KR101692388B1 - System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound - Google Patents

System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound Download PDF

Info

Publication number
KR101692388B1
KR101692388B1 KR1020150039971A KR20150039971A KR101692388B1 KR 101692388 B1 KR101692388 B1 KR 101692388B1 KR 1020150039971 A KR1020150039971 A KR 1020150039971A KR 20150039971 A KR20150039971 A KR 20150039971A KR 101692388 B1 KR101692388 B1 KR 101692388B1
Authority
KR
South Korea
Prior art keywords
stem cells
electromagnetic field
cell
cells
mesenchymal stem
Prior art date
Application number
KR1020150039971A
Other languages
Korean (ko)
Other versions
KR20160114215A (en
Inventor
서영권
박정극
윤희훈
박희정
김유미
조상은
김상헌
조현진
정종섭
Original Assignee
동국대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 동국대학교 산학협력단 filed Critical 동국대학교 산학협력단
Priority to KR1020150039971A priority Critical patent/KR101692388B1/en
Priority to PCT/KR2015/006704 priority patent/WO2016153114A1/en
Publication of KR20160114215A publication Critical patent/KR20160114215A/en
Application granted granted Critical
Publication of KR101692388B1 publication Critical patent/KR101692388B1/en

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/51—Umbilical cord; Umbilical cord blood; Umbilical stem cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0613—Cells from endocrine organs
    • C12N5/0616—Pituitary gland

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Virology (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Reproductive Health (AREA)
  • Hematology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Endocrinology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Ophthalmology & Optometry (AREA)

Abstract

The present invention handles 100 to 2000mT high intensity electromagnetic field and from 0.5 to 2kHz, high-intensity electromagnetic field of from 80 to sound waves, or 100 to 2000mT of 200dB and 10 to 2000kHz, 0.1 to about 100 mW / cm 2 ultrasound in mesenchymal stem cells or adult stem cells To a method for differentiating mesenchymal stem cells or adult stem cells into neural cells. The present invention also relates to a composition comprising neurons differentiated by said method. The method and composition for neuron differentiation using high-intensity magnetic field and sonic wave or ultrasonic wave according to the present invention induces differentiation of adult stem cells into neuronal cells by using high-frequency electromagnetic field of high frequency and sonic wave or ultrasonic wave, Treatment alone can easily differentiate neurons or neural stem cells or improve recovery of damaged neurons.

Description

BACKGROUND OF THE INVENTION Field of the Invention [0001] The present invention relates to a nerve regeneration system using a high-intensity electromagnetic field and a sonic or ultrasonic wave,

The present invention relates to a method for differentiating mesenchymal stem cells or adult stem cells into neural cells. More specifically, the present invention relates to a method for irradiating a high-intensity electromagnetic field to mesenchymal stem cells or adult stem cells, and additionally providing physical stimulation (sound waves or ultrasonic waves) to promote differentiation into neurons, thereby improving nerve regeneration efficiency.

Various studies have been reported to promote the differentiation of stem cells using an electromagnetic field. Fregni et al. (Non-Patent Document 1) reported that various electrical and electromagnetic stimuli alleviated the pain of chronic neuralgia due to spinal injuries. Ahmadian S et al. (Non-Patent Document 2) reported that 25 Hz and 2 mT And the amount of collagen in the skin was increased.

Studies on bone regeneration using electromagnetic fields have also been reported. Ceccarelli et al. (Non-Patent Document 3) promoted bone differentiation of various mesenchymal stem cells with electromagnetic fields of 75 Hz and 2 mT, and Sun et al. Cultured bone marrow-derived mesenchymal stem cells at an electromagnetic field of 15 Hz and 1.8 mT, In this study, we investigated the expression of ALP and bone morphogenetic protein (BMP-2) in rat mesenchymal stem cells. And promoted differentiation. The study on the promotion of bone differentiation using such an electromagnetic field used an electromagnetic field of 7.5 to 15 Hz and 0.1 to 5 mT (Non-Patent Document 4).

In recent years, as a method for treating neurological diseases such as Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, and spinal cord injury has been highlighted, methods using electric stimulation have been reported in research for promoting neural differentiation. However, these techniques are accompanied by a pain in the patient by adding an electrode transplantation method in which the electrode is directly implanted, and embryonic stem cells are limited in application to clinic because of the possibility of tumor formation.

Recently, studies have been reported to promote the proliferation and differentiation of cells using low-intensity ultrasound. Wu et al. Reported the results of promoting proliferation and differentiation by irradiating low-intensity ultrasound to mouse bone precursor cells (Non-Patent Document 5). Yang et al. Reported that low-intensity ultrasound (1.5 MHz, 90 mW / cm 2 ) To promote the differentiation into bone cells (Non-Patent Document 6). El-Bialy et al. Reported that differentiation into neurons was enhanced by irradiating low-intensity ultrasound (1.5 MHz, 30 mW / cm 2 ) to periodontal ligament cells (Non-Patent Document 7). However, such low-intensity ultrasound can be directly adhered to the skin and can cause problems such as tissue necrosis to be applied to the actual clinical application as an effect through the medium.

Therefore, there is a need for a next generation technology for differentiating mesenchymal stem cells and adult stem cells into neural cells by a non-invasive method rather than a chemical method. According to this need, the present inventors have found that the efficiency of a cell therapy agent for treating various neural diseases The present invention has been completed by continuing research on mesenchymal stem cells and adult stem cells by simultaneously irradiating low intensity sound waves, ultrasonic waves, and high intensity electromagnetic fields.

