Effect of photobiomodulation therapy on the morphology, intracellular calcium concentration, free radical generation, apoptosis and necrosis of human mesenchymal stem cells-an in vitro study.

The aim of the study was to investigate the impact of multiwave locked system (MLS M1) emitting synchronized laser radiation at 2 wavelength simultaneous (λ = 808 nm, λ = 905 nm) on the mesenchymal stem cells (MSCs). Human MSCs were exposed to MLS M1 system laser radiation with the power density 195-318 mW/cm2 and doses of energy 3-20 J, in continuous wave emission (CW) or pulsed emission (PE). After irradiation exposure in doses of energy 3 J, 10 J (CW, ƒ = 1000 Hz), and 20 J (ƒ = 2000 Hz), increased proliferation of MSCs was observed. Significant reduction of Fluo-4 Direct™ Ca2+ indicator fluorescence over controls after CW and PE with 3 J, 10 J, and 20 J was noticed. A decrease in fluorescence intensity after the application of radiation with a frequency of 2000 Hz in doses of 3 J, 10 J, and 20 J was observed. In contrary, an increase in DCF fluorescence intensity after irradiation with laser radiation of 3 J, 10 J, and 20 J (CW, ƒ = 1000 Hz and ƒ = 2000 Hz) was also shown. Laser irradiation at a dose of 20 J, emitted at 1000 Hz and 2000 Hz, and 3 J emitted at a frequency of 2000 Hz caused a statistically significant loss of MSC viability. The applied photobiomodulation therapy induced a strong pro-apoptotic effect dependent on the laser irradiation exposure time, while the application of a sufficiently high-energy dose and frequency with a sufficiently long exposure time significantly increased intracellular calcium ion concentration and free radical production by MSCs.

Effect of Photobiomodulation Therapy on the Increase of Viability and Proliferation of Human Mesenchymal Stem Cells.

BACKGROUND AND OBJECTIVES: We have investigated how low intensity laser irradiation emitted by a multiwave-locked system (MLS M1) affects the viability and proliferation of human bone marrow mesenchymal stem cells (MSCs) depending on the parameters of the irradiation. STUDY DESIGN/MATERIALS AND METHODS: Cells isolated surgically from the femoral bone during surgery were identified by flow cytometry and cell differentiation assays. For irradiation, two wavelengths (808 and 905 nm) with the following parameters were used: power density 195, 230, and 318 mW/cm 2 , doses of energy 3, 10, and 20 J (energy density 0.93-6.27 J/cm 2 ), and in continuous (CW) or pulsed emission (PE) (frequencies 1,000 and 2,000 Hz). RESULTS: There were statistically significant increases of cell viability and proliferation after irradiation at 3 J (CW; 1,000 Hz), 10 J (1,000 Hz), and 20 J (2,000 Hz). CONCLUSIONS: Irradiation with the MLS M1 system can be used in vitro to modulate MSCs in preparation for therapeutic applications. This will assist in designing further studies to optimize the radiation parameters and elucidate the molecular mechanisms of action of the radiation. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

The effect of MLS laser radiation on cell lipid membrane.

INTRODUCTION: Authors of numerous publications have proved the therapeutic effect of laser irradiation on biological material, but the mechanisms at cellular and subcellular level are not yet well understood. OBJECTIVE: The aim of this study was to assess the effect of laser radiation emitted by the MLS M1 system (Multiwave Locked System) at two wavelengths (808 nm continuous and 905 nm pulsed) on the stability and fluidity of liposomes with a lipid composition similar to that of human erythrocyte membrane or made of phosphatidylocholine. MATERIAL AND METHODS: Liposomes were exposed to low-energy laser radiation at surface densities 195 mW/cm2 (frequency 1,000 Hz) and 230 mW/cm2 (frequency 2,000 Hz). Different doses of radiation energy in the range 0-15 J were applied. The surface energy density was within the range 0.46 - 4.9 J/cm 2. RESULTS: The fluidity and stability of liposomes subjected to such irradiation changed depending on the parameters of radiation used. CONCLUSIONS: Since MLS M1 laser radiation, depending on the parameters used, affects fluidity and stability of liposomes with the lipid content similar to erythrocyte membrane, it may also cause structural and functional changes in cell membranes.

