Effects of Laser Irradiation on Pulp Cells Exposed to Bleaching Agents

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

Bond strength of composite to dentin: effect of acid etching and laser irradiation through an uncured self-etch adhesive system

This study evaluated the effect on micro-tensile bond strength (mu-TBS) of laser irradiation of etched/unetched dentin through an uncured self-etching adhesive. Dentinal surfaces were treated with Clearfil SE Bond Adhesive (CSE) either according to the manufacturer's instructions (CSE) or without applying the primer (CSE/NP). The dentin was irradiated through the uncured adhesive, using an Nd: YAG laser at 0.75 or 1 W power settings. The adhesive was cured, composite crowns were built up, and the teeth were sectioned into beams (0.49 mm(2)) to be stressed under tension. Data were analyzed using one-way ANOVA and Tukey statistics (alpha = 5%). Dentin of the fractured specimens and the interfaces of untested beams were observed under scanning electron microscopy (SEM). The results showed that non-etched irradiated surfaces presented higher mu-TBS than etched and irradiated surfaces (p < 0.05). Laser irradiation alone did not lead to differences in mu-TBS (p > 0.05). SEM showed solidification globules on the surfaces of the specimens. The interfaces were similar on irradiated and non-irradiated surfaces. Laser irradiation of dentin through the uncured adhesive did not lead to higher mu-TBS when compared to the suggested manufacturer's technique. However, this treatment brought benefits when performed on unetched dentin, since bond strengths were higher when compared to etched dentin.

Low-level laser therapy for osteonecrotic lesions: effects on osteoblasts treated with zoledronic acid

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

Effects of low-level laser therapy on the proliferation and apoptosis of gingival fibroblasts treated with zoledronic acid

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

Biomodulation of inflammatory cytokines related to oral mucositis by low-level laser therapy

This study evaluated the effects of LLLT on the expressionof inflammatory cytokines related to the development of oralmucositis by gingival fibroblasts. Primary gingival fibroblastswere seeded on 24-well plates (105cells/well) for 24 h. Freshserum-free culture medium (DMEM) was then added, andcells were placed in contact with LPS (Escherichia coli,1 lgmL1), followed by LLLT irradiation (LaserTABLE—InGaAsP diode prototype—780 nm, 25 mW) delivering 0,0.5, 1.5 or 3 J cm². Cells without contact with LPS werealso irradiated with the same energy densities. Gene expres-sion of TNF- a, IL-1b, IL-6 and IL-8 was evaluated by Real-Time PCR, and protein synthesis of these cytokines wasdetermined by enzyme-linked immunosorbent (ELISA) assay.Data were statistically analyzed by the Kruskal– Wallis test,complemented by the Mann–Whitney test (P < 0.05). LPStreatment increased the gene expression and protein synthesisof TNF-a, IL-6 and IL-8, while the expression of IL-1b wasnot affected. For LPS-treated groups, LLLT promoted signif-icant decreases in the expression of TNF-a, IL-6, and IL-8 at1.5 J cm2and 3 J cm2. These results demonstrate thatLLLT promoted a beneficial biomodulatory effect on theexpression of inflammatory cytokines related to oral mucosi-tis by human gingival fibroblasts.

Effects of copper vapor laser irradiation (λ = 510.6 nm) on the enamel and dentine of human teeth: An ultra-structural morphologic study

Objective: A morphological and ultra-structural study of copper vapor laser (λ = 510.6 nm) effects on enamel and dentine was performed to show the effects of this radiation. Methods: A total of 15 human molars were cut in half; 15 pieces were separated for irradiation on enamel and 15 for dentine. These two groups were further divided into five experimental groups, including a control group, comprised of three half-sections each, irradiated by a CVL laser with a power of 7 W, a repetition rate of 15,000 pulses/sec and exposed at 500, 600, and 800 msec and 1 sec irradiation times with a 5-sec interval between irradiations. Results: In an ultra-structural SEM exam, we observed that on the enamel surfaces irradiated for 1 sec there was morphological alteration that consisted of catering, flaking, and melting on the surfaces. There was no alteration for the other exposure times. On the dentine teeth irradiated for 1 sec, we observed an evident ultra-structural alteration of melted tissue and loss of morphological characteristics. In the dentine group irradiated by 800 msec, we observed ablation and a partial loss of morphological characteristics. In the dentine groups irradiated by 500 and 600 msec, no alteration was observed. Conclusions: The results showed that irradiation with CVL promoted morphologic changes in the enamel as well as in the dentine and demonstrated a need for future studies in order to establish a safe protocol for further use in the odontological practice.

