D-组氨酸抑制变异链球菌生物膜作用的研究
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摘要
目的:研究D-组氨酸对变异链球菌生物膜的抑制作用。方法:通过生长曲线实验评估D-组氨酸对变异链球菌细菌活性的影响。通过结晶紫染色实验评估变异链球菌形成生物膜的量;采用胞外多糖实验检测D-组氨酸对变异链球菌生物膜胞外多糖含量的影响;经扫描电镜观察经D-组氨酸处理后变异链球菌生物膜的结构。结果:经生长曲线实验评估发现D-组氨酸在浓度≥20%时抑制变异链球菌的生物活性且呈浓度依赖性。通过结晶紫染色实验和生物膜胞外多糖实验评估发现,D-组氨酸浓度≥40%时抑制变异链球菌生物膜的形成并影响生物膜胞外多糖的含量。经扫描电镜观察,40%的D-组氨酸即可影响变异链球菌生物膜的形成,同时细菌的粘附量有所下降。结论:D-组氨酸具有良好的抑制变异链球生物膜的作用,并影响变异链球菌的细菌活性。
Objectives:To evaluate the effects of D-Histidine on the Streptococcus mutans biofilmin vitro.Methods:Antimicrobial activity testing of D-Histidine was evaluated by determining the growth curve.Crystal violet biofilm biomass assay and polysaccharide measurement were performed to evaluate the potential activity of D-Histidine effectively inhibiting biofilm formation.SEM was used to observe the morphology and the structure of biofilm.Results:Growth curve showed that D-Histidine at concentration of 20%interfered with the viability of Streptococcus mutans in a dose-dependent manner.CV and EPS assays showed that D-Histidine at concentration of 40%effectively inhibited Streptococcus mutans biofilm formation and the polysaccharide level of Streptococcus mutans biofilm was greatly reduced.SEM results displayed that D-Histidine effectively inhibited Streptococcus mutans bioflim formation at concentration of 40% and bacterial aggregation amount declined with increasing concentrations of D-Histidine.Conclusion:D-Histidine effectively inhibits Streptococcus mutan biofilm formation in a dose-dependent manner and D-Histidine interferes with the viability of Streptococcus mutans.
Objectives:To evaluate the effects of D-Histidine on the Streptococcus mutans biofilmin vitro.Methods:Antimicrobial activity testing of D-Histidine was evaluated by determining the growth curve.Crystal violet biofilm biomass assay and polysaccharide measurement were performed to evaluate the potential activity of D-Histidine effectively inhibiting biofilm formation.SEM was used to observe the morphology and the structure of biofilm.Results:Growth curve showed that D-Histidine at concentration of 20%interfered with the viability of Streptococcus mutans in a dose-dependent manner.CV and EPS assays showed that D-Histidine at concentration of 40%effectively inhibited Streptococcus mutans biofilm formation and the polysaccharide level of Streptococcus mutans biofilm was greatly reduced.SEM results displayed that D-Histidine effectively inhibited Streptococcus mutans bioflim formation at concentration of 40% and bacterial aggregation amount declined with increasing concentrations of D-Histidine.Conclusion:D-Histidine effectively inhibits Streptococcus mutan biofilm formation in a dose-dependent manner and D-Histidine interferes with the viability of Streptococcus mutans.
引文
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[2] Wade WG.The oral microbiome in health and disease[J].Pharmacol Res,2013,69(1):137-143.
[3]史彦,李成杰,张明正,等.龋齿及牙周病患者与健康人群口腔微生态的比较研[J].口腔医学研究,2016,32(12):1265-1268.
[4] Seminario A,Broukal Z,IvancakováR.Mutans streptococci and the development of dental plaque[J].Prague Med Rep,2005,106(4):349-358.
[5] Tamura S,Yonezama H,Motegi M,et al.Inhibiting effects of Streptococcus salivarius on competence-stimulating peptide-dependent biofilm formation by Streptococcus mutans[J].Oral Microbiol Immunol,2009,24(2):152-161.
[6] Krysciak W,Jurczak A,Koscielniak D,et al.The virulence of Streptococcus mutans and the ability to form biofilms[J].Eur J Clin Microbiol Infect Dis,2014,33(4):499-515.
[7] Marsh PD.Microbiology of dental plaque biofilms and their role in oral health and caries[J].Dent Clin North Am,2010,54(3):441-454.
[8] Pensyan A,Gillngs M,Paulsen IT.Antibiotic discovery:combatting bacterial resistance in cells and in biofilm communities[J].Molecules,2015,20(4):5286-5298.
[9] Qi H,Li B,Wang H,et al.Effects of d-valine on periodontal or peri-implant pathogens:Porphyromonas gingivalis biofilm[J].J Periodontol,2018,89(3):303-314.
[10] Kolodkin-Gal I,Romero D,Cao S,et al.D-amino acids trigger biofilm disassembly[J].Science,2010,328(5978):627-629.
[11] Leiman SA,May JM,Lebar MD,et al.D-amino acids indirectly inhibit biofilm formation in Bacillus subtilis by interfering with protein synthesis[J].J Bacteriol,2013,195(23):5391-5395.
[12] She P,Chen L,Liu H,et al.The effects of D-Tyrosine combined with amikacin on the biofilms of Pseudomonas aeruginosa[J].Microb Pathog,2015,86:38-44.
[13] Pitts B,Hamilton MA,Zelver N,et al.A microtiter-plate screening method for biofilm disinfection and removal[J].J Microbiol Methods,2003,54(2):269-276.
[14] Wang L,Melo MA,Weir MD,et al.Novel bioactive nanocomposite for Class-Ⅴrestorations to inhibit periodontitis-related pathogens[J].Dent Mater,2016,32(12):e351-e361.
[15] Yang Y,Sreenivasan PK,Subramanyam R,et al.Multiparameter assessments to determine the effects of sugars and antimicrobials on a polymicrobial oral biofilm[J].Appl Environ Microbiol,2006,72(10):6734-6742.
[16] Takahashi N,Nyvad B.The role of bacteria in the caries process:ecological perspectives[J].J Dent Res,2011,90(3):294-303.
[17] ten Cate JM,Cummins D.Fluoride toothpaste containing1.5%arginine and insoluble calcium as a new standard of care in caries prevention[J].J Clin Dent,2013,24(3):79-87.
[18] Lam H,Oh DC,Cava F,et al.D-amino acids govern stationary phase cell wall remodeling in bacteria[J].Science,2009,325(5947):1552-1555.
[19] Flemming HC,Neu TR,Wozniak DJ.The EPS matrix:the"house of biofilm cells"[J].J Bacteriol,2007,189(22):7945-7947.
[20] Cava F,Lam H,De Pedro MA,et al.Emerging knowledge of regulatory roles of D-amino acids in bacteria[J].Cell Mol Life Sci,2011,68(5):817-831.
[21] Jia R,Li Y,Al-Mahamedh HH,et al.Enhanced biocide treatments with D-amino acid mixtures against a biofilm consortium from a water cooling tower[J].Front Microbiol,2017,8:1538.