Study on the selection of laser wavelengths in the intravascular low-level laser irradiation therapy

According to the absorption spectra of blood and hemoglobin, a photon-bond energy formula is established using physical methods and the effects on hemoglobin of low-level laser at different wavelengths are analyzed. The results show that lasers with the peak wavelengths of 200～240, 275, and 342 nm in the whole blood absorption spectra curve are easy to destroy protein molecules and then lead to hemoglobin lose biological activity. While lasers with wavelengths longer than 800 nm will reduce the oxygen carrying capacity of blood, only lasers with wavelengths between 630 and 670 nm have the best efficacy.     

Clinical implications of hemoglobin as a nitric oxide carrier

Hemoglobin is perhaps the most intensively studied of the biologically important molecules. Much is known of its structure, its function, and its regulation. In addition to well-characterized processes of oxygen, carbon dioxide, and carbon monoxide transport, new data suggest a key role of hemoglobin as a carrier of nitric oxide. In this review, we describe the basis of this interaction, as well as its clinical relevance to such problems as acute respiratory distress syndrome, percutaneous transluminal coronary angioplasty, and transplant allograft survival.     

Increase in the nitric oxide release without changes in cell viability of macrophages after laser therapy with 660 and 808 nm lasers

The aim of this study was to evaluate the influence of low-level laser therapy (LLLT) with different parameters and wavelengths on nitric oxide (NO) release and cell viability. Irradiation was performed with Ga-Al-As laser, continuous mode and wavelengths of 660 and 808 nm at different energy and power densities. For each wavelength, powers of 30, 50, and 100 mW and times of 10, 30, and 60 s were used. NO release was measured using Griess reaction, and cell viability was evaluated by mitochondrial reduction of bromide 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) to formazan. LLLT promoted statistically significant changes in NO release and MTT value only at the wavelength of 660 nm (p?<?0.05). LLLT also promoted an increase in the NO release and cell viability when the energy densities 64 (p?=?0.04) and 214 J/cm² (p?=?0.012), respectively, were used. LLLT has a significant impact on NO release without affecting cell viability, but the significance of these findings in the inflammatory response needs to be further studied.     

Low-level laser irradiation modifies the effect of hyperglycemia on adhesion molecule levels

Endothelium plays a key role in maintaining vascular homeostasis by secreting active factors involved in many biological processes such as hemostasis, angiogenesis, and inflammation. Hyperglycemia in diabetic patients causes dysfunction of endothelial cells. Soluble fractions of adhesion molecules like sE-selectin and vascular cell adhesion molecule (sVCAM) are considered as markers of endothelial damage. The low-level laser therapy (LLLT) effectively supports the conventional treatment of vascular complications in diabetes, for example hard-to-heal wounds in patients with diabetic foot syndrome. The aim of our study was to evaluate the effect of low-energy laser at the wavelength of 635 nm (visible light) and 830 nm (infrared) on the concentration of adhesion molecules: sE-selectin and sVCAM in the supernatant of endothelial cell culture of HUVEC line. Cells were cultured under high-glucose conditions of 30 mM/L. We have found an increase in sE-selectin and sVCAM levels in the supernatant of cells cultured under hyperglycemic conditions. This fact confirms detrimental influence of hyperglycemia on vascular endothelial cell cultures. LLLT can modulate the inflammation process. It leads to a decrease in sE-selectin and sVCAM concentration in the supernatant and an increase in the number of endothelial cells cultured under hyperglycemic conditions. The influence of LLLT is greater at the wavelength of 830 nm.     

