Hydrogel composition for storing acellular dermal matrix, hydrogel, comprising same and exhibiting excellent moisture retention, for storing acellular dermal matrix, and preparation method therefor

Abstract

The present invention relates to a hydrogel composition for storing an acellular dermal matrix, a hydrogel, comprising same and exhibiting excellent moisture retention, for storing an acellular dermal matrix, and a preparation method therefor. The present invention can provide a hydrogel composition for storing an acellular dermal matrix, and a hydrogel for storing an acellular dermal matrix, comprising same, wherein the hydrogel composition comprises a crosslinked hydrophilic polymer and an antibiotic, enabling the packaged acellular dermal matric to exhibit excellent moisture retention even without a preservation solution and also can suppress the activity of the activity of immune rejection-inducing cells remaining in the acellular dermal matrix and minimize bacterial contamination that may occur during storage and distribution. In addition, the composition comprises a crosslinking regulator to enhance crosslinking uniformity of the hydrophilic polymer compound, whereby moisture retention and antibiotic action effects can be improved.

Description

Hydrogel composition for preserving acellular dermal matrix, hydrogel for preserving acellular dermal matrix with excellent moisture retention property including the same, and method for preparing the same

[0001] The present invention relates to a hydrogel composition for storing an acellular dermal matrix, a hydrogel for storing an acellular dermal matrix with excellent moisture retention properties including the same, and a method for manufacturing the same.

[0002] Since the first cross-linked hydrogel using polyhydroxyethyl methacrylate (PHEAM) was introduced by Wichterle and Lim in the 1960s, numerous researchers have continuously studied hydrogels using materials with water-soluble properties and potential biocompatibility. Meanwhile, the production of hydrogels using calcium alginate around the 1980s marked a turning point in the field of biomaterials. Since then, the synthesis of biomaterial hydrogels using natural or synthetic polymers has made rapid progress.

[0003] Hydrogels are polymeric materials capable of retaining water and possess a three-dimensional network structure. They are typically composed of two or more components, one of which is a hydrophilic polymer. A key characteristic of hydrogels is that the hydrophilic polymer chains bind together to form a three-dimensional network structure, rendering them insoluble in water. These hydrogels maintain their original shape and swell in water until they reach equilibrium.

[0004] These hydrogels, due to their excellent biocompatibility, are used in a wide range of medical fields. Their specific applications include wound dressings, surgical sutures, scaffolds, artificial cartilage or membranes, dental materials, contact lenses, and drug delivery systems. Furthermore, they can be modified by various environmental factors within the human body. They can prevent drug denaturation due to enzymes or pH in the intestines, and can also be endowed with specific properties that release drugs in response to various internal stimuli. Hydrogels with stimuli-sensitive properties can undergo reversible volume changes or sol-gel transitions within minutes in response to internal stimuli. External stimuli can be broadly categorized into physical stimuli such as temperature, electricity, solvent changes, light, pressure, sound, and magnetism. The development of stimuli-responsive hydrogels utilizing these stimuli is expected to have potential applications in highly efficient, sustained-release drug delivery systems that minimize adverse drug reactions. Meanwhile, hydrogels have recently become the subject of intensive research due to their limitless applicability in various fields, including not only the pharmaceutical field but also chemical valves, enzyme and cell immobilization, and bio-separation.

[0005] However, despite various studies on the application of hydrogels, there is currently no research on their use for storage of acellular dermal matrix, which is skin tissue for transplantation.

[0006] [Prior Art Literature]

[0007] [Patent Document]

[0008] (Patent Document 1) KR 10-2721032 B1

[0009] The purpose of the present invention is to provide a hydrogel composition for storing an acellular dermal matrix, an acellular dermal matrix hydrogel having excellent moisture retention properties including the same, and a method for preparing the same.

[0010] Another object of the present invention is to provide a hydrogel composition for storing an acellular dermal matrix, which can exhibit excellent moisture retention even in the absence of a preservation solution, including a cross-linked hydrophilic polymer compound and an antibiotic, and can suppress the activity of cells that induce an immune rejection response remaining in the acellular dermal matrix and minimize bacterial contamination that may occur during storage and distribution, and an acellular dermal matrix storage hydrogel comprising the same.

[0011] Another object of the present invention is to provide a hydrogel composition for preserving an acellular dermal matrix, which can improve moisture retention and antibiotic action by improving the crosslinking uniformity of a hydrophilic polymer compound including a crosslinking regulator, and a hydrogel for preserving an acellular dermal matrix comprising the same.

[0012] To achieve the above purpose, a hydrogel composition for storing an acellular dermal matrix according to one embodiment of the present invention may include a hydrophilic polymer compound and an antibiotic.

[0013] Additionally, the hydrophilic polymer compound may be selected from the group consisting of carboxymethyl cellulose (CMC), hyaluronic acid (HA), gelatin, cellulose, biocellulose, gellan gum, agar, agarose, poloxamer, and mixtures thereof.

[0014] Additionally, the antibiotic may be selected from the group consisting of penicillin, streptomycin, kanamycin, neomycin, bacitracin, gentamicin, vancomycin, beta-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, glycopeptide antibiotics, lincomycin antibiotics, quinolone antibiotics, and mixtures thereof.

