In vitro effect of low-level laser on odontoblast-like cells

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

Effects of low-level laser therapy (685 nm) at different doses in osteogenic cell cultures

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Low-level laser therapy (670 nm) on viability of random skin flap in rats

This study investigated the effects of 670 nm laser, at different fluences, on the viability of skin flap in rats. One hundred male animals were used. The animals were divided into control group; group treated with 3 J/cm(2); group treated with 6 J/cm(2); group treated with 12 J/cm(2) and group treated with 24 J/cm(2). The skin flap was made on the backs of all animals studied, with a plastic sheet interposed between the flap and the donor site. Laser irradiation was done immediately after the surgery and on days 1, 2, 3 and 4 after surgery. The percentage of necrosis of the flap was calculated at the 7th postoperative day. Additionally, a sample of each flap was collected to enable us to count the blood vessels. Treated animals showed a statistically significant smaller area of necrosis than did the control group. The necrosis in the treated groups was 41.82% (group 2), 36.51% (group 3), 29.45% (group 4) and 20.37% (group 5). We also demonstrated that laser irradiation at 670 nm, at all doses used, had a stimulatory effect on angiogenesis. Our study showed that the 670 nm laser was efficient to increase the viability of the skin flap, at all fluences used, with a tendency of reaching better results at higher doses.

Quantification of fibrosis and mast cells in the tissue response of endodontic sealer irradiated by low-level laser therapy

Low-level laser therapy (LLLT) accelerates tissue repair. Mast cells induce the proliferation of fibroblasts and the development of local fibrosis. The objective of this study was to quantify fibrosis rate and mast cells in connective tissue after endodontic sealer zinc oxide and eugenol (ZOE) was implanted and submitted to LLLT, immediately after implant and again 24 h later. Sixty mice were distributed into three groups: GI, GII, and GIII (n = 20). In GI, the tubes filled with Endofill were implanted in the animals and were not irradiated with LLLT. In GII, the tubes containing Endofill were implanted in the animals and then irradiated with red LLLT (InGaAIP) 685-nm wavelength, D=72 J/Cm(2), E = 2 J, T=58 s, P=35 mW, and in GIII, the tubes with Endofill were implanted and irradiated with infrared LLLT (AsGaAl) 830-nm wavelength, D=70 J/Cm(2), E = 2 J, T=40 s, P=50 mW. After 7 days and 30 days, the animals were killed. A series of 6-mu m-thick sections were obtained and stained with Toluidine Blue and Picrosirius and analyzed under a standard light microscope using a polarized light filter for the quantification of fibrosis. The statistics were qualitative and quantitative with a significance of 5%. The irradiation with LLLT did not offer improvement in the fibrosis rate, however, it provided a significant decrease in the concentration of independent mast cells for the period studied.

Low-power laser irradiation improves histomorphometrical parameters and bone matrix organization during tibia wound healing in rats

The influence of daily energy doses of 0.03, 0.3 and 0.9 J of He-Ne laser irradiation on the repair of surgically produced tibia damage was investigated in Wistar rats. Laser treatment was initiated 24 h after the trauma and continued daily for 7 or 14 days in two groups of nine rats (n=3 per laser dose and period). Two control groups (n=9 each) with injured tibiae were used. The course of healing was monitored using morphometrical analysis of the trabecular area. The organization of collagen fibers in the bone matrix and the histology of the tissue were evaluated using Picrosirius-polarization method and Masson's trichrome. After 7 days, there was a significant increase in the area of neoformed trabeculae in tibiae irradiated with 0.3 and 0.9 J compared to the controls. At a daily dose of 0.9 J (15 min of irradiation per day) the 7-day group showed a significant increase in trabecular bone growth compared to the 14-day group. However, the laser irradiation at the daily dose of 0.3 J produced no significant decrease in the trabecular area of the 14-day group compared to the 7-day group, but there was significant increase in the trabecular area of the 15-day controls compared to the 8-day controls. Irradiation increased the number of hypertrophic osteoclasts compared to non-irradiated injured tibiae (controls) on days 8 and 15. The Picrosirius-polarization method revealed bands of parallel collagen fibers (parallel-fibered bone) at the repair site of 14-day-irradiated tibiae, regardless of the dose. This organization improved when compared to 7-day-irradiated tibiae and control tibiae. These results show that low-level laser therapy stimulated the growth of the trabecular area and the concomitant invasion of osteoclasts during the first week, and hastened the organization of matrix collagen (parallel alignment of the fibers) in a second phase not seen in control, non-irradiated tibiae at the same period. The active osteoclasts that invaded the regenerating site were probably responsible for the decrease in trabecular area by the fourteenth day of irradiation. (C) 2003 Elsevier B.V. All rights reserved.

