불소 민감성 Streptococcus mutans와 불소 저항성Streptococcus mutans의 우식원성 특성 비교 Comparison of Cariogenic Characteristics between Fluoride-sensitive and Fluoride-resistant Streptococcus mutans원문보기
이 연구의 목적은 불소 민감성(fluoride-sensitive) Streptococcus mutans (FS S. mutans ) 와 불소 저항성(fluoride-resistant) Streptococcus mutans (FR S. mutans )의 우식원성 특성을 비교하는 것이다. S. mutans ATCC 25175 균주를 NaF (70 ppm)를 포함한 trypticase soy broth에서 40일 동안 배양하여 FR S. mutans를 획득하였다. 2% 자당 유무에 따른 FS와 FR S. mutans의 성장과 산 생성 변화를 비교하였고, 타액 세균을 이용하여 FS와 FR S. mutans 바이오필름을 형성하여 공초점 레이저 현미경으로 관찰하고 세균 수를 측정하였다. 또한 RT-PCR을 통해 FS와 FR S. mutans의 gtf 유전자 발현 정도를 비교하였다. FR S. mutans는 FS S. mutans 보다 자당을 이용한 성장과 산 생성이 모두 낮았다. 타액 세균과 단일 균주 바이오필름의 형성 또한 FR S. mutans가 FS S. mutans 보다 적었고, 더 낮은 gtfB, gtfC 및 gtfD 발현을 보였다. 이 연구를 통해 FR S. mutans는 FS S. mutans 보다 감소된 우식원성 특성을 가지고 있음을 관찰하였고, 불소의 주기적 사용은 S. mutans 세균의 활성을 낮추어 우식 발생을 줄이는데 도움이 될 수 있을 것이다.
이 연구의 목적은 불소 민감성(fluoride-sensitive) Streptococcus mutans (FS S. mutans ) 와 불소 저항성(fluoride-resistant) Streptococcus mutans (FR S. mutans )의 우식원성 특성을 비교하는 것이다. S. mutans ATCC 25175 균주를 NaF (70 ppm)를 포함한 trypticase soy broth에서 40일 동안 배양하여 FR S. mutans를 획득하였다. 2% 자당 유무에 따른 FS와 FR S. mutans의 성장과 산 생성 변화를 비교하였고, 타액 세균을 이용하여 FS와 FR S. mutans 바이오필름을 형성하여 공초점 레이저 현미경으로 관찰하고 세균 수를 측정하였다. 또한 RT-PCR을 통해 FS와 FR S. mutans의 gtf 유전자 발현 정도를 비교하였다. FR S. mutans는 FS S. mutans 보다 자당을 이용한 성장과 산 생성이 모두 낮았다. 타액 세균과 단일 균주 바이오필름의 형성 또한 FR S. mutans가 FS S. mutans 보다 적었고, 더 낮은 gtfB, gtfC 및 gtfD 발현을 보였다. 이 연구를 통해 FR S. mutans는 FS S. mutans 보다 감소된 우식원성 특성을 가지고 있음을 관찰하였고, 불소의 주기적 사용은 S. mutans 세균의 활성을 낮추어 우식 발생을 줄이는데 도움이 될 수 있을 것이다.
The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm ...
The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. S. mutans ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR S. mutans. FS and FR S. mutans were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both S. mutans was investigated through RT-PCR. FR S. mutans exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS S. mutans . Both cariogenic and single species biofilm formation was lower in the presence of FR S. mutans, along with reduced number of bacteria. FR S. mutans showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS S. mutans . On the basis of results, FR S. mutans may be less virulent in the induction of dental caries.
The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. S. mutans ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR S. mutans. FS and FR S. mutans were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both S. mutans was investigated through RT-PCR. FR S. mutans exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS S. mutans . Both cariogenic and single species biofilm formation was lower in the presence of FR S. mutans, along with reduced number of bacteria. FR S. mutans showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS S. mutans . On the basis of results, FR S. mutans may be less virulent in the induction of dental caries.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
문제 정의
This study aimed to compare the cariogenic characteristic of FR S. mutans with that of FS S. mutans in the presence of sucrose and to determine its effect on cariogenic biofilm formation using salivary bacteria.
제안 방법
In order to investigate growth and acidogenicity, the concentration of FS and FR S. mutans was adjusted to 0.4 optical density at 660 nm wavelength using fresh medium, and 5 mL of FS and FR S. mutans media were each inoculated into 200 mL of BHI broth with or without 2% sucrose.
mutans for fluoride had been investigated by serial dilution method from 1000 ppm of fluoride. The 50% inhibitory concentration of fluoride against S. mutans growth was found to be between 62.5 and 125 ppm, and 70 ppm of fluoride concentration was decided for the study. 10,000 ppm of NaF solution was filtered with 0.
mutans was analyzed by RT-PCR. The experiment was performed four times, and the gene expression data were shown as the average fold change with means and their standard deviations.
대상 데이터
S. mutans ATCC 25175 was used in the following experiments. The bacteria were cultured in trypticase soy broth (TSB; BD Biosciences, Franklin Lakes, NJ, USA) in the presence and absence of fluoride at 37℃ in anaerobic conditions.
데이터처리
All statistical analyses were performed by Mann-Whitney test using SPSS 21 (SPSS Inc., Chicago, IL, USA). The data are presented as mean ± standard deviation or as median and interquartile range.