1. Fregni. et al., Cranial electrotherapy stimulation and transcranial pulsed current stimulation: A computer based high-resolution modeling study, NeuroImage, Volume 65, Pages 280-287, 15 January 2013. 2. Ahmadian S et al., Effects of extremely low-frequency pulsed electromagnetic fields on collagen synthesis in rat skin, Biotechnol. Appl. Biochem. 2006, 43, 71-75. 3. Ceccarelli et al., A Comparative Analysis of In Vitro Effects of Pulsed Electromagnetic Field Treatment on Osteogenic Differentiation of Two Different Mesenchymal Cell Lineages, BioResearch Open Access, 2013, 2 (4): 283-294. 4. Schwartz Z, Simon BJ, Duran MA, Barabino G, Chaudhri R, Boyan BD. Pulsed electromagnetic fields enhance BMP-2 dependent osteoblastic differentiation of human mesenchymal stem cells. J Orthop Res. 2008; 26 (9): 1250-1255. 5. Wu L, Lin L, Qin YX. Enhancement of cell ingrowth, proliferation, and early differentiation in a three-dimensional silicon carbide scaffold using low-intensity pulsed ultrasound, Tissue Eng Part A. 2015 Jan; 21 (1-2): 53-61. 6. Yang Z, Ren L, Deng F, Wang Z, Song J. Low-intensity pulsed ultrasound induces osteogenic differentiation of human periodontal ligament cells through activation of bone morphogenetic protein-smad signaling. J Ultrasound Med. 2014 May; 33 (5): 865-73. 7. El-Bialy T, Alhadlaqa, Wong B, Kucharski C. Ultrasound effect on neural differentiation of gingival stem / progenitor cells. Ann Biomed Eng. 2014 Jul; 42 (7): 1406-12.

It is an object of the present invention to provide a method for differentiating mesenchymal stem cells or adult stem cells into neural cells by treating a mesenchymal stem cell or adult stem cell with a high intensity electromagnetic field and a low intensity sound or ultrasonic wave.

Another object of the present invention is to provide a composition comprising the neuron differentiated by the method.

It is still another object of the present invention to provide a system for treating neurological diseases by inducing differentiation of neurons.

It is still another object of the present invention to provide a medical device constituting the neurological disease treatment system.

The present invention relates to a method for differentiating mesenchymal stem cells or adult stem cells into neuronal cells by treating the mesenchymal stem cells or the adult stem cells with an electromagnetic field and a sound wave of 100 to 2000 mT or an electromagnetic field and an ultrasonic wave of 100 to 2000 mT .

The electromagnetic field of 100 to 2000 mT corresponds to a high-intensity electromagnetic field, and a high-intensity electromagnetic field can be processed together with a sound wave or an ultrasonic wave to differentiate mesenchymal stem cells or adult stem cells into neuronal cells.

As used herein, the term " electromagnetic field " refers to a phenomenon in which an electromagnetic field periodically changing in intensity propagates into a space. The electromagnetic field used in the present invention includes a pulse wave form, a continuous wave (sinusoidal) And shock wave form.

As used herein, the term " high strength " refers to having an electromagnetic field strength of 100 mT or greater in strength, preferably 100 mT to 2000 mT, and most preferably 150 to 500 mT in the present invention. If it is out of the above range, the conversion efficiency of the stem cells into the neurons decreases, and the efficiency of regeneration of the damaged neurons decreases. In the prior art, since the electromagnetic field irradiation time is 6 to 12 hours or more using a low-intensity electromagnetic field having a strength of less than 100 mT, it is difficult to clinically apply to patients in the outpatient treatment field. In case of using low-intensity electromagnetic fields for a long time, The efficiency of treatment for nerve damage is reduced. However, since the present invention utilizes a "high-strength electromagnetic field ", the electromagnetic field irradiation time is shortened to about 15 minutes to prevent adverse effects in the body, and thus clinical application is possible.

Further, the electromagnetic field of the present invention may be a low-frequency electromagnetic wave having a frequency of 0.01 to 1000 Hz, preferably 1 to 100 Hz, and most preferably 45 to 75 Hz. If it is out of the above range, the conversion efficiency of the stem cells into the neurons decreases, and the efficiency of regeneration of the damaged neurons decreases.

As used herein, the term " electromagnetic wave " refers to a phenomenon in which an electromagnetic field periodically changing in intensity propagates into a space, such as an electromagnetic wave, and a low frequency electromagnetic wave means a low frequency wave. .

As used herein, the term " sound waves " refers to those having an intensity of 0.5 kHz or higher in the present invention, and preferably in the present invention is a frequency of 0.5 to 2 kHz, a frequency of 80 to 200 dB, most preferably a frequency of 1 to 1.5 kHz, And can be treated with an intensity of 80 to 100 dB. If it is out of the above range, the conversion efficiency of the stem cells into the neurons decreases, and the efficiency of regeneration of the damaged neurons decreases. In the present invention, the " sonic wave " treatment uses a method of irradiating air with a medium without direct contact with cells or tissues. Therefore, unlike the conventional vibration type sound wave processing using a speaker or the like, the present invention is an invention optimized for clinical commercialization because the specific frequency of the sound wave region can be uniformly irradiated, and thus the reproducibility is excellent.

As used herein, the term " ultrasound " refers to having a strength of at least 10 kHz, and in the present invention preferably a frequency of 10 to 2000 kHz, a frequency of 0.1 to 100 mW / cm 2 , most preferably 20 to 500 kHz, To 25 mW / cm < 2 >. If it is out of the above range, the conversion efficiency of the stem cells into the neurons decreases, and the efficiency of regeneration of the damaged neurons decreases. In the present invention, the " ultrasonic " treatment uses a method of irradiating air with a medium without direct contact with cells or tissues. Therefore, the conventional contact method can induce tissue necrosis due to tissue contact of the ultrasonic probe, but the present invention can be said to be a clinically applicable application because it can eliminate the problem of non-contact tissue damage or necrosis .

In the embodiment of the present invention, when adult stem cells were cultured using electromagnetic fields and sound waves under high intensity and low frequency conditions or electromagnetic fields and ultrasonic waves under high intensity and low frequency conditions, electromagnetic fields, sound waves, and ultrasound waves it was confirmed that when examining the electromagnetic field and ultrasound with a neuron-related mRNA expression and protein expression still more increased, especially from 1 to 100 Hz low-frequency, high intensity electromagnetic field and from 20 to 500kHz, 0.15 to 25mW / cm 2 ultrasound of 150 to 500 mT The highest expression of neuron - related protein was observed in adult stem cells. Therefore, when the method of the present invention was applied, it was observed that adult stem cells can further improve the differentiation into neurons (FIGS. 2 and 3).

