The influence of growth medium on the photodynamic susceptibility of Aggregatibacter actinomycetemcomitans to antimicrobial blue light.

The aim of this study was to investigate whether antimicrobial blue light (aBL) can cause the death of Aggregatibacter actinomycetemcomitans (A.a) and to determine the influence of different culture media, specifically brain heart infusion and blood agar, on bacterial survival fraction. An LED emitting at 403 ± 15 nm, with a radiant power of 1W, irradiance of 588.2 mW/cm2, and an irradiation time of 0 min, 1 min, 5 min, 10 min, 30 min, and 60 min, was used. The plates were incubated in microaerophilic conditions at 37 °C for 48 h, and the colony-forming units were counted. The photosensitizers were investigated using spectroscopy and fluorescence microscopy. There was no significant difference between the culture media (p > 0.05). However, a statistical reduction in both media was observed at 30 min (1058 J/cm2) (p < 0.05). The findings of this study suggest that aBL has the potential to kill bacteria regardless of the culture media used. Light therapy could be a promising and cost-effective strategy for preventing periodontal disease when used in combination with mechanical plaque control.

Photobiomodulation therapy preconditioning modifies nitric oxide pathway and oxidative stress in human-induced pluripotent stem cell-derived ventricular cardiomyocytes treated with doxorubicin.

Doxorubicin (DOX) is an anthracycline antibiotic that exhibits high heart toxicity. Human-induced pluripotent stem cell-derived ventricular cardiomyocytes (hiPSC-vCMs) are important in vitro models for testing drug cardiotoxicity. Photobiomodulation therapy (PBMT) is a non-invasive therapy that stimulates cells growth and self-repair using light irradiation. This study aimed to investigate the in vitro effects of PBMT preconditioning on cardiotoxicity induced by DOX. HiPSC-vCMs were treated with PBMT for 500 s, followed by the addition of 2 µM DOX. LED irradiation preconditioning parameters were at 660 nm with an irradiance of 10 mW/cm2, performing 5 J/cm2, followed by 24-h DOX exposure (2 µM). Human iPSC-vCMs treated with 2 µM DOX or irradiated with PBMT composed the second and third groups, respectively. The control group did neither receive PBMT preconditioning nor DOX and was irradiated with a white standard lamp. Cells from all groups were collected to perform mRNA and miRNA expressions quantification. PBMT, when applied before the DOX challenge, restored the viability of hiPSC-vCMs and reduced ROS levels. Although downregulated by DOX, myocardial UCP2 mRNA expression presented marked upregulation after PBMT preconditioning. Expression of eNOS and UCP2 mRNA and NO production were decreased after DOX exposure, and PBMT preconditioning before the DOX challenge reversed these changes. Moreover, our data indicated that PBMT preconditioning lowered the miR-24 expression. Our data suggested that PBMT preconditioning ameliorated in vitro DOX-induced cardiotoxicity on transcription level, restoring NO levels and reducing oxidative stress.

Effects of photobiomodulation in salivary glands of chronic kidney disease patients on hemodialysis.

This randomized placebo-controlled trial evaluates the impact of photobiomodulation (PBMT) on the salivary flow and biochemistry of patients with chronic kidney disease (CKD) on hemodialysis. Forty-four patients on hemodialysis self-responded two questionnaires for oral health and salivary gland function perception. The subjects were evaluated for function of salivary glands and randomly allocated to two groups: PBMT group (three irradiations at 808 nm, 100 mW, 142 J/cm2, and 4 J per site); and placebo group. Patients were submitted to non-stimulated and stimulated sialometry and after the treatment at baseline and 14 days. Salivary volume and biochemical of the saliva were analyzed. At baseline, most subjects had self-perception of poor oral health (52.6%) and salivary dysfunction (63.1%). Clinical exam revealed that 47.3% of subjects presented dry mucosa. PBMT promoted increase of the non-stimulated (p = 0.027) and stimulated saliva (p = 0.014) and decrease of urea levels in both non-stimulated (p = 0.0001) and stimulated saliva (p = 0.0001). No alteration was detected in total proteins and calcium analysis. Patients with kidney disease can present alteration in flow, concentrations, and composition of saliva, affecting oral health, but our findings suggest that PBMT is effective to improve hyposalivation and urea levels in saliva of patients with CKD.

Antimicrobial photodynamic therapy with Bixa orellana extract and blue LED in the reduction of halitosis-A randomized, controlled clinical trial.

