In th is paper, an Extremely Low Frequency Multi-waveform Electromagnetic Field Generator has been developed with single chip computer, which provides pulsed, rectangular, triangular and sinusoidal magnetic field with the frequency range from 0-150 Hz and the intensity range from 0-50 mT. The application shows that the device is easy to operate and the parameters of magnetic fields are stable. It has provided successful application in the experiments on biological effects of magnetics and has obtained valuable result.
Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Equipment Design ; Humans ; Magnetics ; instrumentation ; Osteoblasts ; radiation effects

This study investigated that whether a 2 mT, 60 Hz, sinusoidal electromagnetic field (EMF) alters the structure and function of cells. This research compared the effects of EMF on four kinds of cell lines: hFOB 1.19 (fetal osteoblast), T/G HA-VSMC (aortic vascular smooth muscle cell), RPMI 7666 (B lymphoblast), and HCN-2 (cortical neuronal cell). Over 14 days, cells were exposed to EMF for 1, 3, or 6 hours per day (hrs/d). The results pointed to a cell type-specific reaction to EMF exposure. In addition, the cellular responses were dependent on duration of EMF exposure. In the present study, cell proliferation was the trait most sensitive to EMF. EMF treatment promoted growth of hFOB 1.19 and HCN-2 compared with control cells at 7 and 14 days of incubation. When the exposure time was 3 hrs/d, EMF enhanced the proliferation of RPMI 7666 but inhibited that of T/G HA- VSMC. On the other hand, the effects of EMF on cell cycle distribution, cell differentiation, and actin distribution were unclear. Furthermore, we hardly found any correlation between EMF exposure and gap junctional intercellular communication in hFOB 1.19. This study revealed that EMF might serve as a potential tool for manipulating cell proliferation.
Signal Transduction ; Microfilaments/radiation effects ; Humans ; Gap Junctions/metabolism/radiation effects ; *Electromagnetic Fields ; Cell Proliferation/radiation effects ; Cell Physiology/*radiation effects ; Cell Line ; Cell Differentiation/radiation effects ; Cell Cycle/radiation effects

This study investigated that whether a 2 mT, 60 Hz, sinusoidal electromagnetic field (EMF) alters the structure and function of cells. This research compared the effects of EMF on four kinds of cell lines: hFOB 1.19 (fetal osteoblast), T/G HA-VSMC (aortic vascular smooth muscle cell), RPMI 7666 (B lymphoblast), and HCN-2 (cortical neuronal cell). Over 14 days, cells were exposed to EMF for 1, 3, or 6 hours per day (hrs/d). The results pointed to a cell type-specific reaction to EMF exposure. In addition, the cellular responses were dependent on duration of EMF exposure. In the present study, cell proliferation was the trait most sensitive to EMF. EMF treatment promoted growth of hFOB 1.19 and HCN-2 compared with control cells at 7 and 14 days of incubation. When the exposure time was 3 hrs/d, EMF enhanced the proliferation of RPMI 7666 but inhibited that of T/G HA- VSMC. On the other hand, the effects of EMF on cell cycle distribution, cell differentiation, and actin distribution were unclear. Furthermore, we hardly found any correlation between EMF exposure and gap junctional intercellular communication in hFOB 1.19. This study revealed that EMF might serve as a potential tool for manipulating cell proliferation.
Signal Transduction ; Microfilaments/radiation effects ; Humans ; Gap Junctions/metabolism/radiation effects ; *Electromagnetic Fields ; Cell Proliferation/radiation effects ; Cell Physiology/*radiation effects ; Cell Line ; Cell Differentiation/radiation effects ; Cell Cycle/radiation effects

OBJECTIVETo explore the effect of static magnetic field (SMF) on the differentiation and proliferation of rat embryonic midbrain neurons cells.

METHODSThe micromass culture of rat embryonic midbrain neurons cell was applied to study the effect of varieties of SMF (1.0, 10.0, 50.0, 100.0, 200.0 mT) and FACS.

RESULTSSMF inhibited the differentiation of the cell without affecting cell proliferation. The concentration of 50% inhibition of cell differentiation (ICD(50)) was 25 mT. The concentration of 50% inhibition of cell differentiation (IVD(50)) was 45 mT.

CONCLUSIONSThe inhibition of SMF on embryonic midbrain neurons cells may be associated with impact of protein syntheses, and lipid peroxidation.

