Response of genes related to iron and porphyrin transport in Porphyromonas gingivalis to blue light.

BACKGROUND: Antimicrobial blue light (aBL) kills a variety of bacteria, including Porphyromonas gingivalis. However, little is known about the transcriptomic response of P. gingivalis to aBL therapy. This study was designed to evaluate the selective cytotoxicity of aBL against P. gingivalis over human cells and to further investigate the genetic response of P. gingivalis to aBL at the transcriptome level. METHODS: Colony forming unit (CFU) testing, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM) were used to investigate the antimicrobial effectiveness of blue light against P. gingivalis. The temperatures of the irradiated targets were measured to prevent overheating. Multiple fluorescent probes were used to quantify reactive oxygen species (ROS) generation after blue-light irradiation. RNA sequencing (RNA-seq) was used to investigate the changes in global gene expression. Following the screening of target genes, real-time quantitative polymerase chain reaction (RT-qPCR) was performed to confirm the regulation of gene expression. RESULTS: A 405 nm aBL at 100 mW/cm2 significantly killed P. gingivalis within 5 min while sparing human gingival fibroblasts (HGFs). No obvious temperature changes were detected in the irradiated surface under our experimental conditions. RNA-seq showed that the transcription of multiple genes was regulated, and RT-qPCR revealed that the expression levels of the genes RgpA and RgpB, which may promote heme uptake, as well as the genes Ftn and FetB, which are related to iron homeostasis, were significantly upregulated. The expression levels of the FeoB-2 and HmuR genes, which are related to hydroxyl radical scavenging, were significantly downregulated. CONCLUSIONS: aBL strengthens the heme uptake and iron export gene pathways while reducing the ROS scavenging pathways in P. gingivalis, thus improving the accumulation of endogenous photosensitizers and enhancing oxidative damage to P. gingivalis.

Clinical Evaluation of 532-nm Green Laser on Dentin Hypersensitivity: A Randomized Double-Blind Clinical Trial.

Objective: The aim of this study was to evaluate the therapies of low-level green laser and chemical desensitizer in the treatment of dentin hypersensitivity (DH). Methods: Forty-eight patients with 96 sensitive teeth were invited to participate in this clinical trial and were randomly divided into three groups. One group was treated with low-level green laser, the second group was treated with desensitizer [sodium fluoride (NaF)], and the third group acted as the placebo group and was treated with distilled water and placebo laser. The wavelength of green laser was 532 nm and the irradiance was 15 J/cm2 per treatment site. Hypersensitivity was assessed by visual analogue scale (VAS) according to cold test and probing at baseline. Immediately, 2 weeks, and 3 months after the application of green laser, NaF, and placebo, the participants' sensitivity level was accessed by new VAS analysis. Results: Forty-five patients with 90 teeth (n = 15 patients/group; 30 teeth/group) were followed up for 2 weeks and 3 months after treatment. There were significant differences in VAS scores between the placebo group and intervention group (green laser group and NaF group; analysis of variance, p < 0.05) at all three time points. The mean pain scores in DH reduced significantly immediately after treatment in the green laser group and NaF group when stimulated by cold and probing, whereas no significant difference was observed with these two therapies after 2 weeks (p > 0.05). After 3 months, mean VAS scores of the NaF group were higher than those of the green laser group (p < 0.01). Conclusions: Therefore, the green laser displayed similar effectiveness as NaF in treatment of DH and could be a promising new therapy to reduce DH.

Effect of Photobiomodulation on Periodontal Ligament Cells Under Inflamed and Nutrient-Deficient Conditions Simulating Damaged Cells of Avulsed Teeth: An In Vitro Study.

Background: The prognosis of replanted teeth is depended on the vitality of periodontal ligament cells residual on the root surface. Photobiomodulation has photobiological effects that can promote cell vitality. The study aimed to explore the effect of photobiomodulation on the periodontal ligament cells under inflamed or starved conditions mimicking clinically damaged periodontal ligament cells of avulsed teeth and provide the adjuvant procedure for tooth replantation. Materials and methods: Normal, starved, or inflamed periodontal ligament cells were irradiated with an 808 nm laser at densities of 0, 1, 3, 5, or 10 J/cm2. The cell counting kit-8 (CCK-8) assay and scratch test were applied to determine the effects on the proliferation and migration of cells. Anti-inflammatory effects were assessed according to the mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) measured by reverse-transcription polymerase chain reaction. Osteogenic capacity was evaluated by alkaline phosphatase (ALP) staining, ALP activity assay, Alizarin Red S staining, and ALP and osteocalcin (OCN) mRNA expression. Results: The CCK-8 assay and scratch test demonstrated that the 808 nm laser significantly promoted proliferation and migration of normal condition periodontal ligament cells at a density of 3 J/cm2 versus 5 J/cm2 under the starved and inflamed conditions. Moreover, the 808 nm laser had anti-inflammatory effects and promoted osteogenesis of periodontal ligament cells at 3 J/cm2 under normal conditions, while photobiomodulation at 5 J/cm2 upregulated the osteogenesis of periodontal ligament cells under starved and inflamed conditions. Conclusions: The photobiomodulation of 808 nm laser reduced inflammation and improved the proliferation, migration, and osteogenesis of normal, starved, and inflamed periodontal ligament cells. These effects required a higher energy density under starved or inflamed conditions compared with normal conditions. The photobiomodulation of 808 nm has a potential application in root surface treatment for replanted teeth.

