静磁场对小鼠黑色素瘤细胞B16生长和氧化应激的影响

目的:利用小鼠黑色素瘤细胞B16,研究静磁场对肿瘤细胞生长和氧化应激的影响,探讨氧化应激介导静磁场影响肿瘤细胞生长的机制,为磁场在肿瘤疾病的治疗中的应用提供理论依据。方法:采用MTT法测定磁场对B16细胞活力的影响;利用流式细胞仪测定静磁场暴露对B16细胞周期分布的影响;利用生物化学方法测定磁场暴露对细胞氧化防御系统相关蛋白酶活性的影响。结果:24 h内50 m T-200 m T静磁场暴露可以抑制B16生长,但超过24 h的磁场暴露可以促进B16生长;100 m T和200 m T静磁场暴露对B16的细胞周期分布没有影响;B16暴露于100 m T和200 m T静磁场48 h,GST活性和GSH/GSSG水平表现为先上升后下降,SOD活性和T-AOC水平先下降后上升,CAT活性没有受到影响。结论:50 m T-200 m T静磁场可以抑制小鼠黑色素瘤细胞B16的生长,诱导肿瘤细胞产生氧化应激。     

The effect of 100 mT SMF on activation of the hsp70 promoter in a heat shock/luciferase reporter system

Human exposure to magnetic fields, increased through use of new technologies like magnetic resonance imaging (MRI), has prompted investigations into possible effects of static magnetic fields (SMFs) on cellular processes. However, controversy still remains between many studies, which likely results from a lack of uniformity across experimental parameters, including the length of magnetic field exposure, the strength of the magnetic field, and the cell type or organism under investigation. The purpose of this research was to monitor effects of SMF exposure using real‐time luminescence photometry. The study investigated the potential interaction of a 100 mT SMF on a heat shock protein (hsp70)/luciferase reporter construct in stably transfected NIH3T3 cells. Changes in heat shock promoter activation following 100 mT SMF exposure were analyzed and detected as bioluminescence in real‐time. Two heat parameters were considered in combination with sham‐ and 100 mT‐exposed experiments: no heat or 1,800 s heat. As expected, there was a significant increase in bioluminescence in response to 1,800 s of heat alone. However, no significant difference in average hsp70 promoter activation between sham and 100 mT experiments was observed for no heat or 1,800 s heat experiments. Therefore, a 100 mT SMF was shown to have no effect on the activation of the heat shock protein promoter during SMF exposure or when SMF exposure was combined with a heat insult. J. Cell. Biochem. 108: 956–962, 2009. © 2009 Wiley‐Liss, Inc.     

Chronic exposure to a 2 mT static magnetic field affects the morphology,the metabolism and the function of in vitro cultured swine granulosa cells

In recent years, the exposure of organisms to static magnetic fields (SMFs) is continuously increasing. Thus, we investigated the effect of chronic exposure to a 2 mT SMF on in vitro cultured swine granulosa cells (GCs). In particular, the culture expansion (cell viability and doubling time), the cell phenotype (cell morphology and orientation, actin and α-tubulin cytoskeleton), the cell metabolism (intracellular Ca²⁺ concentration [Ca²⁺]_i and mitochondrial activity) and the cell function (endocrine activity) were assessed. It has been found that the exposure to the field did not affect the cell viability, but the doubling time was significantly reduced (p < 0.05) in exposed samples after 72 h of culture. At the same time, the cell length and thickness significantly changed (p < 0.05), while the cell orientation was unaffected. Evident modifications were induced on actin and α-tubulin cytoskeleton after 3 days of exposure and, simultaneously, a change in [Ca²⁺]_i and mitochondrial activity started to become evident. Finally, the SMF exposure of GCs longer than 72 h determined a significant alteration of progesterone and estrogen production (p < 0.05). In conclusion, our results demonstrate that the chronic exposure of swine GCs to a 2 mT SMF exerts a negative effect on cell proliferation, morphology, biochemistry and endocrine function in an in vitro model.     

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.     