성능/효과
As biofilm formation exhibited a difference between FS and FR S. mutans , Gtfs production as a biofilm formation related factor in FS and FR S. mutans was compared (Fig. 5). The expression of gtfB, gtfC, and gtf D was significantly reduced to about 30%, in FR S.
mutans was observed. FR S. mutans formed reduced mass of salivary biofilm and single species biofilm compared to FS S. mutans , and the number of total bacteria and S. mutans was also significantly reduced. Oral biofilms are complex polymicrobial communities embedded within an extracellular matrix, present as a highly organized 3D structure adhered to the tooth[27,28].
mutans can be changed in the presence of other oral bacteria[27], and fluoride did not seem to have an inhibitory effect on the Gtf activity of Streptococcus species in previous studies[29,30]. However, our results showed that FR S. mutans expressed significantly lower levels of gtf gene expression and formed a reduced mass of cariogenic biofilm compared to FS S. mutans with the salivary bacteria.
mutans have been evaluated[20-22]. In this study, the cariogenic properties of FR S. mutans were compared with those of FS S. mutans in the presence of sucrose, and the effect of FR S. mutans on salivary biofilm formation was assessed.
mutans show stress mechanisms, such as alteration in F-ATPase activity, catabolic pathways, and gene expressions[25]. The results of the present study suggest the modification of sucrose metabolic activity in FR S. mutans , resulting in a shift to decreased cariogenicity. This is in line with a previous study that lower cariogenicity of FR S.
Moreover, decreased salivary biofilm formation was observed along with decreased gtf gene expression. These findings suggest that FR S. mutans may be less virulent than FS S. mutans in inducing dental caries and may be effective in maintaining oral microbial homeostasis and reducing dental caries.
후속연구
mutans in the oral environment is not easy[10]. Also, investigation of the stability of the resistance acquired form long term exposure of fluoride, and the affects of FR S. mutans to other microbial species should be considered in the future studies.
참고문헌 (35)
Takahashi N, Nyvad B : Caries ecology revisited: microbial dynamics and the caries process. Caries Res, 42:409-418, 2008.
Marsh PD : Dental plaque as a biofilm and a microbial community-implications for health and disease. BMC Oral Health, 6:14, 2006.
Krzysciak W, Jurczak A, Skalniak A, et al. : The virulence of Streptococcus mutans and the ability to form biofilms. Eur J Clin Microbiol Infect Dis, 33:499-515, 2014.
Fitzgerald RJ, Adams BO, Sandham HJ, Abhyankar S : Cariogenicity of a lactate dehydrogenase-deficient mutant of Streptococcus mutans serotype c in gnotobiotic rats. Infect Immun, 57:823-826, 1989.
Banas JA : Virulence properties of Streptococcus mutans. Front Biosci, 9:1267-1277, 2004.
Sturr MG, Marquis RE : Comparative acid tolerances and inhibitor sensitivities of isolated F-ATPases of oral lactic acid bacteria. Appl Environ Microbiol, 58:2287-2291, 1992.
Bowen WH, Koo H : Biology of Streptococcus mutans - derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res, 45:69-86, 2011.
Brown LR, White JO, Streckfuss JL, et al. : Effect of continuous fluoride gel use on plaque fluoride retention and microbial activity. J Dent Res 62:746-751, 1983.
Lee SH, Choi BK, Kim YJ : The cariogenic characters of xylitol-resistant and xylitol-sensitive Streptococcus mutans in biofilm formation with salivary bacteria. Arch Oral Biol, 57:697-703, 2012.
Bowden GH : Effects of fluoride on the microbial ecology of dental plaque. J Dent Res, 69:653-659, 1990.
Brussock SM, Kral TA : In vitro flouride resistance in a cariogenic Streptococcus. Ohio J Sci, 86:73-79, 1986.
Liao Y, Chen J, Deng DM, et al. : Identification and functional analysis of genome mutations in a fluoride-resistant Streptococcus mutans strain. PLoS One, 10:0122630, 2015.
Streckfuss JL, Perkins D, Graves L, et al. : Fluoride resistance and adherence of selected strains of Streptococcus mutans to smooth surfaces after exposure to fluoride. J Dent Res, 59:151-158, 1980.
Xiao J, Klein MI, Koo H, et al. : The exopolysaccharide matrix modulates the interaction between 3D architecture and virulence of a mixed-species oral biofilm. PLoS Pathog, 8:1002623, 2012.
Bowen WH, Burne RA, Wu H, Koo H : Oral biofilms: pathogens, matrix, and polymicrobial interactions in microenvironments. Trends Microbiol, 26:229-242, 2018.
Pandit S, Kim JE, Jeon JG, et al. : Effect of sodium fluoride on the virulence factors and composition of Streptococcus mutans biofilms. Arch Oral Biol, 56:643-649, 2011.
Shani S, Friedman M, Steinberg D : The anticariogenic effect of amine fluorides on Streptococcus sobrinus and glucosyltransferase in biofilms. Caries Res, 34:260-267, 2000.
Marinho VC, Chong LY, Worthington HV, Walsh T : Fluoride mouthrinses for preventing dental caries in children and adolescents. Cochrane Database Syst Rev, 7:CD002284, 2016.
AI-Helper
※ AI-Helper는 부적절한 답변을 할 수 있습니다.