Biostimulatory effect of low-level laser therapy on keratinocytes in vitro

Epithelial cells play an important role in reparative events. Therefore, therapies that can stimulate the proliferation and metabolism of these cells could accelerate the healing process. To evaluate the effects of low-level laser therapy (LLLT), human keratinocytes were irradiated with an InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm; 40 mW) using 0.5, 1.5, 3, 5, and 7 J/cm2 energy doses. Irradiations were done every 24 h totaling three applications. Evaluation of cell metabolism (MTT assay) showed that LLLT with all energy doses promoted an increase of cell metabolism, being more effective for 0.5, 1.5, and 3 J/cm2. The highest cell counts (Trypan blue assay) were observed with 0.5, 3, and 5 J/cm2. No statistically significant difference for total protein (TP) production was observed and cell morphology analysis by scanning electron microscopy revealed that LLLT did not promote morphological alterations on the keratinocytes. Real-time polymerase chain reaction (qPCR) revealed that LLLT also promoted an increase of type I collagen (Col-I) and vascular endothelial growth factor (VEGF) gene expression, especially for 1.5 J/cm2, but no change on fibroblast growth factor-2 (FGF-2) expression was observed. LLLT at energy doses ranging from 0.5 to 3 J/cm2 promoted the most significant biostimulatory effects on cultured keratinocytes. © 2012 Springer-Verlag London Ltd.

In vitro transdentinal effect of low-level laser therapy

Low-level laser therapy (LLLT) has been used for the treatment of dentinal hypersensitivity. However, the specific LLL dose and the response mechanisms of these cells to transdentinal irradiation have not yet been demonstrated. Therefore, this study evaluated the transdentinal effects of different LLL doses on stressed odontoblast-like pulp cells MDPC-23 seeded onto the pulpal side of dentin discs obtained from human third molars. The discs were placed in devices simulating in vitro pulp chambers and the whole set was placed in 24-well plates containing plain culture medium (DMEM). After 24 h incubation, the culture medium was replaced by fresh DMEM supplemented with either 5% (simulating a nutritional stress condition) or 10% fetal bovine serum (FBS). The cells were irradiated with doses of 15 and 25 J cm-2 every 24 h, totaling three applications over three consecutive days. The cells in the control groups were removed from the incubator for the same times as used in their respective experimental groups for irradiation, though without activating the laser source (sham irradiation). After 72 h of the last active or sham irradiation, the cells were evaluated with respect to succinic dehydrogenase (SDH) enzyme production (MTT assay), total protein (TP) expression, alkaline phosphatase (ALP) synthesis, reverse transcriptase polymerase chain reaction (RT-PCR) for collagen type 1 (Col-I) and ALP, and morphology (SEM). For both tests, significantly higher values were obtained for the 25 J cm-2 dose. Regarding SDH production, supplementation of the culture medium with 5% FBS provided better results. For TP and ALP expression, the 25 J cm-2 presented higher values, especially for the 5% FBS concentration (Mann-Whitney p < 0.05). Under the tested conditions, near infrared laser irradiation at 25 J cm -2 caused transdentinal biostimulation of odontoblast-like MDPC-23 cells. © 2013 Astro Ltd.

Effect of diamond-like carbon thin film coated acrylic resin on candida albicans biofilm formation

Purpose: The purpose of this study was to evaluate the effect of diamond-like carbon thin films doped and undoped with silver nanoparticles coating poly(methyl methacrylate) (PMMA) on Candida albicans biofilm formation. The control of biofilm formation is important to prevent oral diseases in denture users. Materials and Methods: Forty-five PMMA disks were obtained, finished, cleaned in an ultrasonic bath, and divided into three groups: Gc, no surface coating (control group); Gdlc, coated with diamond-like carbon film; and Gag, coated with diamond-like carbon film doped with silver nanoparticles. The films were deposited using a reactive magnetron sputtering system (physical vapor deposition process). The specimens were characterized by optical profilometry, atomic force microscopy, and Rutherford backscattering spectroscopy analyses that determined differences in chemical composition and morphological structure. Following sterilization of the specimens by γ-ray irradiation, C. albicans (ATCC 18804) biofilms were formed by immersion in 2 ml of Sabouraud dextrose broth inoculated with a standardized fungal suspension. After 24 hours, the number of colony forming units (cfu) per specimen was counted. Data concerning biofilm formation were analyzed using ANOVA and the Tukey test (p < 0.05). Results: C. albicans biofilm formation was significantly influenced by the films (p < 0.00001), reducing the number of cfu, while not affecting the roughness parameters (p > 0.05). The Tukey test showed no significant difference between Gdlc and Gag. Films deposited were extremely thin (∼50 nm). The silver particles presented a diameter between 60 and 120 nm and regular distribution throughout the film surface (to Gag). Conclusion: Diamond-like carbon films, doped or undoped with silver nanoparticles, coating the base of PMMA-based dentures could be an alternative procedure for preventing candidosis in denture users. © 2013 by the American College of Prosthodontists.

