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Comparative Study
. 2015 Apr;94(4):622-9.
doi: 10.1177/0022034515571416. Epub 2015 Feb 24.

Effect of antibacterial dental adhesive on multispecies biofilms formation

K Zhang  1 , S Wang  2 , X Zhou  2 , H H K Xu  3 , M D Weir  3 , Y Ge  2 , M Li  1 , S Wang  2 , Y Li  1 , X Xu  2 , L Zheng  2 , L Cheng  4
Affiliations
Comparative Study

Effect of antibacterial dental adhesive on multispecies biofilms formation

K Zhang et al. J Dent Res. 2015 Apr.

Abstract

Antibacterial adhesives have favorable prospects to inhibit biofilms and secondary caries. The objectives of this study were to investigate the antibacterial effect of dental adhesives containing dimethylaminododecyl methacrylate (DMADDM) on different bacteria in controlled multispecies biofilms and its regulating effect on development of biofilm for the first time. Antibacterial material was synthesized, and Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis were chosen to form multispecies biofilms. Lactic acid assay and pH measurement were conducted to study the acid production of controlled multispecies biofilms. Anthrone method and exopolysaccharide (EPS):bacteria volume ratio measured by confocal laser scanning microscopy were performed to determine the EPS production of biofilms. The colony-forming unit counts, scanning electron microscope imaging, and dead:live volume ratio decided by confocal laser scanning microscopy were used to study the biomass change of controlled multispecies biofilms. The TaqMan real-time polymerase chain reaction and fluorescent in situ hybridization imaging were used to study the proportion change in multispecies biofilms of different groups. The results showed that DMADDM-containing adhesive groups slowed the pH drop and decreased the lactic acid production noticeably, especially lactic acid production in the 5% DMADDM group, which decreased 10- to 30-fold compared with control group (P < 0.05). EPS was reduced significantly in 5% DMADDM group (P < 0.05). The DMADDM groups reduced the colony-forming unit counts significantly (P < 0.05) and had higher dead:live volume ratio in biofilms compared with control group (P < 0.05). The proportion of S. mutans decreased steadily in DMADDM-containing groups and continually increased in control group, and the biofilm had a more healthy development tendency after the regulation of DMADDM. In conclusion, the adhesives containing DMADDM had remarkable antimicrobial properties to serve as "bioactive" adhesive materials and revealed its potential value for antibiofilm and anticaries clinical applications.

Keywords: antibacterial material; bonding agent; caries inhibition; controlled multispecies biofilm; dimethylaminododecyl methacrylate; tooth restoration.

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Conflict of interest statement

The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article.

Figures

Figure 1.
Figure 1.
Acid production of multispecies biofilms. (A) The lactic acid production of 16-, 48-, and 72-h multispecies biofilms and (B) the supernatant pH of 3 groups—Clearfil SE Bond without dimethylaminododecyl methacrylate (DMADDM), Clearfil SE Bond with 2.5% DMADDM, and Clearfil SE Bond with 5% DMADDM. Data are presented as mean ± standard deviation. *P < 0.05.
Figure 2.
Figure 2.
Extracellular polysaccharides (EPS) of multispecies biofilms. (A) The water-insoluble glucan of each disk from different groups, measured by anthrone method; (B) the 3-dimensional reconstruction of multispecies biofilms (bacteria, stained green; EPS, stained red); (C) the volume of EPS and bacteria, calculated according to 5 random sights of controlled multispecies biofilms. Data are presented as mean ± standard deviation. *P < 0.05. This figure is available in color online at http://jdr.sagepub.com.
Figure 3.
Figure 3.
The antibacterial effect of dimethylaminododecyl methacrylate (DMADDM) on multispecies biofilms. (A) The scanning electron microscopy images multispecies biofilms; (B) colony-forming unit counts of biofilms formed on each disk from 3 groups—Clearfil SE Bond without DMADDM, Clearfil SE Bond with 2.5% DMADDM, and Clearfil SE Bond with 5% DMADDM; (C) the 3-dimensional reconstruction of multispecies biofilms (live bacteria, stained green; dead cells, stained red); (D) the ratio between dead and live bacteria computed in line with 5 random sights of multispecies biofilms. All these data supported the antibacterial effect of DMADDM on multispecies biofilms. Data are presented as mean ± standard deviation. *P < 0.05. This figure is available in color online at http://jdr.sagepub.com.
Figure 4.
Figure 4.
The composition shift of multispecies biofilms. (A) The ratio of Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis in multispecies biofilms, conducted by TaqMan real-time polymerase chain reaction (dimethylaminododecyl methacrylate [DMADDM] content: 0%, 2.5%, 5%); (B) fluorescent in situ hybridization images of multispecies biofilms (S. mutans, stained green; S. sanguinis, stained red; S. gordonii, stained blue). This figure is available in color online at http://jdr.sagepub.com.
Figure 5.
Figure 5.
The antibacterial effect of dimethylaminododecyl methacrylate (DMADDM) on single-species biofilms. (A) The scanning electron microscopy images of Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis biofilms; (B) colony-forming unit counts of S. mutans, S. gordonii, and S. sanguinis biofilms formed on each disk from different DMADDM content groups. Data are presented as mean ± standard deviation. *P < 0.05.
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