Effects of Violet and IR LED Light on mast cell degranulation: in vivo study in a murine model.

The aim of this study was to evaluate the influence of IR (λ850 ± 10 nm) and violet (λ405 ± 10 nm) LED phototherapy on total mast cells counts and its ability to influence mast cell degranulation. For this, 27 Wistar rats were used and were randomly distributed into three groups: control, IR LED, and violet LED. When indicated, irradiation done and they were sacrificed, had their tongue removed immediately, 20-min, 45-min, and 2-h after irradiation. Samples were processed to wax, cut, and stained with Toluidine Blue. Intact and degranulated mast cells were counted under light microscopy, and statistical analysis was carried out. In the superficial connective tissue and muscular tissues, violet LED light caused a significant increase in both total number and degranulated mast cells when compared to the control group immediately after irradiation. The degranulation indexes were higher in the groups irradiated with Violet light, both in superficial connective tissue and muscular tissues in relation to the timing. Irradiation with IR LED caused immediate increase in the total number and degranulated of mast cells when compared to the control group only in the superficial connective tissue. In all times observed, the highest total amount of mast cells was seen immediately after irradiation, except in the muscular tissue, which presented the highest amount after 20-min. It was concluded that IR and violet LED light were able to increase the number of mast cells and inducing degranulation in oral mucosa. However, considering that violet LED light can be harmful in periodontal disease, it seems that the use of IR LED light could be the best option in Dentistry.

Effect of low-power diode laser on infected root canals

Abstract This study evaluated the effect of photodynamic therapy (PDT) on infected root canals. Twenty-one human teeth were selected, and 18 were infected by E. faecalis for 60 days. The antimicrobial strategies tested were: G1. Root canal preparation (RCP) using Niquel-Titanium (NiTi) rotary instruments, 2.5% NaOCl, and final irrigation with 17% EDTA, followed by PDT with methylene blue photosensitizer and laser diode low power; G2. RCP using stainless steel files and the same irrigation and PDT protocols as G1; G3. Same RCP protocol as G1 without PDT; G4. Only irrigation with 2.5% NaOCl; G5. Same PDT protocol as G1 without RCP; G6. Negative control; G7. Positive control. Samples for microbiological tests were collected initially (S1), after RCP (S2), and after PDT (S3). Subsequently, the roots were sectioned and prepared for Scanning Electron Microscopy (SEM) analysis. Bacterial growth was analyzed according to the turbidity of the culture medium, followed by spectrophotometric optical density (nm). The effect of PDT on the dentinal structure was evaluated at magnifications 1,600X and 5,000X and described qualitatively. The Wilcoxon test was used for the comparisons from the same specimens, and the Mann-Whitney test was used to compare groups ((=5%). Bacteria were found in all experimental groups' microbiological samples (S1, S2 and S3). The optical density of culture media was lower in S2 than in S1 of G1, 2, 3, and 4 (p> 0.05). After PDT (S3) in G1 and 2, there was an additional reduction in optical density of the culture medium, respectively (p>0.05). In Group 5, the analysis of culture media at S2 revealed an increase in optical density compared to S1(p>0.05). In SEM images of G1, 2, and 5, dentin with melting and recrystallization areas were evidenced. After preparation of the root canal with the rotary system or manually associated with 2.5% NaOCl, PDT was not able to completely eliminate E. faecalis present in the root canal.

Resumo Este estudo avaliou o efeito da terapia fotodinâmica (PDT) em canais radiculares infectados com E. faecalis. Vinte e um dentes humanos extraídos foram selecionados, e 18 foram infectados por E. faecalis por 60 dias. As estratégias antimicrobianas testadas foram: G1. Preparo do canal radicular (PCR) com instrumentos rotatórios de NiTi, NaOCl 2,5% e irrigação final com EDTA 17%, seguido de PDT com fotossensibilizador azul de metileno e laser diodo de baixa potência; G2. PCR usando limas de aço inoxidável e os mesmos protocolos de irrigação e PDT do G1; G3. Protocolo de PCR similar que G1 sem PDT; G4. Somente irrigação com NaOCl 2,5%; G5. Protocolo similar ao G1, sem PCR; G6. Controle negativo; G7. Controle positivo. Amostras para exames microbiológicos foram coletadas inicialmente (S1), após PCR (S2) e após PDT (S3). Na sequência, as raízes foram seccionadas e preparadas para análise em microscopia eletrônica de varredura (MEV). O crescimento bacteriano foi analisado de acordo com a turbidez do meio de cultura seguida pela densidade óptica espectrofotométrica (nm). O efeito da PDT na estrutura dentinária foi avaliado em aumentos de 1.600X e 5.000X, e descrito qualitativamente. O teste de Wilcoxon foi utilizado para as comparações dos mesmos espécimes e o teste de Mann-Whitney para as comparações entre os grupos ((=5%). Bactérias foram encontradas em todos os grupos experimentais, e em todas as coletas microbiológicas (S1, S2 e S3). A densidade óptica dos meios de cultura foi menor em S2 do que em S1 de G1, 2, 3 e 4 (p>0,05). Após a PDT (S3) em G1 e 2, houve redução adicional na densidade óptica do meio de cultura de 90,0% e 92,0%, respectivamente (p>0,05). No Grupo 5, a análise dos meios de cultura em S2 revelou um aumento de 3,2% na densidade óptica em comparação com S1(p>0,05). Nas imagens de MEV do G1, 2 e 5 foram evidenciadas dentina com áreas de fusão e recristalização. O PDT utilizado após preparo do canal radicular com sistema rotatório ou manual, associado ao NaOCl 2,5%, não foi capaz de eliminar completamente o E. faecalis em biofilme maduro presente no canal radicular.

