KR101630017B1 - Method of osteogenesis for bone healing using nano magnetic particle and electromagnetic field system - Google Patents

Method of osteogenesis for bone healing using nano magnetic particle and electromagnetic field system Download PDF

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KR101630017B1
KR101630017B1 KR1020130118475A KR20130118475A KR101630017B1 KR 101630017 B1 KR101630017 B1 KR 101630017B1 KR 1020130118475 A KR1020130118475 A KR 1020130118475A KR 20130118475 A KR20130118475 A KR 20130118475A KR 101630017 B1 KR101630017 B1 KR 101630017B1
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bone
electromagnetic field
magnetic nanoparticles
hydrogel
group
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KR20150039997A (en
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서영권
박희정
김유미
박정극
김상헌
김민옥
윤희훈
정현
이종호
김수찬
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동국대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/446—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with other specific inorganic fillers other than those covered by A61L27/443 or A61L27/46
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

Abstract

The present invention relates to a method for promoting bone regeneration at damaged bone and bone defect sites using magnetic nanoparticles and low-frequency electromagnetic fields, and the magnetic nanoparticle-containing hydrogel according to the present invention uses only natural collagen without cross- It has excellent biocompatibility because it does not induce an immune response in the body. It can be injected into the bone defect site after mixing with other bone inducing materials. After the injection of hydrogel, the electromagnetic field can control the irradiation time and the strength of 1 mT to 1.5 T is used Thereby promoting bone regeneration.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for promoting bone regeneration using magnetic nanoparticles and an electromagnetic field,

The present invention relates to a method of using magnetic nanoparticles and an electromagnetic field to induce bone regeneration, particularly healing or regeneration of injured alveolar bone and bone tissue.

Recently, a variety of external stimulation therapies such as ultrasound, electromagnetic fields and LEDs have been used to improve the treatment of injured tissue. In particular, electromagnetic fields have been used for neurogenesis, bone differentiation, and pain treatment by using a low intensity of less than 20 G (Gauss) in the low frequency range of 7.5 Hz to 100 Hz.

Freigni et al. (NeuroImage, 2013) reported that a variety of electrical and electromagnetic stimuli alleviated the pain of chronic neuralgia due to spinal injuries. Ahmadian S et al. (Biotechnol. Appl. Biochem. 2006, 43, 71-75) And skin collagen increased when the skin was irradiated at 25 Hz and 2 mT per day for 2.5 hours. In addition, Bae et al. (Cytotherapy. 2013 15 (8): 961-970) reported that mesenchymal stem cells were exposed to electromagnetic fields of 50 Hz and 5 mT for 60 minutes every day for 12 days.

Recently, studies on bone regeneration using electromagnetic fields have also been reported. Nascimento et al. (Gerodontlogy, 2012, 29: e1249-1251) reported that osseointegration was promoted by implantation of 1.5-MHz and 0.8-mT electromagnetic fields for 20 weeks after implant placement in animals. Sun et al. Reported 15 Hz and 1.8 mT (ALP) and bone morphogenetic protein (BMP-2) are promoted by culturing bone marrow-derived mesenchymal stem cells in an electromagnetic field of a mouse, (BioResearch Open Access, 2013, 2 (4): 283-294) promoted osteogenic differentiation of various mesenchymal stem cells with electromagnetic fields of 75 Hz and 2 mT. In this study, the electromagnetic field of 7.5 ~ 75 Hz and 0.1 ~ 5 mT was used for the study of bone differentiation promotion using this electromagnetic field.

In order to promote osteoclast activation and differentiation of mesenchymal stem cells into osteocytes using electromagnetic fields, the inventors of the present invention have found that when magnetic nanoparticles are injected or implanted into a bone defect site, The present invention has been accomplished by developing a technique for maximizing the effect of bone treatment on an electromagnetic field by irradiating an electromagnetic field with a strength and a high intensity within one hour.

It is an object of the present invention to provide a bone marrow treatment method and a bone marrow treatment method in which a hydrogel or an implant containing magnetic nanoparticles is applied to a bone defect area and then an electromagnetic field of a specific frequency and intensity is irradiated to thereby improve treatment efficiency of a bone defect area, Method.

In order to solve the above object,

1) injecting a hydrogel containing magnetic nanoparticles into a cell or tissue; And

2) a step of irradiating the cell or tissue with an electromagnetic field to promote bone differentiation and bone regeneration.