[0015] According to another embodiment of the present invention, an acellular dermal matrix storage hydrogel with excellent moisture retention may include the hydrogel composition of the present invention.

[0016] In addition, the hydrogel for storing the acellular dermal matrix can exhibit excellent moisture retention properties by including a cross-linked hydrophilic polymer compound.

[0017] In addition, the hydrogel for storing the acellular dermal matrix is ​​in the form of a sheet, and the thickness of the hydrogel sheet may be 1 mm to 5 mm.

[0018] A method for preparing a hydrogel for preserving an acellular dermal matrix having excellent moisture retention according to another embodiment of the present invention may include the steps of: 1) preparing a hydrogel solution including a cross-linked hydrophilic polymer compound by mixing purified water, a hydrophilic polymer compound, and a cross-linking agent; 2) curing the hydrogel solution to form a hydrogel; and 3) immersing the formed hydrogel in an impregnation solution including an antibiotic.

[0019] The present invention relates to a hydrogel composition for storing an acellular dermal matrix, an acellular dermal matrix storage hydrogel having excellent moisture retention properties including the same, and a method for preparing the same. The present invention provides an acellular dermal matrix storage hydrogel composition and an acellular dermal matrix storage hydrogel containing the same, which can not only exhibit excellent moisture retention properties without a preservation solution when packaged, including a cross-linked hydrophilic polymer compound and an antibiotic, but can also suppress the activity of cells that induce an immune rejection response remaining in the acellular dermal matrix and minimize bacterial contamination that may occur during storage and distribution.

[0020] In addition, by improving the crosslinking uniformity of a hydrophilic polymer compound including a crosslinking regulator, the moisture retention and antibiotic effect can be improved.

[0021] Figure 1 shows changes in thickness and weight of a hydrogel before and after water absorption according to one embodiment of the present invention.

[0022] Figure 2 shows the moisture evaporation rate of an acellular dermal matrix wrapped with a hydrogel according to one embodiment of the present invention.

[0023] Figure 3 shows the moisture content of an acellular dermal matrix wrapped with a hydrogel according to one embodiment of the present invention.

[0024] Figure 4 illustrates a hydrogel for storing an acellular dermal matrix with excellent moisture retention according to one embodiment of the present invention.

[0025] The present invention relates to a hydrogel composition for preserving an acellular dermal matrix comprising a hydrophilic polymer compound and an antibiotic.

[0026] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0027] The term "acellular dermal layer" or "acellular dermal matrix" in the specification of the present invention may refer to an acellular dermal matrix (ADM) that is a skin tissue in which cells in the epidermis or dermis that may cause an immune response in a patient are substantially removed after transplantation from skin tissue separated from an individual, and which can provide a three-dimensional structural support for the influx of fibroblasts, nerve generation, and blood vessel regeneration after transplantation. The term "substantially removed" as described above means "substantially not including" because it is substantially removed, and "substantially" may refer to a state in which cells, etc. are removed by 90% or more, for example, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more, compared to the raw material. Meanwhile, the acellular allogeneic dermis of the present invention is processed using allogeneic skin tissue derived from a human body.

[0028] Meanwhile, the skin tissue needs to be regenerated when there is severe tissue damage due to burns, trauma, ulcers, bedsores, etc., and many studies are currently being conducted on methods of using processed skin substitutes for this purpose.

[0029] The above skin substitutes can be classified into human-derived allogeneic dermis, animal-derived xenogeneic dermis, and synthetic ones. While allogeneic dermis-derived skin substitutes (acellular allogeneic dermis) are relatively expensive, they have the advantage of not causing immune rejection or inflammatory reactions after transplantation, and are therefore evaluated as the optimal skin substitute.

[0030] Acellular Dermal Matrix is ​​a product that selectively removes only the cellular antigens that are the target of the immune response while maintaining various structural proteins and components without damaging the three-dimensional structure of the dermal layer of the skin tissue provided by cadaveric skin from donors. It has important characteristics such as very low antigenicity, rapid blood vessel formation ability, and stability as a dermal layer, so it can be used in areas where skin tissue is severely damaged due to burns, trauma, or other diseases.

[0031] In order to store and distribute the acellular allogeneic dermal matrix (ADM) that has undergone specific processing or treatment steps from donated cadaveric skin tissue for transplantation and engraftment in a defective skin tissue area, techniques for freeze-drying, freezing, or hydrating the acellular dermal matrix have been used in the past.

[0032] However, the freeze-drying method has the problem of not only being cumbersome in the process but also worsening the prognosis of patients requiring urgent surgery, as it requires a process of restoring the acellular dermal matrix by immersing it in water for a certain period of time prior to the surgery.

[0033] In addition, the above freezing method also has limitations in long-term storage and transportation because it is stored and transported under very low temperature conditions, and there is a problem that it must be thawed before surgery.

[0034] Meanwhile, the above-mentioned hydration method is a method of storing an acellular dermal matrix (ADM) at room temperature while immersed in a "preservation solution" containing specific ingredients. Therefore, it is relatively easy to store at room temperature compared to the two methods using low-temperature conditions. However, during the process of storing the acellular dermal matrix in the preservation solution, the dermal layer may come off in the form of floating matter, which may induce an inflammatory reaction upon engraftment. In addition, the acellular dermal matrix may cluster on the preservation solution, which reduces the storage properties.