Evaluation of the Antimicrobial Effect of Photodynamic Therapy and Er:YAG Laser Irradiation on Root Canals Infected with Enterococcus faecalis.

Objective: To carry out a histological and morphometric analysis of the antimicrobial effect of Er:YAG laser irradiation combined with photodynamic therapy (PDT) on root canals infected with Enterococcus faecalis. Background: PDT and Er:YAG laser irradiation may be alternatives for effective endodontic disinfection but there are no data on the combination of these therapies. Materials and methods: Forty single-rooted bovine teeth had their roots contaminated with E. faecalis for 72 h. The teeth were randomly divided into four groups (n = 10): group 1, irrigation with 2.5% sodium hypochlorite (NaOCl); group 2, Er:YAG laser (λ2940 nm, 15 Hz, 100 mJ); group 3, PDT with 0.07% methylene blue as photosensitizer and laser irradiation (λ660 nm, power 40 mW, 5 min); and group 4, Er:YAG laser + PDT. After treatment, the teeth were examined by confocal laser scanning microscopy to verify bacterial viability, and morphometric analysis of the images was performed. Results: The PDT and Er:YAG + PDT treatments promoted the greatest reduction in bacteria among the proposed therapies, whereas 2.5% NaOCl was the least effective in bacterial elimination. A statistically significant difference (p < 0.05) was observed among the groups studied, except between the group combining Er:YAG and PDT and the group treated with PDT alone. Conclusions: PDT combined or not with Er:YAG laser was found to be more effective in root canal disinfection when compared with the other groups.

Effect of low-power diode laser on infected root canals.

This study evaluated the effect of photodynamic therapy (PDT) on infected root canals. Twenty-one human teeth were selected, and 18 were infected by E. faecalis for 60 days. The antimicrobial strategies tested were: G1. Root canal preparation (RCP) using Niquel-Titanium (NiTi) rotary instruments, 2.5% NaOCl, and final irrigation with 17% EDTA, followed by PDT with methylene blue photosensitizer and laser diode low power; G2. RCP using stainless steel files and the same irrigation and PDT protocols as G1; G3. Same RCP protocol as G1 without PDT; G4. Only irrigation with 2.5% NaOCl; G5. Same PDT protocol as G1 without RCP; G6. Negative control; G7. Positive control. Samples for microbiological tests were collected initially (S1), after RCP (S2), and after PDT (S3). Subsequently, the roots were sectioned and prepared for Scanning Electron Microscopy (SEM) analysis. Bacterial growth was analyzed according to the turbidity of the culture medium, followed by spectrophotometric optical density (nm). The effect of PDT on the dentinal structure was evaluated at magnifications 1,600X and 5,000X and described qualitatively. The Wilcoxon test was used for the comparisons from the same specimens, and the Mann-Whitney test was used to compare groups ((=5%). Bacteria were found in all experimental groups' microbiological samples (S1, S2 and S3). The optical density of culture media was lower in S2 than in S1 of G1, 2, 3, and 4 (p> 0.05). After PDT (S3) in G1 and 2, there was an additional reduction in optical density of the culture medium, respectively (p>0.05). In Group 5, the analysis of culture media at S2 revealed an increase in optical density compared to S1(p>0.05). In SEM images of G1, 2, and 5, dentin with melting and recrystallization areas were evidenced. After preparation of the root canal with the rotary system or manually associated with 2.5% NaOCl, PDT was not able to completely eliminate E. faecalis present in the root canal.

Raman spectroscopy and sciatic functional index (SFI) after low-level laser therapy (LLLT) in a rat sciatic nerve crush injury model.

