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

The present in vitro study evaluated parameters of osteogenesis under the influence of low-level laser therapy (LLLT) at different doses. Osteogenic cells originated from rat calvaria were cultivated in polystyrene plates and exposed to a laser irradiation using an indium-gallium-aluminum phosphide therapeutic laser (InGaAIP), at wavelength of 685 nm, power of 35 mW, 600-μm-diameter optical fiber, and continuous wave. In the attempt of observing the existence of a dose response and its effects, laser irradiation was performed at 25, 77, and 130 J/cm² (7, 22, and 37 s, respectively). The following parameters were assessed: growth curve (4, 7, and 11 days), cell viability (24 h), and nodular formation of mineralized matrix (14 days). The results did not show significant differences related to the growth curve (4, 7, and 11 days) and cell viability (24 h). Within 14 days, osteogenic cultures showed nodular areas with well-defined calcified matrix. The total area stained with Alizarin Red did not show any differences between doses of 25 and 130 J/cm². However, the percentage of stained area was significantly higher in the 25 J/cm² group when compared to the group of 77 J/cm² (Kruskal-Wallis test, p < 0.05). It was possible to conclude that the 685-nm laser irradiation (at 25, 77, and 130 J/cm²) did not influence cell growth and proliferation, although the extracellular mineralization process may have its pattern altered by the LLLT on osteogenic cell cultures.     

A prospective randomized study of 980 nm diode laser‐assisted venous ulcer healing on 34 patients

Venous ulcers are chronic wounds affecting up to 1% of adults in developed countries. Considering that noncontact normothermic therapy has been shown to modify the wound healing process, we conducted a prospective comparative clinical trial aimed at evaluating 980 nm diode laser in laser‐assisted venous ulcer healing. Thirty‐four Caucasian patients with venous leg ulcers were included in the study and separated into two homogenous groups based on age, sex ratio, size, and etiologies of the ulcers. In the laser group, 980 nm InGasAs diode laser (power 15 W, spot size 8 mm, time 3 seconds, fluence 90 J/cm²) was applied weekly for 9 weeks to the ulcers in a homogenous standardized manner, resulting in a local temperature of 45–50 °C, which was controlled with a thermal infrared camera. Complete healing, reduction in size, and pain during and between each procedure were evaluated each time. Of the 18 patients in the laser group, three (16.7%) were completely healed during laser treatment. In the control group (16 patients), the healing was complete for four patients (25%). This difference was not significant (p=0.62). At the ninth follow‐up visit, in the control group, the ulcers had decreased on an average to 94.3% of the original area of the ulcers. In the laser group, the decrease was to 74.2% of the original area of the ulcers. Again, this difference was not significant (p=0.60). The mean VAS score between each treatment was 2.7 (0.5–4.4) in the laser group compared with 3.8 (2.3–5.0) in the control group (0.13<p<0.86). During the treatment, the mean VAS score was 1.8 (0.2–3.8) in the laser group compared with 3.8 (2.1–6.0) in the control group (0.08<p<0.67). 980 nm diode laser‐assisted venous ulcer healing was easy to perform and very well tolerated. However, there were no statistically significant differences in reduction of ulcer size between the two groups, suggesting that this particular laser regimen does not promote wound healing. Additional studies involving larger patient populations and an increased frequency of treatment should be performed to confirm our initial conclusions.     

Influence of laser (660 nm) on functional recovery of the sciatic nerve in rats following crushing lesion

With the aim of accelerating the regenerative processes, the objective was to study the influence of gallium–aluminum–arsenide (GaAlAs) laser (660 nm) on functional and histomorphological recovery of the sciatic nerve in rats. The sciatic nerves of 12 Wistar rats were crushed divided into two groups: control and laser therapy. For the latter, GaAlAs laser was utilized (660 nm, 4 J/cm², 26.3 mW and 0.63 cm² beam), at three equidistant points on the lesion, for 20 days. Comparison of the sciatic functional index (SFI) showed that there was a significant difference only between the pre-lesion value of the laser therapy group and that after the 21st day in the control group. It was concluded that the parameters and methods utilized demonstrated positive results regarding the SFI over the time period evaluated.     

