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Introduction: Successful treatment of infected or inflamed endodontic tissues requires chemomechanical debridement of the canal spaces and proper sealing of the coronal and apical canal openings. Only a few methods are available to further disinfect areas or initiate regeneration of local tissues. In this study, we assessed the ability of 255 nm and 405 nm light emitting diode (LED) treatment to kill planktonic cultures of Enterococcus faecalis (E. faecalis) and induce the production of cellular biomarkers related to endodontic tissue regeneration. Methods: We determined the antimicrobial effects of 255 nm and 405 nm LED treatment on E. faecalis and the effects of 255 nm and 405 nm LED treatment on the production of osteoinductive, angiogenic, proliferative, and proinflammatory biomarkers from human embryonic palatal mesenchyme (HEPM) cells and gingival fibroblasts. Results: We demonstrated that 1) 255 nm LED, but not 405 nm LED, treatment killed E. faecalis, 2) 255 nm LED and sodium hypochlorite (NaOCl) more efficiently killed E. faecalis, 3) neither 255 nm nor 405 nm LED treatment affected the viability of HEPM cells and gingival fibroblasts, and 4) 255 nm LED treatment, alone or in combination with 405 nm LED treatment, of HEPM cells and gingival fibroblasts induced the production of biomarkers related to endodontic tissue regeneration. Conclusions: The results in this study suggest a new treatment modality using short periods of 255 nm LED treatment as an adjunct to chemo-mechanical debridement for the disinfection of inflamed sites and the production of biomarkers related to endodontic tissue regeneration.

Antibiotics are often used to treat oral infections. Unfortunately, excessive antibiotic use can adversely alter oral microbiomes and promote the development of antibiotic-resistant microorganisms, which can be difficult to treat. An alternate approach could be to induce the local transcription and expression of endogenous oral antimicrobial peptides (AMPs). To assess the feasibility and benefits of this approach, we conducted literature searches to identify (i) the AMPs expressed in the oral cavity; (ii) the methods used to induce endogenous AMP expression; and (iii) the roles that expressed AMPs may have in regulating oral inflammation, immunity, healing, and pain. Search results identified human neutrophil peptides (HNP), human beta defensins (HBD), and cathelicidin AMP (CAMP) gene product LL-37 as prominent AMPs expressed by oral cells and tissues. HNP, HBD, and LL-37 expression can be induced by micronutrients (trace elements, elements, and vitamins), nutrients, macronutrients (mono-, di-, and polysaccharides, amino acids, pyropeptides, proteins, and fatty acids), proinflammatory agonists, thyroid hormones, and exposure to ultraviolet (UV) irradiation, red light, or near infrared radiation (NIR). Localized AMP expression can help reduce infection, inflammation, and pain and help oral tissues heal. The use of a specific inducer depends upon the overall objective. Inducing the expression of AMPs through beneficial foods would be suitable for long-term health protection. Additionally, the specialized metabolites or concentrated extracts that are utilized as dosage forms would maintain the oral and intestinal microbiome composition and control oral and intestinal infections. Inducing AMP expression using irradiation methodologies would be applicable to a specific oral treatment area in addition to controlling local infections while regulating inflammatory and healing processes.

Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faecalis and has the potential to kill other oral microorganisms associated with endodontic infections. This same bacteriocidal device shows great promise in the stimulation of periapical healing and pain reduction resulting from inflammation in root canals. Previously, we found that 255 nm UVC LED killed E. faecalis and induced the production of cellular biomarkers in HEPM cells and gingival fibroblasts (Morio et al., 2019). Here, we extend those findings and hypothesize that UVC LED at other wavelengths and power levels kill microorganisms associated with root canal infections. Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) wavelenths were assembled and the energy levels of their emissions were measured. The energy doses in millijoules (mJ) were calculated from the power readings of the meter (µW) × time of exposure (seconds). Ex vivo models of root canals were prepared in extracted, instrumented, single canal human premolars. Five cultures of microorganisms were treated with 265 nm (12 mW), 265 nm (22.5 mW), or 280 nm (8 mW) UVC LED on discs in laboratory assays and 4 cultures of microorganisms were treated with 265 nm (22.5 mW) UVC LED in root canals of extracted, instrumented teeth. After UVC LED treatment, all microorganisms were cultivated on microbiological media. Colony forming units (CFU) of viable microorganisms treated with UVC LED were counted and compared with those of viable microorganisms not treated with UVC LED as controls. Tukey's Honestly Significant Difference was used to determine statistical significances (0.05). Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) killed Candida albicans, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), E. faecalis, and Streptococcus sanguinis after 30-90 seconds of exposure in laboratory assays ( p < 0.05). Microbial killing differed among treatment times, UVC LED wavelengths, power levels of each unit, and specific microorganism. The unit emitting UVC LED at 265 nm (22.5 mW) killed C. albicans, S. aureus, MRSA, and E. faecalis in 30 s in root canals of extracted, instrumented teeth ( p < 0.05). This dataset can be reused to assess the ability of other wavelengths and power levels to kill microorganisms as well as improve procedures for treating endodontic infections and inflammation in root canals.

Background: Ultraviolet (UV) irradiation can modulate host immune responses and this approach is a novel application for treating endodontic infections and inflammation in root canals. Methods: A dataset of UV-induced molecules was compiled from a literature search. A subset of this dataset was used to calculate expression log2 ratios of endodontic tissue molecules from HEPM cells and gingival fibroblasts after 255, 405, and 255/405 nm UV irradiation. Both datasets were analyzed using ingenuity pathway analysis (IPA, Qiagen, Germantown, MD, USA). Statistical significance was calculated using Fisher’s exact test and z-scores were calculated for IPA comparison analysis. Results: The dataset of 32 UV-induced molecules contained 9 antimicrobial peptides, 10 cytokines, 6 growth factors, 3 enzymes, 2 transmembrane receptors, and 2 transcription regulators. These molecules were in the IPA canonical pathway annotations for the wound healing signaling pathway (9/32, p = 3.22 × 10−11) and communication between immune cells (6/32, p = 8.74 × 10−11). In the IPA disease and function annotations, the 32 molecules were associated with an antimicrobial response, cell-to-cell signaling and interaction, cellular movement, hematological system development and function, immune cell trafficking, and inflammatory response. In IPA comparison analysis of the 13 molecules, the predicted activation or inhibition of pathways depended upon the cell type exposed, the wavelength of the UV irradiation used, and the time after exposure. Conclusions: UV irradiation activates and inhibits cellular pathways and immune functions. These results suggested that UV irradiation can activate innate and adaptive immune responses, which may supplement endodontic procedures to reduce infection, inflammation, and pain and assist tissues to heal.