The electromagnetic field, sonic waves and ultrasonic waves of the present invention can be treated for 5 to 60 minutes / day for 1 to 180 days, preferably 5 to 60 minutes / day for 3 to 180 days, more preferably 5 to 30 minutes / More preferably 5 to 30 minutes / day, more preferably 3 to 30 days, more preferably 10 to 30 minutes / day, and most preferably 15 to 20 minutes / day, for 5 to 15 days . Since the present invention utilizes a "high-intensity electromagnetic field, " the electromagnetic field irradiation time can be shortened to about 15 minutes, thereby preventing side effects such as inhibition of the therapeutic effect. In addition, it is possible to uniformly irradiate a specific frequency of a sound wave region, which is not a vibration type, and it is possible to eliminate the problem that a tissue is damaged or necrotized by using a non-contact ultrasonic wave. In addition, electromagnetic fields and sound waves can be processed together or electromagnetic fields and ultrasonic waves can be processed together to produce a synergistic effect for differentiating stem cells into neurons, rather than treating them individually. Therefore, it can be expected that the patient will recover quickly with short treatment time and excellent neuronal differentiation effect when clinical commercialized.

The term " stem cell " refers to a cell capable of dividing for self-renewal for a long period of time as an undifferentiated cell and capable of differentiating into various kinds of cells when given a certain condition. Stem cells are divided into embryonic stem cells, adult stem cells, iPS cells, and stem cells, depending on the tissue from which they originate, and neural stem cells include neural progenitor cells.

The term " adult stem cells " also includes mesenchymal stem cells, and adult stem cells include periodontal ligament cells, mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, umbilical-derived mesenchymal stem cells, . The adult stem cells may be commercially available stem cells or stem cells isolated from living tissues without limitation.

As used herein, the term " nerve cell " includes all of Schwann cells, Astrocytes, Oligodendrocytes, Neurons, and neurons differentiated by the method of the present invention May include astrocytes, schwann cells, or rarely projecting glial cells.

In the examples of the present invention, the results of treatment of electromagnetic fields, sound waves, ultrasonic waves of high intensity and low frequency, or electromagnetic fields and sound waves, electromagnetic fields and ultrasonic waves were analyzed. In addition, The exercise recovery ability was evaluated and the organization was analyzed.

As a result, the mesenchymal stem cells cultured using electromagnetic field, sonic wave, and ultrasonic wave showed homogeneous cell morphology in all experimental groups and no saponinization phenomenon such as liquid formation was observed in cytoplasm. As a result of mRNA analysis, NeroD1 and NF-1 expression was increased in ultrasound and high-intensity EM irradiated group compared with the control group. In particular, NeroD1 expression was significantly increased when high-intensity electromagnetic field and ultrasound were irradiated together. After 7 days, western analysis showed that the expression of NeroD1, p-CREB and p-ERK was increased in the acoustic and ultrasound-irradiated groups compared to the control group. The expression of MAP2, NF-1, p-CREB, and p-ERK was better than that of the high-intensity EMF alone when high-intensity electromagnetic field and sonic or high-intensity electromagnetic field and ultrasonic wave were examined together. That is, as the nerve protein is most strongly expressed in the cultured adult stem cells, the electromagnetic field of 60 Hz low frequency of the present invention, 100 to 2500 mT of the present invention is processed, and a sound wave of 1 to 2 kHz, 80 to 100 dB, 0.1 to 100 mW / cm 2 It was found that an electromagnetic field treated with ultrasonic waves for 5 to 30 minutes / day actively induces neural differentiation of adult stem cells (FIGS. 3, 4 and 5).

The present invention also provides a composition comprising the neuron differentiated by the method. Accordingly, the present invention provides a system for improving the treatment efficiency of a damaged nerve tissue including neurons differentiated by the above method. The composition may be administered in a suitable manner in the body according to a conventional method, and the cell contains an effective dose capable of maximizing therapeutic effect by one or several administrations. The cells may be mixed with an injection solution immediately before use. As the injection solution, physiological saline, glucose, mannitol, Ringer's solution and the like may be used.

The damaged nerve tissue may be derived from one or more diseases selected from the group consisting of Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, spinal cord injury and peripheral nerve, preferably neurological diseases, Differentiated neurons or neural stem cells can function as a system for the treatment of neurological diseases by restoring the function of nerve cells in neuronal diseases.

delete

delete

delete

In addition, the present invention provides a method for treating a subject, comprising injecting mesenchymal stem cells or adult stem cells into a subject; And irradiating the mesenchymal stem cells or adult stem cells with an electromagnetic field and a sound wave of 100 to 2000 mT or irradiating an electromagnetic field and an ultrasonic wave of 100 to 2000 mT to the mesenchymal stem cells or adult stem cells, Lt; RTI ID = 0.0 > cells. ≪ / RTI >

The subject is a vertebrate including a human, preferably a mammal, and more preferably a human, an ape, a pig, a mouse (mouse or rat), a rabbit, a guinea pig, a hamster, a dog or a cat But is not limited thereto.

It is preferable that the electromagnetic field is processed at a frequency of 0.01 to 1000 Hz. If the range is out of the above range, the efficiency of nerve cell conversion of stem cells is decreased and a side effect of deteriorating the regeneration efficiency of the damaged nerve region occurs. Since the present invention utilizes a "high-intensity electromagnetic field ", the electromagnetic field irradiation time is shortened to about 15 minutes, and thus clinical application is possible.

It is preferable that the sound wave is processed at a frequency of 0.5 to 2 kHz and an intensity of 80 to 200 dB. If the sound wave is out of the range, the efficiency of nerve cell conversion of the stem cell is lowered and the efficiency of regeneration of damaged nerve area is decreased. In the present invention, the treatment of "sound waves" uses a method of irradiating the air with a medium without direct contact with the cells or tissues. Therefore, the present invention is capable of uniformly irradiating a specific frequency of a sound wave region, so that the reproducibility is excellent.