[0035] Accordingly, the present invention is intended to overcome the problems of conventional techniques used for storage and distribution of the acellular dermal matrix (ADM), and is characterized by utilizing a "hydrogel sheet" to excellently maintain the moisture preservation effect of the packaged acellular dermal matrix even under conditions that do not include a conventional "preservation solution," and the "hydrogel sheet" may include a cross-linked hydrogel.

[0036] A hydrogel composition for storing an acellular dermal matrix according to one embodiment of the present invention may include a hydrophilic polymer compound and an antibiotic.

[0037] The above hydrophilic polymer compound is a polymer containing a hydrophilic functional group and is a major component for forming a hydrogel by reacting with a crosslinking agent described later, and can perform moisture absorption and retention functions, thereby exhibiting long-term moisture retention for storage of the acellular dermal matrix desired by the present invention.

[0038] Specifically, the hydrophilic polymer compound may be selected from the group consisting of carboxymethyl cellulose (CMC), hyaluronic acid (HA), gelatin, cellulose, biocellulose, gellan gum, agar, agarose, poloxamer, and mixtures thereof.

[0039] In addition, the hydrophilic polymer compound may include the hydrophilic polymer compound itself or a derivative thereof listed above, and the derivative may also include a salt of each hydrophilic polymer compound.

[0040] In addition, the molecular weight of the hydrophilic polymer compound may be 100,000 to 3,000,000 Daltons (Da), 100,000 to 2,700,000 Daltons (Da), 100,000 to 2,400,000 Daltons (Da), 500,000 to 3,000,000 Daltons (Da), 500,000 to 2,700,000 Daltons (Da), 500,000 to 2,400,000 Daltons (Da), 800,000 to 3,000,000 Daltons (Da), 800,000 to 2,700,000 Daltons (Da), or 800,000 to 2,400,000 Daltons (Da).

[0041] Additionally, the concentration of the hydrophilic polymer compound in the composition of the present invention may be 0.1 to 30 wt%, 0.1 to 25 wt%, 0.1 to 20 wt%, 1 to 30 wt%, 1 to 25 wt%, 1 to 20 wt%, 5 to 30 wt%, 5 to 25 wt%, or 5 to 20 wt%.

[0042] If the concentration of the hydrophilic polymer compound in the above composition is less than 0.1 wt%, the content of the hydrophilic polymer compound may be too low, which may lower the efficiency of manufacturing a hydrogel. If the concentration of the hydrophilic polymer compound exceeds 30 wt%, the crosslinking reaction may not occur properly due to mixing of the crosslinking agent.

[0043] Meanwhile, the hydrogel composition for storing the acellular dermal matrix may additionally include a cross-linking agent.

[0044] Meanwhile, the cross-linking agent is a substance that enables the formation of a network structure through chemical cross-linking between the hydrophilic polymer compounds, and the hydrogel composition of the present invention can include a hydrophilic polymer compound having a cross-linked structure by including the cross-linking agent as an active ingredient.

[0045] More specifically, when using the hydrophilic polymer compound of the above cross-linked structure, as described above, the hydrophilic polymer compound can be formed into a network structure, thereby further improving the moisture retention capacity or water holding capacity of the manufactured hydrogel, thereby improving the storage properties of the acellular dermal matrix aimed at by the present invention.

[0046] Specifically, the crosslinking agent is genipin, glutaraldehyde, 1,4-butandiol diglycidyl ether (BDDE), EDC-NHS, 1, 2, 7, 8-diepoxyoctane (DEO), ethylene glycol diglycidyl ether (EGDGE), 1,6-hexanediol diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, aluminum hydroxide, aluminum hydroxide gel, hydrous aluminum silicate, kaolin, It may be selected from the group consisting of aluminum acetate, aluminum lactate, aluminum stearate, calcium chloride, magnesium chloride, aluminum chloride, magnesium aluminometasilicate, and magnesium aluminosilicate. However, the present invention is not limited thereto, and any compound capable of inducing crosslinking between the above-mentioned friend polymer chains may be used without limitation.

[0047] Specifically, the hydrophilic polymer compound and the crosslinking agent may be included in a molar ratio of 5:1 to 20:1, and may be included in a molar ratio of 5:1 to 10:1.

[0048] When the molar ratio of the hydrophilic polymer compound and the cross-linking agent is within the above range, efficient cross-linking between the hydrophilic polymer compounds can be induced, thereby not only improving the durability of the manufactured hydrogel but also maximizing the moisture retention capacity. However, when the molar ratio is below the above range, the content of the hydrophilic polymer compound may be relatively low, which may lower the manufacturing efficiency of the hydrogel or reduce the physical durability of the manufactured hydrogel. When the molar ratio exceeds the above range, the cross-linking agent may be included in a large amount, which may prevent the cross-linking reaction from occurring properly.

[0049] Meanwhile, the hydrogel composition of the present invention is characterized in that it can be used for the storage of an acellular dermal matrix used in skin transplant surgery, etc.