Regulation of Osteoblast Differentiation and Iron Content in MC3T3-E1 Cells by Static Magnetic Field with Different Intensities

Many studies have indicated that static magnetic fields (SMFs) have positive effects on bone tissue, including bone formation and bone healing process. Evaluating the effects of SMFs on bone cell (especially osteoblast) function and exploring the mechanism, which is critical for understanding the possible risks or benefits from SMFs to the balance of bone remodeling. Iron and magnetic fields have the natural relationship, and iron is an essential element for normal bone metabolism. Iron overload or deficiency can cause severe bone disorders including osteoporosis. However, there are few reports regarding the role of iron in the regulation of bone formation under SMFs. In this study, hypomagnetic field (HyMF) of 500 nT, moderate SMF (MMF) of 0.2 T, and high SMF (HiMF) of 16 T were used to investigate how osteoblast (MC3T3-E1) responses to SMFs and iron metabolism of osteoblast under SMFs. The results showed that SMFs did not pose severe toxic effects on osteoblast growth. During cell proliferation, iron content of osteoblast MC3T3-E1 cells was decreased in HyMF, but was increased in MMF and HiMF after exposure for 48 h. Compared to untreated control (i.e., geomagnetic field, GMF), HyMF and MMF exerted deleterious effects on osteoblast differentiation by simultaneously retarding alkaline phosphatase (ALP) activity, mineralization and calcium deposition. However, when exposed to HiMF of 16 T, the differentiation potential showed the opposite tendency with enhanced mineralization. Iron level was increased in HyMF, constant in MMF and decreased in HiMF during cell differentiation. In addition, the mRNA expression of transferrin receptor 1 (TFR1) was promoted by HyMF but was inhibited by HiMF. At the same time, HiMF of 16 T and MMF of 0.2 T increased the expression of ferroportin 1 (FPN1). In conclusion, these results indicated that osteoblast differentiation can be regulated by altering the strength of the SMF, and iron is possibly involved in this process.

     

ENHANCEMENT OF INVERTASE PRODUCTION BY Aspergillus niger OZ-3 USING LOW-INTENSITY STATIC MAGNETIC FIELDS

The aim of this study is to investigate the effect of low-intensity static magnetic fields (SMFs) on invertase activity and growth on different newly identified molds. The most positive effect of SMFs on invertase activity and growth was observed for Aspergillus niger OZ-3. The submerged production of invertase was performed with the spores obtained at the different exposure times (120, 144, 168, and 196 hr) and magnetic field intensities (0.45, 3, 5, 7, and 9 mT). The normal magnetic field of the laboratory was assayed as 0.45 mT (control). Optimization of magnetic field intensity and exposure time significantly increased biomass production and invertase activity compared to 0.45 mT. The maximum invertase activity (51.14 U/mL) and biomass concentration (4.36 g/L) were achieved with the spores obtained at the 144 hr exposure time and 5 mT magnetic field intensity. The effect of low-intensity static magnetic fields (SMFs) on invertase activities of molds was investigated for the first time in the present study. As an additional contribution, a new hyper-invertase-producing mold strain was isolated.

Supplemental materials are available for this article. Go to the publisher's online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.     

Seed orientation and magnetic field strength have more influence on tomato seed performance than relative humidity and duration of exposure to non-uniform static magnetic fields

Different factors (e.g., light, humidity, and temperature) including exposure to static magnetic fields (SMFs), referred here as critical factors, can significantly affect horticultural seed performance. However, the link between magnetic field parameters and other interdependent factors affecting seed viability is unclear. The importance of these critical factors affecting tomato (Solanum lycopersicum L.) var. MST/32 seed performance was assessed after performing several treatments based on a L₉ (3⁴) (four factors at three levels) orthogonal array (OA) design. The variable factors in the design were magnetic flux density (R₁ = 332.1 ± 37.8 mT; R₂ = 108.7 ± 26.9 mT; and R₃ = 50.6 ± 10.5 mT), exposure time (1, 2, and 24 h), seed orientation (North polarity, South polarity, and control – no magnetic field), and relative humidity (RH) (7.0, 25.5, and 75.5%). After seed moisture content stabilisation at the different chosen RH, seeds were exposed in dark under laboratory conditions to several treatments based on the OA design before performance evaluation. Treatments not employing magnetic field exposure were used as controls. Results indicate that electrolyte leakage rate was reduced by a factor of 1.62 times during seed imbibition when non-uniform SMFs were employed. Higher germination (∼11.0%) was observed in magnetically-exposed seeds than in non-exposed ones, although seedlings emerging from SMF treatments did not show a consistent increase in biomass accumulation. The respective influence of the four critical factors tested on seed performance was ranked (in decreasing order) as seed orientation to external magnetic fields, magnetic field strength, RH, and exposure time. This study suggests a significant effect of non-uniform SMFs on seed performance with respect to RH, and more pronounced effects are observed during seed imbibition rather than during later developmental stages.     

