KR20150137416A - Cell therapy composition for healing wounds comprising mesenchymal stem cell or the culture medium treated with silver nano particle - Google Patents

Abstract

The present invention relates to a cell therapy composition or an external application composition for healing wounds containing mesenchymal stem cells (MSC) treated with silver nanoparticles or a culture medium thereof as an active ingredient. In addition, the present invention relates to a method for preparing an MSC with increased generation of HIF-1α or IL-8, which comprises the following steps: treating an MSC before, during, and after culturing the MSC with silver nanoparticles; and culturing the same again. The present invention also relates to a method for preparing a culture medium thereof. The cell therapy composition or the external application composition prepared by the preparation method of the present invention facilitates effective wound healing, thereby providing a new wound healing method.

Description

 TECHNICAL FIELD [0001] The present invention relates to a mesenchymal stem cell or a culture medium for treating a wound, which contains mesenchymal stem cells treated with silver nanoparticles or a culture thereof as an active ingredient.

The present invention relates to a mesenchymal stem cell (MSC) treated with a silver nanomaterial or a cell therapeutic composition for wound healing comprising the culture medium as an active ingredient.

Silver nanoparticles have been reported to promote skin regeneration and differentiation of fibroblasts, promote collagen accumulation during wound healing, and promote skin wound healing by antimicrobial action (Nanomedicine, 7: 497-504 , 2011; ChemMedChem, 2: 129-136, 2007; Journal of nanoscience and nanotechnology, 12: 3766-3774, 2012). Clinical studies have reported that wound healing is enhanced by treating silver sulphadiazine and nanocrystalline silver dressings containing gauze at the burn site (Burns, 33: 161-166, 2007) (Toxicol Appl Pharmacol, 233: 404-410, 2008).

Human mesenchymal stem cells are pluripotent cells, and they are present in fat, bone marrow, etc. (Science, 284: 143-147, 1999), endothelial growth factor, interleukin-6, (J et al., 1998; J Invest Dermatol, 131: 1559-1567, 2004). In the present study, we investigated the effect of various cytokines and chemokines such as interleukin-8 2011; Stem Cells, 25: 2896-2902, 2007).

IL-8 is secreted in a variety of cells such as monocytes, endothelial cells, fibroblasts, and keratinocytes. IL-8 is known to be chemotactic, activates neutrophils, increases mobility of hematopoietic stem cells, and induces keratinocyte proliferation during wound healing (Science, 246: 1601-1603, 1989 ; Clin Exp Immunol, 64: 214-222, 1986; Immunology, 68: 31-36, 1989; Blood 85: 2269-2275, 1995; J Surg Res, 93: 41-54, 2000).

 Hypoxia-inducible factor-1α (HIF-1α) is a major transcription factor that regulates cell proliferation, differentiation and metabolism in hypoxic conditions (Fish physiology and biochemistry, 2013; European Tissue Repair Society, 15: 628-635, 2007).

It has been reported that mesenchymal stem cells are effective in wound healing (Stem cells translational medicine, 2: 33-42, 2013), and cytotoxicity is observed when mesenchymal stem cells are present at high concentrations, It has been reported that low levels of nano-materials in cells promote the proliferation of mesenchymal stem cells (Langenbecks Arch Surg, 394: 495-502, 2009).

However, the present invention has not been disclosed to date on a cell therapy composition for treating a wound containing mesenchymal stem cells treated with silver nanoparticles or a culture thereof as an active ingredient.

Accordingly, the present inventors have made intensive efforts to clarify that mesenchymal stem cells treated with silver nanoparticles or a culture thereof have an effect on wound healing, and as a result, they have experimentally proved this experimentally, Thereby preparing a cell therapy composition for wound healing. In addition, the inventors of the present invention have completed the present invention by discovering the mechanism of action that mesenchymal stem cells containing silver nanoparticles increase HIF-1α and have wound healing efficacy.