[0050] Specifically, in order to overcome the problems associated with the use of conventional preservative solutions, which have been used for the purpose of enabling the acellular dermal matrix to continuously maintain a certain amount of moisture as described above, it is important to demonstrate moisture retention ability equivalent to or greater than that achieved when using a preservative solution without using the solution.

[0051] In this way, in order for the hydrogel including the hydrogel composition of the present invention to exhibit excellent moisture retention, it is necessary to control the crosslinking reaction between the hydrophilic polymer compound and the crosslinking agent.

[0052] Furthermore, as will be described later, the composition of the present invention includes an antibiotic as an active ingredient, and since the antibiotic can be included by being adsorbed or bound between cross-linked polymer compounds forming the manufactured hydrogel, the manufactured hydrogel should not only have excellent moisture retention capacity, but also have the antibiotic effectively adsorbed or bound between the cross-linked hydrophilic polymer compounds, and for this purpose, it is necessary to control the cross-linking rate of the hydrophilic polymer and / or the cross-linking agent.

[0053] To this end, the hydrogel composition for storing an acellular dermal matrix of the present invention may additionally include a cross-linking regulator.

[0054] The above crosslinking regulator is included to control the crosslinking rate of the polymer and / or crosslinking agent to achieve the above-described effect, and may include glycerol, a compound represented by the following chemical formula 1, and a compound represented by the following chemical formula 2:

[0055] [Chemical Formula 1]

[0056]

[0057] [Chemical Formula 2]

[0058]

[0059] Here,

[0060] m and o are equal to or different from each other and are each independently an integer from 1 to 4,

[0061] n is an integer from 1 to 3,

[0062] p and q are equal to or different from each other and are each independently an integer from 1 to 5,

[0063] r, s and t are equal to or different from each other and are each independently an integer from 1 to 2,

[0064] R1 to R 10 are the same or different from each other, and are each independently selected from the group consisting of hydrogen, a halogen group, a cyano group, a hydroxy group, a nitro group, a substituted or unsubstituted alkyl group having 1 to 30 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 24 carbon atoms, a substituted or unsubstituted heteroalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted aralkyl group having 6 to 30 carbon atoms, a substituted or unsubstituted aryl group having 5 to 30 carbon atoms, and a substituted or unsubstituted heteroaryl group having 2 to 30 carbon atoms,

[0065] The above R1 to R 10 When substituted, it is substituted with a substituent selected from the group consisting of hydrogen, a cyano group, a nitro group, a halogen group, a hydroxy group, a carboxyl group, an alkyl group having 1 to 30 carbon atoms, an alkenyl group having 2 to 30 carbon atoms, an alkynyl group having 2 to 24 carbon atoms, a heteroalkyl group having 2 to 30 carbon atoms, an aralkyl group having 6 to 30 carbon atoms, an aryl group having 5 to 30 carbon atoms, a heteroaryl group having 2 to 30 carbon atoms, a heteroarylalkyl group having 3 to 30 carbon atoms, an alkoxy group having 1 to 30 carbon atoms, an alkylamine group having 1 to 30 carbon atoms, an arylamine group having 6 to 30 carbon atoms, an aralkylamine group having 6 to 30 carbon atoms, and a heteroarylamine group having 2 to 24 carbon atoms, and when substituted with multiple substituents, they are the same or different from each other.

[0066] More preferably, the compound represented by the above chemical formula 1 may be a compound represented by the following chemical formula 3, and the compound represented by the above chemical formula 2 may be a compound represented by the following chemical formula 4:

[0067] [Chemical Formula 3]

[0068]

[0069] [Chemical Formula 4]

[0070]

[0071] The above glycerol is included as a gelation rate-controlling agent capable of controlling the crosslinking rate of the crosslinking agent. By controlling the viscosity within the hydrogel using its high viscosity, the speed of the gelation reaction can be controlled. This allows for modification of the interactions between polymer chains during crosslinking between hydrophilic polymer compounds, thereby controlling the crosslinking rate.

[0072] Meanwhile, the compound represented by the above chemical formula 1 and the compound represented by the chemical formula 2 can form hydrogen bonds with hydrogen atoms contained in the hydrophilic polymer compound during cross-linking between hydrophilic polymer compounds by including a nitrogen element, thereby controlling the cross-linking reaction between the hydrophilic polymer compounds.

[0073] Furthermore, these compounds contain specific functional groups or substituents, and can form intermolecular bonds or interactions with hydrophilic polymer compounds during the process of crosslinking between the hydrophilic polymer compounds. This allows for controlling the crosslinking reaction or forming adsorption or binding sites for moisture molecules and / or antibiotics, thereby enhancing the moisture retention capacity desired by the present invention and effectively adsorbing or binding antibiotics.

[0074] More preferably, the crosslinking regulator may include 30 to 50 parts by weight of the compound represented by the chemical formula 1 and 30 to 50 parts by weight of the compound represented by the chemical formula 2, per 100 parts by weight of glycerol.

[0075] When the crosslinking regulator is mixed within the above weight range, the moisture retention capacity of the hydrogel targeted by the present invention can be further maximized, and the antibiotic adsorption effect can also be maximized. However, when the weight range is exceeded, the additional additive is included in an excessive amount, which may inhibit the formation of crosslinking bonds between hydrophilic polymer compounds or between the hydrophilic polymer compound and the crosslinking regulator. When the weight range is less than the above weight range, the crosslinking control effect targeted by the present invention may not be sufficiently exhibited.

