Effects of a moderate-intensity static magnetic field and adriamycin on K562 cells

The aim of this study was to investigate whether a moderate‐intensity static magnetic field (SMF) can enhance the killing effect of adriamycin (ADM) on K562 cells, and to explore the effects of SMF combined with ADM on K562 cells. We analyzed the metabolic activity of cells, cell cycle distribution, DNA damage, change in cell ultrastructure, and P‐glycoprotein (P‐gp) expression after K562 cells were exposed continuously to a uniform 8.8 mT SMF for 12 h, with or without ADM. Our results showed that the SMF combined with ADM (25 ng/ml) significantly inhibited the metabolic activity of K562 cells (P < 0.05), while neither ADM nor the SMF alone affected the metabolic activity of these cells. Cell ultrastructure was altered in the SMF + ADM group. For example, cell membrane was depressed, some protuberances were observable, and vacuoles in the cytoplasm became larger. Cells were arrested at the G2/M phase and DNA damage increased after cells were treated with the SMF plus ADM. ADM also induced the P‐gp expression. In contrast, in the SMF group and SMF + ADM group, the P‐gp expression was decreased compared with the ADM group. Taken together, our results showed that the 8.8 mT SMF enhanced the cytotoxity potency of ADM on K562 cells, and the decrease in P‐gp expression may be one reason underlying this effect. Bioelectromagnetics 32:191–199, 2011. © 2010 Wiley‐Liss, Inc.     

Spatial gradient effects of 120 mT static magnetic field on endothelial tubular formation in vitro

This study investigated the spatial magnetic gradient effects of static magnetic fields (SMF) on endothelial tubular formation by applying the maximum spatial gradient to a target site of culture wells for cell growth. The respective maximum values of magnetic flux density (B(max)), magnetic flux gradient (G(max)) and the magnetic force product of the magnetic flux density and its gradient (a parameter of magnetic force) were 120 mT, 28 mT/mm and 1428 mT(2)/mm. The effects of gradient SMF on tubular formation were compared with those of uniform SMF that has no spatial gradients on the entire bottom area of culture wells. Five experimental groups of 25 samples each were examined: (1) sham exposure (control); (2) peak gradient exposure in the peripheral part; (3) peak gradient exposure in the central part; (4) uniform exposure to 20 mT; (5) uniform exposure to 120 mT. The SMF or sham exposure was carried out for 10 days. Photomicrographs of tubular cells, immunostained with an anti-platelet-endothelial cell adhesion molecule-1 (PECAM-1 [CD31]) antibody as a pan-endothelial marker, were analyzed after the 10-day culture. Gradient SMF in the peripheral or central part was found to significantly promote tubular formation in terms of the area density and length of tubules in each peak gradient/force part of the wells, compared with the sham exposure. In contrast, uniform SMF did not induce any significant change in the tubular formation. These findings suggest that tubule formation can be promoted by applying the peak gradient/force to a target site of culture wells.     

Contribution of a 300 kHz alternating magnetic field on magnetic hyperthermia treatment of HepG2 cells

We investigated the relative contributions of temperature and a 300 kHz alternating magnetic field (AMF) on magnetic hyperthermia treatment (MHT). Our system consisted of an induction coil, which generated AMF by electric current flow, and a newly developed, temperature‐controlled circulating water‐jacketed glass bottle placed inside the coil. The AMF generator operated at a frequency of 300 kHz with variable field strength ranging from 0 to 11 mT. Four treatment conditions were employed: (A) control (37 °C, 0 mT), (B) AMF exposure (37 °C, 11 mT), (C) hyperthermia (46 °C, 0 mT), and (D) hyperthermia plus AMF exposure (46 °C, 11 mT) for 30 min. Cell viability and apoptotic death rate were estimated. The relative contributions or interactions of hyperthermia (46 °C) and AMF (11 mT) on MHT were evaluated using 2 × 2 factorial experiment analysis. Group A was statistically different (P < 0.05) from each of the other treatments. The observed effects on both cell viability and apoptotic cell death were influenced by temperature (97.36% and 92.15%, respectively), AMF (1.78% and 4.99%, respectively), and the interactions between temperature and AMF (0.25% and 2.36%, respectively). Thus, the effect of hyperthermia was significant. Also, AMF exposure itself might play a role in MHT, although these observations were made in vitro. These findings suggest a possible presence of an AMF effect during clinical magnetic hyperthermia. Bioelectromagnetics 34:95–103, 2013. © 2012 Wiley Periodicals, Inc.     

