Doxycycline-functionalized polymeric nanoparticles inhibit Enterococcus faecalis biofilm formation on dentine

Aim To evaluate in a laboratory setting the antimicrobial properties and the potential to inhibit biofilm formation of novel remineralizing polymeric nanoparticles (NPs) when applied to dentine surfaces and to ascertain the effect of the functionalization of these NPs with zinc, calcium or doxycycline. Methodology The antimicrobial activity and inhibition of biofilm formation of polymeric NPs were analysed on human dentine blocks that were infected with Enterococcus faecalis before or after application of NPs. LIVE/DEAD (R) testing under Confocal Laser Scanning Microscopy and bacterial culturing were employed to analyse biofilm biovolume and bacterial viability. Field Emission Scanning Electron Microscopy was also employed to assess biofilm morphology. One-way anova with Welch's correction and post hoc comparison by the Games-Howell test were performed for comparisons between groups. Results The un-functionalized NPs displayed the greatest antimicrobial activity against E. faecalis biofilms as they provided the lowest biovolume (3865.7 +/- 2926.97 mu m(3); P < 0.001) and the highest dead/injured cells percentage (79.93 +/- 18.40%; P < 0.001), followed by Dox-NPs (biovolume: 19,041.55 +/- 17,638.23 mu m(3), dead/injured cells: 45.53 +/- 26.50%; P < 0.001). Doxycycline-loaded NPs had the largest values of inhibition of biofilm formation with the lowest biofilm biovolume (8517.65 +/- 7055.81 mu m(3); P < 0.001) and a high dead/injured bacterial percentage (68.68 +/- 12.50%; P < 0.001). Un-functionalized NPs did not reduce biomass growth (P > 0.05), but attained the largest percentage of compromised cells (93 +/- 8.23%; P < 0.001), being able to disrupt biofilm formation. It also produced occlusion of dentinal tubules, potentially interfering with bacterial tubule penetration. Conclusions A new generation of bioactive nano-fillers (doxycycline-functionalized polymeric NPs) had antibacterial activity and occluded dentinal tubules. Incorporating these NPs into endodontic sealers may have the potential to enhance the outcome of root canal treatment.

Automated summary

Un-functionalized NPs exhibited strong antimicrobial activity against E. faecalis biofilms, reducing biovolume and increasing the percentage of dead/injured cells. Doxycycline-loaded NPs showed the highest inhibition of biofilm formation, with reduced biofilm biovolume and a high percentage of dead/injured bacterial cells. The un-functionalized NPs did not reduce biomass growth but had the highest percentage of compromised cells, capable of disrupting biofilm formation.