It is an object of the present invention to provide a cell therapy composition for treating a wound comprising mesenchymal stem cells (MSC) treated with a silver nanomaterial or a culture thereof as an active ingredient.

It is still another object of the present invention to provide a composition for external application for skin for wound healing, which comprises mesenchymal stem cells treated with silver nanoparticles or a culture thereof as an active ingredient.

The present invention also relates to a method for producing HIF-1α or IL-8, which comprises treating a mesenchymal stem cell with a silver nanoparticle before, during, or after culturing the mesenchymal stem cell, And to provide a method for producing a culture solution.

It is still another object of the present invention to provide a method for producing HIF-1α or IL-8-enhanced mesenchymal stem cells or culture thereof, which comprises culturing mesenchymal stem cells with a silver nanoparticle followed by culture.

In order to achieve the above object, the present invention provides a cell therapy composition for treating a wound comprising an mesenchymal stem cell treated with a silver nanoparticle or a culture thereof as an active ingredient.

The present invention also provides a composition for external application for skin comprising mesenchymal stem cells treated with the silver nanomaterial or a culture thereof as an active ingredient.

The present invention also relates to a method for producing HIF-1α or IL-8, which comprises treating a mesenchymal stem cell with a silver nanoparticle before, during, or after culturing the mesenchymal stem cell, Thereby producing a culture solution.

The present invention also provides a method for producing HIF-1α or IL-8-enhanced mesenchymal stem cells or a culture thereof, characterized in that the mesenchymal stem cells are treated with a silver nanomaterial and then cultured.

The present invention provides an effective cell therapy composition for treating wounds and an external preparation for skin by treating a mesenchymal stem cell with a silver nanomaterial at an appropriate concentration and time to provide a cell therapeutic agent optimized for wound treatment. The present invention also relates to a method for producing HIF-1α or IL-8, which comprises culturing mesenchymal stem cells before, during, or after culturing mesenchymal stem cells, By providing a method for producing stem cells or a culture thereof, a new wound treatment strategy different from the existing methods can be presented.

Figure 1 shows the results of treatment of human mesenchymal stem cells with silver nanoparticles as a result of proliferation of human mesenchymal stem cells (A: human mesenchymal stem cells were treated with each concentration of silver nanomaterials, B: Human mesenchymal stem cells were treated with silver nanomaterials for each hour, and the degree of cell proliferation in human mesenchymal stem cells was measured by BrdU assay * P < 0.05).
FIG. 2 shows that HIF-1α expression was increased in human mesenchymal stem cells as a result of treatment with silver nanomaterials on human mesenchymal stem cells (A: human mesenchymal stem cells were treated with silver nanoparticles at respective concentrations Next, the expression of HIF-1α in human mesenchymal stem cells was confirmed by Western blot analysis. B: Human mesenchymal stem cells were treated with silver nanoparticles for each concentration, The expression of HIF-1α in the cells was confirmed by western blot, which was compared with the expression of HIF-1α (set to 1) in human mesenchymal stem cells without silver nanomaterials. 0.05, * P < 0.01).
FIG. 3 is a graph showing that IL-8 expression is increased in human mesenchymal stem cells as a result of treatment of human mesenchymal stem cells with silver nanomaterials (A: human mesenchymal stem cells, The expression of IL-8 in human mesenchymal stem cells after RT-PCR was confirmed by RT-PCR compared with the expression of IL-8 (set to 1) in human mesenchymal stem cells without silver nanoparticles B: A graph showing the amount of IL-8 secreted by human mesenchymal stem cells after the treatment with silver nanomaterials in human mesenchymal stem cells by an ELISA assay ** P <0.01, *** P < 0.001).
FIG. 4 shows that HIF-1α-dependent cell proliferation and IL-8 expression were increased in human mesenchymal stem cells as a result of treatment with silver nanomaterials in human mesenchymal stem cells (A: siRNA against GFP HIF-1α-treated human mesenchymal stem cells were treated with 0 or 7.5 μg / ml of silver nanomaterials, respectively, and the expression level of HIF-1α in the cells was determined by Western blot analysis. B: C: The amount of IL-8 secreted by the human mesenchymal stem cells treated with silver nanoparticles was determined by an ELISA method, and the amount of IL-8 secreted by the human mesenchymal stem cells was measured by CCK-8 * P <0.05, ** P <0.01).
FIG. 5 is a graph showing an increase in wound healing ability of a mouse after administration of silver nanomaterial-treated human mesenchymal stem cells by a subcutaneous injection method in a mouse. (A: siRNA against GFP and siRNA against HIF- A photograph showing the progress of wound healing over time after subcutaneous injection of a composition prepared by treating each of the treated human mesenchymal stem cells with a silver nanomaterial at a concentration of 0 or 7.5 μg / ml in a 5 mm wound area of a mouse; : A graph showing the wound area area of the mouse; * P> 0.1, ** P <0.05, *** P <0.01).

