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.     

Effect of low-level laser therapy on types I and III collagen and inflammatory cells in rats with induced third-degree burns

Low-level laser therapy (LLLT) has been increasingly used to accelerate wound healing in third-degree burns. This study investigated the effects of lasers on the tissue repair process of third-degree burns. Burns were produced on the backs of male Wistar rats. The animals were divided into four groups (n?=?12): control, injury, LLLT 3 J/cm², and LLLT 4 J/cm². Each group was further divided into two subgroups; the rats in one subgroup were killed on day 8 and those in the other, on day 16 after injury. The animals in LLLT 3 J/cm² and LLLT 4 J/cm² were irradiated 1 h after injury, and irradiation was repeated every 48 h. Laser (660 nm, 35 mW) treatment at fluences of 3 and 4 J/cm² were used. After killing the rats, tissue fragments from the burnt area were removed for histological analysis. The LLLT-treated groups showed a significant decrease (p <0.05) in the number of inflammatory cells and increased collagen deposition compared to the injury group. Laser irradiation (both 3 and 4 J/cm²) resulted in reduction in the inflammatory process and improved collagen deposition, thereby ameliorating the healing of third-degree burns.     

The effects of low-level laser irradiation on cellular viability and proliferation of human skin fibroblasts cultured in high glucose mediums

Delayed wound healing is one of the most challenging complications of diabetes mellitus (DM) in clinical medicine. This study has aimed to evaluate the effects of low-level laser therapy (LLLT) on human skin fibroblasts (HSFs) cultured in a high glucose concentration. HSFs were cultured either in a concentration of physiologic glucose (5.5 mM/l) or high glucose media (11.1 and15?mM/l) for either 1 or 2 weeks after which they were subsequently cultured in either the physiologic glucose or high concentration glucose media during laser irradiation. LLLT was carried out with a helium–neon (He–Ne) laser unit at energy densities of 0.5, 1, and 2 J/cm², and power density of 0.66 mW/cm² on 3 consecutive days. HSFs’ viability and proliferation rate were evaluated with the dimethylthiazol-diphenyltetrazolium bromide (MTT) assay. The LLLT at densities of 0.5 and 1 J/cm² had stimulatory effects on the viability and proliferation rate of HSFs cultured in physiologic glucose (5.5 mM/l) medium compared to their control cultures (p?=?0.002 and p?=?0.046, respectively). All three doses of 0.5, 1, and 2 J/cm² had stimulatory effects on the proliferation rate of HSFs cultured in high glucose concentrations when compared to their control cultures (p?=?0.042, p?=?0.000, and p?=?0.000, respectively). This study showed that HSFs originally cultured for 2 weeks in high glucose concentration followed by culture in physiologic glucose during laser irradiation showed enhanced cell viability and proliferation. Thus, LLLT had a stimulatory effect on these HSFs.     

Effect of low-level laser therapy on odontoblast-like cells exposed to bleaching agent

The aim of the present study was to evaluate the effect of low-level laser therapy (LLLT) on odontoblast-like MDPC-23 cells exposed to carbamide peroxide (CP 0.01 %–2.21 μg/mL of H₂O₂). The cells were seeded in sterile 24-well plates for 72 h. Eight groups were established according to the exposure or not to the bleaching agents and the laser energy doses tested (0, 4, 10, and 15 J/cm²). After exposing the cells to 0.01 % CP for 1 h, this bleaching solution was replaced by fresh culture medium. The cells were then irradiated (three sections) with a near-infrared diode laser (InGaAsP—780?±?3 nm, 40 mW), with intervals of 24 h. The 0.01 % CP solution caused statistically significant reductions in cell metabolism and alkaline phosphate (ALP) activity when compared with those of the groups not exposed to the bleaching agent. The LLLT did not modulate cell metabolism; however, the dose of 4 J/cm² increased the ALP activity. It was concluded that 0.01 % CP reduces the MDPC-23 cell metabolism and ALP activity. The LLLT in the parameters tested did not influence the cell metabolism of the cultured cells; nevertheless, the laser dose of 4 J/cm² increases the ALP activity in groups both with and without exposure to the bleaching agent.     