Effects of Rose Bengal- and Methylene Blue-Mediated Potassium Iodide-Potentiated Photodynamic Therapy on Enterococcus faecalis: A Comparative Study.

BACKGROUND AND OBJECTIVES: This study was performed to compare the use of methylene blue (MB) and rose bengal (RB) in antimicrobial photodynamic therapy (PDT) targeting Enterococcus faecalis (E. faecalis) bacteria in planktonic and biofilm forms with potassium iodide (KI) potentiation. STUDY DESIGN/MATERIALS AND METHODS: E. faecalis bacteria in planktonic form were exposed to antimicrobial PDT protocols activating MB and RB, with or without KI potentiation, following laser irradiation with different exposure times, 60 mW/cm2 laser power, and different photosensitizer agent (PS)/potentiator concentrations to observe relationships among the variables. Two continuous-wave diode lasers were used for irradiation (red light: λ = 660 nm and green light: λ = 565 nm). The pre-irradiation time was 10 minutes. The vitality of E. faecalis biofilm was assessed by confocal laser scanning microscopy, and the morphology was determined by scanning electron microscopy. The effects on the proliferation of stem cells from the apical papilla (SCAPs) were analyzed by cell counting kit-8 assay. The staining effect of antimicrobial PDT on dentin slices was investigated. Statistical analysis using a one-way analysis of variance was done. RESULTS: KI-potentiated RB and MB antimicrobial PDT both effectively eradicated E. faecalis bacteria in planktonic and biofilm forms. The minimum bactericidal concentrations of PSs (±100 mM KI) were obtained through PDT on planktonic E. faecalis, and the optimal light parameters were 60 mW/cm2 , 6 J/cm2 for 100 seconds. KI-potentiated PDT effectively strengthened the ability to inhibit E. faecalis biofilm with 86.50 ± 5.78% for MB (P = 0.0015 < 0.01) and 91.50 ± 1.75% for RB (P = 0.0418 < 0.05) of bactericidal rate, with less toxicity for SCAPs (P < 0.001) and less staining. KI could reduce the staining induced by antimicrobial PDT on dentin slices. CONCLUSION: A combination of KI and antimicrobial PDT may be a useful alternative to conventional disinfection methods in endodontic treatment. MB and RB antimicrobial PDT at much lower concentrations with KI could hopefully achieve disinfection effects comparable with those of 1.5% NaClO while causing few adverse effects on SCAPs. KI helps to avoid staining problems associated with high concentrations of photosensitizer agents. Lasers Surg. Med. © 2020 Wiley Periodicals, LLC.

Antimicrobial Activity of an Implantable Wireless Blue Light-Emitting Diode Against Root Canal Biofilm In Vitro.

Objective: We developed an implantable wireless blue micro light-emitting diode (micro-LED) device and evaluated the utility of continuous antimicrobial blue light (aBL) irradiation emitted from this micro-LED for root canal disinfection. Methods: An implantable wireless blue micro-LED device (peak wavelength: 410 nm, maximum power: 15 mW) was developed to be placed in the root canal. Optical transmission of the device in human dentin tissue was simulated using Monte Carlo ray-tracing method. The bactericidal effect of low-level aBL on planktonic root canal infection-related bacteria [Enterococcus faecalis, methicillin-resistant Streptococcus aureus (MRSA), and Prevotella intermedia] was evaluated by colony counting. The biocompatibility of continuous low-level aBL exposure was evaluated by infrared thermal imaging and cell viability tests. Thirty extracted intact human single-rooted teeth were prepared and the root canals were infected with E. faecalis for 14 days to form biofilm. The infected root canals were randomly divided into three groups (n = 10), and treated with normal saline (group NS), calcium hydroxide (group CH), and micro-LED device (group aBL) for 3 and 7 days. The bactericidal effect of each group was evaluated by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Results: Monte Carlo simulation showed that blue light irradiation of the micro-LED device decreased exponentially with the light transmission distance through human dentin tissue. Planktonic E. faecalis, MRSA, and P. intermedia were significantly eliminated after irradiation with 432, 36, and 1.35 J/cm2 aBL, respectively (p < 0.05). Infrared thermal imaging and cell viability tests showed that continuous aBL exposure is biocompatible in vitro. CLSM and SEM analyses revealed that the micro-LED device had a greater antimicrobial effect than CH on E. faecalis biofilm in the root canal. Conclusions: The wireless blue micro-LED device is a promising and user-friendly approach for root canal disinfection that will facilitate infection control in the root canal using aBL.

