Persisting apical periodontitis is a primary reason for multiple intervention in root canal. Persisting bacteria in root canal is related with the persisting infection. Despite the advancement in treatment strategies the persisting infection is a major challenge for endodontist. Here we tested two newly developed quaternary ammonium methacrylates (QAMs) against endodontic bacteria and their biofilms. Their antibacterial and antibiofilm efficiency were compared with chlorhexidine (CHX) and sodium hypochlorite (NaOCl). We measured the MIC, MBC and MBIC of DMADDM and DMAHDM respectively. We also detected the ratio of live/dead bacteria and bacterial composition in the biofilms treated by DMADDM and DMAHDM. We found that DMADDM and DMAHDM could inhibit the growth of bacteria and biofilms formation. The result showed that novel QAMs were remarkably efficient than CHX against biofilms. In addition, we found that Streptococcus gordonii (S. gordonii) and Enterococcus faecalis (E. faecalis) were frequent isolates after treatment with antimicrobial compounds.

Quaternary ammonium methacrylates (QAMs) are useful antimicrobial compounds against oral bacteria. Here, we investigated the effects of two QAMs, dimethylaminododecyl methacrylate (DMADDM) and dimethylaminohexadecyl methacrylate (DMAHDM), on biofilm formation, survival and development of tolerance by biofilm, and survival and development of tolerance against QAMs after prolonged starvation. Enterococcus faecalis (E. faecalis), Streptococcus gordonii (S. gordonii), Lactobacillus acidophilus (L. acidophilus), and Actinomyces naeslundii (A. naeslundii) were used. Minimum inhibitory concentration (MIC) of QAMs against multispecies biofilm was determined. Biofilm formed under sub-MIC was observed by crystal violet staining and confocal laser scanning microscopy (CLSM). Metabolic activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactic acid production measurement. Development of tolerance was determined by MIC values before and after exposure to QAMs or after prolonged starvation. It was found that E. faecalis and S. gordonii could survive and form biofilm under sub-MIC of QAMs. Lactic acid production from biofilms formed under sub-MIC was significantly higher than control specimens (p < 0.05). The exposure to sub-MIC of QAMs promoted biofilm formation, and prolonged starvation or prolonged contact with sub-MIC helped bacteria develop tolerance against killing by QAMs.

Quaternary ammonium compounds constitute a large group of antibacterial chemicals with a potential for inhibiting dental plaque. The aims of this study were to evaluate short-time antibacterial and regulating effects of dimethylaminododecyl methacrylate (DMADDM) on multispecies biofilm viability, reformation, and bacterial composition in vitro. DMADDM, chlorhexidine (CHX), and sodium fluoride (NaF) were chosen in the present study. Streptococcus mutans, Streptococcus sanguinis, and Streptococcus gordonii were used to form multispecies biofilm. Cytotoxicity assay was used to determine the optimal tested concentration. 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and resazurin test of biofilm were conducted to study the biomass changes and metabolic changes of controlled multispecies biofilm. Scanning electron microscopy (SEM) was used to observe biofilm images. TaqMan real-time polymerase chain reaction was performed to evaluate the proportion change in multispecies biofilm of different groups. Cytotoxicity assay showed that there existed a certain concentration application range for DMADDM, CHX, and NaF. MTT assay and resazurin test results showed that DMADDM and CHX groups decreased multispecies biofilm growth and metabolic activity (p < 0.05), no matter after 1 min or 5 min direct contact killing or after 24 h regrowth. The proportion of S. mutans decreased steadily in DMADDM and CHX groups after 1 min and 5 min direct contact killing and 24 h regrowth, compared to control groups. A novel DMADDM-containing solution was developed, achieving effective short-time antibacterial effects and regulation ability of biofilm formation.