Low-level laser on femoral growth plate in rats

OBJETIVO: Determinar a influência do Laser Terapêutico de Baixa Potência sobre a placa de crescimento de ratos. MÉTODOS: Trinta ratos Wistar machos com 40 dias de idade foram divididos em dois grupos, G1 e G2. O grupo G1 foi submetido à irradiação com laser GaAlAs 830 nm, potência de saída de 40 mW, e densidade de energia de 10 J/cm2. A irradiação foi aplicada diariamente por um período máximo de 21 dias. O mesmo procedimento foi realizado no grupo G2, com a probe desativada. Cinco animais em cada grupo foram sacrificados nos dias 7, 14 e 21 e submetidas à análise histomorfométrica. RESULTADOS: em ambos os grupos, o disco fisário esteve radiograficamente visível em todos os momentos nas incidências craniocaudal e médio-lateral. No 21º dia a porcentagem de comprimento longitudinal do fêmur foi maior em G1 que em G2 em relação ao valor basal, e o número de condrócitos da zona hipertrófica foi maior em G1 que em G2. A zona de cartilagem calcificada estava maior em G1 em relação a G2 em todos os momentos de avaliação. A angiogênese foi maior em G1 que em G2 nos 14º e 21º dias. CONCLUSÃO: A terapia com laser terapêutico de baixa potência influenciou negativamente o disco fisário distal do fêmur de ratos.

Effect of Low-Level Laser Therapy and Calcitonin on Bone Repair in Castrated Rats: A Densitometric Study

Objective: To investigate the healing of bone defects in male rats treated with salmon calcitonin, low-level laser therapy (LLLT), or both. Background: Healing of bone defects still represents a challenge to health professionals in several areas. In this article, the effect of calcitonin in combination with LLLT on bone repair was studied. Densitometry was used as a valuable tool for the measurement of bone regeneration. Methods: Sixty male Wistar rats underwent bilateral castration surgery before the creation of a surgical bone defect. The animals were randomly divided into four groups: control, treated with calcitonin (Ca), treated with LLLT (La), and treated with calcitonin and LLLT (CaLa). Groups Ca and CaLa received 2 IU/kg of synthetic salmon calcitonin intra-muscularly three times a week. Groups La and CaLa received laser therapy using a gallium-aluminum-arsenide laser (10mW, 20 J/cm(2), wavelength 830 nm). Control animals were submitted to sham irradiation. The animals were sacrificed 7, 14, and 21 days after surgery, and bone defects were analyzed using densitometry. Results: The CaLa group had a higher degree of bone regeneration 14 and 21 days after surgery. Conclusions: The La and CaLa had significantly higher bone mineral density than the control and Ca groups.

Comparison between wound healing in induced diabetic and nondiabetic rats after low-level laser therapy

Objective: the aim of this work was to compare the effect of low-level laser therapy (LLLT) on the wound healing process in nondiabetic and diabetic rats. Background Data: Among the clinical symptoms caused by diabetes mellitus, a delay in wound healing is a potential risk for patients. It is suggested that LLLT can improve wound healing. Methods: the tissue used for this study was extracted from animals suffering from diabetes, which was induced by Streptozotocin (R), and from nondiabetic rats. Animals were assembled into two groups of 25 rats each (treated and control) and further subdivided into two groups: diabetic (n = 15) and nondiabetic (n = 10). A full-thickness skin wound was made on the dorsum. area, with a round 8-mm hole-punch. The treated group was irradiated by a HeNe laser at 632.8 nm, with the following parameters: 15 mW, exposition time of 17 sec, 0.025 cm(2) irradiated area, and energy density of 10 J/cm(2). Square full-thickness skin samples (18 mm each side, including both injured and noninjured tissues) were obtained at 4, 7, and 15 days after surgery and analyzed by qualitative and quantitative histological methods. Results: Quantitative histopathological analysis confirmed the results of the qualitative analysis through histological microscope slides. When comparing tissue components (inflammatory cells, vessels and fibroblast/area), we found that treated animals had a less intense inflammatory process than controls. Conclusion: Results obtained by both qualitative and quantitative analyses suggested that irradiation of rats with HeNe (632.8 nm), at the tested dose, promoted efficient wound healing in both nondiabetic and diabetic rats as, compared to the control group.