Thermal imaging during photodynamic therapy (PDT)

This study was designed to measure the PDT associated changes in tumour tissue temperature and to calculate if tumoricidally relevant hyperthermia might be induced by the current clinically used PDT modalities. Syrian golden hamsters with amelanotic melanomas (A-Mel-3) received intravenous injections of Photofrin II (5 mg kg⁻¹ b.w.) or 0.9% sodium chloride (2 ml kg⁻¹ b.w.). Twenty-four hours later tumour surface and tumour centre temperature were quantified by an infra-red sensitive thermal imaging device and an implanted thermal probe during laser treatment at a wavelength of 630 nm. With laser light irradiation using power densities of 100 mW cm⁻² or 200 mW cm⁻², a significant temperature increase was noted in comparison to the baseline values. At total energy delivery of 100 J cm⁻² the tumoricidally relevant threshold of 43°C was exceeded with 200 mW cm⁻² only. However, no significant temperature differences were measured between photosensitized and control tumours receiving 100 mW cm⁻² or 200 mW cm⁻². Calculation of the corresponding applied hyperthermic dose revealed that submaximum hyperthermic effects were involved with irradiation intensities of 200 mW cm⁻².     

Organic light emitting diode improves diabetic cutaneous wound healing in rats

A major complication for diabetic patients is chronic wounds due to impaired wound healing. It is well documented that visible red wavelengths can accelerate wound healing in diabetic animal models and patients. In vitro and in vivo diabetic models were used to investigate the effects of organic light emitting diode (OLED) irradiation on cellular function and cutaneous wound healing. Human dermal fibroblasts were cultured in hyperglycemic medium (glucose concentration 180 mM) and irradiated with an OLED (623 nm wavelength peak, range from 560 to 770 nm, power density 7 or 10 mW/cm² at 0.2, 1, or 5 J/cm²). The OLED significantly increased total adenosine triphosphate concentration, metabolic activity, and cell proliferation compared with untreated controls in most parameters tested. For the in vivo experiment, OLED and laser (635 ± 5 nm wavelength) treatments (10 mW/cm², 5 J/cm² daily for a total of seven consecutive days) for cutaneous wound healing were compared using a genetic, diabetic rat model. Both treatments had significantly higher percentage of wound closure on day 6 postinjury and higher total histological scores on day 13 postinjury compared with control. No statistical difference was found between the two treatments. OLED irradiation significantly increased fibroblast growth factor‐2 expression at 36‐hour postinjury and enhanced macrophage activation during initial stages of wound healing. In conclusion, the OLED and laser had comparative effects on enhancing diabetic wound healing.     

The Influence of Low Intensity Infrared Laser Irradiation on Conduction Characteristics of Peripheral Nerve: A Randomised,Controlled, Double Blind Study on the Sural Nerve

. The aim of this study was to investigate the effects of low intensity laser therapy (LILT) on the conduction characteristics of peripheral sensory nerve. Fifteen healthy subjects participated in this experiment. Six points on the sural nerve of the left leg were irradiated with an 830 nm GaAlAs laser with different parameters. All 15 subjects received at random one of the following treatments: an average power of 400 mW at a dose of 1.5 J per point (7.65 J/cm²) (treatment A); an average power of 140 mW and a dose of 1.0 J per point (5.1 J/cm²) (treatment B); an average power of 30 mW, 0.5 J per point (2.55 J/cm²) (treatment C) and to evaluate possible placebo effects in a fourth moment, no power was given (sham). The influence of skin temperature on conduction characteristics of peripheral nerve was taken into account by using correction factors for conductance velocity (1.47 m/s per °C) and negative peak latency (0.2 ms per °C) to a reference temperature of 32°C. One way analysis of variation with repeated measurements of the data collected within 15 min postirradiation showed a significant decrease in conduction velocity and increase in negative peak latency in treatment B. Those results could explain possible laser-mediated pain relief. Paper received 3 August 1999; accepted after revision 19 October 1999.     