Axonotmesis causes sensorimotor and neurofunctional deficits, and its regeneration can occur slowly or not occur if not treated appropriately. Low-level laser therapy (LLLT) promotes nerve regeneration with the proliferation of myelinating Schwann cells to recover the myelin sheath and the production of glycoproteins for endoneurium reconstruction. This study aimed to evaluate the effects of LLLT on sciatic nerve regeneration after compression injury by means of the sciatic functional index (SFI) and Raman spectroscopy (RS). For this, 64 Wistar rats were divided into two groups according to the length of treatment: 14 days (n = 32) and 21 days (n = 32). These two groups were subdivided into four sub-groups of eight animals each (control 1; control 2; laser 660 nm; laser 808 nm). All animals had surgical exposure to the sciatic nerve, and only control 1 did not suffer nerve damage. To cause the lesion in the sciatic nerve, compression was applied with a Kelly clamp for 6 s. The evaluation of sensory deficit was performed by the painful exteroceptive sensitivity (PES) and neuromotor tests by the SFI. Laser 660 nm and laser 808 nm sub-groups were irradiated daily (100 mW, 40 s, energy density of 133 J/cm2). The sciatic nerve segment was removed for RS analysis. The animals showed accentuated sensory and neurofunctional deficit after injury and their rehabilitation occurred more effectively in the sub-groups treated with 660 nm laser. Control 2 sub-group did not obtain functional recovery of gait. The RS identified sphingolipids (718, 1065, and 1440 cm-1) and collagen (700, 852, 1004, 1270, and 1660 cm-1) as biomolecular characteristics of sciatic nerves. Principal component analysis revealed important differences among sub-groups and a directly proportional correlation with SFI, mainly in the sub-group laser 660 nm treated for 21 days. In the axonotmesis-type lesion model presented herein, the 660 nm laser was more efficient in neurofunctional recovery, and the Raman spectra of lipid and protein properties were attributed to the basic biochemical composition of the sciatic nerve.

Impact of photobiomodulation therapy on the morphological aspects of submandibular gland submitted to excretory duct ligation and hypothyroidism: an animal study.

The aim of this study was to evaluate the impact of photobiomodulation therapy (PBMT) on histomorphological aspects of submandibular gland (SMG) submitted to salivary gland duct obstruction in hypothyroid rats. Fifty-six male Wistar rats (250 to 300 g) were divided into 4 groups (n = 14): euthyroid (EU), EU + PBMT, hypothyroid (HYPO), and HYPO + PBMT. Duct obstruction of the left submandibular gland (LSMG) was performed in all animals by a ligature procedure. For the induction of hypothyroidism, total thyroidectomy was performed. PBMT groups received irradiation with AlGaInP diode laser (808 nm, 0.04 W, 0.04cm2 spot size, 60 s, 2.4 J per point, 60 J/cm2, 1 W/cm2). Irradiation was performed immediately, 24 h, and 48 h after the obstruction of the salivary gland duct, in one point, extra oral and perpendicular to the gland. Animals were sacrificed after 24 h and 72 h after duct ligature. Our results indicated that salivary duct obstruction and hypothyroidism caused negative modifications on the salivary glands' histomorphology, especially acinar atrophy, after 24 h and 72 h. HYPO + PBMT showed a significant reduction of the inflammatory infiltrate, congested blood vessels, and acinar atrophy in the SMG submandibular salivary gland in 72 h compared to 24 h (p < 0.05). In conclusion, obstruction of the salivary gland excretory duct and hypothyroidism causes severe sialoadenitis with expressive atrophy of the glandular parenchyma. However, PBMT was able to modulate the inflammatory process and delaying acinar atrophy. This study provided insights to better understand the role of the PBMT on the altered salivary gland by duct ligation and associate hypothyroidism.

Photobiomodulation and Pain Reduction in Patients Requiring Orthodontic Band Application: Randomized Clinical Trial.