Anti-inflammatory effects of low-level laser therapy (LLLT) with two different red wavelengths (660 nm and 684 nm) in carrageenan-induced rat paw edema

It has been suggested that low-level laser therapy (LLLT) can modulate inflammatory processes. The aim of this experiment was to investigate what effects red laser irradiation with two different wavelengths (660 nm and 684 nm) on carrageenan-induced rat paw edema and histology. Thirty two male Wistar rats were randomly divided into four groups. One group received a sterile saline injection, while inflammation was induced by a sub-plantar injection of carrageenan (1 mg/paw) in the three other groups. After 1 h, LLLT was administered to the paw in two of the carrageenan-injected groups. Continuous wave 660 nm and 684 nm red lasers respectively with mean optical outputs of 30 mW and doses of 7.5 J/cm(2) were used. The 660 nm and 684 nm laser groups developed significantly (P < 0.01) less edema (0.58 ml [SE +/- 0.17] ml and 0.76 ml [SE +/- 0.10] respectively) than the control group (1.67 ml [SE +/- 0.191) at 4 h after injections. Similarly, both laser groups showed a significantly lower number of inflammatory cells in the muscular and conjunctive sub-plantar tissues than the control group.We conclude that both 660 nm and 684 nm red wavelengths of LLLT are effective in reducing edema formation and inflammatory cell migration when a dose of 7.5 J/cm(2) is used. (c) 2007 Elsevier B.V. All rights reserved.

Low-level laser therapy for pain relief after episiotomy: a double-blind randomised clinical trial

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

A randomised clinical trial of the effect of low-level laser therapy for perineal pain and healing after episiotomy: A pilot study

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

In vitro effect of low-level laser therapy on typical oral microbial biofilms

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm 2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts

The aim of this study was to determine adequate energy doses using specific parameters of LLLT to produce biostimulatory effects on human gingival fibroblast culture. Cells (3 10 4 cells/cm 2) were seeded on 24-well acrylic plates using plain DMEM supplemented with 10 fetal bovine serum. After 48-hour incubation with 5 CO2 at 37C, cells were irradiated with a InGaAsP diode laser prototype (LASERTable; 780 3 nm; 40mW) with energy doses of 0.5, 1.5, 3, 5, and 7J/cm 2. Cells were irradiated every 24h totalizing 3 applications. Twenty-four hours after the last irradiation, cell metabolism was evaluated by the MTT assay and the two most effective doses (0.5 and 3J/cm 2) were selected to evaluate the cell number (trypan blue assay) and the cell migration capacity (wound healing assay; transwell migration assay). Data were analyzed by the Kruskal-Wallis and Mann-Whitney nonparametric tests with statistical significance of 5. Irradiation of the fibroblasts with 0.5 and 3J/cm 2 resulted in significant increase in cell metabolism compared with the nonrradiated group (P 0.05). Both energy doses promoted significant increase in the cell number as well as in cell migration (P 0.05). These results demonstrate that, under the tested conditions, LLLT promoted biostimulation of fibroblasts in vitro. Copyright © 2012 Fernanda G. Basso et al.

Biomechanical effect of one session of low-level laser on the bone-titanium implant interface

Low-level laser (LLL) has been used on peri-implant tissues for accelerating bone formation. However, the effect of one session of LLL in the strength of bone-implant interface during early healing process remains unclear. The present study aims to evaluate the removal torque of titanium implants irradiated with LLL during surgical preparation of implant bed, in comparison to non-irradiation. Sixty-four Wistar rats were used. Half of the animals were included in LLL group, while the other half remained as control. All animals had the tibia prepared with a 2 mm drill, and a titanium implant (2.2 × 4 mm) was inserted. Animals from LLL group were irradiated with laser (gallium aluminum arsenide), with a wavelength of 808 nm, a measured power output of 50 mW, to emit radiation in collimated beams (0.4 cm2), for 1 min and 23 s, and an energy density of 11 J/cm2. Two applications (22 J/cm 2) were performed immediately after bed preparation for implant installation. Flaps were sutured, and animals from both groups were sacrificed 7, 15, 30, and 45 days after implant installation, when load necessary for removing implant from bone was evaluated by using a torquimeter. In both groups, torque values tended to increase overtime; and at 30 and 45 days periods, values were statistically higher for LLL group in comparison to control (ANOVA test, p < 0.0001). Thus, it could be suggested that a single session of irradiation with LLL was beneficial to improve bone-implant interface strength, contributing to the osseointegration process. © 2012 Springer-Verlag London Ltd.

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.