Composition containing dry-fibrillated acellular dermal matrix and method for producing same

Abstract

To provide a composition comprising a fibrous acellular dermal matrix having enhanced physical properties, which is easily applicable even to a bent wound site, and a method for producing the same.SOLUTION: A composition comprising 48 to 95.5 wt.% of a fibrous acellular dermal matrix and 0.5 to 52 wt.% of water, wherein a biocompatible polymer is not added, an average tensile strength value of 0.2 N / mm2 or more is satisfied, and the composition is a sheet type.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition containing a fiberized acellular dermal matrix and a method for producing the same, and to a composition containing a fiberized acellular dermal matrix to which a dry method is applied and a method for producing the same. [Background technology]

[0002] A wound is a condition in which external pressure causes a cut or defect in the skin or other tissue, disrupting the continuity of the tissue. It usually occurs when the dermis layer of the skin is damaged, resulting in an opening of the skin. Wound repair surgery basically involves suturing and bandaging the wound to block exposure to the external environment, thereby preventing infection and suppressing inflammatory reactions. Wound dressing materials can be broadly divided into allogeneic dermis and xenogeneic dermis, and wound dressings can be manufactured by extracting specific polymers from the material or by using a dermal matrix.

[0003] Acellular dermal matrix (ADM) is made by removing the epidermis from donated cadaver skin tissue and then removing the cells within the dermis to prevent immune rejection. It is widely used in soft tissue reconstruction surgery and in allografts for burn treatment. Dermal tissue is composed of 80-90% collagen, elastin, and glycosaminoglycans.

[0004] Meanwhile, in 1994, LifeCell developed Alloderm (allograft), an acellular dermal matrix (ADM) made by decellularizing and freeze-drying skin tissue harvested from cadavers, for use in burn treatment and skin reconstruction. Alloderm (allograft) is safer than xenografts and has demonstrated superior graft survival and healing effects. Similar products have been developed, including Bard Dabol's AlloMax and Ethicon's FlexHD. However, these products have issues with their lack of flexibility, making them difficult to use on curved or deep wounds.

[0005] To address these issues, LifeCell developed a technology to granulate acellular dermal matrix and launched the injectable micronized AlloDerm (Cymetra) product in 1999. Wright Medical Group also developed the injectable Graftjacket product and launched the Graftjacket Xpress, while L&C Bio developed a composition in 2014 in which acellular dermal matrix was granulated and then crosslinked with hyaluronic acid. However, while compositions in which acellular dermal matrix particles are crosslinked with biocompatible polymers have high structural stability, they also have high crosslinking, viscoelasticity, hardening, and extrusion force. While these compositions are suitable for use as fillers and shaping agents, they are still insufficient for use as wound dressings. Furthermore, the micronized form has problems with its liquid form, which makes it difficult to adhere to the wound and can easily leak out of the dressing, preventing even distribution across the wound.

[0006] Therefore, there is a pressing need to develop a form of acellular dermal matrix that can be fixed to the wound site. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Republic of Korea Patent No. 10-1523878 (2015.05.21) Summary of the Invention [Problem to be solved by the invention]

[0008] The present invention has been derived to solve the above-mentioned problems, and provides a composition comprising a dried fibrous acellular dermal matrix that is easily applied to curved wound sites and has enhanced physical properties by applying a mold drying process without using a biocompatible polymer, and a method for producing the same. [Means for solving the problem]

[0009] To achieve the above object, a composition containing a dried fiberized acellular dermal matrix according to one embodiment of the present invention comprises 48 to 95.5 wt % of a fiberized acellular dermal matrix and 0.5 to 52 wt % of water, without the addition of a biocompatible polymer.

[0010] According to another embodiment of the present invention, a method for producing a composition comprising a fibrous acellular dermal matrix by drying includes the steps of: (a) pulverizing an acellular dermal matrix to produce a fibrous acellular dermal matrix; (b) adding water to the fibrous acellular dermal matrix to rehydrate it; and (c) drying the rehydrated fibrous acellular dermal matrix. [Effects of the Invention]

[0011] According to one embodiment of the present invention, a composition comprising a dry-fibered acellular dermal matrix and a method for producing the same does not require the addition of a biocompatible polymer, does not experience changes in physical properties due to heat, and does not require a step for heat-treating the biocompatible polymer, thereby simplifying the production process and reducing production costs.

