Percutaneous Exposures of volunteers to polychromatic light (480-3400 nm) trigger systemic mechanism of the human myeloma cells growth delay without any effect on bortezomib cytotoxicity in vitro.

BACKGROUND AND AIM: Polychromatic light is actively used in medicine. However, its oncological safety and effect against cytotoxic therapy remains poorly studied. Multiple myeloma (MM) develops in the bone marrow and therefore malignant plasma cells are inaccessible to direct exposure to light. The aim of our work was to study the influence of polychromatic visible and infra-red light (pVIS + pIR) on growth and sensitivity of the myeloma cells to the cytotoxic effect of drug bortezomib (BTZ) through systemic mechanism. MATERIALS AND METHODS: We explored the effect of volunteers blood sera after their 4-daily irradiations with pVIS + pIR light (480-3400 nm) on growth and viability of the human myeloma RPMI 8226 cells in the presence of BTZ at its application in 2 modes: short-term (1 h, 300 nM) and long-term ( 96 h, 5 nM). Viability and proliferative activity of cells was evaluated by MTT assay. RESULTS: It was found that photomodified blood sera delayed growth of myeloma RPMI 8226 cells (by 25 % in 48 h and 23 % in 96 h) but had no effect on spontaneous and mitogen-induced proliferation of autologous peripheral blood T- and B-lymphocytes. We also revealed that pVIS + pIR did not change RPMI 8226 cells sensitivity to BTZ. CONCLUSION: The results suggest the systemic mechanism of polychromatic light and argue in favor of its oncological safety during/after BTZ therapy of MM patients without effect on the drug cytotoxicity.

Polychromatic light (480-3400nm) similar to the terrestrial solar spectrum without its UV component in post-surgical immunorehabilitation of breast cancer patients.

To this day, two methods of phototherapy (PT) have been successfully used in post-surgical immunorehabilitation of patients with breast cancer (BC): intravenous laser irradiation of the patients' blood and reinfusion of lympholeukosuspension of BC patients after single irradiation with HeNe laser. The objective of this pilot experimental study was to verify the effectiveness of the percutaneous use of polychromatic visible light combined with polychromatic infrared (pVIS+pIR) radiation similar to the major components of natural solar spectrum in post-surgical management of BC patients. Patients with BC (adenocarcinoma) of I-II stages, n=19 who had undergone mastectomy, were divided into 2 groups. The control group of patients (n=8) underwent a conventional course of post-surgical rehabilitation and sham irradiation. Patients of the PT group (n=11) additionally received 7days of daily treatment with polychromatic light on the sacral area, D=15cm. The PT course began on the day after mastectomy (Bioptron-2 device; Switzerland, 480-3400nm, 95% polarization, 40mW/cm2, 24J/cm2). Mastectomy produced many changes in cellular and humoral immunity, which was recorded on the 1st and 8th post-surgical days. The PT course resulted in a faster normalization of post-surgical leukocytosis and activation of cytotoxic CD8+ T-lymphocytes (Lym), reduced the elevated concentration in blood of immune complexes and in parallel promoted cytotoxic activity of CD16+/CD56+ NK-cells. The PT up-regulated the number of NK-cells in patients with its decrease on the 1st post-surgical day and prevented the decrease in the amount of monocytes, CD19+ B-Lym, CD3+ T-Lym, CD4+ T-helpers, activated CD3+/HLADR+ T-Lym, and the decrease of the phagocytotic capability of neutrophils. PT blocked the down-regulation of the IgM, IgA concentration and abnormally sharp increase of the proinflammatory cytokine IFN-γ content. Therefore, a 7-day course with polychromatic light prevented the development of immunosupression in the BC patients at the early post-mastectomy period.

Polychromatic Light (480-3400 nm) Upregulates Sensitivity of Tumor Cells to Lysis by Natural Killers.