Laser phototherapy effect on protein metabolism parameters of rat salivary glands

The aim of this study was to evaluate the effects of infrared diode laser phototherapy (LP) on tissues of the submandibular gland (SMG) and parotid gland (PG). Wistar rats were randomly divided into experimental (A and B) and control (C) groups. A diode laser, 808 nm wavelength, in continuous wave mode, was applied to the PG, SMG and sublingual gland in the experimental groups on two consecutive days. The doses were 4 J/cm² and 8 J/cm², and total energy was 7 J and 14 J, respectively. The power output (500 mW) and power density (277 mW/cm²) were the same for both experimental groups. In order to visualize the area irradiated by the infrared laser, we used a red pilot beam (650 nm) with 3 mW maximum power for the experimental groups. For the control group, the red pilot beam was the only device used. The SMG and PG were removed after 1 week of the first irradiation. Total protein concentration, amylase, peroxidase, catalase and lactate dehydrogenase assays were performed, as well as histological analysis. Statistical tests revealed significant increase in the total protein concentration for groups A and B in the parotid glands (P < 0.05). Based on the results of this study, LP altered the total protein concentration in rats’ parotid glands.     

Low-level 809 nm GaAlAs laser irradiation increases the proliferation rate of human laryngeal carcinoma cells in vitro

The aim of the study was to investigate the effect of low-level 809 nm laser irradiation on the proliferation rate of human larynx carcinoma cells in vitro. Epithelial tumor cells were obtained from a laryngeal carcinoma and cultured under standard conditions. For laser treatment the cells were spread on 96-well tissue culture plates. Sixty-six cell cultures were irradiated with an 809 nm GaAlAs laser. Another 66 served as controls. Power output was 10 mW(cw) and the time of exposure 75–300 s per well, corresponding to an energy fluence of 1.96–7.84 J/cm². Subsequent to laser treatment, the cultures were incubated for 72 h. The proliferation rate was determined by means of fluorescence activity of a redox indicator (Alamar Blue Assay) added to the cultures immediately after the respective treatment. The indicator is reduced by metabolic activity related to cellular growth. Proliferation was determined up to 72 h after laser application. The irradiated cells revealed a considerably higher proliferation activity. The differences were highly significant up to 72 h after irradiation (Mann–Whitney U test, p < 0.001). A cellular responsiveness of human laryngeal carcinoma cells to low-level laser irradiation is obvious. The cell line is therefore suitable for basic research investigations concerning the biological mechanisms of LLLT on cells.     

The photodynamic effect of a pulsed dye laser on human bladder carcinoma cells in vitro

Summary The photodynamic effect of a pulsed flashlamp pumped dye laser on cultured human bladder carcinoma cells was studied. MGH-U1 cells were incubated for 1 h in dihaematoporphyrin ether (DHE) and then exposed to green laser light (504 nm, 20 Hz) for varying laser power densities (50–100 mW/cm² and exposure times (2–15 s), representing incident pulse energy fluences of 2.5–5 mJ/cm² and energy densities of 0.1–1.5 J/cm². The cell survival was measured by clonogenic assay and controls exposed to either laser light alone or DHE in the dark showed no cytotoxicity. Sensitised cells were killed by energy densities of less than 1 J/cm² (LD₉₀=0.54 J/cm²). This demonstrates the probable effectiveness of a pulsed dye laser for photodynamic therapy provided that pulse fluence are below the saturation threshold of the photosensitiser (10 mJ/cm²).     

Effect of the laser and light-emitting diode (LED) phototherapy on midpalatal suture bone formation after rapid maxilla expansion: a Raman spectroscopy analysis

