KR102665695B1 - Composition for wound healing or skin regenerating comprising methylsulfonylmethane and agent for wound healing or skin regenerating using the same and method for manufacturing the same - Google Patents

Abstract

The present invention provides a composition for promoting wound healing or skin regeneration, comprising a cell growth material including methylsulfonylmethane (MSM), gelatin or collagen, a viscosity enhancer, and a lubricant; Wound healing or skin regeneration promoter using it; and a method for producing the same. By adding the optimal concentration of methylsulfonylmethane, wound healing or skin regeneration ability can be improved by promoting fibroblast proliferation.
In addition, the wound healing or skin regeneration promoter prepared according to the present invention is characterized by biocompatibility, high hygroscopicity after application, and appropriate viscosity and modulus values (i.e., excellent applicability).

Description

Composition for wound healing or skin regeneration containing methylsulfonylmethane, wound healing or skin regeneration promoter using the same, and method for manufacturing the same METHOD FOR MANUFACTURING THE SAME}

The present invention relates to a composition for wound healing or skin regeneration containing methylsulfonylmethane, a wound healing or skin regeneration promoter using the same, and a method for producing the same.

Covering agents used for the purpose of promoting general skin regeneration must provide an appropriate moist environment for regeneration and meet the following requirements: infection prevention, thermal insulation effect, ease of use, economic efficiency, absorbency, and moisture permeability.

Gelatin is a type of derived protein obtained by treating collagen, a natural protein that makes up animal skin, tendons, and cartilage, with hot water. The change from collagen to gelatin is said to be caused by hydrolysis of the peptide chains, or by cleavage of salt bonds or hydrogen bonds between peptide chains. As a natural polymer, it has excellent biocompatibility, biodegradability, and high hygroscopicity, and has the advantage of providing the environment necessary for skin regeneration.

Methylsulfonylmethane (MSM) is made up of 34% sulfur and refers to a plant-based natural dietary sulfur extracted from various trees such as pine trees. It forms pentadionic acid with organic substances and has a sterilizing and insecticidal effect by dissolving dead skin cells. It is excellent for increasing the activation of skin cells, forming a skin protective film, and has an excellent effect on moisturizing the skin.

Therefore, methylsulfonylmethane is a non-toxic supplier of sulfur elements that reduces inflammation and has strong sterilizing power to improve dermatitis and is also a widely used raw material for atopic dermatitis (Matthew Butawan et al ., Nutrients , 9(3):290, 2017).

Through prior research, the present inventors have disclosed a bio-ink composition containing a specific content of cell transport material, viscosity enhancer, lubricant, and structural material (Korean Patent Publication No. 10-2017-0012099). Through this, it was confirmed that by adding methylsulfonylmethane using the above-described prior art, it was possible to prepare a composition that not only has anti-inflammatory effects but also promotes wound healing or skin regeneration, and the present invention was completed.

The present invention seeks to provide a composition for wound healing or skin regeneration promotion, which is characterized in that it contains a cell growth material containing methylsulfonylmethane (MSM), gelatin or collagen, a viscosity enhancer, and a lubricant. .

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

To achieve the above purpose,

The present invention provides a composition for wound healing or promoting skin regeneration, characterized in that it contains methylsulfonylmethane (MSM), a cell growth material including gelatin or collagen, a viscosity enhancer, and a lubricant.

The concentration of methylsulfonylmethane in the composition may be 0.1 to 3 w/v%.

The cell growth material may further include gelatin methacrylate.

The concentration of the cell growth material in the composition may be 0.1 to 20 w/v%.

The viscosity increasing agent may be hyaluronic acid or dextran, and the lubricant may be glycerol.

The concentration of the viscosity enhancer in the composition may be 0.01 to 3 w/v%, and the concentration of the lubricant in the composition may be 1 to 30 v/v%.

The composition can promote proliferation of fibroblasts (NIH-3T3).

In one embodiment of the present invention, (a) preparing a composition containing a cell growth material including methylsulfonylmethane, gelatin or collagen, a viscosity enhancer, and a lubricant; and (b) treating the skin to have properties that can be applied to the skin.

(c) may additionally include the step of sterilizing with gamma rays.

In another embodiment of the present invention, a wound healing or skin regeneration promoter comprising the composition is provided.

The present invention provides a composition for promoting wound healing or skin regeneration, comprising a cell growth material including methylsulfonylmethane (MSM), gelatin or collagen, a viscosity enhancer, and a lubricant; Wound healing or skin regeneration promoter using it; and a method for producing the same. By adding the optimal concentration of methylsulfonylmethane, wound healing or skin regeneration ability can be improved by promoting fibroblast proliferation.