The method of promoting bone regeneration using the magnetic nanoparticles and the electromagnetic field according to the present invention can induce rapid bone treatment as a method of post-surgical physical therapy, and the magnetic nanoparticles or the bone formation promoting substance are introduced into a hydrogel or a scaffold , Bone transplantation can be maximized by transplanting to the site of bone injury.

Fig. 1 is a diagram showing microscopic results of bone cells irradiated with electromagnetic fields of various frequencies for 3 days.
FIG. 2 is a graph showing changes in mRNAs of collagen, bonesialoprotein, austenoctin, osteocalcin, osteopontin, vimentin, and type 2 osteogenic protein (BMP-2) The results are shown in FIG.
Fig. 3 is a microscope result of mesenchymal stem cells irradiated with electromagnetic fields at various frequencies for 3 days. Fig.
FIG. 4 shows mRNA expression of collagen, austenocin, osteocalcin, osteopontin, vimentin, and type 2 osteogenic protein (BMP-2) after irradiating mesenchymal stem cells with electromagnetic fields of various frequencies for 3 days Fig.
FIG. 5 shows the results of analysis of protein expression of collagen, austenectin, type 2 osteogenic protein (BMP-2), p-ERK and p-CREB after irradiating electromagnetic frequencies of various frequencies on mesenchymal stem cells for 14 days Fig.
FIG. 6 is a graph showing the results of immunostaining osteopontin after irradiating mesenchymal stem cells with electromagnetic fields of various frequencies for 14 days. FIG.
FIG. 7 is a microscope image of mesenchymal stem cells irradiated with high intensity electromagnetic fields at various frequencies for 3 days. FIG.
FIG. 8 shows the results of analysis of mRNA expression of collagen, type 2 osteogenic protein (BMP-2), osteonectin, osteocalcin and osteopontin, after irradiating mesenchymal stem cells with high frequency electromagnetic fields of various frequencies for 3 days Fig.
FIG. 9 shows the morphology of the experimental group in which the magnetic nanoparticles were injected (MP, 5 / / ml), the electromagnetic field group (EMF, 45 Hz, 8 hours × 2 times / Fig.
FIG. 10 shows the results of the analysis of the mRNAs of the experimental group irradiated with the magnetic nanoparticles (MP, 50 / / ml), the electromagnetic field group (EMF, 45 Hz, 8 hours × 2 times / Fig.
Fig. 11 is a graph showing changes in the concentrations of magnetic nanoparticles (MP, 50 占 퐂 / ml), electromagnetic fields (EMF, 45Hz, 8 hours x 2 times / Fig. 5 shows the result of immunotyping of oretin.
FIG. 12 shows the results of a comparison between the number of magnetic nanoparticles injected (MP, 5 / / ml), the number of magnetic fields (EMF, 45 Hz, 8 hours> 2 times / Fig. 3 shows the morphological results of the cells on day 3 of the experimental group.
Fig. 13 is a graph showing the results of the addition of magnetic nanoparticles to the bone cell line (Saos-2) (MP, 5 占 퐂 / ml), the electromagnetic field irradiation group (EMF, 45Hz, Fig. 3 shows the result of analysis of mRNA on day 3 of the experimental group.
Fig. 14 is a graph showing the results of a comparison between an electromagnetic field irradiation group (EMF, 60 Hz, 8 hours x 2 times / day) and a three-dimensional hydrogel (MP, 20 占 퐂 / ml) containing magnetic nanoparticles in a bone cell line (Saos- , And the morphological results of the cells on the third day of the experimental group irradiated with the electromagnetic field on the magnetic nanoparticle-containing three-dimensional hydrogel.
15 is a graph showing the results of a comparison between an electromagnetic field irradiation group (EMF, 60 Hz, 8 hours x 2 times / day) and a three-dimensional hydrogel (MP, 20 占 퐂 / ml) containing magnetic nanoparticles in a bone cell line (Saos- , And the result of analysis of mRNA on the third day of the experimental group irradiated with the electromagnetic field on the magnetic nanoparticle-containing three-dimensional hydrogel.

Hereinafter, the present invention will be described in detail.

The present invention

1) injecting a hydrogel containing magnetic nanoparticles into a cell or tissue; And

2) irradiating the hydrogel prepared in the step 1) with an electromagnetic field, thereby promoting bone differentiation and bone regeneration.