Effect of photobiomodulation on expression of IL-1β in skeletal muscle following acute injury

Muscle repair is regulated by growth factors and cytokines. Low-level laser therapy (LLLT) seems to influence acute inflammation and accelerate skeletal muscle repair. This study verifies the effect of LLLT on the expression of IL-1β in the tibialis anterior (TA) muscle of rats following acute injury. Wistar rats (n?=?35) were allocated into three groups: control (without lesion and LLLT, n?=?5), injury group (n?=?15), and injury + LLLT group (n?=?15). The acute injury was induced by the contact with a cooled metal probe (3 mm in diameter) during 10 s, twice, in the same muscle area. LLLT was used three times a week using the InGaAlP laser (660 nm; beam spot of 0.04 cm², output power of 20 mW, power density of 500 mW/cm², and energy density of 5 J/cm² during 10 s). The animals were analyzed at 1, 7, and 14 days following injury. TA muscles samples were used for obtaining total RNA and performing cDNA synthesis. Real-time polymerase chain reactions were realized using IL-1β primer. There was a decrease in IL-1β expression after 7 days in LLLT group in comparison with the no treated group. In conclusion, LLLT was able to decrease IL-1β expression during the skeletal muscle repair following an acute injury.     

Comparative analysis of low-level laser therapy (660 nm) on inflammatory biomarker expression during the skin wound-repair process in young and aged rats

The wound-healing process plays an essential role in the protective response to epidermal injury by tissue regeneration. In the elderly, skin functions deteriorate as a consequence of morphological and structural changes. This study aimed to evaluate and compare the effect of low-level laser therapy (LLLT) in cutaneous wound healing in young and aged rats. A total of 60 male rats comprising 30 young (±30 days) and 30 aged (±500 days) was used. The animals were divided into four experimental groups and underwent skin wound and/or treatment with LLLT (660 nm, 30 mW, 1.07 W/cm², 0.028 cm², 72 J/cm², and 2 J). Analyses were conducted to verify the effects of LLLT in the tissue repair process, in the gene expression, and protein expression of TNF-α, IL-1β, and IL-10, obtained in skin wound model. Results showed that there were significant differences between the young control group and the aged control group and their respective treated groups (LLLT young and LLLT aged). We conclude that LLLT has shown to be effective in the treatment of skin wounds in young and aged animals at different stages of the tissue repair process, which suggests that different LLLT dosimetry should be considered in treatment of subjects of different ages. Further clinical trials are needed to confirm these findings in clinical settings.     

Effect of low-level laser therapy on healing of tenotomized Achilles tendon in streptozotocin-induced diabetic rats

Diabetes mellitus (DM) is associated with musculoskeletal damage. Investigations have indicated that healing of the surgically tenotomized Achilles tendon was considerably augmented following low-level laser therapy (LLLT) in non-diabetic, healthy animals. The aim of the present study was to evaluate the effect of LLLT on the Achilles tendon healing in streptozotocin-induced diabetic (STZ-D) rats via a biomechanical evaluating method. Thirty-three rats were divided into non-diabetic (n?=?18) and diabetic (n?=?15) groups. DM was induced in the rats by injections of STZ. The right Achilles tendons of all rats were tenotomized 1 month after STZ injections. The two experimental groups (n?=?6 for each group) of non-diabetic rats were irradiated with a helium–neon (He–Ne) laser at 2.9 and 11.5 J/cm² for ten consecutive days. The two experimental groups of diabetic rats (n?=?5 for each group) were irradiated with a He–Ne laser at 2.9 and 4.3 J/cm² for ten consecutive days. The tendons were submitted to a tensiometric test. Significant improvements in the maximum stress (MS) values (Newton per square millimeter) were found following LLLT at 2.9 J/cm² in both the non-diabetic (p?=?0.031) and diabetic (p?=?0.019) experimental groups when compared with their control groups. LLLT at 2.9 J/cm² to the tenotomized Achilles tendons in the non-diabetic and diabetic rats significantly increased the strength and MS of repairing Achilles tendons in our study.     