Animals ; Cell Differentiation ; radiation effects ; Cell Division ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; adverse effects ; Embryo, Mammalian ; Female ; Lipid Peroxidation ; radiation effects ; Mesencephalon ; cytology ; radiation effects ; Neurons ; cytology ; radiation effects ; Protein Biosynthesis ; radiation effects ; Rats ; Rats, Sprague-Dawley

This study examined the osteogenic effect of electromagnetic fields (EMF) under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells (BMSCs) and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit (CCK-8). For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase (ALP) staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure (2 h/day) could obviously promote proliferation of BMSCs and osteoblasts, while long-term EMF (8 h/day) could promote osteogenic differentiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when cells were exposed to EMF. Compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differentiation of BMSCs.
Alkaline Phosphatase ; biosynthesis ; Animals ; Bone Marrow Cells ; cytology ; radiation effects ; Cell Differentiation ; genetics ; radiation effects ; Cell Proliferation ; radiation effects ; Coculture Techniques ; Electromagnetic Fields ; Mesenchymal Stromal Cells ; radiation effects ; Osteoblasts ; radiation effects ; Osteogenesis ; genetics ; radiation effects ; Rats

OBJECTIVE: To explore the effects of low level laser irradiation (LLLI) on the osteogenic capacity of three-dimensional (3D) structure by 3D bio-printing construct used human adipose-derived stem cells (hASCs) as seed cells. METHODS: Using hASCs as seed cells, we prepared sodium alginate/gelatin/hASCs 3D bio-printing construct, and divided them into four groups: PM (proliferative medium), PM+LLLI, OM (osteogenic medium) and OM+LLLI, and the total doses of LLLI was 4 J/cm². Immunofluorescence microscopy was used to observe the viability of the cells, and analyze the expression of the osteogenesis-related protein Runt-related transcription factor 2 (Runx2) and osteocalcin (OCN). RESULTS: The 3D constructs obtained by printing were examined by microscope. The sizes of these 3D constructs were 10 mm×10 mm×1.5 mm. The wall thickness of the printed gelatin mold was approximately 1 mm, and the pores were round and had a diameter of about 700 μm. The cell viability of sodium alginate/gelatin/hASCs 3D bio-printing construct was high, and the difference among the four groups was not significant. On day 7, the expression of OCN from high to low was group OM+LLLI, PM+LLLI, OM and PM. There were significant differences among these groups (P<0.01), but there was no significant difference between group PM+LLLI and OM. On day 14, the expression of OCN in each group was higher than that on day 7, and there was no significant difference between group OM+LLLI and OM. The expression of Runx2 in group OM+LLLI was more than 90%, significantly higher than that in group OM (P<0.01). But the expression of Runx2 in group PM+LLLI and OM+LLLI were significantly lower than that in the non-irradiated groups. The expression of osteogenesis-related protein Runx2 and OCN were higher in OM groups than in PM groups. Furthermore, the irradiated groups were significantly higher than the non-irradiated groups. CONCLUSION LLLI does not affect the cell viability of sodium alginate/gelatin/hASCs 3D bio-printing construct, and may promote the osteogenic differentiation of hASCs.
Adipocytes/radiation effects* ; Alginates ; Cell Differentiation ; Cell Proliferation ; Gelatin ; Humans ; Lasers ; Osteogenesis ; Printing, Three-Dimensional ; Stem Cells/radiation effects*

OBJECTIVETo study the effects of low frequency pulsed magnetic field on the proliferation and differentiation of HepG2 cells.

METHODS3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetry method and ELISA assay of alpha-fetoprotein (AFP) were used to determine the cell proliferation and differentiation after the cells were exposed to pulsed magnetic fields with different frequency but the same field intensity.

RESULTSThere were no significant differences in cell proliferation between sham and treated groups exposed to the field of 80 Hz, 1.55 mT for 1, 4, 8, 12, 24 h (P > 0.05). There were also no significant differences in cell proliferation and AFP secretion between sham and treated groups exposed to 16 Hz, 1.55 mT pulsed magnetic fields for 1, 4, 8, 24 h (P > 0.05).

CONCLUSIONThere were no "window effects" found in HepG2 cells proliferation or AFP secretion at 16 Hz and 80 Hz pulsed magnetic fields.