The aim of this study was to analyze the effect of laser or light-emitting diode (LED) phototherapy on the bone formation at the midpalatal suture after rapid maxilla expansion. Twenty young adult male rats were divided into four groups with 8 days of experimental time: group 1, no treatment; group 2, expansion; group 3, expansion and laser irradiation; and group 4, expansion and LED irradiation. In groups 3 and 4, light irradiation was in the first, third, and fifth experimental days. In all groups, the expansion was accomplished with a helicoid 0.020″ stainless steel orthodontic spring. A diode laser (λ780 nm, 70 mW, spot of 0.04 cm², t?=?257 s, spatial average energy fluence (SAEF) of 18 J/cm²) or a LED (λ850 nm, 150 mW?±?10 mW, spot of 0.5 cm², t?=?120 s, SAEF of 18 J/cm²) were used. The samples were analyzed by Raman spectroscopy carried out at midpalatal suture and at the cortical area close to the suture. Two Raman shifts were analyzed: ～960 (phosphate hydroxyapatite) and ～1,450 cm^?1 (lipids and protein). Data was submitted to statistical analysis. Significant statistical difference (p?≤?0.05) was found in the hydroxyapatite (CHA) peaks among the expansion group and the expansion and laser or LED groups. The LED group presented higher mean peak values of CHA. No statistical differences were found between the treated groups as for collagen deposition, although LED also presented higher mean peak values. The results of this study using Raman spectral analysis indicate that laser and LED light irradiation improves deposition of CHA in the midpalatal suture after orthopedic expansion.     

A randomized placebo-blind study of the effect of low power laser on pain caused by irreversible pulpitis

This randomized placebo-blind study aimed to evaluate the effect of laser phototherapy (LPT) on pain caused by symptomatic irreversible pulpitis (SIP). Sixty patients diagnosed with SIP were randomly assigned to treatment groups (n?=?15): G1 (control), G2 (laser placebo—sham irradiation), G3 (laser irradiation at 780 nm, 40 mW, 4 J/cm²), and G4 (laser irradiation at 780 nm, 40 mW, 40 J/cm²). Spontaneous pain was recorded using a VAS score before (T0), immediately after (T1), and 15 min after treatment (T2). Local anesthetics failure during emergency endodontic treatment was also assessed. There was no pain difference in T1 and T2 between the experimental laser groups (G3 and G4) and the placebo group (G2). The 4-J/cm² (G3) irradiation resulted in significant increase in the local anesthetics failure in lower jar teeth. This effect could be suggested as consequence of the LPT improvement in local circulation and vasodilatation that would result in the increase of local anesthetic agent absorption. The application of 780-nm diode laser irradiation, at 4 and 40 J/cm², showed no effect in reducing the pain in SIP in comparison to the placebo group. The fluence of 4 J/cm² showed a negative effect in local anesthetics, resulting in significant increase of complimentary local anesthesia during emergency endodontic treatment. This work provides evidence of the consequence of LPT application on teeth with symptomatic irreversible pulpitis. LPT should be avoided in teeth with pain due to irreversible pulpitis.     

The relevance of pulse repetition rate and radiant exposure to the neurophysiological effects of low-intensity laser (820 nm/pulsed wave) irradiation upon skin temperature and antidromic conduction latencies in the human median nerve

The effects of low-intensity near-infra-red laser irradiation (820 nm; 1.5 and 9.0 J cm^–2; pulsed at 12 Hz, 73 Hz and 5 kHz) upon peripheral neurophysiology and skin temperature were investigated using antidromic conduction studies in the human median nerve in vivo. Healthy human volunteers (n = 90) were recruited and allocated randomly to either a control group (n=10) or one of eight experimental groups (two radiant exposures, 1.5 J cm^–2 and 9.0 J cm^–2 at one of three pulse repetition rates, 12 Hz, 73 Hz or 5 kHz, in addition to a placebo group for each radiant exposure;n = 10 all groups). Analysis of variance (ANOVA) demonstrated a significant (p0.05) decrease in skin temperature following irradiation at the lowest radiant exposure (1.5 J cm^–2) combined with pulse repetition rates of 73 Hz and 5 kHz, with the greatest effect at 73 Hz. These changes in skin temperature were coupled with increases in negative peak latency (NPL); ie changes in NPL were inversely related to changes in skin temperature. However, in contrast to the authors' previous findings using continuous wave (CW) laser irradiation, differences in NPL were not found to be significant. These findings, therefore, provide little evidence of the neuro-physiological effects of low-intensity infra-red irradiation at the dosage levels and pulse repetition rates used here.     