Effects of homogeneous and inhomogeneous static magnetic fields combined with gamma radiation on DNA and DNA repair

The aim of this study was to reveal whether static magnetic fields (SMFs) influence the repair of radiation‐damaged DNA on leukocytes or has any effect on DNA. After 4 Gy of ⁶⁰Co‐γ irradiation, some of the samples were exposed to inhomogeneous SMFs with a lateral magnetic flux density gradient of 47.7, 1.2, or 0.3 T/m by 10 mm lateral periodicity, while other samples were exposed to homogeneous SMF of 159.2 ± 13.4 mT magnetic flux density for a time period of 0.5 min, 1, 2, 4, 6, 18, 20, or 24 h. Another set of samples was exposed to the aforementioned SMFs before gamma irradiation. The following three groups were examined: (i) exposed to SMF only, (ii) exposed to SMF following irradiation by ⁶⁰Co‐γ, and (iii) exposed to SMF before ⁶⁰Co‐γ irradiation. The analysis of the DNA damage was made by single‐cell gel electrophoresis technique (comet assay). Statistically significant differences were found at 1 h (iSMF), 4 h (hSMF), and 18 h (hSMF) if samples were exposed to only SMF, compared to control. When the SMF exposure followed the ⁶⁰Co‐γ irradiation, statistically significant differences were found at 1 h (iSMF) and 4 h (hSMF). If exposure to SMF preceded ⁶⁰Co‐γ irradiation, no statistically significant difference was found compared to 4 Gy gamma‐irradiated group. Bioelectromagnetics 31:488–494, 2010. © 2010 Wiley‐Liss, Inc.     

Evaluation of the effects of weak and moderate static magnetic fields on the characteristics of human low density lipoprotein in vitro

It has been suggested that exposure to electromagnetic fields may be a risk factor for cardiovascular disease in humans. Low density lipoprotein (LDL) modifications such as peroxidation and aggregation have been implicated in the pathogenesis of atherosclerosis. The present study investigated the effects of weak (0.125–0.5 mT) and moderate (1–4 mT) static magnetic fields (SMFs) on LDL oxidation, aggregation and zeta potential in vitro. Our results demonstrated that magnetic flux densities of 0.25 and 0.5 mT decreased, and magnetic flux densities of 3 and 4 mT increased the zeta potential and LDL oxidation in comparison with the control samples. All doses of SMFs increased the LDL aggregation in a time‐ and dose‐dependent manner. It is concluded that SMFs can alter the susceptibility of LDL to oxidation and this alteration is dependent on the applied magnetic flux density. The SMF, in addition to its role in the production and stabilization of free radicals and promotion of lipid peroxidation, may influence the metabolism of lipoproteins and their interaction with other molecules such as apolipoproteins, enzymes and receptors through the alteration of the LDL zeta potential and its particles tendency to aggregation. Bioelectromagnetics 34:397–404, 2013. © 2012 Wiley Periodicals, Inc.     

Static magnetic field effect on the arterial baroreflex-mediated control of microcirculation: implications for cardiovascular effects due to environmental magnetic fields

Increasing evidence suggests that time-varying and static magnetic fields in the environment might affect the cardiovascular system. To explore the underlying physiology, the effect of static magnetic fields (SMFs) on the carotid baroreflex control of microcirculation was studied. Twenty-four hemodynamic monitorings were performed in rabbits sedated by pentobarbital infusion (5 mg/kg/h) during experiments that lasted 120 min. Mean femoral artery blood pressure, heart rate, and ear lobe skin microcirculatory blood flow, measured by microphotoelectric plethysmogram (MPPG), were simultaneously recorded before and after a 40 min exposure of the sinocarotid baroreceptors to Nd₂–Fe₁₄–B alloy magnets (n = 14) or sham magnets (n = 10, control series). The local SMF field was 350 mT, at the baroreceptors’ site. Arterial baroreflex sensitivity (BRS) was estimated from heart rate/blood pressure response to intravenous bolus injections of nitroprusside and phenylephrine. A significant positive correlation was found between the SMF-induced increase in BRS (ΔBRS = BRS_afterSMF − BRS_priorSMF) and the increment in microvascular blood flow (ΔMPPG = MPPG_afterSMF − MPPG_priorSMF) (r = 0.66, p < 0.009). The SMF probably modulated the arterial baroreflex-mediated microcirculatory control. This could represent one possible mechanism how environmental magnetic fields act on the cardiovascular system, and a method how to complexly adjust macro- and microcirculation with potential clinical implementation.     