Cell Differentiation ; radiation effects ; Cell Division ; radiation effects ; Cell Line, Tumor ; cytology ; metabolism ; radiation effects ; Electromagnetic Fields ; Humans ; alpha-Fetoproteins ; analysis

To study the genic and non-genic regulation of 50 Hz 0.6 mT pulsed electromagnenic fields (PEMF) on rat calvarial osteoblasts (ROB) differentiation.ROBs were achieved by enzyme digestion, and treated with 50 Hz 0.6 mT PEMFs for 1.5 hours after subculture. The alkaline phosphatase (ALP) activity, mRNA transcription of ALP, Runx2 and OSX and protein expression of Runx2 and OSX were detected at 0, 3, 6, 9 and 12 hours after PEMF treatment.The ALP activity at 3 hours after treatment was significantly higher than that in the control(<0.01), while the mRNA transcription of ALP began to increase at 6 hours after treatment. The mRNA transcription of Runx2 increased immediately after treatment and regressed at 6 hours, then increased again. The protein expression of it corresponded but with a little lag. The mRNA transcription of OSX also raised instantly after treatment, then returned to the level of control at 6 hours, and lower than control at 12 hours significantly. The protein expression of it also corresponded but with a bit delay.There are genic regulation for the protein expression of Runx2 and OSX, and non-genic regulation for the ALP activity on the process of 50 Hz 0.6 mT PEMFs prompts ROBs differentiation.
Alkaline Phosphatase ; metabolism ; radiation effects ; Animals ; Cell Differentiation ; genetics ; radiation effects ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; metabolism ; radiation effects ; Electromagnetic Fields ; Osteoblasts ; chemistry ; radiation effects ; Osteogenesis ; genetics ; radiation effects ; Rats ; Transcription Factors ; metabolism ; radiation effects

The present research was to investigate the time effect of sinusoidal electromagnetic fields (SEMFs) at different exposure time on the proliferation and differentiation of osteoblasts (OB) in vitro. The newborn rat calvarial OB were isolated by enzyme digestion and divided randomly into 7 groups after one passage. The exposure times of the SEMFs were 0.5 h, 1.0 h, 1.5 h, 2.0 h, 2.5 h and 3.0 h, respectively, and the frequency was 50 Hz. The cells were exposed in the SEMFs of 1.8 mT. Those without SEMFs exposure were used as the control group. They were observed under the contrast phase microscope each day. After 48 h, cell proliferation was assayed by MTT method. The alkaline phosphatase (Alkaline Phosphatase, ALP) activities were measured after the exposure of SEMFs for 3 d, 6 d, 9 d and 12 d, respectively. The calcified nodules were stained by Alizarin Bordeaux after 10 d. The cells exposed in the SEMFs were arranged in Spiral appearance after 8 d. The SEMFs exposure time at 2.0 h, 2.5 h and 3.0 h significantly inhibited cell proliferation (P < 0.01) and 0.5 h, 1.0 h, 1.5 h groups more significantly than control groups (P < 0.05). When the 3 d, 6 d and 12 d the ALP activities of the 0.5 h, 1.0 h, 1.5 h and 2.0 h, times group were significantly higher than those in the control group (P < 0.05), and after 9 d the 1.0 h, 1.5 h and 2.0 h activity of ALP higher significantly than control and other groups (P < 0.01). Other groups had no effect on the ALP activity. Alizarin Bordeaux staining result showed the amounts of calcified nodules 1.0 h, 1.5 h and 2.0 h higher than control groups. The SEMFs at 50 Hz, 1.8 mT different time exposure groups inhibits the proliferation of OB, but they enhances the maturation and mineralization of the OB and SEMFs at 1.8 mT of the 1.5 h has the strongest activity.
Animals ; Animals, Newborn ; Calcification, Physiologic ; radiation effects ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Osteoblasts ; cytology ; Osteogenesis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigated the effect of 50-Hz 3.6-mT sinusoidal electromagnetic fields (SEMFs) on the proliferation and differentiation of osteoblasts in vitro.

METHODSThe newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 6 groups after one passage. The treatment groups under 50-Hz 3.6-mT SEMFs and controls without SEMFs treatment. The cells were exposed in the SEMFs for 0.5 h, 1.0 h, 1.5 h, 2.0 h, and 2.5 h. They were observed under the contrast phase microscope each day. The calcified nodules were stained by alizarin red. The SEMFs were arranged in spiral appearance after 3 to 5 days.

RESULTSThe SEMFs showed characteristic distribution 3 to 5 days after SEMFs treatment. On the 9(th) day after treatment, the activity of alkaline phosphatase (ALP) significantly increased in the 0.5-h group, whereas the ALP histochemical straining results and the area of calcified nodules were consistent with ALP activity. In the 48-h and 96-h groups, the genetic expression levels of osteoprotegerin and collagen-1 were significantly higher than that in the control group; particularly, the mRNA expression increased in the 0.5-h group.

CONCLUSIONThe SEMFs at 50-Hz 3.6-mT could suppress the proliferation of osteoblasts maturation but stimulate the differentiation and maturation of osteoblasts in vitro.

Animals ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Male ; Osteoblasts ; cytology ; radiation effects ; Rats ; Rats, Sprague-Dawley