In addition, the wound healing or skin regeneration promoter prepared according to the present invention is characterized by biocompatibility, high hygroscopicity after application, and appropriate viscosity and modulus values (i.e., excellent applicability).

Figure 1 shows changes in viscosity and modulus values depending on the concentration of methylsulfonylmethane for Example 1.
Figure 2 is for Example 2, and cell proliferation according to the concentration of methylsulfonylmethane was confirmed using Cell counting kit-8 (CCK-8).
Figure 3 shows Example 2, using a fluorescence microscope to confirm cell proliferation according to the concentration of methylsulfonylmethane.

Hereinafter, the present invention will be described in detail.

Composition for wound healing or skin regeneration promotion

The present invention provides a composition for wound healing or promoting skin regeneration, characterized in that it contains methylsulfonylmethane (MSM), a cell growth material including gelatin or collagen, a viscosity enhancer, and a lubricant.

First, the composition according to the present invention is characterized by essentially including methylsulfonylmethane (MSM) in order to promote wound healing or skin regeneration. The methylsulfonylmethane is a substance that has an anti-inflammatory effect. Due to the addition of the optimal concentration of the methylsulfonylmethane, it not only improves wound healing or skin regeneration ability by promoting fibroblast proliferation, but also improves the ability of living organisms to heal. It is characterized by compatibility, high hygroscopicity after application, and appropriate viscosity and modulus values (i.e., excellent applicability).

The concentration of methylsulfonylmethane in the composition is preferably 0.1 to 3 w/v%, more preferably 0.5 to 3 w/v%, and most preferably 1 (±0.2) w/v%. However, it is not limited to this. At this time, when the concentration of methylsulfonylmethane is outside the above range, it is confirmed that the fibroblast proliferation effect is rather reduced, and the viscosity and modulus values are also confirmed to be relatively reduced. Therefore, there is a limitation that it is difficult to implement it in a formulation that can be applied to the skin, such as a liquid such as a cream or gel, or an ointment for wound healing or promoting skin regeneration.

Next, the composition according to the present invention is characterized by containing a cell growth material containing gelatin or collagen, a viscosity enhancer, and a lubricant.

The cell growth material may be a biocompatible material and may include gelatin or collagen. In this case, the gelatin or collagen may be animal-derived or human-derived. The cell growth material may further include gelatin methacrylate for structural stability of the formulation. In addition, if necessary, it may additionally contain one or more substances selected from the group consisting of alginate, agar, agarose, pluronic, and polyvinyl alcohol.

The concentration of the cell growth material in the composition is preferably 0.1 to 20 w/v%, and in particular, the concentration of the cell growth material in the composition is maintained at 1 to 20 w/v% to maintain excellent applicability. You can. For example, the concentration of the gelatin or collagen in the composition may be 1 to 10 w/v%, and the concentration of the gelatin methacrylate in the composition may also be 1 to 10 w/v%.

The viscosity enhancer is a substance for increasing viscosity and may be hyaluronic acid or dextran. The hyaluronic acid is a viscosity enhancer and has a viscosity enhancing agent, about 0.3MDa to about 60MDa, for example, about 15MDa to about 60MDa, about 25MDa to about 60MDa, about 35MDa to about 60MDa, about 03MDa to about 50MDa, about 0.3MDa to about 40MDa, or may range in molecular weight from about 0.3 MDa to about 30 MDa. The concentration of the viscosity enhancer in the composition is preferably 0.01 to 3 w/v%, more preferably 0.1 to 2 w/v%, but is not limited thereto. Thereby, excellent applicability can be maintained.

The lubricant is a material that can minimize the shear rate and improve the physical properties of the composition so that it is not sensitive to temperature changes, and may be glycerol. The concentration of the lubricant in the composition is preferably 0.01 to 30 v/v%, but is not limited thereto.

As a result, the composition has the advantage of improving wound healing or skin regeneration ability by promoting the proliferation of fibroblasts (NIH-3T3).

Wound healing or skin regeneration promoter and method of manufacturing the same

The present invention provides a wound healing or skin regeneration promoter formed from the above composition.

In addition, the present invention includes the steps of (a) preparing a composition containing a cell growth material containing methylsulfonylmethane, gelatin or collagen, a viscosity enhancer, and a lubricant; and (b) treating the skin to have properties that can be applied to the skin.

First, the production method according to the present invention is characterized by comprising the step [(a) step] of producing a composition containing a cell growth material including methylsulfonylmethane, gelatin or collagen, a viscosity enhancer, and a lubricant. .

Since the methylsulfonine methane, the cell growth material containing gelatin or collagen, the viscosity enhancer, and the lubricant have been described above, redundant description will be omitted.