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

In one embodiment of the present invention, it has been confirmed that human mesenchymal stem cells treated with a silver nanomaterial increase the expression of HIF-1α or IL-8, which is one of the factors regulating cell proliferation and differentiation (see Example 4) In another embodiment, the human mesenchymal stem cells treated with the silver nanomaterial were administered to the injured area of the mouse by a subcutaneous injection method, and it was found to be effective in wound healing (see Example 5).

Accordingly, the present invention provides, in one aspect, a cell therapy composition for wound healing comprising mesenchymal stem cells (MSC) treated with a silver nanomaterial or a culture thereof as an active ingredient.

In the present invention, the mesenchymal stem cells treated with the silver nanomaterial or the culture thereof may be characterized in that HIF-1α or IL-8 is increased compared with a control group not treated with silver nano materials.

In the present invention, it is known that mesenchymal stem cells are distributed in most tissues of the body. These adult stem cells are pluripotent or pluripotent cells capable of differentiating into adipocytes, chondrocytes, bone cells, etc. Multipotent cells (Meirelles et al., J. Cell Sci, 119: 2204-2213, 2006). Mesenchymal stem cells are known to have similar characteristics and characteristics in general, although there are minor differences depending on the origins of the origins (Sakaguchi et al., Arthritis Rheumat, 52: 2521-2529, 2005). Recently, stem cell therapy has been widely recognized as a breakthrough treatment for various diseases. As compared with embryonic stem cells, mesenchymal stem cells are preferable from an ethical point of view, and minimize the response of immune cells It offers a new paradigm of cell therapy in terms of enabling cell infusion.

In the present invention, the 'cell therapeutic agent' refers to a cell therapeutic agent that can be used to regenerate autologous, allogenic, xenogenic cells in vitro or to change the biological characteristics of cells by other methods And the like, which are used for the purpose of treatment, diagnosis and prevention through a series of acts. Since 1993, the United States has been administering cell therapy products as medicines since 2002. These cell treatments can be divided into two major categories, the first is "stem cell treatment" for tissue regeneration or organ function recovery, the second is the regulation of the immune response, such as inhibition of the in vivo immune response or exaggeration of the immune response Is an immune cell therapeutic agent.

The mesenchymal stem cell used in the present invention may be an mesenchymal stem cell of an animal, preferably a mammal, more preferably a human mesenchymal stem cell. Further, the mesenchymal stem cells of the present invention may be derived from bone marrow, adipose tissue, peripheral blood, liver, lung, amniotic fluid, placenta chorionic membrane or umbilical cord blood, and those derived from human adipocytes are particularly preferable.

Mesenchymal stem cells are present in very small amounts in bone marrow, etc., but the process of isolating and culturing them is well known in the art (see, for example, US 5,486,359). In addition, the mesenchymal stem cells can be obtained from the hematopoietic stem cells of the bone marrow according to a known method by their adhesion characteristics and proliferating without losing their differentiation ability.