Low-level laser irradiation stimulates tenocyte proliferation in association with increased NO synthesis and upregulation of PCNA and cyclins

Low-level laser therapy is commonly used to treat tendinopathy or tendon injury. Tendon healing requires tenocyte migration to the repair site, followed by proliferation and synthesis of the extracellular matrix. There are few evidence to elucidate that low-level laser promote tenocyte proliferation. This study was designed to determine the effect of laser on tenocyte proliferation. Furthermore, the association of this effect with secretion of nitric oxide (NO) and the expressions of proliferating cell nuclear antigen (PCNA) and cyclins D1, E, A, and B1 was investigated. Tenocytes intrinsic to rat Achilles tendon were treated with low-level laser (660 nm). Tenocyte proliferation was evaluated by MTT assay and immunocytochemistry with Ki-67 stain. NO in the conditioned medium was measured by ELISA. Western blot analysis was used to evaluate the protein expressions of PCNA and cyclins D1, E, A, and B1. The results revealed that tenocytes proliferation was enhanced dose dependently by laser. NO secretion was increased after laser treatment. PCNA and cyclins E, A, and B1 were upregulated by laser. In conclusion, low-level laser irradiation stimulates tenocyte proliferation in a process that is mediated by upregulation of NO, PCNA, and cyclins E, A, and B1.     

Carbon monoxide: toxic molecule with antiinflammatory and cytoprotective properties

Carbon monoxide arises during incomplete combustion of organic material, is incorporated into the circulation via the lungs and displaces oxygen from hemoglobin. Consecutively, symptoms of intoxication such as headache, vertigo, nausea, seizures and coma may result in a dose dependent fashion. Carbon monoxide is however also generated endogenously during heme degradation catalysed by heme oxgenase enzymes. The isoform hemeoxygenase-1 is inducible by oxidative stress and may mediate cytoprotection mainly attributable to endogenously produced carbon monoxide. Exogenous applied carbon monoxide has also been shown to confer protection in experimental studies. Meanwhile, in addition to the toxicological properties, antiinflammatory and cytoprotective effects of carbon monoxide have moved into the focus of scientific interest.     

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.     

Cell viability,reactive oxygen species,apoptosis, and necrosis in myoblast cultures exposed to low-level infrared laser

Low-level infrared laser is considered safe and effective for treatment of muscle injuries. However, the mechanism involved on beneficial effects of laser therapy are not understood. The aim was to evaluate cell viability, reactive oxygen species, apoptosis, and necrosis in myoblast cultures exposed to low-level infrared laser at therapeutic fluences. C2C12 myoblast cultures at different (2 and 10 %) fetal bovine serum (FBS) concentrations were exposed to low-level infrared laser (808 nm, 100 mW) at different fluences (10, 35, and 70 J/cm²) and evaluated after 24, 48, and 72 h. Cell viability was evaluated by WST-1 assay; reactive oxygen species (ROS), apoptosis, and necrosis were evaluated by flow cytometry. Cell viability was decreased atthe lowest FBS concentration. Laser exposure increased the cell viability in myoblast cultures at 2 % FBS after 48 and 72 h, but no significant increase in ROS was observed. Apoptosis was decreased at the higher fluence and necrosis was increased at lower fluence in myoblast cultures after 24 h of laser exposure at 2 % FBS. No laser-induced alterations were obtained at 10 % FBS. Results show that level of reactive oxygen species is not altered, at least to those evaluated in this study, but low-level infrared laser exposure affects cell viability, apoptosis, and necrosis in myoblast cultures depending on laser fluence and physiologic conditions of cells.     

Carbon monoxide induces relaxation of human internal thoracic and radial arterial grafts

Carbon monoxide is produced by the degradation of heme by intracellular heme-oxygenase. The aim of our study was to evaluate, in vitro, the vasodilating effect of carbon monoxide and its mechanisms of action on human internal thoracic and radial artery grafts. Segments of human internal thoracic artery and radial artery, obtained from isolated coronary artery bypass surgery patients, were studied in organ chambers. The arterial rings were precontracted with norepinephrine then submitted to carbon monoxide. Inhibitors of nitric oxide synthase and of soluble guanylate cyclase were added to some arterial rings. Carbon monoxide induced significant relaxation in precontracted human internal thoracic artery and radial artery rings. This relaxation was independent of the presence of functional endothelium in internal thoracic artery. Blocking soluble guanylate cyclase partially inhibited this relaxation, while blocking nitric oxide synthase had no effect. Carbon monoxide has a relaxing effect on human internal thoracic artery and radial artery grafts in vitro, partially via cyclic guanylate monophosphate (cGMP) pathway activation. Inducing carbon monoxide production at the cellular level in vivo in human arterial grafts might help prevent vasospasm.     