BACKGROUND: This study aimed to evaluate the reduction of halitosis when using antimicrobial photodynamic therapy (aPDT) with Bixa orellana extract and blue light-emitting diode (LED). METHODS: Forty-four UNINOVE students or employees with a diagnosis of sulfide (H2S) ≥ 112 ppb in gas chromatography were selected. The patients were randomly divided in groups: Group 1 (n = 15): aPDT with annatto and LED; Group 2 (n = 14): tongue scraping; Group 3 (n = 15): tongue scraping and aPDT. For aPDT, a wasBixa orellana extract used in a concentration of 20 % w/v (Fórmula e Ação®, São Paulo, Brazil) on the tongue for 2 min, associated with a blue-violet LED (Valo Cordless Ultradent® Products, Inc., South Jordan, UT, USA) (395-480 nm). Six points were irradiated on the back of the tongue, at wavelength 395-480 nm for 20 s, energy of 9.6 J and radiant energy of 6.37 J/cm2 per point. The results were compared before, immediately after treatment and 7 days after. The Friedman test was used for the intragroup analysis and the Kruskal Wallis test for the intergroup analysis. RESULTS: In all groups, there was a difference between baseline and the value immediately after the treatment. In Groups 1 and 3, there was no difference between the baseline and the 7 days control. CONCLUSION: There was an immediate reduction of halitosis, but the reduction was not maintained after 7days.

Photodynamic antimicrobial chemotherapy has an overt killing effect on periodontal pathogens? A systematic review of experimental studies.

The periodontal disease (PD) etiology is mainly associated with some bacterial strains, such as Porphyromonas gingivalis (P. gingivalis). Nonsurgical root scaling (e.g., antibiotics) may achieve a temporary decrease in the P. gingivalis level, yet it cannot eradicate the microorganism. Moreover, antibiotics can lead to bacterial resistance and undesirable side effects. This systematic review was performed to identify animal data defining antimicrobial photodynamic therapy (PACT) role on experimental PD models in the treatment of P. gingivalis. Embase, MEDLINE, and PubMed were examined for studies published from January 1980 to August 2018. MeSH terms and Scopus data were used to find more related keywords. Four studies were selected and reviewed by two independent researches with a structured tool for rating the research quality. The beneficial effect of PACT included reductions in P. gingivalis counts, bleeding on probing, redness, and inflammation on multiple sites (i.e., first molar, dental implants; subgingival; and mandibular premolars). Although our results suggest that PACT displays antimicrobial action on P. gingivalis, thus improving the PD, a nonuniformity in the PACT protocol and the limited number of studies included lead to consider that the bactericidal efficacy of PACT against periodontal pathogens in PD remains unclear.

Photodynamic therapy with Bixa orellana extract and LED for the reduction of halitosis: study protocol for a randomized, microbiological and clinical trial.

BACKGROUND: Halitosis is an unpleasant breath odour that can interfere with the professional life, social life and quality of life of people who suffer from it. A modality of treatment that has been increasing in dentistry is antimicrobial photodynamic therapy (aPDT). Bixa orellana, popularly known as "urucum" is a plant native to Brazil. The seeds are used to produce a dye that is largely used in the food, textile, paint and cosmetic industries. The aim of this study is to verify whether aPDT with Bixa orellana extract and blue light-emitting diodes (LEDs) is effective in reducing halitosis. This method will also be compared with tongue scraping, the most commonly used conventional method for tongue coating removal, and the association of both methods will be evaluated. METHODS/DESIGN: A randomized clinical trial will be conducted at the dental clinic of the Universidade Nove de Julho. Thirty-nine patients will be divided by block randomization into three groups (n = 13) according to the treatment to be performed. In Group 1, tongue scraping will be performed by the same operator in all patients for analysis of the immediate results. Patients will also be instructed on how to use the scraper at home. Group 2 will be treated with aPDT with Bixa orellana extract and the LED light curing device: Valo Cordless Ultradent®. Six points in the tongue dorsum with a distance of 1 cm between them will be irradiated. The apparatus will be pre-calibrated at wavelength 395-480 nm for 20 s and 9.6 J per point. In Group 3, patients will be submitted to the tongue scraping procedure, as well as to the previously explained aPDT. Oral air collection with the Oral Chroma™ and microbiological collections of the tongue coating shall be done before, immediately after and 7 days after treatment for comparison. DISCUSSION: Halitosis treatment is a topic that still needs attention. The results of this trial could support decision-making by clinicians regarding aPDT using blue LEDs for treating halitosis on a daily basis, as most dentists already have this light source in their offices. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03346460 . Registered on 17 November 2017.

Low Intensity laser therapy in patients with burning mouth syndrome: a randomized, placebo-controlled study.