The process of obtaining mesenchymal stem cells will be described in detail as follows: (1) Mesenchymal stem cells from a mammalian animal, preferably a human mesenchymal stem cell source such as blood or bone marrow, Separating the bone marrow from the tibia, femur, spinal cord or iliac bone; (2) culturing the isolated cells in a suitable medium; And (3) removing floating cells in the culture process and subculturing the cells attached to the culture plate to obtain finally constructed mesenchymal stem cells.

As the medium used in the above process, any medium generally used for culturing stem cells can be used. Preferably, it is a medium containing serum (for example, fetal bovine serum, horse serum and human serum). Mediums which may be used in the present invention include, for example, RPMI series, Eagles ' s MEM (Eagle ' s minimum essential medium, Eagle, H. Science 130: 432,1959), alpha-MEM (Stanner, CP et al. New Biol, 230: 52, 1971), Iscove's MEM (Iscove, N. et al., J. Exp. Med., 147: 923, 1978), 199 medium (Morgan et al., Proc. Med, 73: 1, 1950), CMRL 1066, RPMI 1640 (Moore et al., J. Amer. Med. Assoc, 199: 519, 1967), F12 (Ham, Proc. Natl. (Dulbecco's modification of Eagle's medium, Dulbecco, R. et al., Virology, 8: 396, 1959), DMEM (McCoy ' s 5A (McCoy &lt; / RTI &gt; et al., Anal. Biochem, 102: 255, 1980), Way-mouth's MB752 / 1 (Waymouth, CJ Natl. Cancer Inst. Including, but not limited to, the MCDB series (Ham, RG et al., In Vitro, 14: 11, 1978) no. The medium may contain other components such as antibiotics or antifungal agents (e.g., penicillin, streptomycin) and glutamine.

In another embodiment of the present invention (see Example 1), the human mesenchymal stem cells were treated with silver nanomaterials at various concentrations for 48 hours. As a result, in the range of 1 to 20 μg / ml, It was confirmed that stem cells were increased. Thus, in the present invention, the concentration of the silver nanoparticles treated in human mesenchymal stem cells may be 1 to 20 μg / ml, more preferably 7.5 μg / ml.

As a result of treating the human mesenchymal stem cells with 7.5 μg / ml of the silver nanomaterial in the above-mentioned examples, the group treated with the silver nanomaterials showed that the silver nanomaterials It was confirmed that human mesenchymal stem cells were increased within 5 days. Thus, in the present invention, the processing time of the silver nanoparticles in human mesenchymal stem cells may be 1 to 120 hours, more preferably 48 hours.

 The administration route of the cell therapeutic composition of the present invention can be administered through any conventional route as long as it can reach the target tissue. But are not limited to, parenteral administration, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration. The composition may be formulated in a suitable form together with a pharmaceutical carrier commonly used in cell therapy. &Quot; Pharmaceutically acceptable &quot; refers to compositions which are physiologically acceptable and which, when administered to humans, do not normally cause allergic reactions such as gastrointestinal disorders, dizziness, or the like. Pharmaceutically acceptable carriers include, for example, water, suitable oils, saline, aqueous carriers for parenteral administration such as aqueous glucose and glycols, etc., and may further contain stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. Other pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

The composition may also be administered by any device capable of transferring the cell therapy agent to the target cell.

The cell therapeutic composition of the present invention may comprise a therapeutically effective amount of a cell therapy agent for the treatment of diseases. The therapeutically effective amount refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, The amount that induces the relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the cell therapy agent contained in the composition of the present invention will vary depending on the desired effect. The optimal cell therapy agent content can therefore be readily determined by those skilled in the art and will depend upon a variety of factors including the nature of the disease, the severity of the disease, the amount of other ingredients contained in the composition, the type of formulation, and the age, weight, , The time of administration, the route of administration and the fraction of the composition, the duration of the treatment, the co-administered drug, and the like.