PURPOSE: The aim of this study was to investigate the effect of Photobiomodulation (PBM) in managing orthodontic pain intensity over time in patients requiring band application on upper first molars. METHODS: Maxillary first molars were banded. In the trial group, each molar received single-session PBM on two buccal and two palatal points (λ = 830 ± 10 nm; 150 mW, 7.5 J/cm2; spot of 0.1 cm2; 5 sec per point), while the control group received a placebo treatment. All patients were asked to answer five pain rating scales to assess pain intensity at 5 minutes and 1, 12, 24, and 72 hours and completed a survey describing the type of pain and its temporal course in the next 7 days. RESULTS: 26 patients (mean age 11.8 years) were randomly assigned to a control or a trial group. The trial group showed significantly lower pain intensities (p < 0.05) at 5 min (M = 0.92, SD = 1.32), 1 h (M = 0.77, SD = 1.01), and 12 h (M = 0.77, SD = 1.54) after band application compared to the control group (5 min: M = 1.62, SD = 1.26; 1 h: M = 1.77, SD = 1.92; and 12 h: M = 1.77, SD = 2.17), whereas no difference between groups (p > 0.05) was found at 24 h (trial: M = 0.62, SD = 1.71; control: M = 1.08, SD = 1.75) and 72 h (trial: M = 0.31, SD = 0.75; control: M = 0.15, SD = 0.55). Patients in the control group reported more frequently the presence of "compressive pain" (58.8%, p < 0.05) from the appliance during the week after the application, while the trial group showed higher frequency of "no pain" (46.2%, p < 0.05). However, PBM did not affect the pain onset (trial: M = 10.86, SD = 26.97; control: M = 5.25, SD = 7.86), peak (trial: M = 15.86, SD = 26.29; control: 6.17, SD = 7.96), and end time (trial: 39.57, SD = 31.33; control: M = 22.02, SD = 25.42) reported by the two groups (p > 0.05). CONCLUSIONS: PBM might be considered a promising alternative to decrease general pain intensity, although not affecting the typical pain cycle, in terms of the onset, peak, and ending times.

Photobiomodulation Therapy in the Proliferation and Differentiation of Human Umbilical Cord Mesenchymal Stem Cells: An In Vitro Study.

Introduction: Since photobiomodulation therapy (PBMT) favors in vitro mesenchymal stem cell (MSC) preconditioning before MSC transplantation, increasing the proliferation of these cells without molecular injuries by conserving their characteristics, in the present in vitro study we analyzed the effect of PBMT on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs). Methods: Irradiation with an InGaAIP Laser (660 nm, 10 mW, 2.5 J/cm2 , 0.08 cm2 spot size, and 10 s) was carried out. The cells were divided into four groups: CONTROL [cells grown in Dulbecco's Modified Eagle Medium (DMEM)], OSTEO (cells grown in an osteogenic medium); PBMT (cells grown in DMEM+PBMT), and OSTEO+PBMT (cells grown in an osteogenic medium plus PBMT). The cell proliferation curve was obtained over periods of 24, 48 and 72 hours using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Osteogenic differentiation was analyzed by the formation of calcium nodules over periods of 7, 14 and 21 days. Morphometric analysis was performed to quantify the total area of nodular calcification. Results: The highest cell proliferation and cell differentiation occurred in the OSTEO+PBMT group, followed by the PBMT, OSTEO and CONTROL groups respectively, at the observed times (P <0.05). Conclusion: PBMT enhanced the osteogenic proliferation and the differentiation of hUCMSCs during the periods tested, without causing damage to the cells and preserving their specific characteristics, a fact that may represent an innovative pretreatment in the application of stem cells.

Anti-Trypanosoma cruzi effect of the photodynamic antiparasitic chemotherapy using phenothiazine derivatives as photosensitizers.

Chagas disease is endemic in Latin America and increasingly found in non-endemic countries. Its treatment is limited due to the variable efficacy and several side effects of benznidazole. Photodynamic antimicrobial chemotherapy (PACT) may be an attractive approach for treating Chagas disease. Here, the trypanocidal activity of PACT was investigated in vitro using phenothiazine derivatives. The cytotoxicity of both, methylene blue (MB) and toluidine blue (TBO), was determined on macrophages cultures using AlamarBlue method. The trypanocidal activity of the two photosensitizers was initially evaluated by determining their IC50 values against trypomastigote forms. After this, the trypanocidal effect was evaluated in cultures of infected macrophages using an automatized image analysis protocol. All experiments were performed in the dark and in the clear phase (after a photodynamic exposure). The compounds showed no cytotoxicity in both phases at the tested concentrations. The IC50 values for the sole use of MB and TBO were 2.6 and 1.2 µM, respectively. The photoactivation of the compounds using a fixed energy density (J/cm2) caused a reduction of the IC50 values to 1.0 and 0.9 µM, respectively. It was found that, on infected macrophage, the use of TBO significantly reduced the number of infected cells and parasitic load, and this effect was increased in the presence of light. The results of the present study are indicative that PACT may be considered as both selective and effective therapeutic intervention for treating Chagas disease.

Photobiomodulation Therapy in Bone Repair Associated with Bone Morphogenetic Proteins and Guided Bone Regeneration: A Histomorphometric Study.