It is preferable that the ultrasonic wave is treated at a frequency of 10 to 2000 kHz, 0.1 to 100 mW / cm 2. If the ultrasonic wave is out of the range, the efficiency of nerve cell conversion of the stem cells is lowered and the regeneration efficiency of damaged nerve sites is decreased. In the present invention, the " ultrasonic " treatment uses a method of irradiating air with a medium without direct contact with cells or tissues. Therefore, the present invention can eliminate the problem of damage or necrosis of the tissue in a non-contact manner, thus securing safety.

The mesenchymal stem cells or adult stem cells may also be injected into the body of the subject, more preferably the brain of the subject, most preferably the damaged neural tissue of the subject.

In one embodiment of the present invention, human mesenchymal stem cells were injected into a mouse stroke model, and after electromagnetic waves, ultrasonic waves, electromagnetic fields and sound waves were irradiated, tissues were collected and subjected to western blotting. As a result, The expression of Neuro D1 was greatly increased, and the ability to recover exercise was significantly increased in the experimental group in which electromagnetic field, sound, electromagnetic field and ultrasonic wave were examined together.

The electromagnetic field and the sonic wave treatment or the electromagnetic field and the ultrasonic wave treatment can be treated for 5 to 60 minutes / day for 1 to 180 days, preferably 5 to 60 minutes / day for 3 to 180 days, more preferably 5 to 30 minutes / Preferably 3 to 180 days, more preferably 5 to 30 minutes / 3 to 30 days, more preferably 10 to 30 minutes / day 3 to 21 days, most preferably 15 to 20 minutes / day 5 to 15 days ≪ / RTI > The present invention can maximize the therapeutic efficiency of injured neural tissue by injecting undifferentiated mesenchymal stem cells or differentiated neural cells into the body and irradiating the affected area with electromagnetic fields, sound waves, electromagnetic fields, and ultrasound every 15 to 30 minutes. As described above, the present invention utilizes the "high-strength electromagnetic field", so that the electromagnetic field irradiation time is greatly shortened and there is no side effect, the specific frequency of the sound wave area can be uniformly irradiated not by the vibration type, There is a characteristic that it is possible to eliminate the problem. In addition, electromagnetic fields and sound waves can be processed together or electromagnetic fields and ultrasonic waves can be processed together to produce a synergistic effect for differentiating stem cells into neurons, rather than treating them individually. Therefore, it can be expected that the patient will recover quickly with short treatment time and excellent neuronal differentiation effect when clinical commercialized.

The adult stem cells may be periodontal ligament cells, mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, umbilical cord blood stem cells, cord blood-derived mesenchymal stem cells, Mesenchymal stem cells, and adipose-derived mesenchymal stem cells. Preferably, periodontal ligament cells, mesenchymal stem cells, or neural progenitor cells in adult stem cells can be differentiated into neurons by culturing them using electromagnetic fields and sound waves or electromagnetic fields and ultrasonic waves at specific frequencies. The adult stem cells may be commercially available stem cells or stem cells isolated from living tissues without limitation. In addition, the nerve cells include Schwann cells, Astrocytes, Oligodendrocytes, and Neurons, and the neurons differentiated by the method of the present invention are either astrocytes or rare prominent glue Cells.

The present invention also provides a medical device to which the above method is applied. The medical device may be used to treat damaged nerve tissue. The damaged nerve tissue may be derived from one or more diseases selected from the group consisting of Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, spinal cord injury and peripheral nerve, preferably neurological diseases. The differentiated neural cells or neural stem cells according to the present invention can be used as a constitution of medical devices by restoring the function of nerve cells against neurological diseases.

The present invention relates to a complex system for damaged nerve regeneration, and relates to a method for promoting differentiation of mesenchymal stem cells or adult stem cells into nerve-related cells. Specifically, it is a system for promoting differentiation into neuron-like cells by irradiating an electromagnetic field, a sound wave, or an ultrasonic wave together in vitro, or promoting the regeneration of injured neurons along with stem cells injected into the body. More specifically, it is possible to expose mesothelial stem cells or adult stem cells together with a high-intensity electromagnetic field of a certain low frequency and a sound or an ultrasonic wave to promote differentiation into neuron-like cells, or to inject mesenchymal stem cells or adult stem cells into the body And then exposing low frequency electromagnetic fields and sound waves or ultrasound waves together to promote recovery of damaged nerve tissue. The present invention relates to a system capable of improving the efficiency of differentiation of an existing electromagnetic field by applying an electromagnetic field and a sound wave or an ultrasonic wave together. The electromagnetic field of the present invention can promote the differentiation into neural cells only when the sonic and ultrasonic waves are treated together with mesenchymal stem cells or adult stem cells. The inventors of the present invention have found out that the present invention can provide a high-intensity electromagnetic field, And the technique of inducing neural differentiation of adult stem cells was obtained.

According to one embodiment of the present invention, as shown in FIG. 3, the mesenchymal stem cells cultured using an electromagnetic field, a sound wave, and an ultrasonic wave have homogeneous cell morphology in all experimental groups, It was not. mRNA analysis showed that the expression of NeroD1 and NF-1 was increased in the ultrasound and high-intensity electromagnetic field irradiation groups when compared with the control group, and the NeroD1 expression was remarkably increased especially when the high intensity electromagnetic field and the ultrasonic wave were irradiated ). After 7 days, western analysis showed that the expression of NeroD1, p-CREB and p-ERK was increased in the acoustic and ultrasound-irradiated groups compared to the control group. Furthermore, when high-intensity electromagnetic fields and sound waves or high-intensity electromagnetic fields and ultrasonic waves were simultaneously examined, it was observed that the expression of MAP2, NF-1, p-CREB and p-ERK was better than that of high-intensity electromagnetic field alone (FIG.

In addition, in one embodiment of the present invention, FIG. 8 is a graph showing the effect of the high-intensity electromagnetic field irradiation group after cell transplantation, the high intensity electromagnetic field and the sound wave irradiation group after cell transplantation, the cell transplantation group (control group) After 27 days of high - intensity electromagnetic field and ultrasonic irradiation, exercise recovery ability was increased in control group and all experimental group compared to non - treatment group. Especially, high recovery of electromagnetic field and sound wave group, high intensity electromagnetic field and ultrasound group I was able to observe the ability.