LLLT for the management of patients with ankylosing spondylitis

This study aimed to compare the effectiveness of the combined low-level laser therapy (LLLT) and passive stretching with combined placebo LLLT laser and the same passive stretching exercises in patients suffering from Αnkylosing spondylitis. Forty-eight patients suffering from Αnkylosing spondylitis participated in the study and were randomized into two groups. Group A (n?=?24) was treated with a λ?=?820 Ga-Al-As laser CW, with power intensity?=?60 mW/cm², energy per point in each session?=?4.5 J, total energy per session?=?27.0 J, in contact with specific points technique, plus passive stretching exercises. Group B (n?=?24), received placebo laser plus the same passive stretching exercises. Both groups received 12 sessions of laser or placebo within 8 weeks; two sessions per week (weeks 1–4) and one session per week (weeks 5–8). Pain and function scales were completed before the treatment, at the end of the fourth and eighth week of treatment, and 8 weeks after the end of treatment (follow-up). Group A revealed a significant improvement after 8 weeks of treatment in all pain and function scales. At 8-week follow-up, the improvement remained only for the pain, while for all other function outcomes the differences were not statistically significant. The results suggested that after an 8-week treatment and after a follow-up, the combination of LLLT and passive stretching exercises decreased pain more effectively than placebo LLLT along with the same passive stretching exercises in patients with Αnkylosing spondylitis. Future studies are needed to establish the relative and absolute effectiveness of the above protocol.     

The influence of low-level laser therapy on parameters of oxidative stress and DNA damage on muscle and plasma in rats with heart failure

In heart failure (HF), there is an imbalance between the production of reactive oxygen species and the synthesis of antioxidant enzymes, causing damage to the cardiovascular function and increased susceptibility to DNA damage. The aim of this study was to evaluate the influence of low-level laser therapy (LLLT) on parameters of oxidative stress and DNA damage in skeletal muscle and plasma of rats with HF. Wistar rats were allocated into six groups: “placebo” HF rats (P-HF, n?=?9), “placebo” Sham rats (P-sham, n?=?8), HF rats at a dose 3 J/cm² of LLLT (3 J/cm²-HF, n?=?8), sham rats at a dose 3 J/cm² of LLLT (3 J/cm²-sham, n?=?8), HF rats at a dose 21 J/cm² of LLLT (21 J/cm²-HF, n?=?8) and sham rats at a dose 21 J/cm² of LLLT (21 J/cm²-sham, n?=?8). Animals were submitted to a LLLT protocol for 10 days at the right gastrocnemius muscle. Comparison between groups showed a significant reduction in superoxide dismutase (SOD) activity in the 3 J/cm²-HF group (p?=?0.03) and the 21 J/cm²-HF group (p?=?0.01) compared to the P-HF group. 2′,7′-Dihydrodichlorofluorescein (DCFH) oxidation levels showed a decrease when comparing 3 J/cm²-sham to P-sham (p?=?0.02). The DNA damage index had a significant increase either in 21 J/cm²-HF or 21 J/cm²-sham in comparison to P-HF (p?=?0.004) and P-sham (p?=?0.001) and to 3 J/cm²-HF (p?=?0.007) and 3 J/cm²-sham (p?=?0.037), respectively. Based on this, laser therapy appears to reduce SOD activity and DCFH oxidation levels, changing the oxidative balance in the skeletal muscle of HF rats. Otherwise, high doses of LLLT seem to increase DNA damage.     

808-nm laser therapy with a flat-top handpiece photobiomodulates mitochondria activities of <Emphasis Type="Italic">Paramecium primaurelia</Emphasis> (Protozoa)

Photobiomodulation is proposed as a non-linear process, and only low-level laser therapy (LLLT) is assumed to stimulate exposed cells, whereas high powered laser and fluences can cause negative effects, exhausting the cell’s energy reserve as a consequence of excessive photon-based stimulation. In our work, we investigated and compared the effects of 808-nm diode laser (CW) with a new flat-top handpiece. To this purpose, we tested the photobiomodulation effects of 1 and 3 J/cm² fluence, both generated by 100 mW or 1 W of laser power and of 64 J/cm² of fluence generated by 100 mW, 1 W, 1.5 W or 2 W, as expressed through oxygen consumption and ATP synthesis of Paramecium. Data collected indicates the incremental consumption of oxygen through irradiation with 3 J/cm²–100 mW or 64 J/cm²–1 W correlates with an increase in Paramecium ATP synthesis. The Paramecium respiration was inhibited by fluences 64 J/cm²–100 mW or 64 J/cm²–2 W and was followed by a decrease in the endogenous ATP concentration. The 1 J/cm²–100 mW or 1 W and 3 J/cm²–1 W did not affect mitochondrial activity. The results show that the fluence of 64 J/cm²–1 W more than the 3 J/cm²–100 mW causes greater efficiency in Paramecium mitochondria respiratory chain activity. Our results suggest that thanks to flat-top handpiece we used, high fluences by high-powered laser have to be reconsidered as an effective and non-invasive therapy. Possible associated benefits of deeper tissue penetration would increase treatment effectiveness and reduced irradiation time.     