OBJECTIVE: To evaluate the efficacy of photobiomodulation for bone repair of critical surgical wounds with implants of bone morphogenetic proteins (BMPs) and bovine biological membranes, using histological and histomorphometric analysis. BACKGROUND: Tissue engineering has been developing rapidly through the use of various biomaterials for the treatment of bone defects, such as mechanical barriers consisting of biological membranes and implants of biomaterials for bone supply. MATERIALS AND METHODS: Thirty-two male rats were divided into four groups (n = 8): group I-C: control; group II-PT: photobiomodulation therapy; group III-PM: Gen-Pro® BMPs+Gen-Derm® membrane; and group IV-PMPT: Gen-Pro® BMPs+Gen-Derm® membrane+photobiomodulation therapy. A 3 mm bone cavity was performed in the upper third of the lateral surface of the right rat femur to obtain a bone defect considered to be critical. The irradiated groups received seven applications of AlGaAs diode laser 830 nm, P = 40 mW, continuous wave (CW) emission mode, f ∼ 0.6 mm, 4 J/cm2 per point (north, south, east, and west) at 48 h intervals, for a total of 16 J/cm2 per session (final dose: 112 J/cm2). Bone repair was evaluated at sacrifice 15 and 30 days after the procedure. The specimens were embedded in paraffin and stained with hematoxylin and eosin and Picrosirius for analysis by light microscopy and by the Leica interactive measurement module software. Statistical analysis was performed (p < 0.05%). RESULTS: Histological analysis confirmed the histomorphometric results, with the experimental groups showing bone neoformation of significantly higher quality and quantity at the end of 30 days compared with the control group. CONCLUSIONS: Photobiomodulation therapy was effective for bone repair mainly when associated with BMPs and a biological membrane. The results of this study are promising and stimulate further scientific and clinical research.

Does laser phototherapy influence the proliferation of myoepithelial cells in the salivary gland of hypothyroid rats?

Thyroid hormones influence both development and growth of organs and tissues and guarantee metabolic demands that interfere with the quality of digestive secretions, including those of the salivary glands. Laser phototherapy - LPT can modulate various biological phenomena and its diverse effects permit the action on different cell types. The aim of this study was to evaluate the influence of laser phototherapy on myoepithelial cells of salivary glands of hypothyroid rats. Forty-two albino Wistar rats were divided into two main groups: euthyroid (EU) and hypothyroid (HYPO). Hypothyroidism was induced using propylthiouracil (PTU) for 4weeks. Each group was divided into subgroups: control (without laser) and laser groups (Red/infrared - IR). LPT was used on the submandibular gland and was carried out using a diode laser (λ660 or λ780nm, 40mW, spot size 0.04cm2, irradiation area 1cm2, 300s, 6J/cm2 per gland, 12J/cm2 per session) and started two weeks after PTU treatment. LPT was repeated every other day for two weeks. After animal death, the glands were removed, dissected and processed for immunohistochemical analysis. It was observed an increase in the number of myoepithelial cells of hypothyroid control rats in comparison to euthyroid controls (p=0.001). Visible LPT (λ660nm) caused significant higher proliferation of myoepithelial cells in EU rats when compared to IR LPT (λ 780nm)(p≤0.001).It is concluded that, despite the LPT protocol used did not influence myoepithelial proliferation on hypothyroid rats it significantly increased the proliferation on euthyroid animals.

Biochemical changes on the repair of surgical bone defects grafted with biphasic synthetic micro-granular HA + ß-tricalcium phosphate induced by laser and LED phototherapies and assessed by Raman spectroscopy.

This work aimed the assessment of biochemical changes induced by laser or LED irradiation during mineralization of a bone defect in an animal model using a spectral model based on Raman spectroscopy. Six groups were studied: clot, laser (λ = 780 nm; 70 mW), LED (λ = 850 ± 10 nm; 150 mW), biomaterial (biphasic synthetic micro-granular hydroxyapatite (HA) + ß-tricalcium phosphate), biomaterial + laser, and biomaterial + LED. When indicated, defects were further irradiated at a 48-h interval during 2 weeks (20 J/cm2 per session). At the 15th and 30th days, femurs were dissected and spectra of the defects were collected. Raman spectra were submitted to a model to estimate the relative amount of collagen, phosphate HA, and carbonate HA by using the spectra of pure collagen and biomaterials composed of phosphate and carbonate HA, respectively. The use of the biomaterial associated to phototherapy did not change the collagen formation at both 15 and 30 days. The amount of carbonate HA was not different in all groups at the 15th day. However, at the 30th day, there was a significant difference (ANOVA, p = 0.01), with lower carbonate HA for the group biomaterial + LED compared to biomaterial (p < 0.05). The phosphate HA was higher in the groups that received biomaterial grafts at the 15th day compared to clot (significant for the biomaterial; p < 0.01). At the 30th day, the phosphate HA was higher for the group biomaterial + laser, while this was lower for all the other groups. These results indicated that the use of laser phototherapy improved the repair of bone defects grafted with the biomaterial by increasing the deposition of phosphate HA.

Laser and LED phototherapy on midpalatal suture after rapid maxilla expansion: Raman and histological analysis.