According to one embodiment of the present invention (see Example 5), it was confirmed that the cells treated with 2x10 ⁴ silver nano-particles per mouse in the wound area of the mouse were subcutaneously injected into 25 μl of PBS to be effective in wound healing there was. Therefore, considering the above factors and setting the amount to obtain the maximum effect in a minimal amount without side effects, the composition of the present invention may contain a cell therapy agent of 8x10 ⁴ cells / ml to 8x10 ⁷ cells / ml, more preferably 8x10 ⁵ cells / cells / ml.

In the treatment method of the present invention, the composition comprising the cell treatment agent of the present invention as an active ingredient can be administered orally through the rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, percutaneous, topical, &Lt; / RTI &gt;

The composition comprising the mesenchymal stem cells treated with the silver nanomaterial of the present invention as an active ingredient can be used as an external preparation for skin for wound healing. In this case, depending on the body part, the formulation is not particularly limited. Specifically, it may be a cosmetic composition having a formulation of a softening agent, a nutritional lotion, a massage cream, a nutritional cream, a pack, a gel or a skin adhesive type cosmetic composition and may be a cosmetic composition such as a lotion, ointment, gel, cream, patch or spray It may be a transdermal dosage form. Further, in the composition for external application according to each formulation, the components other than the mesenchymal stem cells treated with the silver nanomaterial of the present invention can be selected and mixed without difficulty by those skilled in the art depending on the formulation or use purpose of other external preparation for skin. , In which case synergistic effects may occur if applied simultaneously with other raw materials.

Meanwhile, in one embodiment of the present invention, the mesenchymal stem cells obtained by culturing human mesenchymal stem cells and then culturing the cells after the treatment of silver nanomaterials or cultures thereof increase HIF-1α or IL-8 , And it was confirmed that it was effective in wound healing.

Therefore, the present invention provides, in still another aspect, a method for producing HIF-1α or IL-8, which comprises culturing mesenchymal stem cells before, during, or after culture with a silver nanomaterial, And a method for producing the same. In one embodiment of the present invention (see Example 1), human mesenchymal stem cells are seeded in a cell culture dish and cultured in a 5% carbon dioxide incubator at 37 ° C. for 18 hours. Then, But the method of culturing the mesenchymal stem cells is not necessarily limited thereto as it is well known in the art (see, for example, US Pat. No. 5,486,359).

 In another aspect of the present invention, there is provided a method for producing HIF-1α or IL-8-enhanced mesenchymal stem cells or a culture thereof, comprising culturing mesenchymal stem cells with a silver nanoparticle followed by culturing have.

In the present invention, the treatment concentration of the silver nanoparticles may be 1 to 20 μg / ml, preferably 7.5 μg / ml, and the treatment time of the silver nanoparticles may be 1 to 120 hours, preferably 48 hours .

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are for illustrating the present invention specifically, and the scope of the present invention is not limited to these examples.

Example 1: Correlation between silver nanomaterial treatment and human mesenchymal stem cell proliferation

      To determine the degree of cell proliferation of human mesenchymal stem cells (ATCC, USA) following treatment with silver nanoparticles (nanoComposix, San Diego, USA), human mesenchymal stem cells were seeded in a cell culture dish, And cultured in a carbon dioxide incubator for 18 hours. The mesenchymal stem cells of the human mesenchymal stem cells were treated with the respective concentrations for 48 hours and the degree of cell proliferation was examined by CCK-8 (CCK-8; Dojindo, Kumamoto, Japan) assay.

As a result, it was confirmed that the proliferation of human mesenchymal stem cells was increased depending on the concentration of the silver nanomaterial treated within a concentration range of 1 to 20 μg / ml (FIG. 1A).

BrdU (Roche Applied Science, Germany) method was used to determine the cell proliferation of human mesenchymal stem cells over time according to the treatment of silver nanomaterials.

