Evaluation of bacterial anti-adhesion and sustained antimicrobial abilities of chlorhexidine gluconate-loaded phase-transited lysozyme nanoparticle suspension on dentine: An ex vivo study

Abstract

Aim: To evaluate the effect of chlorhexidine gluconate-loaded phase-transited lysozyme (CHG@PTL) coating on inhibiting bacterial adhesion and biofilm formation in an ex vivo root canal dentine model.

Methodology: The physicochemical and structural characteristics of CHG@PTL nanoparticle suspension and its coating formed on the dentine surface were analysed by thioflavin T fluorescence assay, transmission electron microscopy and confocal laser scanning microscopy (CLSM). The sustained chlorhexidine release profile of the CHG@PTL coating on the dentine surface was compared with that of the 2% CHG solution. By comparing with phosphate-buffered saline, 1% sodium hypochlorite and 2% CHG solutions, the sustained antibacterial ability of the CHG@PTL coating and its effects on adhesion and biofilm formation of three types of bacteria (E. faecalis, S. mutans, and A. viscous) were analysed in ex vivo root canal dentine models using the serial plate transfer test (SPTT) and CLSM with live/dead bacterial staining, respectively.

Results: CHG promoted the lysozyme protein to form a higher proportion of β-sheet structure during phase transition. In the CHG@PTL nanoparticle suspension, characteristic drug-loaded nanospheres with a high concentration of CHG molecules inside and an outer PTL nanofilm were observed, and they formed a thinner and tighter coating on the dentine surface. The CHG@PTL coating on the dentine surface showed a significantly higher cumulative release amount of chlorhexidine than that of 2% CHG (p < .05). The results of SPTT showed that the CHG@PTL coating had a longer antibacterial duration than the control groups. After 12 h of incubation, a higher number of bacteria were agglutinated on the CHG@PTL coating surface compared to the control groups (p < .05). After 7 days of incubation, the number of agglutinated bacteria significantly decreased. At two time points, the percentage of dead bacteria on the CHG@PTL coating surface was the highest among all experimental groups based on CLSM observation (over 99.9% for all three bacteria, p < .001).

Conclusions: CHG@PTL nanoparticle suspension could form an antimicrobial coating on the surface of dentine with a novel 'agglutinating bacteria and sterilizing' mode.

Keywords: antibacterial coating; bacteria agglutination; chlorhexidine gluconate; drug‐loaded nanospheres; phase‐transited lysozyme.