Effect of soft laser and bioactive glass on bone regeneration in the treatment of infra-bony defects (a clinical study)

This study aimed to investigate the influence of low-power 830 nm gallium–aluminium–arsenide (GaAlAs) laser [continuous wave (CW) 40 mW and fluence 4 J/cm², with total energy density of 16 J/cm²] on the healing of human infra-bony defects treated with bioactive glass graft material. Twenty patients with chronic periodontitis and bilateral infra-bony defects were included. Using a split mouth design, we treated 20 defects with bioactive glass plus laser irradiation during surgical procedures and on days 3, 5, 7 postoperatively; 20 contra-lateral defects were treated with bioactive glass only. Clinical probing pocket depths, clinical attachment levels and standardized periapical radiographs were recorded at baseline and at 3 months and 6 months postoperatively. At 3 months there was a statistically significant difference between the laser and non-laser sites in the parameters investigated. However, at 6 months, no difference was observed. Our results have confirmed the positive effect of soft laser in accelerating periodontal wound healing.     

Effect of Helium-Neon (He-Ne) Laser Irradiation on Dog Neoplasm Cells in Culture

The effects of laser light on the cellular proliferation have been extensively characterised. Low-power laser sources, such as the helium–neon (He-Ne) laser irradiation with a wavelength of 632.8 nm, have been found to produce photobiological and photodamaging effects with evidence of interference with cell proliferation functions. The present study has investigated the in vitro effect of He-Ne laser irradiation on the proliferative action of dog tumour cells in culture. Dose–response studies showed that repeated He-Ne irradiation (irradiance 12.8 mW/cm²) once a day for 4 consecutive days in a dose range between 0.13 and 2.08 J/cm² significantly increased with increasing energy density up to a laser dose of 0.26 J/cm², whereas at >1.04 J/cm², the cell proliferation decreased with increasing energy densities. It is concluded that the application of He-Ne laser irradiation at energy densities ranging from 0.13 J/cm² to 2.08 J/cm² produced different effects on cell proliferation in dog tumour cells in culture. Paper received for publication 27 June 1997; accepted following revision 6 February 1998.     

Inhibitory effects of low-level laser therapy on skin-flap survival in a rat model

BACKGROUND:Although several studies have demonstrated the effects of low-level laser therapy (LLLT) on skin flap viability, the role of higher doses has been poorly investigated.OBJECTIVE:To investigate the inhibitory effect of the LLLT (λ=670 nm) on the viability of random skin flaps in a rat model using an irradiation energy of 2.79 J at each point.METHODS:Sixteen Wistar rats were randomly assigned into two groups: sham laser irradiation (n=8); and active laser irradiation (n=8). Animals in the active laser irradiation group were irradiated with a 670 nm diode laser with an energy of 2.79 J/point, a power output 30 mW, a beam area of 0.028 cm², an energy density of 100 J/cm², an irradiance of 1.07 W/cm² for 93 s/point. Irradiation was performed in 12 points in the cranial skin flap portion. The total energy irradiated on the tissue was 33.48 J. The necrotic area was evaluated on postoperative day 7.RESULTS:The sham laser irradiation group presented a mean (± SD) necrotic area of 47.96±3.81%, whereas the active laser irradiation group presented 62.24±7.28%. There was a significant difference in skin-flap necrosis areas between groups (P=0.0002).CONCLUSION:LLLT (λ=670 nm) increased the necrotic area of random skin flaps in rats when irradiated with an energy of 2.79 J (100 J/cm²).     

Effects of low-power laser irradiation (LPLI) at different wavelengths and doses on oxidative stress and fibrogenesis parameters in an animal model of wound healing