[0076] Meanwhile, the hydrogel composition of the present invention may include a hydrophilic polymer compound and a cross-linking agent in a molar ratio of 5:1 to 20:1, and may include 30 to 50 parts by weight of an antibiotic and 20 to 40 parts by weight of a cross-linking regulator for 100 parts by weight of the hydrophilic polymer compound and the cross-linking agent. When the effective ingredients of the hydrogel composition of the present invention are mixed in the above weight range, not only can the moisture retention effect be maximized due to the synergistic effect resulting from the mixing of each effective ingredient, but also the activity inhibition effect of cells that induce immune rejection response remaining in the acellular dermal matrix by the antibiotic can be maximized. However, when the weight range is exceeded or below the above weight range, the effect may not be sufficient.

[0077] Additionally, the antibiotic may be selected from the group consisting of penicillin, streptomycin, kanamycin, neomycin, bacitracin, gentamicin, vancomycin, beta-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, glycopeptide antibiotics, lincomycin antibiotics, quinolone antibiotics, and mixtures thereof.

[0078] The hydrogel composition of the present invention is used for the storage of an acellular dermal matrix. If cells that cause an immune rejection reaction remain in the acellular dermal matrix or bacterial contamination occurs during the storage and distribution of the acellular dermal matrix, there is a problem that the prognosis of a patient who has received the dermal matrix may be poor.

[0079] Accordingly, the present invention is characterized by including an antibiotic as an active ingredient, and the antibiotic can be included in the hydrogel manufactured through this. In this case, when the acellular dermal matrix is ​​stored or distributed in contact with the hydrogel, the antibiotic acts on the acellular dermal matrix to suppress the activity of cells that induce an immune rejection response remaining, and by minimizing bacterial contamination that may occur during the storage and distribution process, the problem of the aforementioned side effects can be minimized.

[0080] Meanwhile, antibiotics that may be included in the composition of the present invention include, but are not limited to, penicillin, streptomycin, kanamycin, neomycin, bacitracin, gentamicin, vancomycin, etc., and are defined as including the types of antibiotics that can be applied by a person skilled in the art to achieve the above purpose.

[0081] According to another embodiment of the present invention, an acellular dermal matrix storage hydrogel with excellent moisture retention may include the hydrogel composition of the present invention.

[0082] In addition, the hydrogel for storing the acellular dermal matrix can exhibit excellent moisture retention properties by including a cross-linked hydrophilic polymer compound.

[0083] In addition, the hydrogel for storing the acellular dermal matrix is ​​in the form of a sheet, and the thickness of the hydrogel sheet may be 1 mm to 5 mm.

[0084] That is, the hydrogel for storage of the acellular dermal matrix of the present invention includes the hydrogel composition, and as the hydrophilic polymer and the crosslinking agent are mixed, crosslinking between hydrophilic polymers and / or crosslinking between the hydrophilic polymer and the crosslinking agent can be induced.

[0085] Specifically, the hydrogel of the present invention can further improve moisture retention by including a cross-linked hydrophilic polymer compound, thereby also exerting a moisture retention effect on the acellular dermal matrix during the process of contacting and storing or distributing the acellular dermal matrix. Meanwhile, it is characterized by minimizing bacterial infection problems during the process of storing and distributing the acellular dermal matrix and the activity of cells that induce immune rejection responses remaining in the dermal matrix by additionally including an antibiotic.

[0086] In addition, by further including the aforementioned cross-linking regulator, it has the characteristic of being able to maximize moisture retention and antibiotic adsorption or binding effect by controlling cross-linking between the hydrophilic polymer compounds or cross-linking between the hydrophilic polymer compound and the cross-linking agent.

[0087] Meanwhile, the effective crosslinking ratio (CrR) of the crosslinked hydrophilic polymer crosslinker according to the following formula 1 may be 1.0 to 2.0:

[0088] [Formula 1]

[0089] Effective cross-linking ratio (CrR) = moles of double cross-linked cross-linker / (moles of single cross-linked cross-linker + moles of double cross-linked cross-linker)

[0090] That is, the effective cross-linker ratio (CrR) refers to the effective cross-linking ratio in which both sides of the total cross-linking agent cross-linked to the hydrophilic polymer compound in the cross-linked hydrophilic polymer cross-linker are cross-linked.

[0091] When the effective crosslinking rate (CrR) of the crosslinked hydrophilic polymer is within the above range, the viscosity and elasticity can be adjusted to a desired level to further maximize the moisture retention capacity and antibiotic adsorption or binding effect of the hydrogel of the present invention. When it is outside the above range, the effect may be somewhat reduced.

[0092] More specifically, the effective crosslinking ratio (CrR) of the crosslinked hydrophilic polymer crosslinker according to the above formula 1 may be 1.5 to 2.0, and may be 1.8 to 2.0.

[0093] A method for preparing a hydrogel for preserving an acellular dermal matrix having excellent moisture retention according to another embodiment of the present invention may include the steps of: 1) preparing a hydrogel solution including a cross-linked hydrophilic polymer compound by mixing purified water, a hydrophilic polymer compound, and a cross-linking agent; 2) injecting the hydrogel solution into a mold having a predetermined shape and then hardening it to form a hydrogel; and 3) immersing the formed hydrogel in an impregnation solution including an antibiotic.