Photobiomodulation therapy for management of inferior alveolar nerve injury post-extraction of impacted lower third molars.

PURPOSE: Inferior alveolar nerve (IAN) injury is one of the most serious complications after extraction of impacted lower third molars. Photobiomodulation (PBM) therapy has been noted to reduce pain and inflammation while promoting tissue healing. This study examined the efficacy of PBM therapy tested in a case series of patients with postoperative IAN injury. MATERIAL AND METHODS: 20 patients with post-extraction IAN injury were involved in this study and divided into two groups. In the study group, PBM therapy (808-nm laser, 16 mW, 3 J/cm2) was used every other day for 2 weeks solely on post-extraction sockets in 10 patients diagnosed with IAN injury. In the control group, mecobalamine was prescribed to 10 patients with IAN injury. Objective and subjective recovery of IAN paresthesia was evaluated using clinical neurosensory testing and visual analog score. RESULTS: All patients showed improvement in both objective and subjective examination. Notably, the visual analog score was significantly improved after PBM treatment compared to the mecobalamine treatment (p < 0.05). CONCLUSION: PBM therapy with 808-nm laser appears to be an effective approach to manage paresthesia post-IAN injury following impacted third molar surgery. Given the limited sample size in this study, large-scale, placebo-controlled, multi-center randomized controlled trials are needed for further validation of this innovative treatment.

Potassium iodide enhances the photobactericidal effect of methylene blue on Enterococcus faecalis as planktonic cells and as biofilm infection in teeth.

OBJECTIVE: To explore the effectiveness, biosafety, photobleaching and mechanism of antimicrobial photodynamic therapy (aPDT) using methylene blue (MB) plus potassium iodide (KI), for root canal infections. METHODS: Different combinations and concentrations of MB, KI and 660 nm LED light were used against E. faecalis in planktonic and in biofilm states by colony-forming unit (CFU), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM). Human gingival fibroblasts (HGF) were used for safety testing by Cell Counting Kit-8 (CCK8) and fluorescence microscopy (FLM). The photobleaching effect and mechanisms were analyzed. RESULTS: KI could not only enhance MB aPDT on E. faecalis in both planktonic and biofilm states even in a hypoxic environment, but also produced a long-lasting bactericidal effect after end of the illumination. KI could accelerate photobleaching to reduce tooth staining by MB, and the mixture was harmless for HGFs. Mechanistic studies showed the generation of hydrogen peroxide and free iodine, and iodine radicals may be formed in hypoxia. CONCLUSION: aPDT with MB plus KI could be used for root canal disinfection and clinical studies are worth pursuing.

[Changes of dentin, dental pulp and periodontium tissue in tail-suspended rats].

OBJECTIVE: To investigate the metabolic changes of calcium and phosphorus in dentin, dental pulp and periodontium in tail-suspended rats, and the functions of TGF-beta 1, c-fos, collagen-I and collagen IV in dentin, dental pulp and periodontium. METHOD: Relative percentage contents of Ca, P in dentin, dental pulp and periodontium were measured with scanning electron microscope and energy spectrum analytical system in 3 groups of rats. The expression of TGF-beta 1, c-fos, collagen-I and collagen IV were also observed. RESULT: In the suspension group, the relative percentage content of Ca declined significantly, while P increased slightly. There were no significant differences of Ca, P in alveolar bone. The expressions of TGF-beta 1, c-fos and collagen-I declined, but the expression of collagen-IV in pulp vessel increased. There were no significant changes of expressions of TGF-beta 1, c-fos, collagen-I and collagen-IV in the vicinity of PDL. After adopting artificial countermeasures, the above expressions restored partly. CONCLUSION: Weightlessness might cause abnormal mineralization in dentin, and 1.5 G artificial countermeasures could eliminate the above changes of mineral metabolism. The poor mineralization of dentin might be associated with the reduced secretion of TGF-beta 1, c-fos and collagen-I in tail-suspended rats.