Bactericide effect of methylene blue associated with low-level laser therapy in <Emphasis Type="Italic">Escherichia coli</Emphasis> bacteria isolated from pressure ulcers

The present study analyzed the bactericidal effect of methylene blue associated with low-level lasers on Escherichia coli isolated from a pressure ulcer. Microbiological material from a pressure ulcer was isolated using an aseptic swab, and antimicrobial activity was verified using the diffusion disc method. Methylene blue was used at concentrations of 0.001 and 0.005%, and low-level lasers of 670, 830, and 904 nm, with the energy densities of 4, 8, 10, and 14 J/cm², were tested on three plates each and combined with methylene blue of each concentration. In addition, three control plates were used, with each concentration and energy density separated without any interventions. The results were analyzed using the paired sample t test to determine the bactericidal effect of the methylene blue and using the ANOVA test to compare the effects of the energy densities and wavelengths among the low-level laser treatment protocols. The results showed bacterial reduction at wavelengths of 830 and 904 nm and more proliferation in wavelengths of 670 nm. In wavelength of 830 nm, a bacterial reduction was observed in the conditions with 0.001% methylene blue in all energy density utilized, with 0.005% methylene blue in energy density of 10 J/cm², and without methylene blue in energy density at 10 J/cm². And in a wavelength of 904 nm, all condition showed bacterial reduction with or without methylene blue. We concluded that the low-level lasers of 904 and 830 nm have bactericidal effects and at better energy densities (10 and 14 J/cm²).     

Effects of low-level laser therapy on mitochondrial respiration and nitrosyl complex content

Among the photochemical reactions responsible for therapeutic effects of low-power laser radiation, the photolysis of nitrosyl iron complexes of iron-containing proteins is of primary importance. The purpose of the present study was to compare the effects of blue laser radiation on the respiration rate and photolysis of nitrosyl complexes of iron-sulfur clusters (NO-FeS) in mitochondria, subjected to NO as well as the possibility of NO transfer from NO-FeS to hemoglobin. It was shown that mitochondrial respiration in State 3 (V₃) and State 4 (V₄), according to Chance, dramatically decreased in the presence of 3 mM NO, but laser radiation (λ?=?442 nm, 30 J/cm²) restored the respiration rates virtually to the initial level. At the same time, electron paramagnetic resonance (EPR) spectra showed that laser irradiation decomposed nitrosyl complexes produced by the addition of NO to mitochondria. EPR signal of nitrosyl complexes of FeS-clusters, formed in the presence of 3 mM NO, was maximal in hypoxic mitochondria, and disappeared in a dose-dependent manner, almost completely at the irradiation dose 120 J/cm². EPR measurements showed that the addition of lysed erythrocytes to mitochondria decreased the amount of nitrosyl complexes in iron-sulfur clusters and produced the accumulation of NO-hemoglobin. On the other hand, the addition of lysed erythrocytes to mitochondria, preincubated with nitric oxide, restored mitochondrial respiration rates V₃ and V₄ to initial levels. We may conclude that there are two possible ways to destroy FeS nitrosyl complexes in mitochondria and recover mitochondrial respiration inhibited by NO: laser irradiation and ample supply of the compounds with high affinity to nitric oxide, including hemoglobin.     