Effect of GaAIAs low-level laser therapy on the healing of human palate mucosa after connective tissue graft harvesting: randomized clinical trial

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

Laser terapêutico no reparo ósseo de ratos submetidos à ausência de carga

Low-intensity laser has been used as a physical agent in various fields of medical sciences such as bone and tissue repair. Meanwhile little is known about its effects in adverse conditions such as abolition of load and osteopenic. With the assumption that the laser Ga-Al-As accelerates the process of bone consolidation, goal of this study was to evaluate bone mineral density (BMD) in incomplete transverse osteotomies of tibia in adult rats, treated with low power laser therapy in three different groups: G1 (n = 10), reference 15 days; G2 (n=10), suspended by the tail and, accordingly, treated with laser for 12 days; G3 (n = 10), suspended by the tail by 36 days and that after 21 days, there was laser treatment for 12 days. The right tibia treated with laser and left served as control. The laser was used to Ga-Al-As, DMC - Flash Lase® III, with wavelength 830nm, 100 mW, 4J, 140 J / cm ², 40s of application in 12 sessions. It was used densitometer-Lunar DPX®, with computer program for "small animals", and the analysis of BMD was made in the bone throughout the region and the osteotomy. The results showed no efficacy of laser therapy in the process of bone repair, both in animals of group 1, as in group 2 and 3. It follows that either the low-power laser was not an effective performance or the effects of laser therapy is not only manifested at the site of irradiation as well as the systemic level.

In Vitro Analysis of Bacterial Morphology by Atomic Force Microscopy of Low Level Laser Therapy 660, 830 and 904 nm

Objective: The objective of this study was to analyze the bacterial morphology by atomic force microscopy (AFM) after the application of low-level laser therapy (LLLT) in in vitro culture of Staphylococcus aureus ATCC 29213. Background data: Infections caused by S. aureus are among the highest occurring in hospitals and can often colonize pressure ulcers. LLLT is among the methods used to accelerate the healing of ulcers. However, there is no consensus on its effect on bacteria. Materials and methods: After being cultivated and seeded, the cultures were irradiated using wavelengths of 660, 830, and 904 nm at fluences of 0, 1, 2, 3, 4, 5, and 16 J/cm(2). Viable cells of S. aureus strain were counted after 24 h incubation. To analyze the occurrence of morphological changes, the topographical measurement of bacterial cells was analyzed using the AFM. Results: The overall assessment revealed that the laser irradiation reduced the S. aureus growth using 830 and 904 nm wavelengths; the latter with the greatest inhibition of the colony-forming units (CFU/mL) (331.1 +/- 38.19 and 137.38 +/- 21.72). Specifically with 660 nm, the statistical difference occurred only at a fluence of 3 J/cm(2). Topographical analysis showed small changes in morphological conformity of the samples tested. Conclusions: LLLT reduced the growth of S. aureus with 830 and 904 nm wavelengths, particularly with 904 nm at a fluence of 3 J/cm(2), where the greatest topographical changes of the cell structure occurred.