Effects of 120 mT static magnetic field on TGF-beta1-inhibited endothelial tubular formation in vitro

We designed this study to examine the effects of static magnetic fields (SMF; 120 mT [B(max)] and a maximum spatial magnetic flux gradient of 21 mT/mm) on inhibited tubular formation when treated with human transforming growth factor (TGF)-beta1 at a relatively high concentration (5 ng/ml). Three experimental groups of 25 samples each were examined: (1) sham exposure alone (control); (2) sham-exposure with TGF-beta1; (3) SMF exposure with TGF-beta1. The SMF or sham exposure was carried out for 10 days. Photomicrographs of tubular cells, immunostained with an anti-platelet-endothelial cell adhesion molecule-1 (PECAM-1 [CD31]) antibody as a pan-endothelial marker, were analyzed after the 10-day culture. SMF was found to significantly reverse the inhibition of TGF-beta1 on tubular formation in terms of the area density and length of tubules (arteriogenesis) in the peripheral part of the wells, compared with the TGF-beta1 treatment alone. These findings suggest that one of the possible exogenous factors for arteriogenesis might involve 'magnetic force' (the product of the magnetic flux density, the magnetic gradient, and the volume susceptibility of the cells) because values are much larger in the peripheral part than in the central part.     

Effects of neck exposure to 5.5 mT static magnetic field on pharmacologically modulated blood pressure in conscious rabbits

Static magnetic fields (SMF) in the millitesla (mT) range have been reported to modulate microcirculatory hemodynamics and/or blood pressure (BP) under pharmacologically modified state in mammals. This study was designed to investigate the acute effects of local application of a SMF to neck or pelvic region under pharmacologically modulated BP; norepinephrine (NE)-induced hypertension as well as an L-type voltage-gated Ca(2+) channel blocker, nicardipine (NIC)-induced hypotension in conscious rabbits. Magnetic flux densities were up to 5.5 mT and the spatial magnetic gradient peaked in neck (carotid sinus baroreceptor) region at the level of approximately 0.06 mT/mm. The duration of exposure was 30 min (including 10 min of pretreatment) and the effects on BP were investigated up to 100 min postexposure. Baroreflex sensitivity (BRS) was estimated from invasive recordings of systolic BP and pulse interval. Neck exposure to 5.5 mT significantly attenuated the pharmacologically induced vasoconstriction or vasodilation, and subsequently suppressed the increase or decrease in BP compared with sham exposure. In contrast, pelvic exposure to 5.5 mT did not significantly antagonized NE-elevated BP or NIC-reduced BP. The neck exposure to 5.5 mT has a biphasic and restorative effect on vascular tone and BP acting to normalize the tone and BP. The neck exposure to 5.5 mT caused a significant increase in BRS in NE-elevated BP compared with sham exposure. The buffering effects of the SMF on increased hemodynamic variability under NE-induced high vascular tone and NIC-induced low vascular tone might be, in part, dependent on baroreflex pathways, which could modulate NE-mediated response in conjunction with Ca(2+) dynamics.     

Morphofunctional study of 12‐O‐tetradecanoyl‐13‐phorbol acetate (TPA)‐induced differentiation of U937 cells under exposure to a 6 mT static magnetic field

This study deals with the morphofunctional influence of 72 h exposure to a 6 mT static magnetic field (SMF) during differentiation induced by 50 ng/ml 12‐O‐tetradecanoyl‐13‐phorbol acetate (TPA) in human leukaemia U937 cells. The cell morphology of U937 cells was investigated by optic and electron microscopy. Specific antibodies and/or molecules were used to label CD11c, CD14, phosphatidylserine, F‐actin and to investigate the distribution and activity of lysosomes, mitochondria and SER. [Ca²⁺]_i was evaluated with a spectrophotometer. The degree of differentiation in SMF‐exposed cells was lower than that of non‐exposed cells, the difference being exposure time‐dependent. SMF‐exposed cells showed cell shape and F‐actin modification, inhibition of cell attachment, appearance of membrane roughness and large blebs and impaired expression of specific macrophagic markers on the cell surface. The intracellular localization of SER and lysosomes was only partially affected by exposure. A significant localization of mitochondria with an intact membrane potential at the cell periphery in non‐exposed, TPA‐stimulated cells was observed; conversely, in the presence of SMF, mitochondria were mainly localised near the nucleus. In no case did SMF exposure affect cell viability. The sharp intracellular increase of [Ca²⁺]_i could be one of the causes of the above‐described changes. Bioelectromagnetics 30:352–364, 2009. © 2009 Wiley‐Liss, Inc.     