The cells are preferably bone cells or mesenchymal stem cells. But is not limited thereto.

The hydrogel was prepared using a buffer solution and a collagen-injectable form. A buffer solution of pH 8.0 containing 8.4% sodium bicarbonate or NaOH may be used as the buffer solution, but it may be used in combination with other injection solutions. The buffer solution may also contain magnetic nanoparticles and bone morphogenetic protein (BMP) and / or nanohydroxyapatite.

At this time, the magnetic nanoparticles are mixed at a concentration of 20 ug / ml, and the hydroxyapatite is mixed at a concentration of 0.015 g / ml. In order to promote bone regeneration, the magnetic nanoparticles may have a particle diameter of 10 to 200 nm, and the nanohydroxyapatite may have a particle diameter of 1 to 500 nm.

The magnetic nanoparticles of step 1) are preferably selected from the group consisting of FeO 2 , FeO 3 , and FeO 4 , but are not limited thereto.

The magnetic nanoparticles are preferably attached to the end of the PEG, but are not limited thereto.

The magnetic nanoparticles preferably use SiO 2 as a surface modification derivative, but are not limited thereto.

The modified derivative induces surface modification to control the polarity of the magnetic nanoparticles and induce the introduction of the nanoparticles into the cells and minimizes toxicity, and further induces surface modification to bind the desired functional groups or molecules.

The size of the magnetic nanoparticles is preferably less than 200 nm, but is not limited thereto.

The electromagnetic field in step 2) is preferably continuous or pulsed, but is not limited thereto.

The frequency of the electromagnetic field in step 2) is preferably 45 Hz to 75 Hz, but is not limited thereto. When the frequency of the electric field is more than 75 Hz, the osteogenic differentiation-related osteoectin, osteocalcin, and osteopontin protein expression are somewhat reduced, resulting in less bone differentiation.

The intensity of the electromagnetic field in step 2) is preferably 10 G to 1.5 T, but is not limited thereto.

The pH of the hydrogel of step 1) is preferably 3 to 5, more preferably 4 to 5, most preferably 4, but is not limited thereto.

The magnetic nanoparticles of the step 1) are preferably coated on the surface of the implant, or coated or contained on the surface of the bone, by the hydrogel supporting the magnetic nanoparticles alone or together with the bone formation inducing materials.
The bone formation inducing substance may be at least one component selected from the group consisting of BMP (Bone morphogenetic protein family), hydroxyapatite, TCP (Tricalcium phosphate) and DCP (dicalcium phosphate), but is not limited thereto.
The implant is made of a natural polymer composed of collagen, urnic acid, alginate and chitosan, PLA (poly (lactic acid)), PLGA (poly (glycolic acid)), PGA (poly (glycolic acid)), PCL (polycaprolactone) a synthetic polymer composed of poly (methyl methacrylate), and a synthetic polymer composed of titanium, a titanium alloy, and a nickel-cobalt alloy, but is not limited thereto

According to another aspect of the present invention,

1) coating the surface of the implant with the nano-magnetic nanoparticle-containing hydrogel of the present invention;

2) fixing or filling the implant prepared in step 1) to a bone defect site; And

And 3) irradiating an electromagnetic field to a defective portion fixed or filled in 2) above.

In addition,

1) preparing a bone regeneration scaffold containing the magnetic nanoparticles of the present invention;

2) fixing or filling the scaffold prepared in step 1) to a bone defect site; And

And 3) irradiating an electromagnetic field to a defective portion fixed or filled in 2) above.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Analysis of bone cell activity using low-intensity electromagnetic field

Primary cultured bone cells were inoculated at 1 × 10 5 cells in a 100 mm dish for cell culture, and then exposed to electromagnetic fields (10 gauss) at a frequency of 30, 45, 50, 60, 75 and 100 Hz for 8 hours / The morphological changes and mRNA expression of the cells were analyzed.

As a result, morphological changes of cells due to cytotoxicity were not observed (Fig. 1). As the frequency of the electromagnetic field increased, collagen, bonesialoprotein, austenectin, osteocalcin, osteopontin, MRNA expression of type 2 osteogenic protein (BMP-2) was increased (Fig. 2).