OBJECTIVE: This study evaluates the participation of immunological mechanisms of downregulation of murine hepatoma cells MH22a after direct exposure to polychromatic polarized light. BACKGROUND DATA: Previous studies have shown that exposure to a combination of visible (VIS) and infrared (IR) light leads to decreased tumorigenicity of the murine hepatoma cells MH22a, which correlated with an increase in the amount of cells with reorganized cytoskeleton in the submembrane region. The mechanism of tumor inhibition and elimination has not been determined. MATERIALS AND METHODS: Polychromatic light (480-3400 nm) has been used at doses of 4.8 and 9.6 J/cm(2) to determine the sensitivity of murine MH22a cells and human erythroleukemia cells K562 exposed to this light, to lysis by effector cells of innate immunity (NK cells), and enhancement of the glycocalyx of the studied tumor cells. This was determined using flow cytometry, the H(3)-uridine cytotoxic test followed by spectrophotometry. RESULTS: VIS-IR light increases the sensitivity of MH-22a cells at a dose 4.8 J/cm(2) and K562 cells at 9.6 J/cm(2). The enhancement of sensitivity of tumor cells to NK lysis changed their ability to absorb alcian blue, reflecting a change in the expression of the glycocalyx. CONCLUSIONS: Increasing the sensitivity of the murine tumor cells MH22a and human K562 irradiated VIS-IR light correlated with a change in the expression of their glycocalyx. The results of the present study demonstrate that the reduction of tumorigenicity of irradiated tumor cells is due to their sensitivity to lysis by NK cells of the immune system.

Regulatory systemic effect of postsurgical polychromatic light (480-3400 nm) irradiation of breast cancer patients on the proliferation of tumor and normal cells in vitro.

OBJECTIVE: The aim of this work was to study the effect of phototherapy (PT) with percutaneous exposures to polychromatic visible and IR light (pVIS + pIR) on breast cancer (BC) patients at the early postmastectomy period, on the growth-promoting (GP) properties of their blood serum, by evaluating its capability to support proliferation of normal and tumor human cells in vitro. MATERIAL AND METHODS: After mastectomy, one group of patients was treated daily for 1 week on the sacral area with pVIS + pIR light (480-3400 nm, 40 mW/cm(2), 95% polarization, 24 J/cm(2)). The second group used as a control was sham irradiated. Blood serum samples collected before surgery, and 1 and 8 days after surgery, were added (2.5%) into nutrition media for cells instead of 10% of fetal calf serum. Cell targets were cultures of human fibroblasts (FBs), keratinocytes (KCs), three lines of the human BC cells (BT-474, HBL-100, Hs 578T) and cells of human epidermoid carcinoma (A-431). Cell number was evaluated by staining cell nuclei with crystal violet and a spectrometric assay of the extracted dye. RESULTS: The day after mastectomy there were no significant changes in the GP activity of sera. After a 7-day PT course, an increase of this activity was recorded for normal FBs and KCs by 18% and 24%, respectively, in comparison with presurgical levels. GP activity of the same patients' sera for all tumor cells, BT-474, HBL-100, Hs 578T and A-431, decreased by 32%, 17%, 11%, and 7% respectively. As a result, enhancement of proliferation of KCs and FBs and inhibition of proliferation of tumor cells was seen. CONCLUSIONS: The results suggest an effect at the systemic level where pVIS + pIR light may stimulate growth of human skin cells and simultaneously downregulate the proliferation of tumor cells, including BC cells. This argues in favor of the oncological safety of PT for BC patients postsurgically.

Downregulation of tumorogenicity and changes in the actin cytoskeleton of murine hepatoma after irradiation with polychromatic visible and IR light.