Gallium-arsenide (GaAs) and helium-neon (HeNe) lasers are the most commonly used low-energy lasers in physiotherapy for promoting wound healing and pain modulation. The aim of this study was investigate the effect of low-power laser irradiation (LPLI) at different wavelengths and doses on oxidative stress and fibrogenesis parameters in an animal model of wound healing. The animals were randomly divided into five groups (n = 6): Controls (skin injured animals without local or systemic treatment), skin injury treated with HeNe 1 J/cm² (two seg); skin injury treated with HeNe 3 J/cm² (six seg); skin injury treated with GaAs 1 J/cm² (three seg); skin injury treated with GaAs 3 J/cm² (nine seg). A single circular wound measuring 8 mm in diameter was surgically created on the back of the animal. The rats were irradiated at 2, 12, 24, 48, 72, 96, and 120 h after skin injury. The parameters, namely hydroxyproline content, activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), and lipid (TBARS) and protein oxidation (carbonyl groups) measurements were assessed. In addition, wound size regression was also analyzed. The results showed an improvement in the wound healing reflected by the reduction in wound size and increased collagen synthesis. Moreover, a significant reduction in TBARS levels, carbonyl content, and SOD and CAT activities were observed after laser irradiation, particularly with the treatments HeNe laser 1 and 3 J/cm² dose and GaAs 3 J/cm² dose. The data strongly indicate that LPLI therapy is efficient in accelerating the skin wound healing process after wounding, probably by reducing the inflammatory phase and inducing collagen synthesis.     

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/cm²; group treated with 6 J/cm²; group treated with 12 J/cm² and group treated with 24 J/cm². 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.     

Effect of red and near-infrared wavelengths on low-level laser (light) therapy-induced healing of partial-thickness dermal abrasion in mice

Low-level laser (light) therapy (LLLT) promotes wound healing, reduces pain and inflammation, and prevents tissue death. Studies have explored the effects of various radiant exposures on the effect of LLLT; however, studies of wavelength dependency in in vivo models are less common. In the present study, the healing effects of LLLT mediated by different wavelengths of light in the red and near-infrared (NIR) wavelength regions (635, 730, 810, and 980 nm) delivered at constant fluence (4 J/cm²) and fluence rate (10 mW/cm²) were evaluated in a mouse model of partial-thickness dermal abrasion. Wavelengths of 635 and 810 nm were found to be effective in promoting the healing of dermal abrasions. However, treatment using 730- and 980-nm wavelengths showed no sign of stimulated healing. Healing was maximally augmented in mice treated with an 810-nm wavelength, as evidenced by significant wound area reduction (p?<?0.05), enhanced collagen accumulation, and complete re-epithelialization as compared to other wavelengths and non-illuminated controls. Significant acceleration of re-epithelialization and cellular proliferation revealed by immunofluorescence staining for cytokeratin-14 and proliferating cell nuclear antigen (p?<?0.05) was evident in the 810-nm wavelength compared with other groups. Photobiomodulation mediated by red (635 nm) and NIR (810 nm) light suggests that the biological response of the wound tissue depends on the wavelength employed. The effectiveness of 810-nm wavelength agrees with previous publications and, together with the partial effectiveness of 635 nm and the ineffectiveness of 730 and 980 nm wavelengths, can be explained by the absorption spectrum of cytochrome c oxidase, the candidate mitochondrial chromophore in LLLT.     

Effect of equal daily doses achieved by different power densities of low-level laser therapy at 635 nm on open skin wound healing in normal and corticosteroid-treated rats

Optimal parameters of low-level laser therapy (LLLT) for wound healing are still discussed. Hence, our study was aimed to compare effects of different power densities of LLLT at 635 nm in rats. Four, round, full-thickness, skin wounds were made on the backs of 48 rats that were divided into two groups (non-steroid laser-treated and steroid laser-treated). Three wounds were stimulated daily with a diode laser (daily dose 5 J/cm²) each with different power density (1 mW/cm², 5 mW/cm², and 15 mW/cm²), whereas the fourth wound served as a control. Two days, 6 days, and 14 days after surgery, eight animals from each group were killed and samples were removed for histological evaluation. In the non-steroid laser-treated rats, significant acceleration of epithelization and collagen synthesis 2 days and 6 days after surgery was observed in stimulated wounds. In steroid laser-treated rats, 2 days and 14 days after surgery, a decreased leucocyte/macrophage ratio and a reduction in the area of granulation tissue were recorded, respectively. In conclusion, LLLT, by the method we used, improved wound healing in the non-steroid laser-treated rats, but it was useless after corticosteroid treatment.