The aim of this study was to analyze the effect of laser or LED phototherapy on the acceleration of bone formation at the midpalatal suture after rapid maxilla expansion. Forty-five rats were divided into groups at 7 days (control, expansion, expansion and laser irradiation, and expansion and LED irradiation) and into 14 days (expansion, expansion and laser in the 1st week, expansion and LED in the 1st week, expansion and laser in the 1st and 2nd weeks, expansion and LED in the 1st and 2nd weeks). Laser/LED irradiation occurred every 48 h. Expansion was accomplished with a spatula and maintained with a triple helicoid of 0.020-in stainless steel orthodontic wire. A diode laser (λ780 nm, 70 mW, spot of 0.04 cm2, t = 257 s, SAEF of 18 J/cm2) or a LED (λ850 ± 10 nm, 150 ± 10 mW, spot of 0.5 cm2, t = 120 s, SAEF of 18 J/cm2) was applied in one point in the midpalatal suture immediately behind the upper incisors. Raman spectroscopy and histological analyses of the suture region were carried and data was submitted to statistical analyses (p ≤ 0.05). Raman spectrum analysis demonstrated that irradiation increases hydroxyapatite in the midpalatal suture after expansion. In the histological analysis of various inflammation, there was a higher production of collagen and osteoblastic activity and less osteoclastic activity. The results showed that LED irradiation associated to rapid maxillary expansion improves bone repair and could be an alternative to the use of laser in accelerating bone formation in the midpalatal suture.

Effectiveness of photodynamic therapy on Candida species isolated from oral samples of children exposed and not exposed to HIV / Efetividade da terapia fotodinâmica sobre espécies de Candida isoladas de amostras bucais de crianças expostas e não expostas ao HIV

ABSTRACT Objective: Identify yeast species isolated from unexposed, exposed and HIV-carrier children, and verify the effectiveness of low power laser photodynamic therapy (PDT) on the yeasts species belonging to the Candida genus. Methods: Fifty children assisted by the Public Health Program of the city of Vitória da Conquista, Bahia, were selected and divided into three groups: unexposed to HIV, exposed to HIV during pregnancy, and HIV-carrier. Saliva samples were collected in a disposable sterile universal container and were plated to Sabouraud dextrose agar supplemented with 0.1 mg/mL chloramphenicol. The plates were incubated at 37°C for 48 h. Three strains of each patient were identified by using an API 20 C AUX system. The strains were submitted to photodynamic therapy (PDT) with a 660 nm low power laser and methylene blue dye at different times of irradiation (90, 180 and 282 sec.). Results: The results showed that the most prevalent species was Candida albicans followed by Candida famata (second most prevalent in unexposed to HIV and HIV carriers)) and Candida parapsilosis (second most prevalent in exposed to HIV group). The CFU/mL of Candida spp. decreased significantly (p<0,05) in all groups treated with PDT compared to the controls. Photodynamic therapy treatments at different exposure times (e.g., PS+L90+, PS+L180+, PS+L282+) revealed that the exposure time of 282 sec. gave the highest reduction of the mean logarithmic CFU/mL. Conclusion: Candida albicans was the most prevalent Candida species in these three groups and Candida non-albicans species, when combined, amounted to a significant percentage of Candida isolates. Photodynamic therapy was effective in inactivating the Candida spp. isolated from the oral cavity of children not exposed to HIV, exposed to HIV and HIV-carriers, with the best photodynamic therapy irradiation time being 282 sec.

RESUMO Objetivo: Identificar espécies de leveduras isoladas de crianças não expostas, expostas ao HIV e portadoras de AIDS, e verificar a eficácia da terapia fotodinâmica com laser de baixa potência sobre as espécies de leveduras pertencentes ao gênero Candida. Métodos: Cinquenta crianças atendidas pelo Programa de Saúde Pública da cidade de Vitória da Conquista, Bahia, foram selecionadas e divididas em três grupos: não expostas ao HIV, expostas ao HIV durante a gravidez e portadoras da AIDS. Amostras de saliva foram coletadas em um recipiente universal descartável estéril e foram semeadas em ágar Sabouraud dextrose suplementado com 0,1 mg / mL de cloranfenicol. As placas foram incubadas a 37°C durante 48h. Três cepas de cada paciente foram identificadas utilizando o sistema API 20 C AUX. As cepas foram submetidas à Terapia Fotodinâmica com laser de baixa potência de 660 nm e corante azul de metileno em diferentes tempos de irradiação (90, 180 e 282 segundos). Resultados: Os resultados mostraram que a espécie isolada mais prevalente nos grupos estuddos foi Candida albicans, seguida de Candida famata (segunda mais prevalene nos grupos não expostos ao HIV e com AIDS) e Candida parapsilosis (segunda mais prevalente no grupo exposto ao HIV). Houve diminuição significante de CFU/ml de Candida spp. (p <0,05) em todos os grupos tratados com terapia fotodinâmica, em comparação com os controles. A terapia fotodinâmica, nos diferentes tempos (e.g., PS+L90+, PS+L180+, PS+L282+) mostrou que o tempo de 282 seg. apresentou a maior redução em media de logarítmo de UFC/mL. Conclusão: Candida albicans foi a espécie de Candida mais prevalente isolada nos três grupos e as espécies de Candida não-albicans, quando combinadas, contribuíram com porcentagem significativa dos isolados de Candida. A PDT foi eficaz na inativação de Candida spp. isoladas a partir da cavidade oral de crianças não expostas, expostas ao HIV e portadoras da AIDS, com o melhor tempo de irradiação sendo o de 282 seg.