OBJECTIVE: This study evaluated the function and structural consequences of direct exposure of murine hepatoma MH-22a cells to polychromatic polarized light, to determine potential risk of malignancy following irradiation. BACKGROUND DATA: Visible (VIS) and infrared (IR) light have been actively used for prevention and treatment of complications developed after conventional tumor therapy. However, the safety associated with this irradiation has not been determined. MATERIALS AND METHODS: Polychromatic light (480-3400 and 385-750 nm), were used at different doses (4.8-38.4 J/cm(2)) to determine the viability, proliferation, and actin cytoskeleton in vitro by flow cytometry and confocal microscopy. Tumorogenic properties of cells were studied in vivo after transplantation in C3HA mice. RESULTS: Polychromatic light of a wide range of doses did not change the viability and proliferation of cells. After transplantation of cells irradiated with VIS-IR light (4.8 and 9.6 J/cm(2)) and VIS light (38.4 J/cm(2)) the tumor volume was lower in the treated group than in the control group in vivo. Transplantability of the irradiated cells also decreased, whereas survival of tumor-bearing mice increased. Three cell populations with different cytoskeleton structure were identified. After irradiation, the reorganized part of the actin cytoskeleton changed its localization to the submembranous area. CONCLUSIONS: A decrease of tumorigenicity in cells irradiated with polychromatic light used in non-damaging doses correlated with an increase in the number of cells with reorganized actin in the submembranous area. The results of the present study argue in favor of the oncological safety of polychromatic VIS-IR light (480-3400 nm).

Role of nitric oxide in the visible light-induced rapid increase of human skin microcirculation at the local and systemic level: I. diabetic patients.

OBJECTIVE: This study aimed to reveal the effects of polychromatic visible (pVIS) or pVIS + near IR (nIR) light similar to some components of solar light on skin microcirculation and microvascular response to the vasodilatators acetylcholine (ACh) and nitroglycerine (NG), in the extremities of patients with diabetic microangiopathy. BACKGROUND DATA: The mechanisms behind light-induced increases in microcirculation as well as extracellular effects of terrestrial pVIS and pVIS + nIR light remain unknown. MATERIALS AND METHODS: In 24 subjects with type 2 diabetes mellitus local microcirculation was measured in the skin of the foot before and after exposure to both types of light. In another 26 patients systemic microcirculation was studied in the back of the hand before and after exposure of the lumbar-sacral area to light energy. Two different types of light therapy were performed by using two devises: Q-light, which delivers pVIP (385-750 nm) and pVIS nIR light (385-1700 nm) with a power density of 40 mW/cm2, which is similar to summer sunlight at noon in Central Europe. RESULTS: At 2 min after irradiation (12 J/cm2) of the forefoot with pVIS or pVIS + nIR light, a rise in local blood flow volume (Qas) was observed, on average by 39% and 31%, respectively. The maximal effect (+41-47%) had developed in all patients at 30 min, and it then decreased and disappeared completely 24 h post-irradiation. We obtained similar results after irradiation of the sacral area in Qas of the skin of the hand. Both types of microcirculation also increased following a second exposure to the light sources. Enhancement of microcirculation was accompanied by a decrease in the microvascular response to ACh and NG solutions administered intracutaneously by iontophoresis. CONCLUSION: Both types of irradiation stimulated microcirculation at the local and systemic levels through a mechanism of enhancement of endothelium-dependent and endothelium-independent vasodilation, in which nitric oxide plays a major role.

Role of nitric oxide in the visible light-induced rapid increase of human skin microcirculation at the local and systemic levels: II. healthy volunteers.