As a result, when treated with 7.5 μg / ml of silver nanomaterials in human mesenchymal stem cells, compared with human mesenchymal stem cells not treated with silver nanoparticles, human mesenchymal stem cells It was confirmed that the proliferation was increased (Fig. 1B).

Example 2: Increased expression of HIF-1α in human mesenchymal stem cells by treatment with silver nanoparticles.

The human mesenchymal stem cells cultured in the same manner as in Example 1 were treated with the silver nanomaterials for each concentration for 48 hours, and then the cells were placed in a RIPA buffer (Elpis, Korea) and allowed to stand in ice for 30 minutes. The separator was centrifuged at a speed of 13000 rpm for 30 minutes. Only the supernatant was taken, and 50 μg of the protein was loaded on the SDS-PAGE gel. Then, the gel loaded on PVDF was transferred and blocked with 5% BSA for 1 hour. Then, HIF-1α (BD, USA) or Actin antibody (cell signaling, USA) was diluted 1: 1000 and incubated. After the HRP-conjugated secondary antibody reaction and washing, the protein band was confirmed using ECL (West-Q Chemiluminescent Substrate Kit, Plus GenDEPOT) solution.

As a result, it was confirmed that the expression of HIF-1α was increased depending on the concentration of silver nanomaterial (FIG. 2).

Example 3: Increased expression of IL-8 in human mesenchymal stem cells by treatment with silver nanoparticles

TRIzol (Invitrogen, USA) reagent was added to human mesenchymal stem cells treated in the same manner as in Example 2, and then finely ground using a homogenizer, followed by RNA isolation according to the manufacturer's instructions. CDNA of each tissue was prepared using the reverse transcriptase (Super-bio, Korea). (95 ° C., 30 seconds), an annealing step (56 ° C., 30 seconds) and an extension (extension) using the IL-8 primer of SEQ ID NOs: 1 and 2 and the GAPDH primer of SEQ ID NOs: (72 ° C, 40 seconds) as one cycle and amplified by PCR.

As a result, it was confirmed that expression of IL-8 was increased depending on the concentration of silver nanomaterial (FIG. 3A).

 SEQ ID NO:   gene                     Base sequence One IL-8 forward: 5'-CTGGCCGTGGCTCTCTTG-3 ' 2 IL-8 reverse: 5'-TTAGCACTCCTTGGAAAACTG-3 ' 3 GAPDH forward: 5'-GAAGGTGAAGGTCGGAGTC-3 ' 4 GAPDH reverse: 5'- GAAGATGGTGATGGGATTTC-3 '

After culturing the human mesenchymal stem cells with each concentration of silver nanoparticles for 96 hours, the cultured medium was recovered and ELISA (R & D Systems, Germany) was performed according to the manufacturer's test method to measure the amount of IL-8 secreted in the culture. The culture was left to stand at 37 ° C for 1 hour on a plate coated with IL-8 antibody, washed three times, treated with horseradish-peroxidase conjugated antibody, left at 37 ° C for 1 hour, washed three times, (ELISA reader MR 5000, Dynatech, Guernsey, UK).

As a result, it was confirmed that the expression of IL-8 was increased depending on the concentration of the processed silver nanomaterial (FIG. 3B).

Example 4: HIF-1α-dependent cell proliferation and IL-8 expression increase in human mesenchymal stem cells by treatment with silver nanoparticles

Human mesenchymal stem cells were transfected with siRNA for GFP (Bioneer, Daejeon, Korea) or HIF-1α (GenePharma, Shanghai, China) by Lipofectamin 2000 (Invitrogen, USA) After the silver nanomaterials were treated for 48 hours, the expression level of HIF-1α in the cells was confirmed by Western blotting. The degree of cell proliferation at each time was confirmed by the CCK-8 method and the amount of IL-8 secreted from the cells was measured by ELISA Respectively.