Low-Level Laser Therapy Decreases Renal Interstitial Fibrosis

Objective: the purpose of this study was to investigate the effect of low-level laser therapy (LLLT) on chronic kidney disease (CKD) in a model of unilateral ureteral obstruction (UUO). Background data: Regardless of the etiology, CKD involves progressive widespread tissue fibrosis, tubular atrophy, and loss of kidney function. This process also occurs in kidney allograft. At present, effective therapies for this condition are lacking. We investigated the effects of LLLT on the interstitial fibrosis that occurs after experimental UUO in rats. Methods: The occluded kidney of half of the 32 Wistar rats that underwent UUO received a single intraoperative dose of LLLT (AlGaAs laser, 780 nm, 22.5 J/cm(2), 30mW, 0.75W/cm(2), 30 sec on each of nine points). After 14 days, renal fibrosis was assessed by Sirius red staining under polarized light. Immunohistochemical analyses quantitated the renal tissue cells that expressed fibroblast (FSP-1) and myofibroblast (alpha-SMA) markers. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the mRNA expression of interleukin (IL)-6, monocyte chemotactic protein-1 (MCP-1), transforming growth factor (TGF)-beta 1 and Smad3. Results: The UUO and LLLT animals had less fibrosis than the UUO animals, as well having decreased expression inflammatory and pro-fibrotic markers. Conclusions: For the first time, we showed that LLLT had a protective effect regarding renal interstitial fibrosis. It is conceivable that by attenuating inflammation, LLLT can prevent tubular activation and transdifferentiation, which are the two processes that mainly drive the renal fibrosis of the UUO model.

Nd:YAG Laser Irradiation of Etched/Unetched Dentin Through an Uncured Two-step Etch-and-Rinse Adhesive and its Effect on Microtensile Bond Strength

Purpose: To evaluate whether Nd:YAG laser irradiation of etched and unetched dentin through an uncured adhesive affected the microtensile bond strength (pTBS). Materials and Methods: Flat dentin surfaces were created in 19 extracted human third molars. Adper Single Bond (SB) adhesive was applied over etched (groups 1 to 3) or unetched dentin (groups 4 to 6). The dentin was then irradiated with a Nd:YAG laser through the uncured adhesive, using 0.75 or 1 W power settings, except for the control groups (groups 1 and 4). The adhesive was light cured and composite crowns were built up. After 24 h, the teeth were sectioned into beams, with cross-sectional areas of 0.49 mm(2), and were stressed under tension. Data were statistically analyzed using two-way ANOVA and Tukey's test (alpha = 5%). Dentin surfaces of fractured specimens and the interfaces of untested beams were observed under scanning electron microscopy (SEM). Results: Acid etching, laser irradiation, and their interaction significantly affected bonding (p < 0.05). Laser irradiation did not improve bonding of etched dentin to resin (p > 0.05). However, higher pTBS means were found on unetched lased dentin (groups 5 and 6), but only in comparison to group 4, where neither lasing nor etching was performed. Groups 4 to 6 showed the lowest pTBS means among all groups tested (p < 0.05). Laser irradiation did not change the characteristics of the hybrid layers created, while solidification globules were observed on lased dentin surfaces under SEM. Conclusion: Laser irradiation of dentin through the uncured adhesive did not significantly improve the pTBS in comparison to the suggested manufacturer's technique.

Low-Level Laser Therapy Influences Mouse Odontoblast-Like Cell Response in Vitro

Objective: The purpose of this study was to analyze the influence of two different irradiation times with 85mW/cm(2) 830nm laser on the behavior of mouse odontoblast-like cells. Background data: The use of low-level laser therapy (LLLT) to stimulate pulp tissue is a reality, but few reports relate odontoblastic responses to irradiation in in vitro models. Methods: Odontoblast-like cells (MDPC-23) were cultivated and divided into three groups: control/nonirradiated (group 1); or irradiated with 85mW/cm(2), 830nm laser for 10 sec (0.8 J/cm(2)) (group 2); or for 50 sec (4.2 J/cm(2)) (group 3) with a wavelength of 830 nm. After 3, 7, and 10 days, it was analyzed: growth curve and cell viability, total protein content, alkaline phosphatase (ALP) activity, calcified nodules detection and quantification, collagen immunolocalization, vascular endothelial growth factor (VEGF) expression, and real-time polymerase chain reaction (PCR) for DMP1 gene. Data were analyzed by Kruskall-Wallis test (alpha = 0.05). Results: Cell growth was smaller in group 2 (p < 0.01), whereas viability was similar in all groups and at all periods. Total protein content and ALP activity increased on the 10th day with 0.8 J/cm(2) (p < 0.01), as well as the detection and quantification of mineralization nodules (p < 0.05), collagen, and VEGF expression (p < 0.01). The expression of DMP1 increased in all groups (p < 0.05) compared with control at 3 days, except for 0.8 J/cm(2) at 3 days and control at 10 days. Conclusions: LLLT influenced the behavior of odontoblast-like cells; the shorter time/smallest energy density promoted the expression of odontoblastic phenotype in a more significant way.