[0094] Specifically, the step 1) above is to mix and stir a hydrophilic polymer compound and a crosslinking agent in purified water to cause a crosslinking reaction between hydrophilic polymer compounds or between a hydrophilic polymer compound and a crosslinking agent, thereby producing a hydrogel solution including a crosslinked hydrophilic polymer compound.

[0095] Meanwhile, preferably, in the step 1), the crosslinking regulator of the present invention can be additionally mixed, thereby controlling the effective crosslinking rate of the crosslinked product, thereby maximizing moisture retention and minimizing bacterial contamination, thereby maximizing storage and distribution properties.

[0096] Meanwhile, step 2) above is a step of injecting the hydrogel solution manufactured in step 1) into a mold of a predetermined shape and then hardening it to form a sheet-shaped hydrogel. This allows for the production of a sheet-shaped hydrogel that can easily encapsulate the acellular dermal matrix, thereby improving the convenience of distribution and storage.

[0097] In addition, the step 3) above is a step of immersing the sheet-shaped hydrogel manufactured through step 2) in an impregnating solution containing an antibiotic, thereby allowing the antibiotic to be adsorbed or bound between the cross-linked hydrogels, and ultimately, when the sheet-shaped hydrogel comes into contact with an acellular dermal matrix, the antibiotic can directly act on the acellular dermal matrix to minimize the activity of cells that induce an immune rejection response, and also has the characteristic of minimizing contamination by harmful bacteria, thereby allowing the acellular dermal matrix to be distributed and stored for a long period of time.

[0098] Manufacturing example

[0099] Example 1

[0100] Hyaluronic acid (Mw: approximately 4,000,000 g / mol) was prepared as a hydrophilic polymer compound, and butanediol diglycidyl ether (1,4-butandiol diglycidyl ether: BDDE) was prepared as a crosslinking agent.

[0101] The hyaluronic acid and BDDE were mixed in purified water at a molar ratio of 7:1, and stirred for the first time at 50 to 60°C for 20 to 30 minutes at 1500 to 1700 rpm.

[0102] Afterwards, the temperature was increased to 65 to 75 ℃ and the solution was stirred a second time at 2000 to 2500 rpm for 30 to 50 minutes to prepare a hydrogel solution.

[0103] Afterwards, the hydrogel solution was injected into a mold having a predetermined shape, cured at 50 to 60°C for 20 minutes, and then aged at room temperature (20 to 25°C) for 10 hours to form a sheet-shaped hydrogel.

[0104] Thereafter, the formed sheet-shaped hydrogel was immersed in an impregnation solution containing 40 parts by weight of vancomycin per 100 parts by weight of the mixed weight of hyaluronic acid and BDDE at room temperature (20 to 25°C) for 15 to 20 minutes, then taken out and aged a second time at room temperature to produce the hydrogel of Example 1.

[0105] Examples 2 to 6

[0106] Hydrogels of Examples 2 to 6 were prepared in the same manner as in Example 1, except that 30 parts by weight each of crosslinking regulators (CC1 to CC5) according to Table 1 below was additionally mixed with 100 parts by weight of the mixed weight of hyaluronic acid and BDDE in the purified water.

[0107] Meanwhile, the hydrophilic polymer compound, crosslinking agent, glycerol, and compounds represented by the following chemical formulas 3 and 4 used in the present invention were purchased from Tokyo Chemical Co., Ltd.

[0108] CC1CC2CC3CC4CC5Glycerol100100100100100Compound represented by chemical formula 32530405055Compound represented by chemical formula 42530405055

[0109] (Unit: parts by weight)

[0110] [Chemical Formula 3]

[0111]

[0112] [Chemical Formula 4]

[0113]

[0114] Experimental example

[0115] Thickness measurement of sheet-shaped hydrogels

[0116] Two types of outer packaging materials (Sample 1, Sample 2) containing sheet-shaped hydrogels of different thicknesses were manufactured using the method of Example 1. The thicknesses of each sample were measured at three or more locations using thickness gauges (n=5). The results are shown in Table 2 below:

[0117] Sample ThicknessMVTR (g / m 2 ·day)1100-200um1.62200-300um5.2

[0118] Referring to Table 2 above, it can be confirmed that each sample has a thickness suitable for external packaging material, and the water vapor permeability (MVTR) is also confirmed to be within an appropriate range, confirming that it can be used as a packaging material for the purpose of storing an acellular dermal matrix with excellent moisture retention, which is the purpose of the present invention.

[0119] Confirmation of thickness and weight changes before and after water absorption of sheet-shaped hydrogels

[0120] The sheet-shaped hydrogel manufactured in Example 1 was immersed in purified water for 10 minutes, then taken out and allowed to absorb moisture. Thereafter, changes in thickness and weight before and after moisture absorption were measured using thickness gauges and a precision balance, respectively, and the results are shown in Fig. 1 (n=5).

[0121] Referring to Figure 1, it can be confirmed that the thickness and weight before and after water absorption are greatly increased, respectively, and it can be seen that the hydrogel of the present invention exhibits excellent water absorption ability by including a cross-linked hydrophilic polymer compound.