In an embodiment of the present invention, NF200 protein was analyzed to observe tissue regeneration at the lesion site of stroke. As a result, tissue regeneration was actively observed due to the deepest brown staining observed in the high-intensity electromagnetic field and the ultrasound-irradiated experimental group after cell transplantation (Fig. 9). In addition, the expression of Tau and NeuroD1 was increased and the expression of nestin, Tau and NeuroD1 was significantly increased as a result of electromagnetic and ultrasonic irradiation (FIG. 10).

In addition, the present invention can maximize the therapeutic efficiency of damaged nerve tissue by injecting undifferentiated mesenchymal stem cells or differentiated neurons into the body, irradiating the affected part with a high-intensity electromagnetic field, a sound wave, a high-intensity electromagnetic field and an ultrasonic wave for 15 minutes a day .

The method for differentiating stem cells using an electromagnetic field, a sound wave, an electromagnetic field and an ultrasonic wave according to the present invention can improve the differentiation efficiency of nerve cells or neural stem cells by improving the cell differentiation efficiency using only existing electromagnetic fields, It is possible to easily differentiate nerve cells or neural stem cells only by processing time. In addition, the differentiated stem cells can be useful for treating neurological diseases such as Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, spinal cord injury, and peripheral nerve damage.

FIG. 1 shows a sonic wave generator and a sonic wave generator (a function generator, an amplifier, and a sonic / ultrasonic generator) for cell culture according to an embodiment of the present invention.
FIG. 2 shows a high-intensity electromagnetic field apparatus for use in animal experiments, a sound wave apparatus, and an ultrasonic apparatus according to an embodiment of the present invention.
FIG. 3 is a graph showing morphological changes of cells after 3 days after irradiation with sonic waves, ultrasonic waves, high intensity electromagnetic fields, high intensity electromagnetic fields, sound waves, high intensity electromagnetic fields and ultrasonic waves on cells cultured in vitro according to an embodiment of the present invention.
FIG. 4 shows the result of mRNA analysis after 3 days after the irradiation of the cells cultured in vitro according to an embodiment of the present invention with sonic waves, ultrasonic waves, high intensity electromagnetic fields, high intensity electromagnetic fields, sound waves, high intensity electromagnetic fields and ultrasonic waves.
FIG. 5 shows results of western analysis after 7 days of irradiation of sonicated cells, ultrasonic waves, high intensity electromagnetic fields, high intensity electromagnetic fields, sound waves, high intensity electromagnetic fields and ultrasonic waves to cells cultured in vitro according to an embodiment of the present invention.
FIG. 6 is a graph showing the results of histopathologic examinations after staining of a mouse with a stroke mouse according to an embodiment of the present invention, MAP-2, and MMP-9.
FIG. 7 is a graph showing the results of immunohistochemical staining of a stroke mouse according to an embodiment of the present invention in a non-treatment group (negative control group), a cell transplantation group (control group) (Cont: negative control, cell: control (cell), cell + sound: sonic wave group after cell transplantation, cell + ultrasound: ultrasonic wave after cell transplantation group).
FIG. 8 is a graph showing the results of a comparison between a non-treatment group (negative control), a cell transplantation group (control group), a high-intensity electromagnetic field irradiation group after cell transplantation, It is the result of evaluating the exercise recovery power for 27 days after the high intensity electromagnetic field and the ultrasonic irradiation.
FIG. 9 is a graph showing the results of a comparison between a non-treatment group (negative control group), a cell transplantation group (control group), a high-intensity electromagnetic field group after cell transplantation, After the transplantation, high-intensity electromagnetic fields and ultrasound were irradiated and biopsied, and HE staining, prussian blue, NF200 and nestin staining results were shown.
FIG. 10 is a graph showing the results of a comparison between a non-treatment group (negative control), a cell transplantation group (control group), a high-intensity electromagnetic field irradiation group after cell transplantation, a high- After high-intensity electromagnetic field and ultrasonic irradiation, the protein was separated from the tissue by biopsy, and the result was a western analysis.

Hereinafter, the present invention will be described in more detail in the following examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Example  One. Mesenchymal stem cells  Evaluation of neuronal differentiation efficacy of high intensity electromagnetic field, sonic wave, and ultrasonic using

Example  1.1 Of mesenchymal stem cells  culture

Mesenchymal stem cells were purchased from Lonza (Wailersville, MD) and cultured in DMEM medium supplemented with 10% (v / v) FBS. The cells were cultured in a CO 2 incubator maintained at 37 ° C while being inoculated on a culture plate and the medium was changed every 3 days.

Example  1.2 High intensity electromagnetic field, sound wave, using ultrasonic wave Of mesenchymal stem cells  Differentiation confirmation

The cultured mesenchymal stem cells were inoculated into a 60 mm culture dish at 0.3 × 10 5 / dish for RNA analysis and 0.15 × 10 5 / dish for protein analysis, and triplicate experiments were performed for each experimental group.

Electromagnetic fields were irradiated in a clean bench at a frequency of 60 Hz and 500 mT for 15 minutes once a day. Ultrasonic waves were irradiated at 40 kHz, 0.4 mW / cm 2 once a day for 15 minutes and sound waves at 1 kHz once a day for 1 minute and 81 dB for 60 minutes The cells were irradiated in a non-contact state at 37 ° C in a CO 2 incubator at a distance of 1 cm from the culture dish. The culture media used were Dulbecco's Modified Eagle's Medium (DMEM), 5% fetal bovine serum (FBS), 10 ng / mL basic fibroblast growth factor (bFGF) and 10 uM forskolin.

Under the above conditions, 3 days for PCR analysis and 7 days for protein analysis were analyzed.

A sonic and ultrasonic generator for cell culture is shown in FIG. 1, and the frequency is adjusted through a function generator and the intensity is controlled through an amplifier.

FIG. 3 shows the result of observation using an optical microscope to confirm morphological changes of mesenchymal stem cells after culturing. As shown in FIG. 3, the mesenchymal stem cells cultured using electromagnetic fields, sonic waves, and ultrasonic waves showed homogeneous cell morphology in all experimental groups and no saponinization phenomenon such as vacuolization was observed in the cytoplasm.