For example, for the above production, as the cell growth material, gelatin or collagen may be mixed with water and then heated to a temperature of 45 to 65° C. to dissolve the gelatin or collagen before use. Additionally, when using hyaluronic acid as the viscosity enhancer, hyaluronic acid can be mixed with water and then heated to a temperature of 35 to 40°C to dissolve the hyaluronic acid before use. According to a specific embodiment of the present invention, deionized water, glycerol, and hyaluronic acid are mixed, and then gelatin and/or gelatin mecarrylate and methylsulfonylmethane are stirred at 50 to 70° C. for 1 to 10 hours to form a uniform mixture. The composition can be prepared by mixing well.

Afterwards, a process to remove the characteristic odor of gelatin or collagen may be added.

Next, the manufacturing method according to the present invention is characterized by including a step [step (b)] of treating the product to have physical properties that can be applied to the skin.

In order to implement the composition prepared in step (a) into a formulation that can be applied to the skin, such as a liquid such as a cream or gel, or an ointment, it needs to be appropriately processed through a known method to have physical properties that can be applied to the skin.

Thereafter, (c) sterilization with gamma rays may be further included. The gamma ray may include an irradiation dose ranging from 5 kGy to 25 kGy, and is preferably irradiated at a dose of 25 kGy, but is not limited thereto. Meanwhile, the method of sterilizing the gelatin sol composition further includes the step of packaging a wound healing or skin regeneration promoter, and the gamma ray irradiation step may be performed after the packaging step. Additionally, the source of gamma rays can be cobalt-60 (Co-60) or cesium-137 (Se-137).

The wound healing or skin regeneration promoter prepared according to the present invention can improve wound healing or skin regeneration ability by promoting fibroblast (NIH-3T3) proliferation due to the addition of the optimal concentration of methylsulfonylmethane. In addition, it is characterized by biocompatibility, high hygroscopicity after application, and appropriate viscosity and modulus values (i.e., excellent applicability).

The wound healing or skin regeneration promoter may be administered in the form of an external skin preparation, and specifically, may be administered in a formulation that can be applied to the skin, such as a liquid such as a cream or gel, or an ointment. At this time, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as the carrier component.

The appropriate dosage of the wound healing or skin regeneration promoter varies depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. may be prescribed. The dosage of the wound healing or skin regeneration promoter is preferably 0.1 to 1 mg/kg (body weight) per day.

Hereinafter, the present invention will be described in more detail through examples.

These examples are only for illustrating the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Referring to Table 1 below, hyaluronic acid (Sigma-Aldrich) was added to deionized water and glycerol (Sigma-Aldrich), and then stirred at 37°C for 1 hour to mix evenly. Next, gelatin (Sigma-Aldrich, USA) and methylsulfonylmethane (source: Sigma-Aldrich, USA) were added and stirred at 60°C for 4 hours to prepare a uniformly mixed composition.

Sample glycerol
(mL/mL) Hyaluronic acid
(mg/mL) gelatin
(mg/mL) Methylsulfonylmethane
(mg/mL) 1-1 5 15 15 0 1-2 10 1-3 20 1-4 30 1-5 40 1-6 50

To confirm the complex viscosity and modulus of the composition, the viscoelasticity was measured at 25°C using a rheometer (MCR 302, Anton Paar, Germany) equipped with a temperature control system. The results are shown in Figure 1.

As shown in Figure 1, it was confirmed that samples 1-2 and 1-3 showed high viscosity and modulus values, and samples 1-4, 1-5, and 1-6 showed similarly low viscosity and modulus values. did. That is, samples 1-2 and 1-3 with high viscosity and modulus values (i.e., when the concentration of MSM is 1 to 2 (w/v)%) can be considered to have excellent applicability.

Afterwards, a process of sterilizing various germs or viruses present in the composition is required. Gamma rays are electromagnetic waves with a shorter wavelength than x-rays and are a form of electromagnetic radiation with high energy generated by atomic nuclear transitions. While gamma rays have low ionizing and photosensitizing effects, they have excellent penetrating power. Using these gamma rays, the energy of gamma rays can be transferred to a specific part of the target object to kill all microorganisms remaining on the target object to a sterilizing level. Considering this, gamma rays can be irradiated at a dose of 5 kGy to 25 kGy. Gamma rays have a very strong penetrating power to the target object, so they can sufficiently sterilize various germs or viruses present in the gel.