The pattern of exposure to static magnetic field of nurses involved in activities related to contrast administration into patients diagnosed in 1.5 T MRI scanners

Static magnetic fields (SMFs) and time-varying electromagnetic fields exposure is necessary to obtain the diagnostic information regarding the structure of patient's tissues, by the use of magnetic resonance imaging (MRI) scanners. A diagnostic procedure may also include the administration of pharmaceuticals called contrast, which are to be applied to a patient during the examination. The nurses involved in administering contrast into a patient during the pause in examination are approaching permanently active magnets of MRI scanners and are exposed to SMF. There were performed measurements of spatial distribution of SMF in the vicinity of MRI magnets and parameters of personal exposure of nurses (i.e. individual exposimetric profiles of variability in time of SMF affecting the nurse who is performing tasks in the vicinity of magnet, characterized by statistical parameters of recorded magnetic flux density affecting the nurse). The SMF exposure in the vicinity of various MRI magnets depends on both magnetic flux density of the main field B ₀ (applicable to a patient) and the construction of the scanner, but the most important factor determining the workers' exposure is the work practice. In the course of a patient's routine examination in scanners of B ₀ = 1.5 T, the nurses are present over ～0.4–2.9 min in SMF exceeding 0.03% of B ₀ (i.e. 0.5 mT), but only sometimes they are present in SMF exceeding 5% of B ₀ (i.e. 75 mT). When patients need more attention because of their health status/condition, the nurses' exposure may be significantly longer – it may even exceed 10 min and 30% of B ₀ (i.e. 500 mT). We have found that the level of exposure of nurses to SMF may vary from < 5% of the main field (a median value: 0.5–1.5%; inter-quartile range: 0.04–8.8%; max value: 1.3–12% of B ₀) when a patient is moved from the magnets bore before contrast administration, up to the main field level (B ₀) when a patient stays in the magnets bore and nurse is crawling into the bore.     

Geomagnetic field modulates artificial static magnetic field effect on arterial baroreflex and on microcirculation

Spreading evidence suggests that geomagnetic field (GMF) modulates artificial magnetic fields biological effect and associated with increased cardiovascular morbidity. To explore the underlying physiological mechanism we studied 350 mT static magnetic field (SMF) effect on arterial baroreflex-mediated skin microcirculatory response in conjunction with actual geomagnetic activity, reflected by K and K _p indices. Fourteen experiments were performed in rabbits sedated by pentobarbital infusion (5 mg/kg/h). Mean femoral artery blood pressure, heart rate, and the ear lobe skin microcirculatory blood flow, measured by microphotoelectric plethysmogram (MPPG), were simultaneously recorded before and after 40 min of NdFeB magnets local exposure to sinocarotid baroreceptors. Arterial baroreflex sensitivity (BRS) was estimated from heart rate/blood pressure response to intravenous bolus injections of nitroprusside and phenylephrine. We found a significant positive correlation between SMF-induced increase in BRS and increment in microvascular blood flow (ΔBRS with ΔMPPG, r=0.7, p<0.009) indicated the participation of the arterial baroreflex in the regulation of the microcirculation and its enhancement after SMF exposure. Geomagnetic disturbance, as opposed to SMF, decreased both microcirculation and BRS, and counteracted SMF-induced increment in microcirculatory blood flow (K-index with ΔMPPG; r _s=−0.55, p<0.041). GMF probably affected central baroreflex pathways, diminishing SMF direct stimulatory effect on sinocarotid baroreceptors and on baroreflex-mediated vasodilatatory response. The results herein may thus point to arterial baroreflex as a possible physiological mechanism for magnetic-field cardiovascular effect. It seems that geomagnetic disturbance modifies artificial magnetic fields biological effect and should be taken into consideration in the assessment of the final effect. An erratum to this article can be found at     