Fig. 4 shows the result of mRNA analysis after 3 days after the irradiation of the cells cultured in vitro with sonic waves, ultrasonic waves, high intensity electromagnetic fields, high intensity electromagnetic fields and sound waves, high intensity electromagnetic fields and ultrasonic waves. In comparison with the control group, NeroD1 and NF-1 expression was increased in the ultrasonic and high-intensity electromagnetic fields. Especially, NeroD1 expression was significantly increased when high-intensity electromagnetic fields and ultrasonic waves were applied together.

Fig. 5 is a western analysis result after 7 days after the irradiation of the cells cultured in vitro with sonic waves, ultrasonic waves, high intensity electromagnetic fields, high intensity electromagnetic fields and sound waves, high intensity electromagnetic fields and ultrasonic waves. Compared with the control group, expression of NeroD1, p-CREB and p-ERK was increased in the sonic and ultrasound-irradiated groups. The expression of MAP2, NF-1, p-CREB, and p-ERK was better than that of the high-intensity EMF alone when high-intensity electromagnetic field and sonic or high-intensity electromagnetic field and ultrasonic wave were examined together.

That is, as the nerve protein is most strongly expressed in the cultured adult stem cells, an electromagnetic field of 60 Hz low frequency and 500 mT intensity is applied once a day for 15 minutes, and 1 kHz, 81 dB sound wave or 1 It was found that when ultrasound was applied at 40 kHz and 0.4 mW / cm 2 for 15 minutes, the adult stem cells were actively induced.

Example  2. Evaluation of nerve regeneration effect of sonic or ultrasonic using stroke animal model

Example  2.1 Small animal  Stroke modeling

Three weeks old (45-50 g) SD rats were used for the preparation of stroke animal models. The anesthetics were 0.1 cc / 100 g (50 mg / kg) of zoletil (250 mg / 5 cc, Virbac) (Rompun 2%, Bayer) 0.025 ~ 0.04 / 100g (5 ~ 10mg / kg) were anesthetized by intraperitoneal administration. Stroke models were produced by photochemical methods as follows. 300 ㎕ of systemic photoactive dye substance Rose Bengal (10 mg / ml) was administered and light was projected for 15 minutes through a skull with a light beam (KL 1500 LCD (SCHOTT), 532 nm wavelength) to induce stroke .

Example  2.2 Cell Implantation and Sonic or Ultrasound in Stroke Model

To select the mice for the experiment, all mice were subjected to a rotarod (20 rpm) for 1 week before the stroke model was manufactured, and mice having a motor force of 80 seconds or more were used. Three days after the stroke formation, the mice were again subjected to the rotarod. Only the mice falling from the rotarod at 40 seconds or less were selected for the experiment.

After 3 days of stroke, human mesenchymal stem cells were injected through penis vein at a concentration of 1 × 10 5 / mouse in 10 mice in each experimental group. Dulbeco's Modified Eagle's Medium (DMEM), 5% Fetal bovine Cells were cultured with serum (FBS), 10 ng / mL basic fibroblast growth factor (bFGF), and 10 uM forskolin. Twenty-four hours prior to cell injection, 50 ug / 10 ml of nanomagnetic particles were added to the culture medium. After 24 hours from the injection of the cells, sonic or ultrasonic waves were irradiated.

A sound wave device for an animal experiment or an ultrasonic wave device is shown in Fig. Sound wave was irradiated after which lighten the sound wave generator probe for mouse light head contact once a day 1kHz, 81dB, 20 bun during ultrasound with the 3 ~ 5 cm distance from the light head 40kHz, 0.7 mW / cm 2 per day And examined for 20 minutes. Sonic wave or ultrasonic irradiation was performed for 27 days.

Example  2.3 Small animal  Immunostaining and Westernization after sonic or sonic irradiation on stroke model western ) analysis

After 27 days, the mice were euthanized and the tissue of the stroke site was collected, and hematoxylin & eosin staining and immunochemical examination were performed.

FIG. 6 shows the results of HE staining, prussian blue, MAP-2, and MMP-9 after biopsy of the untreated group (negative control), cell transplantation group (control group) Was stained. In the non-treated group, the necrotic area of the stroke site fell off and holes were formed, and the remaining experimental group showed that the damaged tissue remained (HE staining). The presence of prussian blue cells was observed, and blue stained nanomagnetic particles were observed in the lesion of the stroke, suggesting that the cells migrated (prussian blue). MAP-2 protein was analyzed in order to observe the tissue regeneration at the lesion site of the stroke, and it was observed that the tissue regeneration was progressing due to the brown staining observed in the sonic wave group (MAP-2). MMP-9 staining was performed to observe the inflammatory response in the area of brain injury. MMP-9 was stained with a small amount of MMP staining (MMP-9) .

FIG. 7 shows the results of Western blot analysis after isolating proteins from tissues in a non-treated group (negative control group), a cell transplantation group (control group), a post-transplantation sonography group, The results are shown in Fig. As shown in Western analysis, nestin, Tau, p-CREB and p-ERK expression were increased after transplantation of the cells after stroke formation, and nestin, Tau, MAP-2, NF-L, p-CREB, and p-ERK.

Example  3. Small animal  Assessment of nerve regeneration efficacy of electromagnetic field, sonic or electromagnetic field and ultrasonic using stroke animal model

Example  3.1 Electro-magnetic field, sonic or electromagnetic field and ultrasonic irradiation on stroke model

To select the mice for the experiment, all mice were subjected to a rotarod (20 rpm) for 1 week before the stroke model was manufactured, and mice having a motor force of 80 seconds or more were used. Three days after the stroke formation, the mice were again subjected to the rotarod. Only the mice falling from the rotarod at 40 seconds or less were selected for the experiment.

To evaluate the nerve regeneration efficacy of high intensity electromagnetic fields, sonic waves, and ultrasound using a mouse stroke model, experimental groups were classified into five categories. First, the adult model was applied to the stroke model (negative control group), second, the adult model was applied to the adult stem cell group (cell number: 1 × 10 5 , control group), and the adult stem cell was administered to the stroke model (40 kHz, 15 min / day) after the injection of adult stem cells into the stroke model, and high intensity electromagnetic fields and sound waves after the injection of adult stem cells into the stroke model (1 kHz, 81 dB, 15 min / day).

