Apical periodontitis, an inflammatory lesion causing bone resorption around the apex of teeth, is treated by eradicating infectious bacteria from the root canal. However, it has a high recurrence rate and often requires retreatment. We investigated the bactericidal effect of antimicrobial photodynamic therapy (aPDT)/photodynamic antimicrobial chemotherapy (PACT) using indocyanine green (ICG)-loaded nanospheres coated with chitosan and a diode laser on a biofilm of Enterococcus faecalis, a pathogen of refractory apical periodontitis. Biofilm of E. faecalis was cultured in a porcine infected root canal model. ICG solution was injected into the root canal, which was then irradiated with a laser (810 nm wavelength) from outside the root canal. The bactericidal effect was evaluated by colony counts and scanning electron microscopy. The result of the colony counts showed a maximum 1.89 log reduction after irradiation at 2.1 W for 5 min. The temperature rise during aPDT/PACT was confirmed to be within a safe range. Furthermore, the light energy transmittance through the root was at a peak approximately 1 min after the start of irradiation, indicating that most of the ICG in the root canal was consumed. This study shows that aPDT/PACT can suppress E. faecalis in infected root canals with high efficiency.

Bacteroides forsythus produces species‐specific major proteins with high molecular weights of 270 and 230‐kDa (270K and 230K). A specific antibody raised against 270K was used for Western blot analysis and immunoelectron microscopy. Western blot analysis showed that the 270K and 230K proteins were immunologically similar. Immunogold labeling of ultrathin‐sectioned bacterial cells and biochemical fractionation revealed that these proteins were localized at the outermost cell surface, not in the cytoplasm. These results suggest that major proteins ubiquitous to this species may form the S‐layer.

Streptococcus pneumoniae is an important causative organism of respiratory tract infections. Although periodontal bacteria have been shown to influence respiratory infections such as aspiration pneumonia, the synergistic effect of S. pneumoniae and Porphyromonas gingivalis, a periodontopathic bacterium, on pneumococcal infections is unclear. To investigate whether P. gingivalis accelerates pneumococcal infections, we tested the effects of inoculating P. gingivalis culture supernatant (PgSup) into S. pneumoniae-infected mice. Mice were intratracheally injected with S. pneumoniae and PgSup to induce pneumonia, and lung histopathological sections and the absolute number and frequency of neutrophils and macrophages in the lung were analyzed. Proinflammatory cytokine/chemokine expression was examined by qPCR and ELISA. Inflammatory cell infiltration was observed in S. pneumoniae-infected mice and S. pnemoniae and PgSup mixed-infected mice, and mixed-infected mice showed more pronounced inflammation in lung. The ratios of monocytes/macrophages and neutrophils were not significantly different between the lungs of S. pneumoniae-infected mice and those of mixed-infected mice. PgSup synergistically increased TNF-α expression/production and IL-17 production compared with S. pneumoniae infection alone. We demonstrated that PgSup enhanced inflammation in pneumonia caused by S. pneumoniae, suggesting that virulence factors produced by P. gingivalis are involved in the exacerbation of respiratory tract infections such as aspiration pneumonia.

Antimicrobial photodynamic therapy (aPDT) is considered a potential treatment for biofilm infections, which have become an increasing health issue because of the rise in antimicrobial resistance. This study aimed to evaluate the bactericidal effect of aPDT using indocyanine green-loaded nanospheres with chitosan coating (ICG-Nano/c) against polymicrobial periodontal biofilms. Composite biofilms of Porphyromonas gingivalis and Streptococcus gordonii were constructed in 96-well plates, and aPDT with ICG-Nano/c and an 810 nm diode laser was performed either by direct irradiation or transmitting irradiation through a 3-mm-thick gingival model. The efficacy of ICG-Nano/c-based aPDT was compared with antibiotics (minocycline and amoxicillin). Additionally, attenuated aPDT under sublethal conditions was used to investigate gene expression related to the antioxidant response (oxyR and sod of P. gingivalis) and biofilm formation via quorum sensing (luxS of both species) with real-time polymerase chain reaction. ICG-Nano/c-based aPDT significantly reduced the bacterial load in the biofilm, achieving at least a 2 log₁₀ reduction in colony-forming units within 5 minutes for both irradiation methods. After 6 hours of treatment, the bactericidal effects of aPDT and antibiotics were similar, but after 32 hours, antibiotics were more effective, particularly against P. gingivalis. Attenuated aPDT showed a slight increase in sod expression in P. gingivalis, while luxS expression decreased in both bacteria. The ICG-Nano/c-based aPDT system exerted a certain degree of bactericidal activity against a composite biofilm of periodontal bacteria. Therefore, it has potential as an alternative option or adjunctive therapy to conventional antibiotics in periodontal treatment.