Effect of antimicrobial photodynamic therapy on periodontally infected tooth sockets in rats

The aim of the present study was to evaluate the antimicrobial effect of antimicrobial photodynamic therapy (aPDT) in alveolar treatment of areas with induced periodontitis. Thirty male Wistar rats were subjected to ligature-induced periodontal disease (PD) in the first left inferior molars, while the right side molars did not receive ligatures. After 7 days of PD evolution, ligatures were removed from the left side, and the first left and right mandibular molars were extracted. Afterwards, animals were divided into groups according to the following treatments: control (C)-no treatment; mechanical debridement (MD)-mechanical debridement and irrigation with saline solution; and aPDT-mechanical debridement, irrigation with toluidine blue O (TBO), and 1 min of laser irradiation (GaAlAs, 660 nm, 30 mW, 32 J/cm2, 60 s). Ligatures were removed and samples of the alveolar content after extraction and after each treatment were collected for microbial processing by real-time polymerase chain reaction with specific primers for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Treponema denticola. Data were submitted to statistical analysis by multiple comparison tests (McNemar test; p < 0.05). T. denticola was not found in the collected samples. A. actinomycetemcomitans and P. gingivalis were found in ligature samples. Tooth socket samples without periodontitis induction presented lesser microbial charge than samples with induced periodontitis (p < 0.05). aPDT significantly reduced A. actinomycetemcomitans levels on the left side (p < 0.05). It was concluded that aPDT was an effective antimicrobial treatment for tooth sockets in areas affected by induced periodontitis. © 2013 Springer-Verlag London.

Comparison of the effect of rose bengal- and eosin Y-mediated photodynamic inactivation on planktonic cells and biofilms of Candida albicans

Candida albicans is an opportunistic yeast that can cause oral candidosis through the formation of a biofilm, an important virulence factor that compromises the action of antifungal agents. The objective of this study was to compare the effect of rose bengal (RB)- and eosin Y (EY)-mediated photodynamic inactivation (PDI) using a green light-emitting diode (LED; 532 ± 10 nm) on planktonic cells and biofilms of C. albicans (ATCC 18804). Planktonic cultures were treated with photosensitizers at concentrations ranging from 0.78 to 400 μM, and biofilms were treated with 200 μM of photosensitizers. The number of colony-forming unit per milliliter (CFU/mL) was compared by analysis of variance and Tukey's test (P ≤ 0.05). After treatment, one biofilm specimen of the control and PDI groups were examined by scanning electron microscopy. The photosensitizers (6.25, 25, 50, 200, and 400 μM of EY, and 6.25 μM of RB or higher) significantly reduced the number of CFU/mL in the PDI groups when compared to the control group. With respect to biofilm formation, RB- and EY-mediated PDI promoted reductions of 0.22 log10 and 0.45 log10, respectively. Scanning electron microscopy showed that the two photosensitizers reduced fungal structures. In conclusion, EY- and RB-mediated PDI using LED irradiation significantly reduced C. albicans planktonic cells and biofilms. © 2013 Springer-Verlag London.

Critical Water Effect on the Plasmon Band and Visible Light Activity of Au/ZnO Nanocomposites

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of Laser Irradiation on Pulp Cells Exposed to Bleaching Agents

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Infrared LED irradiation photobiomodulation of oxidative stress in human dental pulp cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of an Er,Cr:YSGG laser on external adaptation of healthy and decayed cavities

The aim of this study was to evaluate the influence of chlorhexidine and Er, Cr:YSGG laser irradiation on the bond strength and external adaptation in mixed healthy and caries-affected class V cavities before and after thermal cycling. Thirty-six cavity preparations were made in mixed class V buccal human molars, half of them being artificially caries-induced. Any remaining affected dentin was removed from the cavity with a round burr at low speed. The teeth were divided into six groups, according to cleaning agent for both healthy and caries-induced dentin: no treatment, chlorhexidine and erbium, chromium-doped: yttrium, scandium, gallium, garnet (Er,Cr:YSGG) laser irradiation. A Filtek P90 (3M ESPE, St Paul, MN, USA) silorane adhesive restorative system was used. The specimens were subjected to 5000 thermal cycles (5-55 degrees C 60 min). Epoxy replicas were obtained to characterize the external adaptation under scanning electron microscopy. The average percentages of non-continuous margins were 5.41% and 6.49% in enamel dentin before thermal cycling and 25% and 33.7% after thermal cycling, respectively. The caries-affected and laser irradiated cavities showed higher non-continuous margins. Thermal cycling was able to raise the percentage of non-continuous margin for all groups. Chlorhexidine did not affect the marginal adaptation results, and the Er,Cr:YSGG laser irradiation showed significantly worse results compared with the control group.