Effect of low-level laser therapy after implantation of poly-<Emphasis Type="SmallCaps">L</Emphasis>-lactic/polyglycolic acid in the femurs of rats

This study evaluated the use of red and infrared lasers on tissue surrounding the femurs of 60 rats randomly divided into three groups after implantation of bioabsorbable plates. The control group were not subjected to laser irradiation; group A was treated with red laser [indium–gallium–aluminum–phosphide (InGaAlP) laser, wavelength 685 nm, 35 mW, continuous wave (CW), Ø?=?0.06 cm, 2.23 min], and group B was subjected to infrared laser [gallium–aluminum–arsenium (GaAlAs) laser, wavelength 830 nm, 50 mw, CW, Ø?=?0.06 cm, 1.41 min], both at 10 J/cm². Samples were stained with hematoxylin and eosin (H&;E) and examined microscopically. Results showed that the laser irradiation had had a positive photobiomodulation effect on inflammation, confirmed by a better histologic pattern than that of the control group at 3 days and 7 days. Semiquantitative analysis revealed that groups A and B had a histologic score significantly greater than that of the control group at 3 days. At 21 days, histomorphometric analysis revealed a more intense inflammation in the red laser group than in the other groups. We concluded that low-level laser therapy (LLLT) has positive effects on the photobiomodulation of inflammation in the tissues surrounding the poly-L-lactic/polyglycolic acid (PLLA/PGA) bioabsorbable plate. It stimulated vascularization, fibroblast proliferation, and collagen deposition.     

Effect of low-level laser irradiation and epidermal growth factor on adult human adipose-derived stem cells

The study investigated the effects of low-level laser radiation and epidermal growth factor (EGF) on adult adipose-derived stem cells (ADSCs) isolated from human adipose tissue. Isolated cells were cultured to semi-confluence, and the monolayers of ADSCs were exposed to low-level laser at 5 J/cm² using 636 nm diode laser. Cell viability and proliferation were monitored using adenosine triphosphate (ATP) luminescence and optical density at 0 h, 24 h and 48 h after irradiation. Application of low-level laser irradiation at 5 J/cm² on human ADSCs cultured with EGF increased the viability and proliferation of these cells. The results indicate that low-level laser irradiation in combination with EGF enhances the proliferation and maintenance of ADSCs in vitro.     

Ablation efficiency and relative thermal confinement measurements using wavelengths 1,064, 1,320, and 1,444 nm for laser-assisted lipolysis

Laser-assisted lipolysis is routinely used for contouring the body and the neck while modifications of the technique have recently been advocated for facial contouring. In this study, wavelength-dependence measurements of laser lipolysis effect were performed using different lasers at 1,064, 1,320, and 1,444 nm wavelengths that are currently used clinically. Fresh porcine skin with fatty tissue was used for the experiments with radiant exposure of 5–8 W with the same parameters (beam diameter?=?600 μm, peak power?=?200 mJ, and pulse rate?=?40 Hz) for 1,064, 1,320 and 1,444 nm laser wavelengths. After laser irradiation, ablation crater depth and width and tissue mass loss were measured using spectral optical coherence tomography and a micro-analytical balance, respectively. In addition, thermal temporal monitoring was performed with a thermal imaging camera placed over ex vivo porcine fat tissue; temperature changes were recorded for each wavelength. This study demonstrated greatest ablation crater depth and width and mass removal in fatty tissue at the 1,444 nm wavelength followed by, in order, 1,320 and 1,064 nm. In the evaluation of heat distribution at different wavelengths, reduced heat diffusion was observed at 1,444 nm. The ablation efficiency was found to be dependent upon wavelength, and the 1,444 nm wavelength was found to provide both the highest efficiency for fatty tissue ablation and the greatest thermal confinement.     