LLLT improves tendon healing through increase of MMP activity and collagen synthesis

The Achilles tendon has a high incidence of rupture, and the healing process leads to a disorganized extracellular matrix (ECM) with a high rate of injury recurrence. To evaluate the effects of different conditions of low-level laser (LLL) application on partially tenotomized tendons, adult male rats were divided into the following groups: G1, intact; G2, injured; G3, injured + LLL therapy (LLLT; 4 J/cm² continuous); G4, injured + LLLT (4 J/cm², 20 Hz); G5, injured; G6, injured + LLLT (4 J/cm² continuous); and G7, injured + LLLT (4 J/cm², 20 Hz until the 7th day and 2 kHz from 8 to 14 days). G2, G3, and G4 were euthanized 8 days after injury, and G5, G6, and G7 were euthanized on the 15th day. The quantification of hydroxyproline (HOPro) and non-collagenous protein (NCP), zymography for matrix metalloproteinase (MMP)-2 and MMP-9, and Western blotting (WB) for collagen types I and III were performed. HOPro levels showed a significant decrease in all groups (except G7) when compared with G1. The NCP level increased in all transected groups. WB for collagen type I showed an increase in G4 and G7. For collagen type III, G4 presented a higher value than G2. Zymography for MMP-2 indicated high values in G4 and G7. MMP-9 increased in both treatment groups euthanized at 8 days, especially in G4. Our results indicate that the pulsed LLLT improved the remodeling of the ECM during the healing process in tendons through activation of MMP-2 and stimulation of collagen synthesis.     

Evaluation of low-level laser therapy,platelet-rich plasma,and their combination on the healing of Achilles tendon in rabbits

Tendon repair is still one of the challenges for rehabilitation. Various treatments for tendon injuries have been used in recent decade. This study was established to investigate the effects of low-level laser therapy (LLLT), platelet-rich plasma (PRP) treatment alone, and using combined method on the healing of Achilles tendon in rabbits. Seventy-two healthy mature male white New Zealand rabbits were divided randomly into four groups of 18 animals each: control: partial tenotomy with no treatment, only 1 mL normal saline was injected on days 1, 8, and 15 at the site of splitting; PRP: partial tenotomy with PRP treatment on days 1, 8, and 15 at the site of splitting; LLLT: partial tenotomy with LLLT (K30 hand-held probe, AZOR, Technica, Russia, 650 nm, 30 mW, surface area?=?1 cm², 60 S/cm², energy density?=?1.8 J/cm²) for 15 consecutive days; LLLT?+?PRP: partial tenotomy with LLLT?+?PRP. At the end of trial, the rabbits were euthanatized and tendon specimens were harvested and were submitted for histopathological evaluation, hydroxyproline levels, and biomechanical measurement. The Tukey post hoc test was performed. The results for these parameters showed that PRP or LLLT alone has significant advantages over untreated animals (P?<?0.05). Furthermore, it was found that the combined treatment with PRP and LLLT is even more efficient. There was no significant difference (P?>?0.05) between the two groups of LLLT and PRP. However, the treatments combining PRP and LLLT showed significant results in comparison of PRP or LLLT alone (P?<?0.05). Our results demonstrate that the healing time of injured tendon decreases by using the two therapies combined.     

Effects of different fluences of low-level laser therapy in an experimental model of spinal cord injury in rats

The aim of this study was to evaluate the in vivo response of different fluences of low-level laser therapy (LLLT) on the area of the injury, inflammatory markers, and functional recovery using an experimental model of traumatic spinal cord injury (SCI). Thirty two rats were randomly divided into four experimental groups: control group (CG), laser-treated group 500 J/cm² (L-500), laser-treated group 750 J/cm² (L-750), and laser-treated group 1000 J/cm² (L-1000). SCI was performed by an impactor equipment (between the ninth and tenth thoracic vertebrae), with a pressure of 150 kdyn. Afterwards, the injured region was irradiated daily for seven consecutive sessions, using an 808-nm laser, at the respective fluence of each experimental groups. Motor function and tactile sensitivity were performed on days 1 and 7 post-surgery. Animals were euthanized on the eighth day after injury, and the samples were retrieved for histological and immunohistochemistry analyses. Functional evaluation and tactile sensitivity were improved after LLLT, at the higher fluence. Additionally, LLLT, at 750 and 1000 J/cm², reduces the lesion volume and modulates the inflammatory process with decrease of CD-68 protein expression. These results suggest that LLLT at higher doses was effective in promoting functional recovery and modulating inflammatory process in the spinal cord of rats after SCI.     