Assessment of visual perception in adolescents with a history of central coordination disorder in early life - 15-year follow-up study.

INTRODUCTION: Central nervous system damage in early life results in both quantitative and qualitative abnormalities of psychomotor development. Late sequelae of these disturbances may include visual perception disorders which not only affect the ability to read and write but also generally influence the child's intellectual development. This study sought to determine whether a central coordination disorder (CCD) in early life treated according to Vojta's method with elements of the sensory integration (S-I) and neuro-developmental treatment (NDT)/Bobath approaches affects development of visual perception later in life. MATERIAL AND METHODS: The study involved 44 participants aged 15-16 years, including 19 diagnosed with moderate or severe CCD in the neonatal period, i.e. during the first 2-3 months of life, with diagnosed mild degree neonatal encephalopathy due to perinatal anoxia, and 25 healthy people without a history of developmental psychomotor disturbances in the neonatal period. The study tool was a visual perception IQ test comprising 96 graphic tasks. RESULTS: The study revealed equal proportions of participants (p < 0.05) defined as very skilled (94-96), skilled (91-94), aerage (71-91), poor (67-71), and very poor (0-67) in both groups. These results mean that adolescents with a history of CCD in the neonatal period did not differ with regard to the level of visual perception from their peers who had not demonstrated psychomotor development disorders in the neonatal period. CONCLUSIONS: Early treatment of children with CCD affords a possibility of normalising their psychomotor development early enough to prevent consequences in the form of cognitive impairments in later life.

Influence of various laser therapy methods on knee joint pain and function in patients with knee osteoarthritis.

BACKGROUND: The aim of the study was to estimate the influence of various laser therapy methods on knee joint pain and function in patients with knee osteoarthritis. MATERIAL AND METHODS: 125 patients were randomly assigned to 4 groups: • group I received one-wave laser irradiation (wave length 810 nm, dose 8 J/point) • group II received two-wave MLS laser irradiation (power 1100 mW, frequency 2000 Hz, dose 12.4 J/point) • group III received a similar regimen of two-wave MLS laser irradiation, but at a dose of 6.6 J per point • group IV was a placebo group where laser therapy procedures were simulated without actual irradiation. The effectiveness of the therapy was evaluated by means of Lequesne's scale, a modified Laitinen questionnaire and a visual analogue scale (VAS). Statistical analysis utilised non-parametric Wilcoxon's and Mann-Whitney's tests. Calculations were carried out with MedCalc v. 11.6.1.0. RESULTS: Statistically significant improvements in knee joint function and pain relief were seen in all groups (I, II and III). When groups I, II and III were compared, the largest improvement was found in group II (MLS laser, dose 12.4 J/point). The degrees of improvement in groups I and III were similar. CONCLUSIONS: One-wave laser irradiation at a dose of 8 J per point and two-wave laser irradiation with doses of 12.4 J and 6.6 J per point significantly improved knee joint function and relieved knee pain in patients with osteoarthritis.

Comparative evaluation of the direct analgesic efficacy of selected physiotherapeutic methods in subjects with knee joint degenerative disease - preliminary report.

