The effects of high-intensity pulsed electromagnetic stimulation （HIPEMS） on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal ra...The effects of high-intensity pulsed electromagnetic stimulation （HIPEMS） on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5–10 Tesla （T） [8 groups of B–I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance （A） value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of the growth of neural stem cells （P〈0.05）. The A values of neural stem cells in the 6.0 T, 8.0 T, and 10.0 T groups were lower than the sham exposure control group, indicating a restraint of the growth of neural stem cells. The rate of neuron-specific enolase-positive neurons revealed by flow cytometry in HPEMS groups was the same as that in control group （P〉0.05）. It was suggested that 0.1 Hz, 5 pulses stimulation of HPEMS within certain scale of intensity （0.5–10.0 T）, significantly promoted the growth of neural stem cells with the rational intensity being 4.0 T.展开更多
Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal d...Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal differentiation of BMSCs. Low-frequency PEMF can induce the neuronal differentiation of BMSCs in the absence of nerve growth factors. This study was designed to investigate the effects of low-frequency PEMF pretreatment on the proliferation and function of BMSCs and the effects of low-frequency PEMF pre-treated BMSCs on the regeneration of injured peripheral nerve using in vitro and in vivo experiments.In in vitro experiments, quantitative DNA analysis was performed to determine the proliferation of BMSCs, and reverse transcription-polymerase chain reaction was performed to detect S100 （Schwann cell marker）, glial fibrillary acidic protein （astrocyte marker）, and brain-derived neurotrophic factor and nerve growth factor （neurotrophic factors） mRNA expression. In the in vivo experiments, rat models of crush-injured mental nerve established using clamp method were randomly injected with low-frequency PEMF pretreated BMSCs, unpretreated BMSCs or PBS at the injury site （1 × 106 cells）. DiI-labeled BMSCs injected at the injury site were counted under the fluorescence microscope to determine cell survival. One or two weeks after cell injection, functional recovery of the injured nerve was assessed using the sensory test with von Frey filaments. Two weeks after cell injection, axonal regeneration was evaluated using histomorphometric analysis and retrograde labeling of trigeminal ganglion neurons. In vitro experiment results revealed that low-frequency PEMF pretreated BMSCs proliferated faster and had greater mRNA expression of growth factors than unpretreated BMSCs. In vivo experiment results revealed that compared with injection of unpretreated BMSCs, injection of low-frequency PEMF pretreated BMSCs led to higher myelinated axon count and axon density and more DiI-labeled neurons in the trigeminal ganglia, contributing to rapider functional recovery of injured mental nerve. These findings suggest that low-frequency PEMF pretreatment is a promising approach to enhance the efficacy of cell therapy for peripheral nerve injury repair.展开更多
The contrast agent concentration, the time of repetition (TR) and magnetic field strength are significant parameters that influence for the accurate signal intensity (SI) in quantitative Magnetic Resonance Imaging (MR...The contrast agent concentration, the time of repetition (TR) and magnetic field strength are significant parameters that influence for the accurate signal intensity (SI) in quantitative Magnetic Resonance Imaging (MRI). Therefore, this study was conducted to investigate and refine the dependence and the optimal effect of Time of Repetition (TR) on the relationship between signal intensity and Gd-DTPA (Gadolinium-diethylene-triaminepenta-acetic acid) concentration, after applying two-dimensional (2D) Spin Echo (SE) pulse sequence under low-field MRI. In addition to that, the optimal concentration of Gd-DTPA at given sequence parameters at low-field MRI was also evaluated. A water-filled phantom was constructed for a range of Gd-DTPA concentrations (0 - 6 mmol/L) and the mean signal intensities (SIs) were assessed in the defined region of interest on T1-weighted images with different TR values (40 - 2000 ms). The generated signal-concentration curves for Gd-DTPA revealed that increasing TR was associated with the increase of the overall SIs and the maximum relationship between SI to concentration. Moreover, the required Gd-DTPA concentration to produce the maximum SI was associated to decrease with the increase of TR. In addition to this, the application of beyond 100 ms TR values in this study with relatively higher concentrations (beyond 1 - 2 mmol/L) has resulted predominantly non-linear patterns in the signal-concentration curves and it appears the saturation or decay of the SIs due to T2 effect. From these results, it can be suggested that the selection of relatively lower Gd-DTPA concentration ( mmol/L) with less than 800 ms (<800 ms) TR values can produce a better linear relationship between the concertation and SIs in T1-weighted SE low field contrast-enhanced MRI. Furthermore, this study also outlined the significance and necessity of the optimization of TR in SE sequence in low field MRI prior to a particular examination.展开更多
文摘The effects of high-intensity pulsed electromagnetic stimulation （HIPEMS） on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5–10 Tesla （T） [8 groups of B–I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance （A） value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of the growth of neural stem cells （P〈0.05）. The A values of neural stem cells in the 6.0 T, 8.0 T, and 10.0 T groups were lower than the sham exposure control group, indicating a restraint of the growth of neural stem cells. The rate of neuron-specific enolase-positive neurons revealed by flow cytometry in HPEMS groups was the same as that in control group （P〉0.05）. It was suggested that 0.1 Hz, 5 pulses stimulation of HPEMS within certain scale of intensity （0.5–10.0 T）, significantly promoted the growth of neural stem cells with the rational intensity being 4.0 T.