Blockade of sensory neuron action potentials by a static magnetic field in the 10 mT range

To characterize the inhibitory effect of a static magnetic field, action potentials (AP) were elicited by intracellular application of 1 ms depolarizing current pulses of constant amplitude to the somata of adult mouse dorsal root ganglion neurons in monolayer dissociated cell culture. During the control period, <5% of stimuli failed to elicit AP. During exposure to an ?11 mT static magnetic field at the cell position produced by an array of four permanent center-charged neodymium magnets of alternating polarity (MAG-4A), 66% of stimuli failed to elicit AP. The number of failures was maximal after about 200-250 s in the field and returned gradually to baseline over 400–600 s. A direct or indirect effect on the conformation of AP generating sodium channels could account for these results because (I) failure was preceded often by reduction of maximal rate of rise, an indirect measure of sodium current; (2) recovery was significantly prolonged in more than one-half of neurons that were not stimulated during exposure to the MAG-4A field; and (3) resting membrane potential, input resistance, and chronaxie were unaffected by the field. The effect was diminished or prevented by moving the MAG-4A array along the X or Z axis away from the neuron under study and by increasing the distance between magnets in the XY plane. Reduction of AP firing during exposure to the ?0.1 mT field produced by a MAG-4A array of micromagnets was about the same as that produced by a MAG-4A array of the large magnets above. The ?28 mT field produced at cell position by two magnets of alternating polarity and the ?88 mT field produced by a single magnet had no significant effect on AP firing. These findings suggest that field strength alone cannot account for AP blockade. © 1995 Wiley-Liss, Inc.     

Raman spectroscopic evidence for structural changes in poly-L-lysine induced by an approximately 50 mT static magnetic field

We have explored the mechanism of coupling of an approximately 50 mT static magnetic field with the α helices of poly-L-lysine. Structural changes in poly-L-lysine were determined by Raman spectroscopy. Our testable hypothesis is that static magnetic fields of this magnitude can couple with the α-helical segments of the polypeptide, and, as a result, the structure of the polypeptide is significantly altered. Our model further suggests that a static magnetic field can promote protein unfolding and can prevent refolding. © 1996 Wiley-Liss, Inc.     

Effects of weak static magnetic fields on endothelial cells

Pulsed electromagnetic fields (PEMFs) have been used extensively in bone fracture repairs and wound healing. It is accepted that the induced electric field is the dose metric. The mechanisms of interaction between weak magnetic fields and biological systems present more ambiguity than that of PEMFs since weak electric currents induced by PEMFs are believed to mediate the healing process, which are absent in magnetic fields. The present study examines the response of human umbilical vein endothelial cells to weak static magnetic fields. We investigated proliferation, viability, and the expression of functional parameters such as eNOS, NO, and also gene expression of VEGF under the influence of different doses of weak magnetic fields. Applications of weak magnetic fields in tissue engineering are also discussed. Static magnetic fields may open new venues of research in the field of vascular therapies by promoting endothelial cell growth and by enhancing the healing response of the endothelium. Bioelectromagnetics 31:296–301, 2010. © 2010 Wiley‐Liss, Inc.     

Effects of 12 mT static magnetic field on sympathetic agonist-induced hypertension in Wistar rats