< Example  2> Using low-intensity electromagnetic fields Of mesenchymal stem cells  Bone differentiation analysis

Bone marrow mesenchymal stem cells were inoculated at 1 × 10 5 cells in a 100 mm dish for cell culture and then subjected to electrophoresis (10 gauss) at a frequency of 7.5, 30, 45, 50, 60, After 3 days of irradiation, morphological changes, mRNA expression and protein expression of the cells were analyzed. Bone marrow mesenchymal stem cells cultured on cover slides (diameter 12 mm) are fixed with 10% formalin for 30 minutes and washed three times with pH 7.2 PBS. Treated with an osteopontin antibody, incubated at room temperature for 24 hours, and then developed with EnVision Plus reagent.

As a result, morphological changes of cells due to cytotoxicity were not observed (FIG. 2), and collagen, austenectin, visentin, type 2 osteogenic protein (BMP-2 ) MRNA expression was increased. However, mRNA expression of austenectin, osteocalcin, and osteopontin was somewhat reduced in electromagnetic fields above 75 Hz (Fig. 4).

In addition, the protein expression of collagen, austenectin, and type 2 osteogenic protein (BMP-2) was increased as the electromagnetic field frequency of 30 Hz or more increased, and this result was confirmed as a result of activation of pERK and p-CREB there was. However, it was confirmed that this decrease in the expression at a 100 Hz electromagnetic field is effective at frequencies of 45 to 75 Hz (FIG. 5).

In addition, immunostaining with osteopontin revealed that protein secretion was increased from 50 Hz or more (Fig. 6).

< Example  3> Analysis of bone morphology using high-intensity electromagnetic field

The bone marrow mesenchymal stem cells were inoculated at 1 × 10 5 cells in a 100 mm dish for cell culture, and then subjected to electromagnetic field irradiation at frequency of 30, 45, 50, 60, 75 and 100 Hz three times a day, . The intensities were 1.12, 0.89, 0.68, 0.63, 0.57 and 0.4 T, respectively. After 3 days of irradiation, morphological changes and mRNA expression of the cells were analyzed.

As a result, morphological changes of cell death such as vacuolization were not observed when irradiated with various frequencies of high intensity (0.4 to 1.3 T) electromagnetic field for 3 days, so that mesenchymal stem cells were not damaged (FIG. 7) In bone marrow differentiation analysis of mesenchymal stem cells, mRNA expression of collagen, type 2 osteogenic protein (BMP-2), austenectin, osteocalcin, and osteopontin was most increased at 45 to 60 Hz 8).

< Example  4> Using magnetic nanoparticles and low-intensity electromagnetic field Of mesenchymal stem cells  Bone differentiation analysis

The bone marrow mesenchymal stem cells were inoculated into a 100 mm dish for cell culture at 1 × 10 5 cells, and then replaced with a bone-dividing medium, and the following experiment was carried out.

Magnetic nanoparticle injections (5 μg magnetic nanoparticles per ml of medium, diameter approx. 40 nm), EMF (electromagnetic field): electromagnetic field irradiation (45 Hz, 8 hours × 2 times / day) and MP + EMF: Magnetic resonance analysis and immunohistochemical staining were performed after 3 days of each experiment.

As a result, the magnetic nanoparticle injected group (MP, 50 ㎍ / ml), the electromagnetic field irradiating group (EMF, 45 Hz, 8 hours × 2 times / day) When morphological changes were observed, no cytotoxicity was observed by the magnetic nanoparticles and the electromagnetic field (FIG. 9). As a result of analyzing the mRNA after 3 days of experiment, when the magnetic nanoparticles and the electromagnetic field were examined together, The expression of austenocetin, osteocalcin, osteopontin and basal alloprotein was significantly increased, which was induced by an increase in calcium channel and an increase in expression of Cbfa (FIG. 10).

Also, After 10 days of exposure to the electromagnetic field after magnetic nanoparticles were injected, magnetic fields of osteocalcin and osteotypes were observed after the addition of the magnetic nanoparticles (MP, 50 ㎍ / ml), the electromagnetic field irradiation group (EMF, 45 Hz, 8 hours × 2 times / As a result of immunotyping of oretin, protein secretion was promoted in most experimental groups as compared with the control group, and protein expression was markedly increased in the group in which the magnetic napus and the electromagnetic field were examined together (FIG. 11).

< Example  5> Analysis of activity of osteocyte line using magnetic nanoparticles and low-intensity electromagnetic field

The bone cell line (Saos-2) was inoculated in a 100 mm dish for cell culture at 1 × 10 5 cells.