Laser phototherapy improves early stage of cutaneous wound healing of rats under hyperlipidic diet

The aim of this study was to evaluate the influence of laser photobiomodulation in cutaneous healing of rats under a hyperlipidic diet. Forty-eight Wistar Albinus rats, weaned, received standard diet (SD) or hyperlipidic diet (HD) for 20 weeks. The groups were divided into SD rats and HD rats, SD-irradiated rats (LSD), and HD-irradiated rats (LHD). Standard cutaneous wound (1 cm²) was created on the dorsum of each rat. The irradiation started immediately after surgery and every 48 h for 7 or 14 days (λ660 nm, 40 mW, 6 J/cm², ? 0,04 cm², CW), when they were killed under deep anesthesia. The specimens were removed, routinely processed, stained with hematoxylin/eosin (H/E), and evaluated by light microscopy. Rats fed with hyperlipidic diet had greater intensity in the inflammatory process and prolonged hyperemia. At day 7, the intensity of inflammation was reduced in LSD and LHD groups when compared to their control groups, SD (p?=?0.002) and HD (p?=?0.02). There was an increase in fibroblast proliferation and collagen deposition, especially in the LHD group. At day 14, the HD group presented more intensive hyperemia than the SD group. It can be concluded that the hyperlipidic diet modified the inflammation pattern in wound healing and that laser light has a positive biomodulative effect on the healing process only in early stages.     

Effect of low-level laser therapy (LLLT) on orthodontic tooth movement

The aim of this study is to evaluate the effects of low-level laser therapy (LLLT) on (1) the velocity of orthodontic tooth movement and (2) the nitric oxide levels in gingival crevicular fluid (GCF) during orthodontic treatment. The sample consisted of 20 patients (14 girls, six boys) whose maxillary first premolars were extracted and canines distalized. A gallium-aluminum-arsenide (Ga-Al-As) diode laser was applied on the day 0, and the 3rd, 7th, 14th, 21st, and 28th days when the retraction of the maxillary lateral incisors was initiated. The right maxillary lateral incisors composed the study group (the laser group), whereas the left maxillary lateral incisors served as the control. The teeth in the laser group received a total of ten doses of laser application: five doses from the buccal and five doses from the palatal side (two cervical, one middle, two apical) with an output power of 20 mW and a dose of 0.71 J /cm². Gingival crevicular fluid samples were obtained on the above-mentioned days, and the nitric oxide levels were analyzed. Bonferroni and repeated measures variant analysis tests were used for statistical analysis with the significance level set at p ≤ 0.05. The application of low-level laser therapy accelerated orthodontic tooth movement significantly; there were no statistically significant changes in the nitric oxide levels of the gingival crevicular fluid during orthodontic treatment.     

Effect of low-level laser therapy on repair of the bone compromised by radiotherapy

Radiotherapy (RDT) is commonly used for cancer treatment, but high doses of ionizing radiation can directly affect healthy tissues. Positive biological effects of low-level laser therapy (LLLT) on bone repair have been demonstrated; however, this effect on surgical defects of bone previously compromised by radiotherapy has not been evaluated. The aim of this study was to investigate the influence of LLLT (λ?=?830 nm) in femur repair after ionizing radiation. Twenty Wistar rats were divided into four groups: control group (GC, n?=?5) creation of bone defects (BDs) only; laser group (GL), with BD and LLLT (n?=?5); radiotherapy group (GR), submitted to RDT and BD (n?=?5); and radiotherapy and laser group (GRL), submitted to RDT, BD, and LLLT (n?=?5). GL and GRL received punctual laser application (DE?=?210 J/cm², P?=?50 mW, t?=?120 s, and beam diameter of 0.04 cm²) immediately after surgery, with 48-h interval during 7 days. Animals were euthanized at 7 days after surgery, and bone sections were evaluated morphometrically with conventional microscopy. Bone repair was only observed in nonirradiated bone, with significant improvement in GL in comparison to GC. GR and GRL did not present any bone neoformation. The result demonstrated a positive local biostimulative effect of LLLT in normal bone. However, LLLT was not able to revert the bone metabolic damage due to ionizing radiation.     