We investigated the combined effects of a moderate-intensity static magnetic field (SMF) and two different sympathetic agonists, an alpha(1)-adrenoceptor agonist, phenylephrine and a beta(1)-adrenoceptor agonist, dobutamine, which induced hypertension and different hemodynamics in Wistar rats. Five-week-old male rats were continuously exposed to the SMF intensity of 12 mT (B(max)) with the peak spatial gradient of 3 mT/mm for 10 weeks. A loop-shaped flexible rubber magnet was adjusted to fit snugly around the neck region of a rat (diameter-adjustable to an animal size). Sham exposure was performed using a dummy magnet. Six experimental groups of six animals each were examined: (1) sham exposure with intraperitoneal (ip) saline injection (control); (2) SMF exposure with ip saline injection (SMF); (3) sham exposure with ip phenylephrine (1.0 microg/g) injection (PE); (4) SMF exposure with ip phenylephrine injection (SMF + PE); (5) sham exposure with ip dobutamine (4.0 microg/g) injection (DOB); (6) SMF exposure with ip dobutamine injection (SMF + DOB). Fifteen minutes after the injection of each agent, the first set of parameters, arterial blood pressure (BP) and heart rate (HR), the second set of parameters, skin blood flow (SBF) and skin blood velocity (SBV), or the third set of parameters, the number of rearing (exploratory behavior) responses and body weight was monitored. Each agent was administered three times a week for 10 weeks, and each set of parameters was monitored on different days, once a week. The dose of phenylephrine significantly increased BP and decreased HR, SBF, SBV, and the number of rearing responses in the PE group compared with those in the respective age-matched control group. The dose of dobutamine significantly increased BP and HR, increased SBF, SBV, and the number of rearing responses in the DOB group compared with those in the control group. Continuous neck exposure to the SMF alone for up to 10 weeks induced no significant changes in any of the measured cardiovascular and behavioral parameters. The SMF exposure for at least 2 weeks (1) significantly depressed phenylephrine effects on BP, SBF, SBV, and rearing activity (SMF + PE group vs. PE group); (2) significantly depressed dobutamine effects on BP, SBF, and SBV, and suppressed dobutamine-induced increase in the rearing activity (SMF + DOB group vs. DOB group). These results suggest that continuous neck exposure to 12 mT SMF for at least 2 weeks may depress or suppress sympathetic agonists-induced hypertension, hemodynamics, and behavioral changes by modulating sympathetic nerve activity.     

Effects of static magnetic fields on the growth of various types of human cells

The effects of a static magnetic field (SMF) on the proliferation of various types of human cells were determined. All cultures were maintained at 37 °C throughout the experiment. SMF was generated by placing two magnets oppositely oriented on either side of a T25 flask. The flux density in the flask ranged from 35 to 120 mT. Growth curves were constructed by plotting cell number at 18 h and 4, 7, 11, and 14 days after seeding, with the 18‐h point being a measure of attachment efficiency. Exposure to SMF significantly decreased initial attachment of fibroblasts and decreased subsequent growth compared to sham‐exposed control. Significant effects were observed in both fetal lung (WI‐38) and adult skin fibroblasts, but they were generally larger in the fetal lung fibroblast line. SMF did not affect attachment of human melanoma cells, but inhibited their growth by 20% on day 7. SMF produced no effects in a human adult stem cell line. Oxidant production increased 37% in WI‐38 cells exposed to SMF (230–250 mT) during the first 18 h after seeding, when cell attachment occurs. Conversely, no elevation in oxidant levels was observed after a prolonged 5‐day exposure. These results indicate that exposure to SMF has significant biological effects in some, but not all types of human cells. Bioelectromagnetics 32:140–147, 2011. © 2010 Wiley‐Liss, Inc.     

Effect of 6 mT static magnetic field on the bcl-2, bax, p53 and hsp70 expression in freshly isolated and in vitro aged human lymphocytes

An increasing number of evidence indicates that static magnetic fields (SMFs) are capable of altering apoptosis, mainly through modulation of Ca²⁺ influx. Here we present data that suggest apoptotic-related gene expression as an alternative pathway, through which exposure to 6 milliTesla (mT) SMF can interfere with apoptosis. Exposure to 6 mT SMF affects the apoptotic rate (spontaneous and drug-induced) and [Ca²⁺]_i in isolated human lymphocytes; the aged cells are more susceptible to exposure than fresh ones. The exposure to 6 mT exerted a protective effect on chemical or physical-induced apoptosis, irrespective of the age of the cells.The investigation of the gene expression of bcl-2, bax, p53 and hsp70 in freshly isolated and in culture-aged human lymphocytes indicates that these genes are modulated by SMF exposure in the experimental conditions used, in a gene-, age- and time-dependent manner. The exposure of isolated lymphocytes to SMF for up to 24 h modulated increased bax and p53 and decreased hsp70, and bcl-2. The amount of increment and/or decrement of the proteins varied for each gene examined and was independent of the apoptotic inducers. Finally, the same stress applied to freshly isolated or aged lymphocytes resulted in different modulation of bcl-2, bax and hsp70.     