OBJECTIVE: The aim of this study is to evaluate the skin microcirculation increase seen in healthy volunteers after a single exposure to polychromatic visible (pVIS) light, and to prove the role of nitric oxide (NO) in the development of this effect. BACKGROUND DATA: Improvement of microcirculation is one of the most important effects of laser and pVIS light therapy; however, its mechanism of action remains unknown. A main role in the regulation of vascular tone is known to be played by NO. It is produced by NO-synthase (NOS) located in membranes of many cells, including endothelial and blood cells. NOS, a biopteroflavohemoprotein, absorbs pVIS light, resulting in its activation. MATERIALS AND METHODS: The central area of the dorsal side of the right hand (24 cm2) of 42 volunteers was irradiated for 5 min with pVIS light from a Q-light (385-750 nm, 95% polarization, 40 mW/cm2, 12 J/cm2). Then for 90 min, the blood flow rate (Qas) was measured eight times, both in the area of the irradiation (local effect) and in the non-irradiated left hand (systemic effect) by using a high-frequency ultrasound Doppler device, recording Qas in human skin to a depth up to 5 mm. In the central area of the right hand of 14 volunteers an NOS inhibitor, N-monomethyl-L-arginine (L-NMMA, 0.1% solution), was iontophoretically administered prior to exposure, whereas in 10 other subjects it was administered to the left hand with subsequent exposure of the right hand. RESULTS: As soon as 2 min after exposure, Qas in the irradiated area rose on average by 32%, and in 20 min by 45%; it then decreased and in 90 min returned to the initial level. A statistically significant Qas increase in the non-irradiated hand was recorded in 5 min (+9%), and in 20 min it reached a maximum level (+39%), and 90 min later it decreased to the initial values. The presence of L-NMMA in the light-exposed area completely blocked the photoinduced rise of microcirculation, both in the irradiated and in non-irradiated hand; however, its administration to the non-irradiated hand did not prevent these effects. CONCLUSION: The increase in skin microcirculation produced by pVIS light at the local and systemic levels is due to activation of NO synthesis in the irradiated area.

Polychromatic light similar to the terrestrial solar spectrum without its UV component stimulates DNA synthesis in human peripheral blood lymphocytes in vivo and in vitro.

Immunosuppressive effects of the minor component of the terrestrial solar spectrum, UV radiation, have been substantiated over the past several years. This raises the question of what influence the dominant part of the solar spectrum--visible and IR light-would have on the human immune system. In the present randomized, placebo-controlled double-blind study a small area of the body surface of volunteers was irradiated with polychromatic light (480-3400 nm), simulating the significant part of the terrestial sunlight irradiance spectrum and its power density. An average 2.5-fold to three-fold increase in spontaneous and phytohemagglutinin-induced DNA synthesis in peripheral blood lymphocytes (Lym) was revealed at 0.5-24 h after irradiation at a therapeutic dose (12 J/cm(2)) in subjects with low preirradiation levels of both processes. The in vivo findings were echoed in parallel in vitro experiments, when blood drawn from the same subjects was directly irradiated (2.4 J/cm(2)), or when the irradiated blood was mixed 1:10 with nonirradiated autologous blood to model events in the circulation following transcutaneous blood photomodification. Our data suggest that exposure of the human body to polychromatic visible + IR light may photomodify blood in the dermal vasculature of the irradiated area to lead to an immediate transfer of the light-induced effects to Lym of the entire circulating blood, which can result in modulation of Lym functional state at the systemic level.

Pro- and anti-inflammatory cytokine content in human peripheral blood after its transcutaneous (in vivo) and direct (in vitro) irradiation with polychromatic visible and infrared light.