What is better in TRAM flap survival: LLLT single or multi-irradiation?

Low-level laser therapy (LLLT) has been used with the aim of improving vascular perfusion of the skin and musculocutaneous flaps. This study evaluated the effect of LLLT on transverse rectus abdominis musculocutaneous flap (TRAM) viability, vascular angiogenesis, and VEGF release. Eighty-four Wistar rats were randomly divided into seven groups with 12 rats in each group. Group 1 received sham laser treatment; group 2, 3 J/cm² at 1 point; group 3, 3 J/cm² at 24 points; group 4, 72 J/cm² at 1 point; group 5, 6 J/cm² at 1 point; group 6, 6 J/cm² at 24 points; and group 7, 144 J/cm² at 1 point. All experimental groups underwent LLLT immediately after the TRAM operation and on the following 2 days; thus, animals underwent 3 days of treatment. The percentage of skin flap necrosis area was calculated on the fourth postoperative day using the paper template method, and two skin samples were collected using a 1-cm² punch to evaluate alpha smooth muscle actin (1A4) and VEGF levels in blood vessels. Significant differences were found in necrosis percentage, and higher values were seen in group 1 than in the other groups. Statistically significant differences were not found among groups 3 to 7 (p?<?0.292). Groups 5 and 7 showed significantly higher VEGF levels compared to other groups. Groups 3 and 5 had an increase in levels of blood vessels compared to other groups. LLLT at energy densities of 6 to 144 J/cm² was efficient to increase angiogenesis and VEGF levels and promote viability in TRAM flaps in rats.     

Analgesic effect of a low-level laser therapy (830 nm) in early orthodontic treatment

The aim of this study was to evaluate the pain sensation that orthodontic patients experience when elastic separators are placed between molars and premolars and to determine the degree of analgesic efficacy of low-level laser therapy (LLLT) compared to a placebo treatment. The study was conducted with 20 volunteers who were fitted with elastic separators between the maxillary molars and premolars. One quadrant was randomly chosen to be irradiated with an 830-nm laser, 100 mW, beam diameter of 7 mm, 250 mW/cm² applied for 20 s per point (5 J/cm²). Three points were irradiated in the buccal face and three were irradiated in the palate. The same procedure was applied in the contralateral quadrant with a placebo light. A visual analogue scale was used to assess pain 5 min, 6 h, 24 h, 48 h, and 72 h after placement of the separators. Maximum pain occurred 6–24 h after placement of the elastic separators. Pain intensity was significantly lower in the laser-treated quadrant (mean, 7.7 mm) than in the placebo-treated quadrant (mean, 14.14 mm; p?=?0.0001). LLLT at these parameters can reduce pain in patients following placement of orthodontic rubber separators.     

High doses of laser phototherapy can increase proliferation in melanoma stromal connective tissue

It is well established that laser phototherapy (LP) is contraindicated directly over cancer cells, due to its bio modulatory effects in cell and blood vessel proliferation. The aim of the present study was to analyze the influence of typical low-level laser therapy (LLLT) and high intensity laser therapy (HILT) and an in-between dose of 9 J on collagen fibers and blood vessels content in melanoma tumors (B16F10) implanted in mice. Melanoma tumor cells were injected in male Balb C mice which were distributed in four groups: control (no irradiated) or irradiated by 3, 9, or 21 J (150; 450, or 1050 J/cm²). LP was performed in daily sessions for 3 days with a InGaAlP—660 nm (mean output: 50 mW, spot size: 2 mm²). Tumor volume was analyzed using (1) picrosirius staining to quantify collagen fibers content and (2) Verhoeff’s method to quantify blood vessels content. Tumor growth outcome measured in the 3-J group was not significantly different from controls. Nine and 21-J groups, presented significant and dose-dependent increases in tumor volume. Quantitative analysis of the intensity of collagen fibers and their organization in stroma and peri-tumoral microenvironment showed significant differences between irradiated and control group. Blood vessels count of 21-J group outnumbered the other groups. High doses (≥ 9 J) of LP showed a dose-dependent tumor growth, different collagen fibers characteristics, and eventually blood vessel growth, while a typical LLLT dose (3 J) appeared harmless on melanoma cell activity.