基金supported by a grant of the Korea Health Technology R & D Project through the Korea Health Industry Development Institute(KHIDI)funded by the Ministry of Health&Welfare,Republic of Korea(grant number:HI15C1535)
文摘Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal differentiation of BMSCs. Low-frequency PEMF can induce the neuronal differentiation of BMSCs in the absence of nerve growth factors. This study was designed to investigate the effects of low-frequency PEMF pretreatment on the proliferation and function of BMSCs and the effects of low-frequency PEMF pre-treated BMSCs on the regeneration of injured peripheral nerve using in vitro and in vivo experiments.In in vitro experiments, quantitative DNA analysis was performed to determine the proliferation of BMSCs, and reverse transcription-polymerase chain reaction was performed to detect S100 （Schwann cell marker）, glial fibrillary acidic protein （astrocyte marker）, and brain-derived neurotrophic factor and nerve growth factor （neurotrophic factors） mRNA expression. In the in vivo experiments, rat models of crush-injured mental nerve established using clamp method were randomly injected with low-frequency PEMF pretreated BMSCs, unpretreated BMSCs or PBS at the injury site （1 × 106 cells）. DiI-labeled BMSCs injected at the injury site were counted under the fluorescence microscope to determine cell survival. One or two weeks after cell injection, functional recovery of the injured nerve was assessed using the sensory test with von Frey filaments. Two weeks after cell injection, axonal regeneration was evaluated using histomorphometric analysis and retrograde labeling of trigeminal ganglion neurons. In vitro experiment results revealed that low-frequency PEMF pretreated BMSCs proliferated faster and had greater mRNA expression of growth factors than unpretreated BMSCs. In vivo experiment results revealed that compared with injection of unpretreated BMSCs, injection of low-frequency PEMF pretreated BMSCs led to higher myelinated axon count and axon density and more DiI-labeled neurons in the trigeminal ganglia, contributing to rapider functional recovery of injured mental nerve. These findings suggest that low-frequency PEMF pretreatment is a promising approach to enhance the efficacy of cell therapy for peripheral nerve injury repair.
文摘The contrast agent concentration, the time of repetition (TR) and magnetic field strength are significant parameters that influence for the accurate signal intensity (SI) in quantitative Magnetic Resonance Imaging (MRI). Therefore, this study was conducted to investigate and refine the dependence and the optimal effect of Time of Repetition (TR) on the relationship between signal intensity and Gd-DTPA (Gadolinium-diethylene-triaminepenta-acetic acid) concentration, after applying two-dimensional (2D) Spin Echo (SE) pulse sequence under low-field MRI. In addition to that, the optimal concentration of Gd-DTPA at given sequence parameters at low-field MRI was also evaluated. A water-filled phantom was constructed for a range of Gd-DTPA concentrations (0 - 6 mmol/L) and the mean signal intensities (SIs) were assessed in the defined region of interest on T1-weighted images with different TR values (40 - 2000 ms). The generated signal-concentration curves for Gd-DTPA revealed that increasing TR was associated with the increase of the overall SIs and the maximum relationship between SI to concentration. Moreover, the required Gd-DTPA concentration to produce the maximum SI was associated to decrease with the increase of TR. In addition to this, the application of beyond 100 ms TR values in this study with relatively higher concentrations (beyond 1 - 2 mmol/L) has resulted predominantly non-linear patterns in the signal-concentration curves and it appears the saturation or decay of the SIs due to T2 effect. From these results, it can be suggested that the selection of relatively lower Gd-DTPA concentration ( mmol/L) with less than 800 ms (<800 ms) TR values can produce a better linear relationship between the concertation and SIs in T1-weighted SE low field contrast-enhanced MRI. Furthermore, this study also outlined the significance and necessity of the optimization of TR in SE sequence in low field MRI prior to a particular examination.