OBJECTIVE: The aim of this randomized, placebo-controlled, double-blind trial was to investigate changes in the content of 10 cytokines in the human peripheral blood after transcutaneous and in vitro irradiation with polychromatic visible and infrared (IR) polarized light at therapeutic dose. BACKGROUND DATA: The role of cytokines in development of anti-inflammatory, immunomodulatory, and wound-healing effects of visible and IR light remains poorly studied. METHODS: The sacral area of volunteers was exposed (480-3400 nm, 95% polarization, 12 J/cm(2)); in parallel, the blood samples of the same subjects were irradiated in vitro (2.4 J/cm(2)). Determination of cytokine content was performed using enzyme-linked immunosorbent assay (ELISA). RESULTS: A dramatic decrease in the level of pro-inflammatory cytokines TNF-alpha, IL-6, and IFN-gamma was revealed: at 0.5 h after exposure of volunteers (with the initial parameters exceeding the norm), the cytokine contents fell, on average, 34, 12, and 1.5 times. The reduced concentrations of TNF-alpha and IL-6 were preserved after four daily exposures, whereas levels of IFN-gamma and IL-12 decreased five and 15 times. At 0.5 h and at later times, the amount of anti-inflammatory cytokines was found to rise: that of IL-10 rose 2.7-3.5 times (in subjects with normal initial parameters) and of TGF-beta1 1.4-1.5 times (in the cases of its decreased level). A peculiarity of the light effect was a fast rise of IFN-gamma at 3.3-4.0 times in subjects with normal initial values. The content of IL-1beta, IL-2, IFN-alpha, and IL-4 did not change. Similar regularities of the light effects were recorded after in vitro irradiation of blood, as well as on mixing the irradiated and non-irradiated autologous blood at a volume ratio 1:10 (i.e., at modeling the events in a vascular bed of the exposed person when a small amount of the transcutaneously photomodified blood contacts its main circulating volume). CONCLUSION: Exposure of a small area of the human body to light leads to a fast decrease in the elevated pro-inflammatory cytokine plasma content and to an increase in the the anti-inflammatory factor concentration, which may be an important mechanism of the anti-inflammatory effect of phototherapy. These changes result from transcutaneous photomodification of a small volume of blood and a fast transfer of the light-induced changes to the entire pool of circulating blood.

The regulatory effect of polychromatic (visible and infrared) light on human humoral immunity.

The immunological effects of visible and infrared light from laser and non-laser sources have remained insufficiently studied, which has restricted the use of light in the treatment of diseases associated with immune system disorders. The present randomised, placebo-controlled double-blind trial was designed to study changes in the humoral immunity of a large group of volunteers after exposure of a small body area to polychromatic visible and infrared polarized (VIP) and non-polarized (VInP) light (400-3400 nm, 95% polarization, 40 mW cm(-2), 12 J cm(-2) and 400-3400 nm, no polarization, 38 mW cm(-2), 11.2 J cm(-2), respectively). Serum immunoglobulins (Ig) M, A, and G were determined turbidimetrically, and the immune complexes (ICs) by precipitation with 5% polyethylene glycol and subsequent spectrophotometric analysis. A single VIP irradiation induced an average rise in serum IgM levels of 13% (p < 0.05). By the end of the 10 day course, it has exceeded the baseline level by 26%, with an increase in IgA levels of 17 and 12% (p < 0.04) one week after the last session. In subjects with a high IC content, it decreased rapidly to the normal level. A single exposure of volunteers to VInP light rapidly produced changes similar to those observed on VIP irradiation, but with an increase in IgM 2.3 to 3 times lower, independent of the initial levels. On the other hand, VInP light exposure decreased the IC content more than VIP light.

Enhancement of the blood growth promoting activity after exposure of volunteers to visible and infrared polarized light. Part I: stimulation of human keratinocyte proliferation in vitro.

The systemic mechanisms of the wound healing effect of low intensity lasers remain largely uninvestigated. The goal of this randomized, placebo controlled, double blind study is to prove that irradiation of a small area of the human body with visible and infrared polarized (VIP) light (400-3400 nm, 95% polarization, 40 mW cm(2), 12 J cm(2)) leads to an increase of the growth promoting (GP) activity of the entire circulating blood for primary cultures of human keratinocytes (KCs). Thirty minutes after the VIP-irradiation of a sacral area of volunteers, the GP activity of circulating blood was seen to increase through the elevation of the number of KCs cultured with the isolated plasma by 20 +/- 3%, p < 0.001. A similar increase in GP activity was seen in plasma derived from the in vitro irradiated blood of each volunteer, and from the mixture of irradiated and non-irradiated autologous blood 1:10. Enhanced GP activity was also recorded at 24 h after the 1st and 4-9th daily phototherapeutic sessions. Hence, exposure of volunteers to VIP light leads to a fast increase in the GP activity of the entire circulating blood for human KCs in vitro, which is a consequence of the transcutaneous photomodification of blood and its effect on the rest of the circulating blood volume.