Specifically, control: comparison group, magnetic particle: magnetic nanoparticle input group (5 μg magnetic nanoparticles per 1 ml of medium), EMF (electromagnetic field): electromagnetic field irradiation group (45 Hz, 8 hours × 2 times / Day) and MP + EMF: bone morphology mRNA analysis was performed after 3 days of each experiment in electromagnetic field group after magnetic nanoparticle input.

As a result, the magnetic nanoparticle injected group (MP, 5 ㎍ / ml), the electromagnetic field group (EMF, 45 Hz, 8 hours × 2 times / day), and the magnetic nanoparticles were injected into the bone cell line (Saos- (Fig. 12). As a result, there was no cytotoxicity like cell death in all experimental groups (Fig. 12) As a result of analysis of mRNA on day 3, the expression of osteocalcin, collagen, and allialoprotein was increased in the group in which the magnetic nanoparticles were administered and the electromagnetic field was irradiated, which was confirmed to be due to the activation of the calcium channel and Cbfa1 (FIG. 13) .

< Example  6> Magnetic nanoparticle content With hydrogel  Electric field Bone cell line  Activity analysis

Collagen hydrogel was prepared by mixing 1% collagen, 5-fold concentrated medium and buffer solution (pH 8.0) at a ratio of 7: 2: 1. 1.0 x 10 6 cells of bone cell line (Saos-2) After mixing with 1 ml of gel, 1 ml was inoculated into a 100 mm culture dish, and 3-dimensional artificial bone tissue was prepared by inducing gelation in a 37 ° C incubator for 30 minutes. Then, about 15 ml of the medium was added and the experiment was carried out under the following conditions.

Magnetic nanoparticle mixing (mixing of 20 μg magnetic nanoparticles per 1 ml of hydrogel), EMF (electromagnetic field): Electro magnetic field after hydrogel preparation 60 Hz, 8 hours × 2 times / day) and MP + EMF: After the preparation of the hydrogel containing magnetic nanoparticles, the electromagnetic field irradiation group was subjected to bone-related mRNA analysis after 3 days of each condition.

As a result, three-dimensional hydrogels (MP, 20 ㎍ / ml) containing magnetic nanoparticles in the osteocyte line (Saos-2) and an electromagnetic field irradiation group (EMF, 60 Hz, 8 hours × 2 times / , And the morphological photographs of the cells on the third day of the experimental group irradiated with the magnetic nanoparticle-containing three-dimensional hydrogel showed that all the cells of the experimental group were well alive (FIG. 14) As a result of analyzing mRNA on the third day, it was confirmed that the expression of native alloprotein and bone morphogenic protein (BMP-2) was increased in the experimental group irradiated with the electromagnetic field in the magnetic nanoparticle-containing hydrogel (FIG.

Claims (11)

1) injecting a hydrogel containing magnetic nanoparticles into cells or tissues of an animal other than a human; And
2) A method for promoting bone differentiation and bone regeneration comprising irradiating the hydrogel prepared in step 1) with an electromagnetic field,
The frequency of the electromagnetic field is 45 Hz to 75 Hz, and
Wherein the intensity of the electromagnetic field is from 10 G to 1.5 T. A method for promoting bone differentiation and bone regeneration.
2. The method according to claim 1, wherein the cell is a bone cell or mesenchymal stem cell.
The method of claim 1, wherein the magnetic nanoparticles are selected from the group consisting of FeO 2 , FeO 3 , and FeO 4 .
2. The method of claim 1, wherein the size of the magnetic nanoparticles is less than 200 nm.
2. The method of claim 1, wherein the electromagnetic field is continuous or pulsed.
delete delete The method of claim 1, wherein the pH of the hydrogel is 3 to 5. The method of claim 1,
[Claim 2] The method according to claim 1, wherein the magnetic nanoparticles are coated on the surface of an implant or contained in bone defects by being loaded on hydrogel alone or together with bone formation inducing materials, How to promote.
The bone formation inducer according to claim 9, wherein the bone formation inducing substance is at least one component selected from the group consisting of Bone morphogenetic protein family, hydroxyapatite, TCP (Tricalcium phosphate) and DCP (Dicalcium phosphate) / RTI &gt; to promote bone formation and bone regeneration.
delete
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