The source of gamma rays may be one selected from cobalt-60 (Co-60) or cesium-137 (Se-137). In an embodiment of the present invention, when gamma rays are irradiated, there is almost no temperature rise and the change in radiation dose is small, so the object of the present invention can be easily achieved by being irradiated very uniformly. Gamma irradiation is an economically useful method because it can be irradiated under conditions where no chemical substances are added and is almost unaffected by temperature, humidity, and pressure, and the sterilization method is simple and the processing cost is low. In addition, since there is almost no temperature rise, even products that are sensitive to heat can be sterilized.

Referring to Table 2 below, hyaluronic acid (Sigma-Aldrich) was added to deionized water and glycerol (Sigma-Aldrich), and then stirred at 37°C for 1 hour to mix evenly. Next, gelatin (Sigma-Aldrich, USA), chemically modified gelatin (methacrylated gelatin, self-produced), and methylsulfonylmethane (source: Sigma-Aldrich) were added and stirred at 60°C for 4 hours. A uniformly mixed composition was prepared. Afterwards, 1X10 ⁵ Cells/ml of fibroblasts (NIH 3T3) were mixed and seeded at 200 ㎕ each in a 48 well plate. Afterwards, gelation was performed at 4°C for 30 minutes, followed by UV irradiation for 1 minute each at the top and bottom. Next, 500 ㎕ of medium was dispensed and cultured.

Sample glycerol
(mL/mL) Hyaluronic acid
(mg/mL) gelatin
(mg/mL) Chemically modified gelatin
(mg/mL) Methylsulfonylmethane
(mg/mL) 2-1
: Gel4Cell+MSM 0.5% 0.1 3 30 20 5 2-2
: Gel4Cell+MSM 1% 10 2-3
: Gel4Cell+MSM 2% 20 2-4
: Gel4Cell+MSM 3% 30 2-5
: Gel4Cell+MSM 4% 40 2-6
: Gel4Cell+MSM 5% 50

First, Cell counting kit-8 (CCK-8) was performed to check cell proliferation. Specifically, the medium was aspirated and washed with PBS. Next, 450 ㎕ of medium and 50 ㎕ of CCK-8 were dispensed, and reacted for 3 hours in an incubator at 37°C and 5% CO ₂ after blocking light, and the absorbance at 450 nm was measured. The results are shown in Figure 2.

Next, fluorescence microscopy was observed to confirm cell proliferation. Specifically, the medium was aspirated and washed with PBS. Next, 2 ㎖ of PBS and Live (1 ㎕) & Dead (4 ㎕) solution were dispensed, and the mixture was allowed to react for 40 minutes at room temperature after blocking light, and observed under a fluorescence microscope. The results are shown in Figure 3.

As shown in Figures 2 and 3, it was confirmed that samples 2-2, 2-3, and 2-4 showed a high cell proliferation rate, and sample 2-1 where the concentration of MSM was too low, or sample 2 where the concentration of MSM was too high. In sample -5, it was confirmed that the cell proliferation rate was relatively reduced. That is, samples 2-2, 2-3 and 2-4 with high cell proliferation rates for fibroblasts (i.e., when the concentration of MSM is 1 to 3 (w/v)%), especially sample 2-2 ( In other words, when the concentration of MSM is 1 (w/v)%), it can be considered to have excellent wound healing or skin regeneration ability.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.

Claims (10)

methylsulfonylmethane (MSM);
As a cell growth material, gelatin or gelatin methacrylate;
As a viscosity enhancing agent, hyaluronic acid; and
As a lubricant, it contains glycerol,
The concentration of the methylsulfonylmethane in the composition is 1 (±0.2) w/v%, the concentration of the cell growth material is 0.1 to 20 w/v%, and the concentration of the viscosity enhancer is 0.01 to 3 w/v. v%, and the concentration of the lubricant is 0.01 to 30 v/v%. A composition for wound healing or promoting skin regeneration.
delete delete delete delete delete According to paragraph 1,
A composition for promoting wound healing or skin regeneration, characterized in that the composition promotes the proliferation of fibroblasts (NIH-3T3).
(a) methylsulfonylmethane; As a cell growth material, gelatin or gelatin methacrylate; As a viscosity enhancing agent, hyaluronic acid; and as a lubricant, preparing a composition comprising glycerol; and
(b) comprising the step of treating the skin to have properties that can be applied to the skin,
In step (a), the concentration of the methylsulfonylmethane in the composition is 1 (±0.2) w/v%, the concentration of the cell growth material is 0.1 to 20 w/v%, and the concentration of the viscosity enhancer is 0.01 to 3 w/v%, and the concentration of the lubricant is 0.01 to 30 v/v%. A method for producing a wound healing or skin regeneration promoter.
According to clause 8,
(c) A method for producing a wound healing or skin regeneration promoter, further comprising the step of sterilizing with gamma rays.
A wound healing or skin regeneration promoter prepared from the composition according to claim 1 or 7.