Impact of pre-sowing magnetic field exposure of seeds to stationary magnetic field on growth, reactive oxygen species and photosynthesis of maize under field conditions

Impact of pre-sowing exposure of seeds to static magnetic field were studied on 1 month old maize [Zea mays _. var: HQPM.1] plants under field conditions. Pre-standardized magnetic field strength of 100 mT (2 h) and 200 mT (1 h), which were proven best for improving different seedling parameters under laboratory condition, were used for this study. Magnetic field treatment altered growth, superoxide radical level, antioxidant enzymes and photosynthesis. Among the different growth parameters, leaf area and root length were the most enhanced parameters (78–40%, respectively), over untreated plants. Electron paramagnetic resonance spectroscopy study showed that superoxide radical was reduced and hydroxyl radical was unaffected after magnetic field treatment. With decrease in free radical content, antioxidant enzymes like superoxide dismutase and peroxidase were also reduced by 43 and 23%, respectively, in plants that emerged from magnetically treated seeds. Measurement of Chlorophyll a fluorescence by plant efficiency analyzer showed that the potential of processing light energy through photosynthetic machinery was enhanced by magnetic field treatment. Performance index of the plant enhanced up to two-fold and phenomenological leaf model showed more active reaction centers after magnetic field treatment. Among the two field strengths used, 200 mT (1 h) was more effective in altering all these parameters. It is concluded that pre-sowing magnetic field treatment can be effectively used for improving plant growth and development under field conditions.     

Slow Freezing Coupled Static Magnetic Field Exposure Enhances Cryopreservative Efficiency—A Study on Human Erythrocytes

The aim of this study was to assess the cryoprotective effect of static magnetic fields (SMFs) on human erythrocytes during the slow cooling procedure. Human erythrocytes suspended in 20% glycerol were slowly frozen with a 0.4-T or 0.8-T SMF and then moved to a −80°C freezer for 24 hr. The changes in survival rate, morphology, and metabolites of the thawed erythrocytes were examined. To understand possible cryoprotective mechanisms of SMF, membrane fluidity and dehydration stability of SMF-exposed erythrocytes were tested. For each test, sham-exposed erythrocytes were used as controls. Our results showed that freezing coupled with 0.4-T or 0.8-T SMFs significantly increased the relative survival ratios of the frozen-thawed erythrocytes by 10% and 20% (p<0.001), respectively. The SMFs had no effect on erythrocyte morphology and metabolite levels. However, membrane fluidity of the samples exposed to 0.8-T SMF decreased significantly (p<0.05) in the hydrophobic regions. For the dehydration stability experiments, the samples exposed to 0.8-T SMF exhibited significantly lower (p<0.05) hemolysis. These results demonstrate that a 0.8-T SMF decreases membrane fluidity and enhances erythrocyte membrane stability to resist dehydration damage caused by slow cooling procedures.     

Static magnetic field treatment enhanced photosynthetic performance in soybean under supplemental ultraviolet-B radiation

Photosynthesis Research - The study was performed to analyze the impact of seed pretreatment by static magnetic field (SMF) of 200 mT for 1&nbsp;h on photosynthetic performance of soybean...     

Changes in Saccharomyces cerevisiae development induced by magnetic fields

Saccharomyces cerevisiae cultures exposed to 110 mT and 220 mT steady magnetic fields (SMF) were studied to observe eventual induced growth alterations and changes in metabolic activity. Cell mass (biomass) growth was evaluated by light spectrometry, and metabolic alterations were estimated on the basis of the CO2 pressure produced and by the culture media pH changes, measured at the beginning and the end of the observation. The yeast strain DAUFPE-1012, cultivated in a nonaerated liquid agar Sabouraud glucose medium, was exposed to SMF generated by NdFeBr magnets. Results showed alterations induced by 220 mT SMF as an increment in cell proliferation (1.84%) and an increased CO2 production (36.1%) as compared to control groups. Furthermore, the initial-to-final pH difference in 220 mT SMF exposed cultures was higher than the 110 mT SMF and the control values. The whole acidification and the rise in CO2 production observed after 220 mT SMF exposure did not correspond to the biomass growth values, as compared to the other cultures, and was apparently provoked by a enhancement in the cellular metabolic rate. This technique becomes very promising for future biotechnological applications in fermentative processes.     

