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아저해농도(亞沮害濃度)의 항균물질이 mutans streptococci의 세포표면성질과 독력인자에 미치는 영향

Effects of subinhibitory concentrations of antimicrobial agents on cell surface properties and virulence factors of mutans streptococci

이상훈|김종철|장기택|한세현|김영재
대한소아치과학회 (The Korean Academy Of Pediatric Dentistry)|2004.12|대한소아치과학회지 (Journal of the Korean academy of pediatric dentistry)|제31권 제4호|pp. 605-616

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참고문헌

키워드

Mutans streptococci아저해농도세포표면성질독력인자Subinhibitory concentrationCell surface propertiesVirulence factors
주제분류치의학

초록

아저해농도(亞沮害濃度)란 세포의 성장을 저해하는 최소 저해 농도(minimum inhibitory concentrations; MIC)이하의 농도를 말하는데, 적용된 화학물질의 동력학에 따라 항균물질은 반드시 이러한 농도로 존재하는 시기를 거치게 되며, 아저해농도의 항균물질은 세포의 성장을 저해하지는 않지만 세균의 대사와 성장에 스트레스로 작용하게 되어 세포의 성질과 독력인자 발현에 영향을 줄 수 있을 것이다. 본 연구는 여러 항균물질의 최소저해농도를 결정하고 아저해농도의 항균물질이 존재 시 세포 성질과 독력 인자에 미치는 영향을 알아보고, 치아우식증의 예방에 대한 화학적 접근법의 효용성을 조사하기 위해 계획되었다. 연구대상 세균으로는 mutans그룹 streptococci중 두 가지 대표적인 세균종인 S. mutans (S. mutans Ingbritt) 와 S. sobrinus (S. sobrinus 6715-7) 를 사용하였고 화학물질 8종 (Sanguinaria extract; SG, Chlorhexidine digluconate; CHX, Fluoride; F, Propolis; PP, Hydrogen peroxide; HP, Triclosan; TC, Sodium dodecyl sulfate; SDS, Cetylpyridinium chloride; CC) 을 단계 희석하여 배지에 첨가하여 최소저해농도를 구하였으며 아저해농도의 항균물질을 배지에 첨가하여 5% CO2 존재 하에 37℃에서 배양하여 이러한 항균물질을 첨가하지 않은 배지에서 배양한 세균과 성장속도, 산생성능, 체표소수성, 수산화인회석에의 부착능, glucan생성능, 세균응집능 등을 비교 관찰하였다. 아저해농도의 항균물질은 세포의 성장과 체표소수성, 타액으로 처리한 수산화인회석에 대한 세균의 부착과 glucan합성에 영향을 주는 것이 관찰되었다. 또한 12시간 배양 후 S. mutans와 S. sobrinus 모두에서 대조군에 비해 통계적으로 유의하게 최종 pH가 높게 나타나 산생성능이 억제되는 것이 관찰되었다 (p〈0.05). 세균을 proteinase K로 전처리한 경우 처리하지 않았을 때 보다 세균응집역가가 증가하거나 응집이 관찰되지 않았다. 본 연구의 결과에 따르면, 각각의 항균물질은 아저해농도에서도 알려진 작용기전과 유사하게 세균의 성질에 영향을 주었고 이러한 농도에서도 지속적으로 세균을 억제할 수 있었다. 따라서 이러한 항균물질의 사용은 치아우식증의 효과적 예방법의 하나가 될 수 있다.

Abstract

Subinhibitory concentrations (sub-MICs) refer to concentrations below minimum inhibitory concentrations (MICs). The antimicrobial agents may be present at relatively high concentration, at least higher than bacterial MIC and thereafter be desorbed off a surface and function at sub-MICs, perhaps by interfering with bacterial metabolism. Consequently, the aim of this study was to determine the effects of growth, in the presence of sub-MICs of antimicrobial agents, on the cell surface properties and virulence factors of mutans streptococci and to investigate the efficacy of a chemical approach in vitro. Streptococcus mutans Ingbritt and Streptococcus sobrinus 6715-7 were used. Eight antimicrobial agents (Sanguinaria extract;SG, Chlorhexidine digluconate;CHX, Fluoride;F, Propolis;PP, Hydrogen peroxide;HP, Triclosan;TC, Sodium dodecyl sulfate;SDS, Cetylpyridinium chloride; CC) were diluted serially in broth to determine MICs and to compare the growth rate, acid production, hydrophobicity, adhesion activity to saliva coated hydroxyapatite, glucan synthesis and cellular aggregation of experiment groups (in the presence of sub-MICs) with those of control (in the absence of antimicrobial agents). Sub-MICs of antimicrobial agents affected the growth of cells, hydrophobicity, and adhesion of bacteria to saliva coated hydroxyapatite and glucan synthesis. They also resulted in a significant reduction in pH after 12 hours (p〈0.05). By cells pretreated with proteinase K, either the aggregation induced by antimicrobial agents was completely inhibited or the aggregation titers were markedly increased. According to the results of the present study, each antimicrobial agent at sub-MICs could affect similar as its known action mechanism and could continually inhibit cariogenic bacteria at such concentrations. Thus, the use of these antimicrobial agents would be one of the effective methods to prevent dental caries.

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