Red (660 nm) and infrared (830 nm) low-level laser therapy in skeletal muscle fatigue in humans: what is better?

In animal and clinical trials low-level laser therapy (LLLT) using red, infrared and mixed wavelengths has been shown to delay the development of skeletal muscle fatigue. However, the parameters employed in these studies do not allow a conclusion as to which wavelength range is better in delaying the development of skeletal muscle fatigue. With this perspective in mind, we compared the effects of red and infrared LLLT on skeletal muscle fatigue. A randomized double-blind placebo-controlled crossover trial was performed in ten healthy male volunteers. They were treated with active red LLLT, active infrared LLLT (660 or 830 nm, 50 mW, 17.85 W/cm(2), 100 s irradiation per point, 5 J, 1,785 J/cm(2) at each point irradiated, total 20 J irradiated per muscle) or an identical placebo LLLT at four points of the biceps brachii muscle for 3 min before exercise (voluntary isometric elbow flexion for 60 s). The mean peak force was significantly greater (p < 0.05) following red (12.14%) and infrared LLLT (14.49%) than following placebo LLLT, and the mean average force was also significantly greater (p < 0.05) following red (13.09%) and infrared LLLT (13.24%) than following placebo LLLT. There were no significant differences in mean average force or mean peak force between red and infrared LLLT. We conclude that both red than infrared LLLT are effective in delaying the development skeletal muscle fatigue and in enhancement of skeletal muscle performance. Further studies are needed to identify the specific mechanisms through which each wavelength acts.

Infrared (810-nm) low-level laser therapy on rat experimental knee inflammation

Arthritis of the knee is the most common type of joint inflammatory disorder and it is associated with pain and inflammation of the joint capsule. Few studies address the effects of the 810-nm laser in such conditions. Here we investigated the effects of low-level laser therapy (LLLT; infrared, 810-nm) in experimentally induced rat knee inflammation. Thirty male Wistar rats (230-250 g) were anesthetized and injected with carrageenan by an intra-articular route. After 6 and 12 h, all animals were killed by CO(2) inhalation and the articular cavity was washed for cellular and biochemical analysis. Articular tissue was carefully removed for real-time PCR analysis in order to evaluate COX-1 and COX-2 expression. LLLT was able to significantly inhibit the total number of leukocytes, as well as the myeloperoxidase activity with 1, 3, and 6 J (Joules) of energy. This result was corroborated by cell counting showing the reduction of polymorphonuclear cells at the inflammatory site. Vascular extravasation was significantly inhibited at the higher dose of energy of 10 J. Both COX-1 and 2 gene expression were significantly enhanced by laser irradiation while PGE(2) production was inhibited. Low-level laser therapy operating at 810 nm markedly reduced inflammatory signs of inflammation but increased COX-1 and 2 gene expression. Further studies are necessary to investigate the possible production of antiinflammatory mediators by COX enzymes induced by laser irradiation in knee inflammation.

Temperature changes on the root surfaces of mandibular incisors after an 810-nm high-intensity intracanal diode laser irradiation

Temperature changes caused by laser irradiation can promote damage to the surrounding dental tissues. In this study, we evaluated the temperature changes of recently extracted human mandibular incisors during intracanal irradiation with an 810-nm diode laser at different settings. Fifty mandibular incisors were enlarged up to an apical size of ISO No. 40 file. After the final rinse with 17% ethylenediaminetetraacetic acid, 0.2% lauryl sodium sulfate biologic detergent, and sterile water, samples were irradiated with circular movements from apex to crown through five different settings of output power (1.5, 2.0, 2.5, 3.0, and 3.5 W) in continuous mode. The temperature changes were measured on both sides of the apical and middle root thirds using two thermopar devices. A temperature increase of 7 degrees C was considered acceptable as a safe threshold when applying the diode laser. Results: The results showed that only 3.5-W output power increased the outer surface temperature above the critical value. Conclusion: The recommended output power can be stipulated as equal to or less than 3 W to avoid overheating during diode laser irradiation on thin dentin walls. (c) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI: 10.1117/1.JBO.17.1.015006]