BACKGROUND: The goals of the study were to evaluate the efficacy of two physiotherapeutic procedures: low energy laser therapy and low frequency transcutaneous electric nerve stimulation (TENS) and to compare these modalities with regard to their therapeutic effects in patients with knee osteoarthritis. MATERIAL AND METHODS: Fifty (50) subjects were enrolled into the study and divided into two groups of 25 subjects. Group A received 10 MLS laser therapy sessions with a synchronised laser beam at doses of 12 J per treated site. Group B received ten sessions of low frequency TENS. The procedures were carried out every day for two weeks (5 times a week). All patients completed a personal data questionnaire and underwent an examination of knee joint motion range and circumference. Subjective pain intensity was assessed using the VAS pain scale and the modified Laitinen questionnaire. RESULTS: An analysis of the results of the treatment demonstrated statistically significant pain reduction in both groups. This improvement was significantly higher in the two-phase laser therapy group vs. the LF-TENS group. No statistically significant improvement was noted in either of the groups regarding the knee joint range of motion. CONCLUSIONS: 1. Synchronised laser beam (MLS) therapy and low-frequency TENS contribute to direct pain relief effects in subjects with knee osteoarthritis. 2. The study confirmed better analgesic effects of two-phase laser therapy vs. LF-TENS.

Comparative analysis of analgesic efficacy of selected physiotherapy methods in low back pain patients.

BACKGROUND: Low back pain syndromes are one of the most frequent causes of movement limitation in populations of highly industrialized countries. They are listed as the main cause of inability to work among people of working age. Chronic pain and the associated limitation of movement underlie the quest for effective therapies. The use of ultrasound, LLLT, vacuum therapy with Ultra Reiz current in physical therapy of these patients prompts research over their effectiveness in the therapy of patients with low-back pain. The aim of the work was to evaluate the analgesic efficacy of LLLT, ultrasound, and vacuum therapy with Ultra Reiz current in patients with low back pain. MATERIAL AND METHODS: The study involved 94 people divided into three groups (A,B,C). Group A (n=35) received a series of 10 low energy laser therapy sessions (wave length 808 nm, surface density of radiation 510 mW/cm(2), continuous wave form, scanning mode, a dose of 12 J/cm(2) on a surface of 100 cm(2) [10x10cm]). Patients in Group B (n=27) had ultrasound sessions with a wave intensity of 1 W/cm(2) for 3 minutes. Patients in Group C (n=32) underwent vacuum therapy (8 kPa) combined with Ultra Reiz current. Subjective pain assessment was carried out using a modified Latinen questionnaire and a visual analogue scale of pain intensity. Lumbosacral spine mobility was evaluated with the Schober test and the finger-to- floor test. RESULTS: In Group A, following low energy laser therapy, a statistically significant decrease in pain intensity was observed, together with decreased analgesic consumption compared to the other groups. In Group C, following vacuum therapy combined with Ultra Reiz currents, a significant decrease in the frequency of pain was observed together with increased physical activity compared to both Groups A and B, assessed according to a modified Laitinen pain indicator questionnaire. The biggest improvement in global spine mobility and lumbosacral flexion was observed in Group C (vacuum therapy plus Ultra Reiz current) compared to the other groups. However, the most significant improvement in lower spine extension was noted in Group B (ultrasound). CONCLUSIONS: 1. The study showed slightly higher analgesic efficacy of laser biostimulation in comparison to vacuum therapy combined with Ultra Reiz current in patients with low back pain. 2. A more prominent increase in lumbosacral spine mobility was observed after vacuum therapy combined with Ultra Reiz current and ultrasound therapy.

Effect of low-intensity (3.75-25 J/cm2) near-infrared (810 nm) laser radiation on red blood cell ATPase activities and membrane structure.