[0122] Determination of moisture retention or retention capacity of sheet-shaped hydrogels

[0123] In order to confirm the moisture retention or maintenance ability when storing an acellular dermal matrix using the hydrogel of the present invention, an acellular dermal matrix was first prepared as follows.

[0124] After purchasing cadaveric skin tissues from EURO skin bank, Allosource, and CTS, tissues larger than 1 mm were selected and used. Thereafter, 1) the skin tissues were washed for the first time using distilled water at 20 to 27°C for 10 to 12 hours through a shaking incubator. 2) Afterwards, the skin tissues that had completed the first wash were immersed in a composition containing 100 parts by weight of an epidermal separator mixed with 1 M NaCl and 0.1% EDTA in a 1:1 weight ratio, 45 parts by weight of a nonionic surfactant (0.25% Triton X-100), 45 parts by weight of 0.25% sodium deoxycholate, and 25 parts by weight of 0.5% thioglycolic acid for 10 to 12 hours at 35 to 38°C. 3) Afterwards, the processed skin tissue through the above immersion treatment step was washed a second time with PBS buffer at 20 to 27°C for 10 hours to remove impurities, thereby producing a 2 x 2 cm acellular dermal matrix.

[0125] Afterwards, the 6 x 6 cm hydrogel sheets of different thicknesses manufactured in Example 1 were immersed in purified water for 10 minutes, taken out, and after the hydrogel absorbed moisture, the acellular dermal matrix was wrapped with them, and each was sealed with a packaging material.

[0126] Meanwhile, for comparative experiments, acellular dermal matrix not wrapped with the hydrogel sheet was used, sealed with packaging material.

[0127] Afterwards, after storing in an oven at 37℃ and 60℃ for 7 or 14 days, respectively, the moisture evaporation rate was calculated using Equation 2 below, and the results are as shown in Figure 2:

[0128] [Formula 2]

[0129]

[0130] (Here, W7 is the weight of the acellular dermal matrix sealed with packaging material after 7 days, and W 14 is the weight of the acellular dermal matrix sealed with packaging material after 14 days, and W0 is the weight of the acellular dermal matrix sealed with packaging material initially.)

[0131] Meanwhile, the acellular dermal matrix sealed with the above packaging material was stored in ovens at 37°C and 60°C for 14 days, respectively, and the moisture content of the acellular dermal matrix was measured using a moisture meter, and the results are shown in Fig. 3.

[0132] Referring to the above Fig. 2, it was confirmed that when an acellular dermal matrix was wrapped with hydrogel sheets of the present invention manufactured to different thicknesses, compared to a case where the acellular dermal matrix was not wrapped with the hydrogel sheet of the present invention, a relatively small weight loss occurred over time under each temperature condition.

[0133] In addition, referring to FIG. 3, it was confirmed that the moisture content of the acellular dermal matrix was maintained excellently when the acellular dermal matrix was wrapped with the hydrogel sheets of the present invention manufactured with different thicknesses, compared to when the acellular dermal matrix was not wrapped with the hydrogel sheets of the present invention under each temperature condition.

[0134] Through this, it can be seen that the hydrogel sheet of the present invention exhibits excellent moisture retention properties, and when it is used to wrap an acellular dermal matrix, moisture can be supplied to the acellular dermal matrix for a long period of time, and ultimately, the storage and distribution properties of the acellular dermal matrix can be improved.

[0135] Meanwhile, using the same method as the moisture retention test using the hydrogel sheet of Example 1, moisture retention tests of Examples 2 to 6 were conducted, and the results are shown in Table 3 below.

[0136] For relative evaluation, the results were evaluated on an index of 1 to 10, and the results of the moisture retention experiment using the hydrogel sheet of Example 1 were fixed at index 3. Meanwhile, a case in which the acellular dermal matrix not wrapped with the hydrogel sheet of the present invention was sealed with a packaging material was tested in the same manner as above as a comparative example.

[0137] The above index means that the higher the number, the lower the moisture evaporation rate and the higher the moisture content, ultimately indicating excellent moisture retention. The lower the number, the higher the moisture evaporation rate and the lower the moisture content, ultimately indicating poor moisture retention.

[0138] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Comparative Example Moisture Evaporation Rate 369101061 Moisture Content 35910961

[0139] (unit index)

[0140] Referring to Table 3 above, it can be confirmed that in the case of Examples 2 to 6 that additionally include a crosslinking regulator, moisture evaporation is suppressed and the moisture content is maintained excellently, thereby maintaining moisture retention properties excellently for a long period of time, compared to the case that does not include the crosslinking regulator. More specifically, it can be seen that the effect is maximized in the case of Examples 3 to 5. On the other hand, it was confirmed that the moisture retention properties were the poorest in the case of the comparative examples that did not include the hydrogel of the present invention.

[0141] Long-term distribution through inhibition of bacterial contamination

[0142] In the case of acellular dermal matrix contaminated with bacteria, the dermal layer is shed in the form of floating matter.

[0143] Therefore, based on these indicators, an experiment was conducted to confirm the long-term distribution of acellular dermal matrix through inhibition of bacterial contamination.