Estimating the concentration of urea and creatinine in the human serum of normal and dialysis patients through Raman spectroscopy.

Urea and creatinine are commonly used as biomarkers of renal function. Abnormal concentrations of these biomarkers are indicative of pathological processes such as renal failure. This study aimed to develop a model based on Raman spectroscopy to estimate the concentration values of urea and creatinine in human serum. Blood sera from 55 clinically normal subjects and 47 patients with chronic kidney disease undergoing dialysis were collected, and concentrations of urea and creatinine were determined by spectrophotometric methods. A Raman spectrum was obtained with a high-resolution dispersive Raman spectrometer (830 nm). A spectral model was developed based on partial least squares (PLS), where the concentrations of urea and creatinine were correlated with the Raman features. Principal components analysis (PCA) was used to discriminate dialysis patients from normal subjects. The PLS model showed r = 0.97 and r = 0.93 for urea and creatinine, respectively. The root mean square errors of cross-validation (RMSECV) for the model were 17.6 and 1.94 mg/dL, respectively. PCA showed high discrimination between dialysis and normality (95 % accuracy). The Raman technique was able to determine the concentrations with low error and to discriminate dialysis from normal subjects, consistent with a rapid and low-cost test.

Evaluation of laser phototherapy (λ 780 nm) after dental replantation in rats.

BACKGROUND/AIM: Tooth replantation is the treatment of choice in cases of avulsion although the outcomes are variable. The teeth can be lost due to external root resorption. The aim of this study was to histologically assess of the effect of laser phototherapy (λ780 nm) on replanted teeth in rats. MATERIAL AND METHODS: Sixty Wistar Albinus rats had their maxillary right incisors extracted and were then divided into four groups: G1-absence of storage medium; G2-milk as storage medium; G3-milk as storage medium followed by a laser irradiation of the root surfaces and entrance of the alveolus (λ = 780 nm; P = 70 mW; CW; DE = 21 J/cm2 ); G4-milk as storage medium, laser irradiation as in G3 before replantation. After this procedure, laser irradiation was performed on the buccal and palatal mucosa (8.4 J/cm2 per session) every 48 h for 15 days. The animals were euthanized 15, 30, and 60 days after replantation. RESULTS: The histological results showed that after 15 days, G4 exhibited intense chronic inflammation with the presence of clastic cells and moderate external inflammatory root resorption (P < 0.05) when compared with G3, in which these outcomes were not observed. At the 30th day, G1, G2, and G4 showed chronic inflammation varying from discrete to moderate, as well as intense external inflammatory root resorption. G3 remained without any inflammation and external inflammatory root resorption up to the 60th day. CONCLUSIONS: The use of laser phototherapy on the root surface and at the entrance of the alveolus prior to replantation had a positive biomodulative effect on alveolar repair after tooth replantation in rats.

Quantifying creatinine and urea in human urine through Raman spectroscopy aiming at diagnosis of kidney disease.

Due to their importance in the regulation of metabolites, the kidneys need continuous monitoring to check for correct functioning, mainly by urea and creatinine urinalysis. This study aimed to develop a model to estimate the concentrations of urea and creatinine in urine by means of Raman spectroscopy (RS) that could be used to diagnose kidney disease. Midstream urine samples were obtained from 54 volunteers with no kidney complaints. Samples were subjected to a standard colorimetric assay of urea and creatinine and submitted to spectroscopic analysis by means of a dispersive Raman spectrometer (830 nm, 350 mW, 30 s). The Raman spectra of urine showed peaks related mainly to urea and creatinine. Partial least squares models were developed using selected Raman bands related to urea and creatinine and the biochemical concentrations in urine measured by the colorimetric method, resulting in r = 0.90 and 0.91 for urea and creatinine, respectively, with root mean square error of cross-validation (RMSEcv) of 312 and 25.2 mg/dL, respectively. RS may become a technique for rapid urinalysis, with concentration errors suitable for population screening aimed at the prevention of renal diseases.