The aim of this study was to assess the effectiveness of low intensity laser therapy in patients with Burning Mouth Syndrome (BMS). Thirty BMS subjects were randomized into two groups - Laser (LG) and Placebo (CG). Seven patients dropped out, leaving 13 patients in LG and 10 patients in CG. Each patient received 4 irradiations (laser or placebo) twice a week, for two consecutive weeks (blinded to the type of irradiation received). Infrared laser (AsGaAI) irradiations were applied to the affected mucosa in scanning mode, wavelength of 790 nm, output power of 20 mW and fluence of 6 J/cm2. A visual analogue scale (VAS) was used to assess the therapeutic effect before and after each irradiation, and at all the control time periods: 7, 14, 30, 60 and 90 days after the last irradiation. One researcher delivered irradiation and another recorded the results. Both researchers were blinded, the first to the results, and the second to the type of radiation applied. The results were categorized according to the percentage of symptom level variation, and showed a statistically better response in LG in only two categories of the control checkpoints (p=0.02; Fisher's Exact Test). According to the protocol used in this study, low intensity laser therapy is as beneficial to patients with BMS as placebo treatment, indicating a great emotional component of involvement in BMS symptomatology. Nevertheless, there were positive results in some statistical analyses, thus encouraging further research in BMS laser therapy with other irradiation parameters.

Low Intensity laser therapy in patients with burning mouth syndrome: a randomized, placebo-controlled study

Abstract The aim of this study was to assess the effectiveness of low intensity laser therapy in patients with Burning Mouth Syndrome (BMS). Thirty BMS subjects were randomized into two groups - Laser (LG) and Placebo (CG). Seven patients dropped out, leaving 13 patients in LG and 10 patients in CG. Each patient received 4 irradiations (laser or placebo) twice a week, for two consecutive weeks (blinded to the type of irradiation received). Infrared laser (AsGaAI) irradiations were applied to the affected mucosa in scanning mode, wavelength of 790 nm, output power of 20 mW and fluence of 6 J/cm2. A visual analogue scale (VAS) was used to assess the therapeutic effect before and after each irradiation, and at all the control time periods: 7, 14, 30, 60 and 90 days after the last irradiation. One researcher delivered irradiation and another recorded the results. Both researchers were blinded, the first to the results, and the second to the type of radiation applied. The results were categorized according to the percentage of symptom level variation, and showed a statistically better response in LG in only two categories of the control checkpoints (p=0.02; Fisher's Exact Test). According to the protocol used in this study, low intensity laser therapy is as beneficial to patients with BMS as placebo treatment, indicating a great emotional component of involvement in BMS symptomatology. Nevertheless, there were positive results in some statistical analyses, thus encouraging further research in BMS laser therapy with other irradiation parameters.

The influence of red laser irradiation timeline on burn healing in rats.

Low-level laser therapy (LLLT) promotes biomodulation of wound healing and literature reports that light delivery during the inflammation could play a different role compared with latter phases of the healing process. The objective of this study was to investigate whether single dose of a red laser (λ = 660 nm) is different from fractionated delivery protocol in full thickness burns. Two lesions were inflicted on the back of 36 rats. In the fractionated dose group (FG), the lesions were irradiated with 1 J/cm² on days 1, 3, 8, and 10 post-wounding. In the single dose group (SG), the lesions were irradiated with 4 J/cm² on day 1, immediately after injury. Control lesions (CG) received no light and were left to heal spontaneously. Blood flow was measured on days 1, 3, 8, 10, 15, and 21 using laser Doppler flowmetry. Animals were killed on days 3, 8, 10, 15, and 21. Skin specimens were obtained and routinely processed for hematoxylin and eosin. The specimens were evaluated according to differential leukocyte counting and angiogenesis. Statistical analysis was performed, and significance was accepted at p < 0.05. Irradiated groups showed a peak of new vessels on day 15 while, for CG, the peak was on day 21. On day 21, FG exhibited a significantly greater number of cumulative neutrophils while SG showed a higher number of mononuclear cells. Our results confirm that both protocols used accelerate angiogenesis and stimulate leukocyte chemotaxis on burn treatment. In addition, this work suggests that a single-dose LLLT accelerates the inflammatory phase of skin repair.

Antimicrobial photodynamic therapy on drug-resistant Pseudomonas aeruginosa-induced infection. An in vivo study.

Pseudomonas aeruginosa is considered one of the most important pathogens that represent life-threatening risk in nosocomial environments, mainly in patients with severe burns. Antimicrobial photodynamic therapy (aPDT) has been effective to kill bacteria. The purpose of this study was to develop a burn wound and bloodstream infection model and verify aPDT effects on it. In vitro, we tested two wavelengths (blue and red LEDs) on a clinical isolate of P. aeruginosa strain with resistance to multiple antibiotics using HB:La(+3) as photosensitizer. Verapamil(®) associated to aPDT was also studied. In vivo, P. aeruginosa-infected burned mice were submitted to aPDT. Bacterial counting was performed on local infection and bloodstream. Survival time of animals was also monitored. In this study, aPDT was effective to reduce P. aeruginosa in vitro. In addition, Verapamil(®) assay showed that HB:La(+3) is not recognized by ATP-binding cassete (ABC) efflux pump mechanism. In the in vivo study, aPDT was able to reduce bacterial load in burn wounds, delay bacteremia and keep the bacterial levels in blood 2-3 logs lower compared with an untreated group. Mice survival was increased on 24 h. Thus, this result suggests that aPDT may also be a novel prophylactic treatment in the care of burned patients.