Fig. 2 shows a high-intensity electromagnetic field device for animal experiments, a sound wave device, and an ultrasonic wave device. High-intensity electromagnetic field, ultrasonic wave, and sound wave irradiation method were as follows. The high intensity electromagnetic field was investigated by placing the mouse in a 50cc syringe and placing it on a high intensity electromagnetic field coil. The sonography was performed immediately after the completion of the high intensity electromagnetic field irradiation. The sonogram was irradiated at 1 kHz, 81dB, and 15 minutes once a day, And irradiated at 40 kHz and 0.7 mW / cm 2 at 5 cm intervals for 15 minutes once a day. Sonic wave or ultrasonic irradiation was performed for 27 days.

Example  3.2 Assessment of Motor Resilience in Stroke Mice

RotaRoad was performed to evaluate the exercise restoring force of the stroke mouse of Example 3.1 above.

FIG. 8 is a graph showing the effect of the high-intensity electromagnetic field and the ultrasonic wave irradiation group after cell transplantation in the non-treatment group (negative control), the cell transplantation group (control group) Day exercise recovery ability. In the control group and all experimental groups, exercise recovery ability was increased compared with the non - treatment group. In particular, fast recovery ability was observed in the high intensity electromagnetic field and the sound wave irradiation group, and the high intensity electromagnetic field and the ultrasonic irradiation group.

Example  3.3 Stroke Model with Electro-magnetic, Ultrasonic and Ultrasound Examination and Protein Analysis

After 27 days, the mice were euthanized and the tissue of the stroke site was collected, and hematoxylin & eosin staining and immunochemical examination were performed.

FIG. 9 is a graph showing the results of a comparison between a non-treatment group (negative control), a cell transplantation group (control group), a high-intensity electromagnetic field irradiation group after cell transplantation, a high-intensity electromagnetic field and a sound wave irradiation group, And staining with HE, prussian blue, NF200 and nestin. In the non-treated group, the necrotic area of the stroke site fell off and holes were formed (HE staining). The presence of prussian blue cells was observed, and blue stained nanomagnetic particles were observed in the lesion of the stroke, suggesting that the cells migrated (prussian blue). In order to observe tissue regeneration of the lesion, NF200 protein was analyzed. The most intense brown staining was observed in the experimental group irradiated with high intensity electromagnetic field and ultrasound after cell transplantation, and tissue regeneration was actively observed (NF200). Nestin expression was the weakest expression in the negative control group and increased in the cell transplantation group (control group), after the transplantation in the high-intensity electromagnetic field group, after the transplantation in the high-intensity electromagnetic field and the sound wave group, after the cell transplantation, in the high intensity electromagnetic field and the ultrasound- .

FIG. 10 is a graph showing the results of a comparison between a high-intensity electromagnetic field and a high-frequency electromagnetic field group after cell transplantation, a high-intensity electromagnetic field and an ultrasound group after a cell transplantation, a biopsy (a negative control group) And the result of western analysis after separating the protein from the tissue. The expression of Tau and NeuroD1 was increased by electromagnetic fields and sound waves after the injection of cells after stroke formation, and the expression of nestin, Tau and NeuroD1 was significantly increased by electromagnetic field and ultrasound.

That is, as the nerve protein is most strongly expressed in the animal model of stroke, the electromagnetic field of 60 Hz low frequency and 500 mT intensity is treated once a day for 15 minutes, and 1 kHz, 81 dB sound wave or 1 day 1 It was found that ultrasonic treatment of 15 min, 40 kHz, and 0.7 mW / cm 2 actively promotes damaged nerve regeneration.

Claims (19)

A method for differentiating mesenchymal stem cells or adult stem cells into neuronal cells by treating the mesenchymal stem cells or adult stem cells with an electromagnetic field and a sound wave of 100 to 2000 mT, or an electromagnetic field and an ultrasonic wave of 100 to 2000 mT. The method according to claim 1,
Characterized in that the electromagnetic field is processed at a frequency of from 0.01 to 1000 Hz. The method according to claim 1,
Wherein the sound waves are processed at a frequency of 0.5 to 2 kHz, with an intensity of 80 to 200 dB. The method according to claim 1,
Wherein the ultrasonic waves are treated at a frequency of 10 to 2000 kHz, 0.1 to 100 mW / cm < 2 >. The method according to claim 1,
Wherein said electromagnetic field and sonic treatment, or electromagnetic field and ultrasonic treatment are treated for 5 to 30 minutes / day for 3 to 180 days. The method according to claim 1,
Wherein the nerve cell is a Schwann cell, an astrocyte cell, or a rare dendritic cell. The method according to claim 1,
Wherein said mesenchymal stem cells are derived from bone marrow, adipose or cord. The method according to claim 1,
Wherein said adult stem cells are pulmonary stem cells or neural progenitor cells. 9. A composition comprising a neuron differentiated by the method of any one of claims 1-8. delete delete Injecting mesenchymal stem cells or adult stem cells into a subject selected from the group consisting of mammals other than humans; And
Irradiating the mesenchymal stem cells or the adult stem cells with an electromagnetic field and an acoustic wave of 100 to 2000 mT or an electromagnetic field and an ultrasonic wave of 100 to 2000 mT;
A method for differentiating mesenchymal stem cells or adult stem cells into neural cells. 13. The method of claim 12,
Wherein the electromagnetic field is treated at a frequency of 0.01 to 1000 Hz. 13. The method of claim 12,
Wherein the sound waves are treated at a frequency of 0.5 to 2 kHz and an intensity of 80 to 200 dB, wherein the mesenchymal stem cells or adult stem cells are differentiated into neurons. 13. The method of claim 12,
Wherein the ultrasound is treated at a frequency of 10 to 2000 kHz and 0.1 to 100 mW / cm < 2 > to differentiate mesenchymal stem cells or adult stem cells into neuronal cells. 13. The method of claim 12,
Wherein the electromagnetic field and the sonic wave treatment, or the electromagnetic field and the ultrasonic wave treatment are treated for 5 to 30 minutes / day for 3 to 180 days, wherein the mesenchymal stem cells or adult stem cells are differentiated into neurons. 13. The method of claim 12,
Wherein the nerve cell is a Schwann cell, an astrocyte cell, or a spindle-shaped glial cell, wherein the mesenchymal stem cell or the adult stem cell is differentiated into a neuron cell. 13. The method of claim 12,
Wherein the mesenchymal stem cells are derived from bone marrow, adipose, or umbilical cord. 13. The method of claim 12,
Wherein the adult stem cell is a mesenchymal stem cell or a neural progenitor cell, wherein the mesenchymal stem cell or adult stem cell is differentiated into a neural cell.
KR1020150039971A 2015-03-23 2015-03-23 System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound KR101692388B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
KR1020150039971A KR101692388B1 (en) 2015-03-23 2015-03-23 System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound
PCT/KR2015/006704 WO2016153114A1 (en) 2015-03-23 2015-06-30 Nerve regeneration system using high-intensity electromagnetic field and sonic or ultrasonic waves