Chemical composition and lipoxygenase activity in soybeans (Glycine max L. Merr.) submitted to gamma irradiation

Soybeans are an important food due to their functional and nutritional characteristics. However, consumption by western populations is limited by the astringent taste of soybeans and their derivatives which results from the action of lipoxygenase, an enzyme activated during product processing. The aim of this study was to evaluate the effect of gamma irradiation on the chemical composition and specific activity of lipoxygenase in different soybean cultivars. Soybeans were stored in plastic bags and irradiated with doses of 2.5, 5 and 10 kGy. The chemical composition (moisture, protein, lipids, ashes, crude fiber, and carbohydrates) and lipoxygenase specific activity were determined for each sample. Gamma irradiation induced a small increase of protein and lipid content in some soybean cultivars, which did not exceed the highest content of 5% and 26%, respectively, when compared to control. Lipoxygenase specific activity decreased in the three cultivars with increasing gamma irradiation dose. In conclusion, the gamma irradiation doses used are suitable to inactivate part of lipoxygenase while not causing expressive changes in the chemical composition of the cultivars studied. (C) 2014 Elsevier Ltd. All rights reserved.

Effect of Dental Surface Treatment with Nd:YAG and Er:YAG Lasers on Bond Strength to Recently Bleached Dentin

The objective of this study was to evaluate the effect of surface treatment with Er:YAG and Nd:YAG lasers on resin composite bond strength to recently bleached dentin. A total of 120 bovine incisors were distributed into two groups: C- without bleaching; and B- bleached with 35% hydrogen peroxide. Each group was divided into three subgroups: N- without laser treatment; Nd- Nd:YAG laser irradiation; and Er- Er:YAG laser irradiation. The adhesive system (Adper Single Bond 2) was applied and composite build-ups were constructed with Filtek Supreme (3M/ESPE). The teeth were sectioned to obtain dentin-resin sticks (1x1mm(2)) and tested by microtensile bond testing. The bond strength values in group B, subgroup N (16.1 +/- 3.5MPa) presented no significant difference compared with group B, subgroup Er (14.7 +/- 6.1MPa). Group C, subgroup N (26.8 +/- 7.4MPa) presented no significant difference compared with group B, subgroup Nd (28.8 +/- 5.6MPa). Group C, subgroup Nd (36.1 +/- 7.9MPa) presented a significant increase in bond strength compared with the other groups. The Er:YAG laser did not influence the bond strength of bleached specimens, and the use of the Nd:YAG laser on bleached specimens was able to reverse the immediate effects of bleaching, obtaining bond strength values similar to those of the control group.

Laser irradiation and thermal treatment inducing selective crystallization in Sb2O3-Sb2S3 glassy films

The influence of both thermal treatment and laser irradiation on the structural and optical properties of films in the Sb 2 O 3 –Sb 2 S 3 system was investigated. The films were prepared by RF-sputtering using glass compositions as raw materials. Irreversible photodarkening effect was observed after exposure the films to a 458nm solid state laser. It is shown, for the first time, the use of holographic technique to measure “in situ”, simultaneously and independently, the phase and amplitude modulations in glassy films. The films were also photo-crystallized and analysed “in situ” using a laser coupled to a micro-Raman equipment. Results showed that Sb 2 S 3 crystalline phase was obtained after irradiation. The effect of thermal annealing on the structure of the films was carried out. Different from the result obtained by irradiation, thermal annealing induces the crystallization of the Sb 2 O 3 phase. Photo and thermal induced effects on films were studied using UV–Vis and Raman spectroscopy, atomic force microscopy (AFM), thermal analysis (DSC), X-ray diffraction, scanning electron microscopy (MEV) and energy-dispersive X-ray spectroscopy (EDX).

Effect of different bleaching protocols on dental enamel and composite resin

To evaluate changes in microhardness, roughness and surface morphology of dental enamel and composite resin after different tooth bleaching techniques. Material and Methods: Dental fragments from bovine incisors with composite resin restorations were submitted to different bleaching protocols: G1 – daily 8 hours application of a 10% carbamide peroxide (CP) gel during 21 days; G2: 3 applications of 15 minutes of a 38% hydrogen peroxide (H2O2) gel; G3: 38% H2O2 gel associated to irradiation with LED (470nm) during 6 minutes. The Knoop micro hardness of enamel and composite resin were evaluated at 1, 7, 14 and 21 days for G1, and after 1, 2 and 3 sessions for G2 and G3. The roughness and superficial morphology (atomic force microscopy) were evaluated before and at the end of the bleaching treatment. The data were analyzed by Mann-Whitney and Wilcoxon tests (=5%). Results: Significant reduction on enamel hardness was observed after 2 and 3 sessions for G2 and G3. For composite, the reduction occurred after 21 days for G1, and after 3 sessions for G2 and G3 (p<0.05). Significant reduction on roughness and superficial morphology were observed only for enamel of G1 group (p<0.05). Conclusion: The 10% CP gel promoted only superficial alterations on dental enamel, while the 38% H2O2 gel promoted mineral reduction of this dental tissue. All the bleaching protocols promoted reduction on hardness of composite resin.

Effect of Er,Cr:YSGG laser application in the treatment of experimental periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of different light-curing techniques on hardness of a microhybrid dental composite resin

Coordenação de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)