Effects of 180 mT static magnetic fields on diabetic wound healing in rats

Diabetic wound (DW) problems are becoming a formidable clinical challenge due to the sharp increase in the diabetic population and the high incidence of DW. Static magnetic field (SMF) therapy, an inexpensive and accessible noninvasive method, has been proven to be effective on various tissue repairs. However, the issue of the therapeutic effect of SMF on DW healing has never been investigated. The objective of this study was to systematically evaluate the effect of a 180 mT moderate‐intensity gradient SMF on DW healing in streptozotocin‐induced diabetic rats. Forty‐eight 3‐month‐old male Sprague–Dawley rats (32 diabetic and 16 non‐diabetic rats) were assigned to three equal groups: normal wound, DW, and DW + SMF groups. An open circular wound with 1.5 cm diameter was created in the dorsum. The wound was covered with a dressing and the magnet was fixed on top of the dressing. On days 5, 12, and 19, four rats of each group were euthanized and gross wound area, histology and tensile strength were evaluated. The wound area determination suggested that SMF significantly increased the healing rate and reduced the gross healing time. This result was further confirmed by histological observations. The wound tensile strength, reflecting the amount and quality of collagen deposition, increased to a larger extent in the DW + SMF group on days 12 and 19 compared with the DW group. The results indicated that 180 mT SMF presented a beneficial effect on DW healing, and implied the clinical potential of SMF therapy in accelerating DW repair and releasing the psychological and physical burdens of diabetic patients. Bioelectromagnetics 31:640–648, 2010. © 2010 Wiley‐Liss, Inc.     

Effects of a moderate-intensity static magnetic field on VEGF-A stimulated endothelial capillary tubule formation in vitro

Effects of a moderate-intensity static magnetic field (SMF) on the early-stage development of endothelial capillary tubule formation were examined during the initial cell growth periods using co-cultured human umbilical vein endothelial cells and human diploid fibroblasts. The co-cultured cells within a well (16 mm in diameter) were exposed to SMF intensity up to 120 mT (Bmax) with the maximum spatial gradient of 21 mT/mm using a disc-shaped permanent magnet (16 mm in diameter and 2.5 mm in height) for up to 10 days. Control exposure was performed without magnet. Some vascular endothelial cells were treated with vascular endothelial growth factor (VEGF)-A (10 ng/ml) to promote the tubule formation every 2-3 days. Four experimental protocols were performed: (1) non-exposure (control); (2) SMF exposure alone; (3) non-exposure with VEGF-A; (4) SMF exposure with VEGF-A. Photomicrographs of tubule cells immunostained with an anti-platelet-endothelial cell adhesion molecule-1 (PECAM-1 [CD31[) antibody as a pan-endothelial marker, were analyzed after culture at 37 degrees C for 4, 7, and 10 days. The mean values of the area density and the length of tubules (related mainly to arteriogenesis) as well as the number of bifurcations (related mainly to angiogenesis) were determined as parameters of tubule formation and were compared between the groups. After a 10 day incubation, in the peripheral part of the culture wells, SMF alone significantly promoted the tubule formation in terms of the area density and the length of tubules, compared with control group. In the central part of the wells, however, SMF did not cause any significant changes in the parameters of tubule formation. After a 7 day incubation, VEGF-A significantly promoted all the parameters of tubule formation in any part of the wells, compared with control group. With regard to the synergistic effects of SMF and VEGF-A on tubule formation, after a 10 day incubation, SMF significantly promoted the VEGF-A-increased area density and length of tubules in the peripheral part of the wells, compared with the VEGF-A treatment alone. However, SMF did not induce any significant changes in the VEGF-A-increased number of bifurcations in any part of the wells. The tubule cells observed in the wells had elongated, spindle-like shapes, and the direction of cell elongation was random, irrespective of the presence and direction of SMF. These findings suggest that the application of SMF to intact or VEGF-A-stimulated vascular endothelial cells leads mainly to promote or enhance arteriogenesis in the peripheral part of the wells, where the spatial gradient increases relative to the central part. The effects of SMF on the VEGF-A-enhanced tubule formation appear to be synergistic or additive in arteriogenesis but not in angiogenesis.