Low-level laser therapy in burning mouth syndrome patients: a pilot study.

OBJECTIVE: The aim of this study was to investigate the effect of low-level laser therapy (LLLT) on the treatment of burning mouth syndrome (BMS). In addition, the laser effect was compared on the different affected oral sites. MATERIALS AND METHODS: Eleven subjects with a total of 25 sites (tongue, lower lip, upper lip, and palate) affected by a burning sensation were selected. The affected areas were irradiated once a week for three consecutive weeks with an infrared laser (λ = 790 nm). The probe was kept in contact with the tissue, and the mucosal surface was scanned during the irradiation. The exposure time was calculated based on the fluence of 6 J/cm(2), the output power of 120 mW, and the area to be treated. Burning intensity was recorded through a visual analog scale before and after the treatment and at the 6-week follow-up. The percentage of the improvement in symptoms was also obtained. RESULTS: Burning intensity at the end of the laser therapy was statistically lower than at the beginning (p < 0.01). Patients reported an 80.4% reduction in the intensity of symptoms after laser treatment. There was no statistical difference between the end of the treatment and the 6-week follow-up, except for the tongue site. CONCLUSION: Under the investigated parameters, infrared LLLT proved to be a valuable alternative for BMS treatment, providing a significant and lasting reduction in symptoms.

Low-intensity red laser on the prevention and treatment of induced-oral mucositis in hamsters.

OBJECTIVE: The purpose of this study was to investigate the effects of laser phototherapy as preventive and therapeutic regime on induced-oral mucositis in hamsters. DESIGN: The animals were divided into four groups: preventive cryotherapy, preventive laser, therapeutic laser and therapeutic control group. Mucositis was induced in hamsters by intraperitoneal injection of 5-fluorouracil (5-FU) and superficial scratching. All preventive treatment was performed on the right cheek pouch mucosa. The left pouch mucosa was used for a spontaneous development of mucositis and did not receive any preventive therapy. Laser parameters were: lambda=660nm, P= 30mW, D=1.2J/cm(2), Deltat=40s, spot size 3mm(2), I=1W/cm(2). Cryotherapy was done positioning ice packs in the hamster mucosa 5min before 5-FU infusion and 10min afterward. To study the healing of mucositis, the left pouch mucosa of each of the hamsters in the TLG received laser irradiation on the injured area. Irradiation parameters were kept the same as abovementioned. The control hamsters in the TCG did not receive any treatment. The mucositis degree and the animal's body mass were evaluated. An assessment of blood vessels was made based on immunohistochemical staining. RESULTS: The CG animals lost 15.16% of theirs initial body mass while the LG animals lost 8.97% during the first 5 days. The laser treated animals had a better clinical outcome with a faster healing, and more granulation tissue. The quantity of blood vessels at both LG and CG were higher than in healthy mucosa. Regarding the therapeutic analysis, the severity of the mucositis in the TLG was always lower than TCG. TLG presented higher organization of the granulation tissue, parallel collagen fibrils, and increased angiogenesis. CONCLUSION: The results suggest that laser phototherapy had a positive effect in reducing mucositis severity, and a more pronounced effect in treating established mucositis.

Clinical study of the gingiva healing after gingivectomy and low-level laser therapy.

OBJECTIVE: The purpose of this study was to investigate gingival healing after gingivectomy and adjunctive use of low-level laser therapy (LLLT). BACKGROUND DATA: LLLT has been used in animal experiments to examine the influence of laser radiation on the wound healing process since the 1960s. However, clinical trials in dentistry are scarce, and most of them refer to treatment after extraction of the third molars, with only a few reports in the area of periodontics. METHODS: Twenty patients with periodontal disease were selected, and treatment was planned for gingivectomy to bilateral maxillary and mandibular premolar teeth. After surgery, one side was submitted to LLLT using a 685-nm wavelength, output power of 50 mW, and energy density of 4 J/cm(2). The other side was used as the control and did not receive laser irradiation. Healing was evaluated, clinically and biometrically, immediately post-surgery and at days 3, 7, 14, 21, 28, and 35. Results were submitted to statistical analysis. RESULTS: Biometrical evaluation indicated a significant improvement in healing for the laser group at 21 and 28 days. Clinical evaluation showed better repair for the laser group, mainly after the third day. CONCLUSION: LLLT was an effective adjunctive treatment that appeared to promote healing following gingivectomy.