[0144] Specifically, using the same method as the moisture retention test described above, the hydrogel sheets of Examples 1 to 7 were immersed in purified water for 10 minutes, taken out, and allowed to absorb moisture. These sheets were then used to wrap acellular dermal matrices, and each was sealed with packaging material. Afterwards, they were stored at 37°C for 60 days, and the surface of the acellular dermal matrices was visually observed to determine whether or not a dermal layer detached in the form of floating matter was present.

[0145] The results were evaluated on an index of 1 to 10, with a higher number indicating the absence of dermal layers that have fallen off in the form of floating particles. On the other hand, a lower number indicates the presence of a large amount of dermal layers that have fallen off in the form of floating particles, resulting in poor long-term distribution or storage properties.

[0146] For relative comparison, the experimental results using the hydrogel sheet of Example 1 were fixed at index 3, and a case in which the acellular dermal matrix not wrapped with the hydrogel sheet of the present invention was sealed with a packaging material was tested in the same manner as above as a comparative example.

[0147] The results are shown in Table 4.

[0148] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Comparative Example Dermal layer detached in the form of floating matter 35109961

[0149] (unit index)

[0150] Referring to Table 4 above, it can be confirmed that in the case of Examples 2 to 6 that additionally include a cross-linking regulator, the adsorption or binding effect of antibiotics is enhanced compared to the case that does not include the cross-linking regulator, thereby effectively suppressing bacterial contamination during the storage and distribution process of the dermal matrix, and ultimately demonstrating long-term storage or distribution properties. More specifically, it can be seen that the effect is maximized in the case of Examples 3 to 5.

[0151] Meanwhile, in the case of the comparative example in which the hydrogel of the present invention is not wrapped, it can be seen that the long-term storage or distribution property is relatively poor.

[0152] Effective cross-linking ratio (crR) analysis

[0153] The effective crosslinking rate analysis for the hydrogels of Examples 1 to 6 was performed, and specifically, the method described in the literature [Kenne et al., Carbohydrate Polymers 91 (2013) 410-418] was used. The HPLC system was Waters Acquity UPLC I-Class, and the MS / MS system was Xevo TQS micro QQQ. The results are shown in Table 5 below:

[0154] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 CrR 1.0 1.5 1.8 2.0 1.9 1.6

[0155] Referring to Table 5 above, it can be confirmed that the hydrogel of the present invention has an effective crosslinking ratio in the range of 1.0 to 2.0, and more specifically, in the case of Examples 2 to 6, it can be confirmed that the effective crosslinking ratio is 1.5 to 2.0. In addition, in the case of Examples 3 to 5, it can be confirmed that the effective crosslinking ratio is 1.8 to 2.0.

[0156] It can be seen that when the crosslinking regulator of the present invention is included, the effective crosslinking ratio can be adjusted to a range of 1.5 to 2.0 or 1.8 to 2.0, and within the crosslinking ratio range, moisture retention can be maximized, and antibiotic adsorption and binding power can also be maximized, thereby improving the storage and distribution properties of the acellular dermal matrix.

[0157]

[0158] Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0159] The present invention relates to a hydrogel composition for storing an acellular dermal matrix, a hydrogel for storing an acellular dermal matrix with excellent moisture retention properties including the same, and a method for manufacturing the same.

Claims

1. Containing hydrophilic polymer compounds and antibiotics Hydrogel composition for storage of acellular dermal matrix.

2. In paragraph 1, The hydrophilic polymer compound is selected from the group consisting of carboxymethyl cellulose (CMC), biocellulose, hyaluronic acid (HA), gelatin, cellulose, gellan gum, agar, agarose, poloxamer, and mixtures thereof. Hydrogel composition for storage of acellular dermal matrix.

3. In paragraph 1, The above antibiotic is selected from the group consisting of penicillin, streptomycin, kanamycin, neomycin, bacitracin, gentamicin, vancomycin, beta-lactam antibiotics, aminoglycoside antibiotics, macrolide antibiotics, tetracycline antibiotics, glycopeptide antibiotics, lincomycin antibiotics, quinolone antibiotics, and mixtures thereof. Hydrogel composition for storage of acellular dermal matrix.

4. A hydrogel composition comprising any one of claims 1 to 3. Hydrogel for storage of acellular dermal matrix with excellent moisture retention properties.

5. In paragraph 4, The above acellular dermal matrix storage hydrogel exhibits excellent moisture retention properties by including a cross-linked hydrophilic polymer compound. Hydrogel for storage of acellular dermal matrix with excellent moisture retention properties.

6. In paragraph 4, The above acellular dermal matrix storage hydrogel is in sheet form, The thickness of the above hydrogel sheet is 1 mm to 5 mm. Hydrogel for storage of acellular dermal matrix with excellent moisture retention properties. 7.1) A step of preparing a hydrogel solution including a cross-linked hydrophilic polymer compound by mixing purified water, a hydrophilic polymer compound, and a cross-linking agent; 2) a step of forming a hydrogel by hardening the hydrogel solution; and 3) A step of immersing the formed hydrogel in an impregnation solution containing an antibiotic. A method for manufacturing a hydrogel for storing an acellular dermal matrix with excellent moisture retention properties.

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