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020150039971A KR101692388B1 (en) 2015-03-23 2015-03-23 System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound

Publications (2)

Publication Number Publication Date
KR20160114215A KR20160114215A (en) 2016-10-05
KR101692388B1 true KR101692388B1 (en) 2017-01-04

Family

ID=56977479

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020150039971A KR101692388B1 (en) 2015-03-23 2015-03-23 System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound

Country Status (2)

Country Link
KR (1) KR101692388B1 (en)
WO (1) WO2016153114A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024191125A1 (en) * 2023-03-10 2024-09-19 연세대학교 산학협력단 Method for activating endogenous neural stem cells through low-intensity focused ultrasound treatment

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7545714B2 (en) * 2020-07-31 2024-09-05 ニューロテックメディカル株式会社 Treatment for neurological disorders
KR102564040B1 (en) * 2021-03-17 2023-08-04 단국대학교 천안캠퍼스 산학협력단 Methods for enhancing the growth of neural stem cells using low level lasers for neurologic function

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008002917A2 (en) * 2006-06-27 2008-01-03 Cyberkinetics Neurotechnology Systems, Inc. Systems and methods for promoting nerve regeneration
KR101210984B1 (en) * 2010-06-08 2012-12-11 동국대학교 산학협력단 Differentiation method of Mesenchymal stem cells using sonic vibration
KR20130061970A (en) * 2011-12-02 2013-06-12 동국대학교 산학협력단 Differentiation method of mesenchymal stem cells by composite waves and an appratus thereof
KR101423225B1 (en) * 2012-02-29 2014-07-24 서울대학교산학협력단 Method for differentiation of adult stem cell into schwann cell using electromagnetic field

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MENG, D. et al., J Huazhoug Univ Sci Technol [Med Sci], 2009.12, 29권, 6호, 732-736면 (2009.12.29. 온라인 공개)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024191125A1 (en) * 2023-03-10 2024-09-19 연세대학교 산학협력단 Method for activating endogenous neural stem cells through low-intensity focused ultrasound treatment

Also Published As

Publication number Publication date
WO2016153114A1 (en) 2016-09-29
KR20160114215A (en) 2016-10-05

Similar Documents

Publication Publication Date Title
US8753881B2 (en) Method for inducing differentiation of mesenchymal stem cells to nerve cells using sound waves
US20140303525A1 (en) Method and device for non-invasive acoustic stimulation of stem cells and progenitor cells in a patient
Ross et al. The regenerative effects of electromagnetic field on spinal cord injury
US20210402184A1 (en) Klotho modulation
Krueger et al. Effects of low-intensity electrical stimulation and adipose derived stem cells transplantation on the time-domain analysis-based electromyographic signals in dogs with SCI
Wu et al. Implanted spike wave electric stimulation promotes survival of the bone marrow mesenchymal stem cells and functional recovery in the spinal cord injured rats
KR101692388B1 (en) System of Nerve Regeneration Using Electromagnetic field, and sound or ultrasound
Yuan et al. Neuromodulation-based stem cell therapy in brain repair: recent advances and future perspectives
Shi et al. Stem cell-based therapies to treat spinal cord injury: a review
Choi et al. Combined effect of pulsed electromagnetic field and sound wave on In vitro and In vivo neural differentiation of human mesenchymal stem cells
KR101768795B1 (en) System of pulsed electromagnetic field for increasing of the cell activity and inducing of cell-differentiation
Bang et al. Electrical stimulation promotes functional recovery after spinal cord injury by activating endogenous spinal cord-derived neural stem/progenitor cell: an in vitro and in vivo study
KR101743539B1 (en) Method For Differentiation Of Adult Stem Cells Into Nerve Cells Using High Intensity Of Electromagnetic Field
Bersano et al. Clinical studies in stem cells transplantation for stroke: a review
KR20170048111A (en) Method of direct lineage reprogramming for generating neurons using electromagnetic-induced metalilic nanoparticle
EP4252760A1 (en) Therapeutic agent for nerve disorders
CN115348881A (en) Magnetic field exposure system and use thereof
Yu et al. Bioelectronic medicine potentiates endogenous NSCs for neurodegenerative diseases
WO2015152656A1 (en) Method of differentiating adult stem cells into nerve cells by using high-intensity electromagnetic field
Ruan et al. Ultrasound generated mechanical induction of mesenchymal stem cells
Elbaz et al. Effect of electrical stimulation and stem cells on experimentally induced peripheral nerve injury in rats
Zhang et al. Nanoparticles guided precise transplantation of varying numbers of mesenchymal stem cells into post–traumatic syrinx in spinal cord injury rat
JP7125818B1 (en) Composition for treatment of intractable neurological disease and method for producing the same
IL294690A (en) Ultra-wideband micromechanical method for modulation cellular activity
Sumarwotoa et al. Enhancement of Schwann-like cells differentiation from adipose-derived mesenchymal stem cells by administration of platelet-rich plasma: An in vitro study

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant