susceptibility of Mutans streptococci in the Planktonic and Biofilm State to Erythrosine

공정은,서현우,이시영,박호원,이주현 대한소아치과학회 2019 大韓小兒齒科學會誌 Vol.46 No.2

The aim of this study was to investigate the susceptibility of Mutans streptococci in both planktonic and biofilm states to erythrosine. S. mutans was cultured in brain-heart infusion (BHI) broth. Erythrosine was diluted in BHI broth and prepared at a concentration range of 0.02 - 10000 μg/L. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured using the microdilution method. After forming biofilms on 96-well plates, the minimum biofilm inhibitory concentration (MBIC) and minimum biofilm eradication concentration (MBEC) were measured. S. mutans was susceptible to erythrosine in both planktonic and biofilm states. MIC and MBC values were both 19.5 μg/L for the planktonic state, while MBIC and MBEC values were 313 μg/L and 2500 μg/L, respectively, for the biofilm state. Erythrosine (19.5 μg/L) exhibited a bactericidal effect on S. mutans (killing 99.9%) in the planktonic state. For biofilms, erythrosine inhibited biofilm growth and eradicated 99.9% of biofilm bacteria at higher concentrations than MIC and MBC. These MBIC and MBEC concentrations are much lower than known noxious doses, and the MIC, MBC, and MBIC values were even lower than clinical concentrations.

새로운 치면세균막 착색제 개발을 위한 식용색소의 생체적합성 연구

이광희 大韓小兒齒科學會 1999 大韓小兒齒科學會誌 Vol.26 No.1

The purpose of study was to develop new dental plaque disclosants which could replace erythrosine. Three food coloring agents(Red No.40, Blue No.1, and Mixed Green), erythrosine and fluorescein were tested for their color difference, antibacterial property, and biocompatibility. Color difference of Red No,40 was greater than that of erythrosine as concentration of solution increased. Color differences of Blue No.1 and Mixed Green were smaller than that of red dyes. Erythrosine showed obvious antibacterrial property, but food coloring agents showed almost no antibacterial property. The taste and sensation of erythrosine was the worst, and the taste of Red No.40 and the sensation of Mixed Green were the most tolerable. Erythrosine stained dental plaque and oral soft tissue most deeply and long, and Blue No.1 was the next in the depth and longevity of stain.

치면착색제를 이용한 광역동 치료의 항균 효과

고준희,박호원,이주현,서현우,이시영 대한소아치과학회 2020 大韓小兒齒科學會誌 Vol.47 No.2

The aim of this study was to evaluate the antimicrobial effect of photodynamic therapy (PDT) using plaque disclosing agent, 10 - 20 mM erythrosine, as a photosensitizer. Multispecies cariogenic biofilms containing Streptococcus mutans , Lactobacillus casei and Candida albicans were formed on hydroxyapatite disc. 20 μM, 10 mM and 20 mM erythrosine were applied as a photosensitizer for 3 minutes, and then light-emitting diode (LED) irradiated for 24 seconds. Colony-forming unit (CFU) were measured and biofilms were observed using confocal laser scanning microscopy (CLSM). CFU were significantly decreased in the PDT groups using 10 - 20 mM erythrosine (10 mM, 20mM) and the results were also confirmed by CLSM. This study confirms the high antimicrobial effect of photodynamic therapy using plaque disclosing agent as a photosensitizer. 이 연구의 목적은 치면착색제인 10 - 20 mM erythrosine을 광감각제로 사용한 광역동 치료의 항균 효과를 평가하는 것이다. Streptococcus mutans, Lactobacillus casei, Candida albicans 를 포함하는 다종 우식원성 세균막을 hydroxyapatite disc에 형성하였다. 광감각제로 20 μM, 10 mM, 20 mM erythrosine을 3분간 적용 후 24초 동안 광조사를 시행하였다. 각 실험군의 Colony-forming unit (CFU)을 측정하고 세균막을 Confocal laser scanning microscopy (CLSM)을 이용하여 관찰하였다. 10 - 20 mM erythrosine을 광감각제로 사용한 광역동 치료 실험군에서 CFU의 현저한 감소가 관찰되었고, 그 결과는 CLSM 관찰에서도 확인되었다. 이번 연구를 통해 치과 임상에서 사용되는 치면착색제를 광감각제로 이용한 광역동 치료의 높은 항균효과를 확인할 수 있었다.

The effect of photodynamic therapy on the viability of Streptococcus mutans isolated from oral cavity

Ji-Sook Jung,Ho-Won Park,Ju-Hyun Lee,Hyun-Woo Seo,Si-Young Lee 大韓小兒齒科學會 2012 大韓小兒齒科學會誌 Vol.39 No.3

Photodynamic therapy (PDT) is a technique that involves the activation of photosensitizer by light in the presence of tissue oxygen, resulting in the production of reactive radicals capable of inducing cell death. The aim of this study was to evaluate the effect of PDT on Streptococcus mutans in planktonic conditions, previously treated with different photosensitive concentrations of erythrosine, using halogen and LED curing unit as a light source. And we compared the effects of PDT on six strains of S. mutans isolated from oral cavity and reference strain. As a result, S. mutans was susceptible to the combination of hand held photopolymerizer (HHP) and erythrosine. The higher concentration of erythrosine in the presence of light irradiation induced greater effects in reduction of viability of S. mutans. Isolated S. mutans showed a significant reduction in bacterial counts of the groups submitted to PDT compared to the control groups. And they appeared to be similar or slightly lower antimicrobial effect compared with reference strain. However, the difference was not significant (p < 0.05). In conclusion, PDT using erythrosine as a photosensitizing agent and HHP as a light source could be an efficient option for diseases caused by S. mutans. 광역동 치료는 광감각제가 빛에 의해 활성화되면서 발생하는 화학 반응을 이용한 것으로, 치료 원리는 광화학 반응으로 자 유 라디칼 및 반응성 산소가 생성되고 이 산물들에 의한 세포 독성으로 항균 효과를 가지게 되는 것이다. 이 연구의 목적은 치과 임상에서 널리 사용되는 광원(할로겐, LED)과 광감각제(erythrosine)를 이용하여, 치아 우식증과 연관된 세균인 Streptococcus mutans에 대한 광역동 치료의 항균 효과를 알아보고, 광감각제의 농도에 따른 광역동 치료의 효과를 평가하기 위함이다. 또한 임상 분리 균주와 표준 균주에 대한 광역동 치료의 효과를 비교해 보았다. 연구 결과, 표준 및 임상 분리 균주 모두 광감각제 처리 후 광조사를 시행한 군에서만 대조군에 비해 S. mutans의 유의한 감소가 나타났다. 또한 광조사를 시행한 군에서 첨가한 광감각제의 농도가 높을수록 S. mutans의 감소가 증가하는 것으로 나타났다. 표준 균주와 비교 시 임상 분리 균주에서는 표준 균주와 비슷하거나 약간 낮은 S. mutans의 감소가 나타났고, 이 는 통계적으로 유의한 차이는 없었다(p < 0.05). 이상의 결과들로 보아 광감각제로 에리스로신의 사용과 광원으로 치과용 광중합기를 사용한 광역동 치료는 S. mutans 연 관 질병에 대한 효과적인 치료 방법이 될 수 있을 것으로 사료된다.