Involvement of nitrate reductase-dependent nitric oxide production in magnetopriming-induced salt tolerance in soybean

In the present study, experiments were performed to investigate the role of nitric oxide (NO) in magnetopriming-induced seed germination and early growth characteristics of soybean (Glycine max) seedlings under salt stress. The NO donor (sodium nitroprusside, SNP), NO scavenger (2-[4-carboxyphenyl]-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, CPTIO), inhibitors of nitrate reductase (sodium tungstate, ST) or NO synthase (N-nitro-L-Arg-methyl ester, LNAME) and NADPH oxidase inhibitor (diphenylene iodonium, DPI) have been used to measure the role of NO in the alleviation of salinity stress by static magnetic field (SMF of 200 mT, 1 h). Salt stress (50 mM NaCl) significantly reduced germination and early growth of seedlings emerged from non-primed seeds. Pre-treatment of seeds with SMF positively stimulated the germination and consequently promoted the seedling growth. ST, LNAME, CPTIO and DPI significantly decreased the growth of seedling, activities of α-amylase, protease and nitrate reductase (NR), hydrogen peroxide (H₂O₂), superoxide (O₂^•−) and NO content in roots of seedlings emerged from non-primed and SMF-primed seeds. However, the extent of reduction was higher with ST in seedlings of SMF-primed seeds under both conditions, whereas SNP promoted all the studied parameters. Moreover, the generation of NO was also confirmed microscopically using a membrane permanent fluorochrome (4-5-diaminofluorescein diacetate [DAF-2 DA]). Further, analysis showed that SMF enhanced the NR activity and triggered the NO production and NR was maximally decreased by ST as compared to LNAME, CPTIO and DPI. Thus, in addition to ROS, NO might be one of the important signaling molecules in magnetopriming-induced salt tolerance in soybean and NR may be responsible for SMF-triggered NO generation in roots of soybean.     

Magnetostatic Field System for Uniform Cell Cultures Exposure

The aim of the present work has been the design and the realization of a Magnetostatic Field System for Exposure of Cell cultures (MaFiSEC) for the uniform and the reproducible exposure of cell cultures to static magnetic fields (SMFs) of moderate magnetic induction. Experimental and computer-simulated physical measurements show that MaFiSEC: i) generates a SMF with magnetic induction that can be chosen in the range of 3 to 20 mT; ii) allows the uniform SMF exposure of cells growing in adhesion and in suspension; iii) is cheap and easy to use. The efficacy and reproducibility of MaFiSEC has been tested by comparing the biological effects exerted on isolated human lymphocytes by 72 h of exposure to a magnet (i.e. Neodymium Magnetic Disk, NMD) placed under the culture Petri dish. Lymphocytes morphology, viability, cell death, oxidative stress and lysosomes activity were the parameters chosen to evaluate the SMF biological effects. The continuous exposure of cells to a uniform SMF, achieved with MaFiSEC, allows highly reproducible biochemical and morphological data.     

Static magnetic field increases survival rate of dental pulp stem cells during DMSO-free cryopreservation

Abstract

Successful and efficient cryopreservation of living cells and organs is a key clinical application of regenerative medicine. Recently, magnetic cryopreservation has been reported for intact tooth banking and cryopreservation of dental tissue. The aim of this study was to assess the cryoprotective effects of static magnetic fields (SMFs) on human dental pulp stem cells (DPSCs) during cryopreservation. Human DPSCs isolated from extracted teeth were frozen with a 0.4-T or 0.8-T SMF and then stored at ?196?°C for 24?h. During freezing, the cells were suspended in freezing media containing with 0, 3 or 10% DMSO. After thawing, the changes in survival rate of the DPSCs were determined by flow cytometry. To understand the possible cryoprotective mechanisms of the SMF, the membrane fluidity of SMF-exposed DPSCs was tested. The results showed that when the freezing medium was DMSO-free, the survival rates of the thawed DPSCs increased 2- or 2.5-fold when the cells were exposed to 0.4-T or 0.8-T SMFs, respectively (p?<?0.01). In addition, after exposure to the 0.4-T SMF, the fluorescence anisotropy of the DPSCs increased significantly (p?<?0.01) in the hydrophilic region. These results show that SMF exposure improved DMSO-free cryopreservation. This phenomenon may be due to the improvement of membrane stability for resisting damage caused by ice crystals during the freezing procedure.