Modifying effect of intravenous laser therapy on the protein expression of arginase and epidermal growth factor receptor in type 2 diabetic patients

The epidermal growth factor receptor (EGFR) signaling pathway may be involved in cell activation and may influence the neuronal microenvironment, microglia activation, and production of proinflammatory cytokines. Arginase and nitric oxide synthase (NOS) both use l-arginine as a common substrate. Decreasing the arginase expression may increase l-arginine consumption by NOS and increase nitric oxide (NO) synthesis. Intravenous laser blood irradiation (ILBI) is an effective systemic treatment for different pathologies including diabetes mellitus. Previous studies have shown that low-level laser therapy can have an effect on the release of certain cytokines and growth factors. The aim of this study was to evaluate the effects of ILBI on the expression of arginase and epidermal growth factor receptor in type 2 diabetic patients. We used 630 nm red laser light, 1.5 mW, continuous mode, intravenously for 30 min in 13 type 2 diabetic patients and compared their blood samples using the flow cytometry technique, before and after ILBI. The difference between the percentage of cells before and after therapy was analyzed using repeated-measures ANOVA, and the relationship between EGFR and arginase expression in blood and tissue was evaluated by calculating the Pearson correlation coefficient. We found a significant decrease in the expression of both arginase- and EGFR-positive cells after laser therapy (P?<?0.01). In conclusion, laser therapy may have a beneficial effect for diabetic patients via decreasing arginase expression and activation of the NOS/NO pathway which increases NO production and vasodilation, and decreasing EGFR expression which may reduce neuroinflammation and its secondary damages.     

Lack of effect of carbon monoxide inhibitor on relaxation induced by electrical field stimulation in corpus cavernosum

Carbon monoxide has been proposed as a possible neurotransmitter because of its ability to bind to the iron atom of the heme of guanylyl cyclase, which is similar to that of nitric oxide. To determine whether carbon monoxide exerts an effect on the penis, strips of rabbit corpus cavernosum were mounted in an organ bath for isometric tension studies and the effect of zinc deuteroporphyrin, an inhibitor of heme oxygenase which metabolizes hemoprotein and releases carbon monoxide, on relaxation induced by electrical field stimulation (neurally mediated) was determined. Also observed was relaxation induced by electrical field stimulation after incubation with atropine and guanethidine to isolate nonadrenergic noncholinergic neurotransmission. Zinc deuteroporphyrin (10^-6M, 10^-5M, 10^-4M and 3x10^-4M) did not affect relaxation induced by electrical field stimulation in the absence or presence of guanethidine and atropine. Therefore, it appears that carbon monoxide does not contribute to neurally mediated relaxation of the rabbit corpus cavernosum.     

Mitochondrial dependent oxidative stress in cell culture induced by laser radiation at 1265 nm

Photodynamic therapy is the main technique applied for surface carcinoma treatment. This technique employs singlet oxygen generated via a laser excited photosensitizer as a main damaging agent. However, prolonged sensitivity to intensive light, relatively low tissue penetration by activating light the cost of photosensitizer (PS) administration can limit photodynamic therapy applications. Early was reported singlet oxygen generation without photosensitizer induced by a laser irradiation at the wavelength of 1250–1270 nm. Here, we study the dynamics of oxidative stress, DNA damage, changes of mitochondrial potential, and mitochondrial mass induced by a laser at 1265 nm have been studied in HCT-116 and CHO-K cells. Laser irradiation of HCT-116 and CHO-K cells has induced a dose-dependent cell death via increasing intracellular reactive oxygen species (ROS) concentration, increase of DNA damage, decrease of mitochondrial potential, and reduced glutathione. It has been shown that, along with singlet oxygen generation, the increase of the intracellular ROS concentration induced by mitochondrial damage contributes to the damaging effect of the laser irradiation at 1265 nm.     