As a result, it was confirmed by western blotting that expression of HIF-1α protein was inhibited in human mesenchymal stem cells treated with siRNA for HIF-1α (FIG. 4A) (Fig. 4B), and that the secretion of IL-8 was decreased (Fig. 4C) by ELISA. Considering the above results, it was confirmed that human mesenchymal stem cells treated with the silver nanoparticles or the culture medium thereof stimulate HIF-1α-dependent growth and increase secretion of IL-8.

Example 5: Increased wound healing ability by human mesenchymal stem cells treated with in vivo silver nanomaterials

We confirmed that the wound healing ability of human mesenchymal stem cells treated with silver nanoparticles confirmed by in vitro experiments is a phenomenon that occurs in vivo. A 5 mm cut was made in the back of a 6-week-old C57BL / 6 female mouse (Orient bio, Sungnam, Korea) using a punch, and then the human mesenchymal stem cells treated with siRNA for GFP or HIF- Cells treated with 7.5 μg / ml of silver nanoparticles for 48 hours were subcutaneously injected into 25 μl of PBS and treated with 2 × 10 ⁴ silver nanoparticles per mouse.

As a result, it was confirmed that the treatment of the wound of the mouse treated with the human mesenchymal stem cells treated with the silver nanoparticles was the fastest. Mice treated with silver nanomaterial-treated human mesenchymal stem cells treated with siRNA for HIF-1.alpha. With reduced expression of HIF-1.alpha. Treated siRNA against GFP and treated with silver nano- It was confirmed that the wound healing ability was reduced compared with the mouse treated with the human mesenchymal stem cells treated with the material (FIG. 5).

As a result of the above considerations, it was confirmed that the ability of human mesenchymal stem cells treated with silver nanoparticles to have wound healing ability depends on the expression of HIF-1α.

Claims (14)

A mesenchymal stem cell (MSC) treated with silver nanoparticles or a culture solution thereof as an active ingredient.
The method according to claim 1,
Wherein the composition containing the mesenchymal stem cells treated with the silver nanoparticles or the culture thereof has increased HIF-1 alpha or IL-8 as compared to a control not treated with silver nanomaterial.
The method according to claim 1,
Wherein the treatment concentration of the silver nanoparticles is 1 to 20 μg / ml.
The method according to claim 1,
Wherein the treatment time of the silver nanoparticles is 1 to 120 hours.
The method according to claim 1,
Wherein said mesenchymal stem cells are human-derived mesenchymal stem cells.
6. The method according to any one of claims 1 to 5,
Wherein the cell number of the mesenchymal stem cells treated with the silver nanoparticles or the culture thereof is 8x10 ⁴ cells / ml to 8x10 ⁷ cells / ml.
A composition for external application for skin for wound healing comprising mesenchymal stem cells (MSC) treated with silver nanoparticles or a culture thereof as an active ingredient.
8. The method of claim 7,
Wherein the composition containing the mesenchymal stem cells treated with the silver nanoparticles or the culture thereof has increased HIF-1 alpha or IL-8 as compared to the control not treated with the silver nanomaterial.
A method for producing HIF-1.alpha. Or IL-8-producing mesenchymal stem cells or cultures thereof, comprising the step of treating mesenchymal stem cells with silver nanoparticles before, during, or after culturing the mesenchymal stem cells.
10. The method of claim 9,
Wherein the treatment concentration of the silver nanoparticles is 1 to 20 μg / ml.
10. The method of claim 9,
Wherein the treatment time of the silver nanoparticles is 1 to 120 hours.
A method for producing HIF-1.alpha. Or IL-8-producing mesenchymal stem cells or cultures thereof, comprising the step of treating mesenchymal stem cells with a silver nanomaterial and then culturing.
13. The method of claim 12,
Wherein the treatment concentration of the silver nanoparticles is 1 to 20 μg / ml.
13. The method of claim 12,
Wherein the treatment time of the silver nanoparticles is 1 to 120 hours.

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