Inhibitory effect of Streptococcus mutans antisense vicK RNA regulating the cariogenicity of oral streptococci multi-species biofilm
10.3760/cma.j.cn112144-20231031-00229
- VernacularTitle:变异链球菌反义vicK RNA对口腔链球菌多菌种生物膜致龋性的抑制作用
- Author:
Hong CHEN
1
;
Mengmeng XU
;
Yuting SUN
;
Shuang YU
;
Deqin YANG
Author Information
1. 重庆医科大学附属口腔医院北部院区牙体牙髓科 口腔疾病与生物医学重庆市重点实验室 重庆市高校市级口腔生物医学工程重点实验室,重庆 401147
- Keywords:
Dental caries;
Streptococcus mutans;
Antisense vicK RNA;
Streptococcus sanguinis;
Streptococcus gordonii
- From:
Chinese Journal of Stomatology
2024;59(1):64-70
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the regulative effects of Streptococcus mutans (Sm) antisense vicK RNA (ASvicK) on the multi-species biofilm formed by three common oral streptococci (Sm, Streptococcus sanguinis, and Streptococcus gordonii) (Sm+Ss+Sg). Methods:ASvicK over-expression strain was constructed by using a recombinant plasmid, and three-species biofilm UA159+Ss+Sg and ASvicK+Ss+Sg were cultured. The phenotypes of biofilms were detected by scanning electron microscopy (SEM). Crystal violet (CV) assay was used to detect biofilm biomass. Lactate kit and anthrone-sulfuric acid colorimetric assay were used to determine the abilities of lactic acid and exopolysaccharides production, respectively. The proportions of three-species and expression levels of the cariogenic-related genes in biofilms were detected by TaqMan fluorescence quantitative PCR and real-time fluorescence quantitative PCR. A biofilm demineralization model of human enamel slabs was further constructed, and the hardness of enamel surface was detected.Results:Compared to UA159+Ss+Sg, over-expression of ASvicK could inhibit biofilm formation and lactic acid production in ASvicK+Ss+Sg biofilm significantly decreased by 78.93% ( P<0.001) and 62.23% ( P<0.001), respectively. With ASvicK over-expression, the amounts of water-insoluble and-soluble glucoses in ASvicK+Ss+Sg biofilm were reduced respectively by 39.13% ( P<0.001) and 68.00% ( P<0.001). Compared to the UA159+Ss+Sg Group, the proportion of Sm, the cariogenic bacteria, showed 33.00% reduction ( P<0.01) in Sm+Ss+Sg biofilm, and the gene expressions of cariogenic-relative genes vicK/X, gtfB/C/D, and ftf significantly decreased ( P<0.05). The micro-hardness value of enamel slabs after demineralization by ASvicK+Ss+Sg biofilm was significantly increased to 183.84% ( P<0.001). Conclusions:ASvicK over-expression could reduce the Sm proportion and weaken the cariogenicity of oral Streptococcus biofilm, thereby possibly slowing down the progression of caries.