Effect of low-level laser therapy irradiation and Bio-Oss graft material on the osteogenesis process in rabbit calvarium defects: a double blind experimental study

This study aims to assess the effect of low-level laser therapy (LLLT) irradiation and Bio-Oss graft material on the osteogenesis process in the rabbit calvarium defects. Twelve white male New Zealand rabbits were included in this study. Four 8-mm diameter identical defects were prepared on each rabbit's calvarium. One site was left as an untreated control (C), the second site was filled with Bio-Oss (B), the third site was treated with laser irradiation (L), and the fourth site treated with Bio-Oss and laser irradiation (B + L). In the laser group, a diode laser (wavelength 810 nm, output power 300 mW, irradiation mode CW, energy density 4 J/cm2) was applied immediately after surgery and then one other day for the next 20 days. After 4 and 8 weeks, the animals were sacrificed and histological and histomorphometric examinations were performed and the data were subjected to Friedman and repeated measurements ANOVA tests. Significant differences were not found regarding inflammation severity, foreign body reactions, and vitality of newly formed bone on 4th and 8th week after operation. The mean amount of new bone was 15.83 and 18.5 % in the controls on the 4th and 8th week; 27.66 and 25.16 % in the laser-irradiated group; 35.0 and 41.83 % in Bio-Oss and 41.83 and 47.0 % in the laser + Bio-Oss treated specimens with significant statistical differences (p <0.05). Application of LLLT in combination with Bio-Oss® can promote bone healing. Therefore, LLLT may be clinically beneficial in promoting bone formation in skeletal defects.     

Superoxide, NO and CO in liver microcirculation: Physiology and pathophysiology

Superoxide anion (O₂ ⁻), nitric oxide (NO), and carbon monoxide (CO) are metabolites of molecular oxygen endogenously generated through oxygen activation by a variety of oxidases and oxygenases such as xanthine oxidase and NADPH oxidase, NO synthase, and heme oxygenase, respectively. There is an increasing body of evidence showing that these active oxygen metabolites not only exert their cytotoxic properties but also play a modulatory role in regulation of cell function in and around hepatic sinusoidal vessels. Among them, CO generated by heme oxygenase is a novel vasodilatory mediator which can upregulate cGMP in fatstoring Ito cells, liver-specific microvascular pericytes which encircle sinusoidal walls, and thereby control the microvascular tone under control conditions. When exposed to endotoxemia, Kupffer cells and hepatocytes can express inducible NO synthase activity which serves as a NO-dependent cytotoxic mechanisms involving peroxynitrite formation. Disclosure of the whole picture of NO- and CO-dependent mechanisms for regulation of hepatic microcirculation gives a clue to understanding the physiology and pathophysiology of liver function.     

Effect of the transdermal low-level laser therapy on endothelial function

The effect of low-level laser therapy (LLLT) on the cardiovascular system is not fully established. Since the endothelium is an important endocrine element, establishing the mechanisms of LLLT action is an important issue.The aim of the study was to evaluate the effect of transdermal LLLT on endothelial function.In this study, healthy volunteers (n?=?40, age?=?20–40 years) were enrolled. N?=?30 (14 female, 16 male, mean age 30?±?5 years) constituted the laser-irradiated group (LG). The remaining 10 subjects (6 women, 4 men, mean age 28?±?5 years) constituted the control group (CG). Participants were subjected to LLLT once a day for three consecutive days. Blood for biochemical assessments was drawn before the first irradiation and 24 h after the last session. In the LG, transdermal illumination of radial artery was conducted (a semiconductor laser λ?=?808 nm, irradiation 50 mW, energy density 1.6 W/cm² and a dose 20 J/day, a total dose of 60 J). Biochemical parameters (reflecting angiogenesis: vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), angiostatin; antioxidative status: glutathione (GSH) and the nitric oxide metabolic pathway: symmetric dimethylarginine (SDMA), asymmetric dimethylarginine (ADMA) and l-arginine) were assessed. In the LG, a significant increase in GSH levels and considerable decrease in angiostatin concentration following the LLLT were observed. No significant differences in levels of the VEGF, FGF, SDMA, ADMA were observed.LLLT modifies vascular endothelial function by increasing its antioxidant and angiogenic potential. We found no significant differences in levels of the nitric oxide pathway metabolites within 24 h following the LLLT irradiation.