OBJECTIVE: The biostimulation and therapeutic effects of low-power laser radiation of different wavelengths and light doses are well known, but the exact mechanism of action of the laser radiation with living cells is not yet understood. The aim of the present work was to investigate the effect of laser radiation (810 nm, radiant exposure 3.75-25 J/cm(2)) on the structure of protein and lipid components of red blood cell membranes and it functional properties. The role of membrane ATPases as possible targets of laser irradiation was analyzed. BACKGROUND DATA: A variety of studies both in vivo and in vitro showed significant influence of laser irradiation on cell functional state. At the same time another group of works found no detectable effects of light exposure. Some different explanations based on the light absorption by primary endogenous chromophores (mitochondrial enzymes, cytochromes, flavins, porphyrins) have been proposed to describe biological effects of laser light. It was suggested that optimization of the structural-functional organization of the erythrocyte membrane as a result of laser irradiation may be the basis for improving the cardiac function in patients under a course of laser therapy. MATERIALS AND METHODS: Human red blood cells or isolated cell membranes were irradiated with low-intensity laser light (810 nm) at different radiant exposures (3.75-25 J/cm(2)) and light powers (fluence rate; 10-400 mW) at 37 degrees C. As the parameters characterizing the structural and functional changes of cell membranes the activities of Na(+)-, K(+)-, and Mg(2+)-ATPases, tryptophan fluorescence of membrane proteins and fluorescence of pyrene incorporated into membrane lipid bilayer were used. RESULTS: It was found that near-infrared low-intensity laser radiation changes the ATPase activities of the membrane ion pumps in the dose- and fluence rate-dependent manner. At the same time no changes of such integral parameters as cell stability, membrane lipid peroxidation level, intracellular reduced glutathione or oxyhaemoglobin level were observed. At laser power of 10 mW, an increase of the ATPase activity was observed with maximal effect at 12-15 J/cm(2) of light dose (18-26% for the total ATPase activity). At laser power of 400 mW (fluence rate significantly increased), inhibition of ATPases activities mainly due to the inhibition of Na(+)-, K(+)-ATPase was observed with maximal effect at the same light dose of 12-15 J/cm(2) (18-23% for the total ATPase activity). Fractionation of the light dose significantly changed the membrane response to laser radiation. Changes in tryptophan fluorescent parameters of erythrocyte membrane proteins and the increase in lipid bilayer fluidity measured by pyrene monomer/excimer fluorescence ratio were observed. CONCLUSIONS: Near-infrared laser light radiation (810 nm) induced long-term conformational transitions of red blood cell membrane which were related to the changes in the structural states of both erythrocyte membrane proteins and lipid bilayer and which manifested themselves as changes in fluorescent parameters of erythrocyte membranes and lipid bilayer fluidity. This resulted in the modulation of membrane functional properties: changes in the activity of membrane ion pumps and, thus, changes in membrane ion flows.

Low-intensity near-infrared laser radiation-induced changes of acetylcholinesterase activity of human erythrocytes.

OBJECTIVE: The aim of the present study was to investigate the transformations of red blood cells produced by low-intensity infrared laser radiation (810 nm). BACKGROUND DATA: Low-intensity (the output power of a laser device in the milliwatt range) laser radiation as a local phototherapeutic modality is characterized by its ability to induce non-thermic, nondestructive photobiological processes in cells and tissues. However, the exact theory concerning the therapeutic effects of laser biostimulation has not been developed. MATERIALS AND METHODS: The suspensions of human erythrocytes in PBS (10% hematocrit) were irradiated with near-infrared (810 nm) therapy laser at different light doses (0-20 J) and light power (fluence rate; 200 or 400 mW) at 37 degrees C. As the parameters characterizing the cell structural and functional changes membrane acetylcholinesterase (AchEase) activity, the membrane potential, the level of intracellular glutathione, the level of products of membrane lipid peroxidation, and the cell osmotic stability were measured. RESULTS: It was found that near-infrared low-intensity laser radiation produced complex biphasic dose-dependent changes of the parameters of AchEase reaction in the dose-dependent manner: at smaller doses of radiation (6 J) the maximal reaction rate and Michaelis-Menten constant value decreased, and at higher radiation doses these parameters increased. No significant changes of erythrocyte stability, cellular redox state (reduced glutathione or lipid peroxidation product levels), or cell membrane electrochemical potential were observed. CONCLUSION: Low-intensity near-infrared laser radiation (810 mn) produced AchEase activity changes, reflecting the effect of light on the enzyme due to energy absorption. Protein molecule conformational transitions and enzyme activity modifications in cells have been suggested as laser radiation-induced events.