Assessing the biochemical changes of tendons of rats in an experimental model of tenotomy under therapeutic ultrasound and LEDs (625 and 945 nm) by near-infrared Raman spectroscopy.

Given the complexity of the tendon repair process, where the neoformed scar tissue tends to prevent or hamper the tendon from performing its normal functions, ultrasound (US) and light-emitting diode (LED) devices have been applied to modulate the tendon repair process. This study aimed to compare the anti-inflammatory and healing effects of therapeutic US and LED in an experimental model of tenotomy using histomorphometry and Raman spectroscopy. Forty Wistar rats (235 ± 8.3 g) were separated into four groups: tenotomy (TC; control), tenotomy associated with ultrasound (US; 1 MHz, 0.5 W/cm(2)), tenotomy associated with LED at 625 nm (LR; 4 J/cm(2)), and tenotomy associated with LED at 945 nm (LIR; 4 J/cm(2)), in two experimental periods of 7 and 14 days. Histomorphometry showed a decrease of inflammatory cells (macrophages and polymorphonuclear leukocytes) for the group treated with US compared to the TC group (ANOVA, p < 0.0001) and the LIR group compared to the TC group (p < 0.05) at the seventh day, indicating an anti-inflammatory effect. In relation to the LR and TC groups, there was no significant difference. Raman spectroscopy revealed that the LR group presented an increased synthesis of collagen I compared to the other treatments and the TC groups on the 14th day (p < 0.01). US therapy showed an adjuvant effect with anti-inflammatory action, and the LED therapy at 625 nm showed an increase in collagen synthesis, contributing to the process of tendon healing.

Photodynamic Antimicrobial Chemotherapy (PACT) in osteomyelitis induced by Staphylococcus aureus: Microbiological and histological study.

Osteomyelitis is an inflammation either of medullar spaces or of the surface of cortical bones, which represents a bacterial infection. Photodynamic Antimicrobial Chemotherapy (PACT) is a treatment based on a cytotoxic photochemical reaction that induces a series of metabolic reactions and culminates in bacterial suppression. Such effect led to the idea that it could be used as a treatment of osteomyelitis. Following approval by the Animal Experimentation Committee of the School of Dentistry of the Federal University of Bahia, the present randomized study used eighty Wistar rats with the aim to evaluate, by microbiological and histological analysis, the effects of Photodynamic Antimicrobial Chemotherapy - PACT on tibial surgical bone defects in rats infected by Staphylococcus aureus. The animals were divided in groups: Control (non-infected); Control Osteomyelitis Induction; Saline solution; Photosensitizer; Red Laser and PACT - on this group, a diode laser (40mW; λ660nm ∅=0.04cm(2), CW, 10J/cm(2)) was used in combination with 5µg/ml of toluidine blue as the photosensitizer. On the microbiological study, immediately after treatment, the PACT group presented a bacterial reduction of 97.4% (p<0.001). Thirty days after treatment, there was a bacterial reduction of more than 99.9% (p<0.001). In the histological study, it was observed that the PACT group demonstrated an intense presence of osteocytes and absence of bone sequestration and micro-abscesses. The PACT using toluidine blue was effective in reducing the number of S. aureus enabling a better quality bone repair.

Influence of laser photobiomodulation (GaAlAs) on salivary flow rate and histomorphometry of the submandibular glands of hypothyroid rats.

The aim of this study was to analyze the influence of laser photobiomodulation in salivary flow, weight, and histomorphometry of the submandibular glands of hypothyroid rats. Fifty-six male Wistar albino rats were divided in euthyroid group and hypothyroid group, treated with propylthiouracil (PTU) to induce hypothyroidism. Each group was divided into control (without laser) and laser groups (GaAlAs): λ660 nm (40 mW), λ780 nm (40 mW), and λ780 nm (70 mW). The laser application on the submandibular gland occurred after 2 weeks of PTU treatment and repeatedly during 2 weeks every 48 h. The rats were anesthetized, tracheostomized, and the evaluation of the salivary flow rate (µL/min/100 g body weight) was made by the weight of the saliva collected for 15 min from the first drop. After the animals' death, the glands were dissected and processed for histological analysis. There was an evident reduction of the salivary flow of hypothyroid rats in all groups in comparison to euthyroid group (Mann-Whitney test, p < 0.05). No significant difference was found in the salivary flow of rats that received laser photobiomodulation compared with their control groups. Histological analysis revealed a decrease in the parenchyma of the salivary glands of hypothyroid rats, but the laser was not able to reverse this process. Hypothyroid rats irradiated or not with laser showed acini and acinar cells with significantly smaller areas than euthyroid groups. The laser photobiomodulation protocol used was not able to change salivary flow or reverse the acinar atrophy process in the submandibular glands of hypothyroid rats.