[0012] In addition, because it has a structure very similar to the dermal matrix of the wound site of the human body, it has excellent bioadhesion and in-body preservation properties, maintains a moist environment well, has excellent therapeutic effects, and can be easily applied to curved wound sites.

[0013] Additionally, the mold drying step may improve physical properties and tensile strength, making it easier for the practitioner to use during treatment. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a process diagram illustrating a method for producing a composition including a dry-fibered acellular dermal matrix according to another embodiment of the present invention. [Figure 2] 1 is a photograph showing the weight of a composition containing a fibrous acellular dermal matrix according to Example 1 before and after high-temperature drying treatment. [Figure 3] 1 is a photograph showing compositions containing dried fibrous acellular dermal matrices prepared in Example 1 and Comparative Examples 1 to 3. [Figure 4] 10 is a graph showing the results of Experimental Example 2. [Figure 5] 10 is a photograph showing the results of Experimental Example 3. [Figure 6] 10 is a graph showing the results of Experimental Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention will be described in detail below by way of embodiments with reference to the accompanying drawings. However, the following embodiments are presented as examples of the present invention, and if a detailed description of a well-known technology or configuration known to those skilled in the art is deemed to unnecessarily obscure the gist of the present invention, the detailed description may be omitted, and the present invention is not limited thereby. The present invention is susceptible to various modifications and applications within the scope of the claims set forth below and within the equivalent category construed therefrom.

[0016] Furthermore, the terminology used in this specification is used to appropriately describe preferred embodiments of the present invention, and may vary depending on the intention of the user or operator, the practice of the field to which the present invention pertains, etc. Therefore, definitions of these terms should be based on the contents of this specification as a whole. Throughout the specification, when a part is said to "comprise" a certain element, this means that it may further include other elements, rather than excluding other elements, unless otherwise specified.

[0017] Throughout this specification, "%" used to indicate the concentration of a particular substance is % (w / w) for solid / solid, % (w / v) for solid / liquid, and % (v / v) for liquid / liquid, unless otherwise specified.

[0018] A composition containing a fiberized acellular dermal matrix according to one embodiment of the present invention will be described in detail below.

[0019] The composition containing a fiberized acellular dermal matrix of this embodiment comprises a fiberized acellular dermal matrix and water.

[0020] Acellular dermal matrix (ADM) exhibits superior biocompatibility with higher cell adhesion capacity and lower immune response than conventional animal-derived products, making it a widely used component in plastic surgery and orthopedics for the reconstruction, regeneration, and strengthening of skin, muscles, and ligaments. The fibrous acellular dermal matrix used in this embodiment may contain 90% or more of particles with a major axis length of 10-3,000 μm, preferably 100-2,000 μm, and more preferably 50-90% of particles with a major axis length of 200-800 μm. However, if a large number of particles less than 100 μm are included, processing into a sheet type may result in improper formation or only a film-like form that is difficult to flexibly adhere to the wound site. The fibrous acellular dermal matrix may be present in an amount of 48-95.5 wt%.

[0021] Water is an ingredient added for rehydration and can be contained in amounts of 0.5 to 52% by weight; however, if the amount of water is less than 0.5% by weight, it may be difficult to hydrate the acellular dermal matrix, and if the amount of water exceeds 52% by weight, excessive use may make it difficult to achieve the desired formulation.

[0022] Although a biocompatible polymer is not added in this embodiment, the physical properties of the biocompatible polymer are not changed by heat, and a step for heat treatment of the biocompatible polymer is not required, which may simplify the manufacturing process and reduce manufacturing costs. Furthermore, since the structure is very similar to the dermal matrix of the wound site of the human body, it has excellent bioadhesion and in-body preservation properties, maintains a moist environment well, has excellent therapeutic effects, and may be easily applied to curved wound sites.

[0023] In this embodiment, the fibrous acellular dermal matrix may refer to a granulated acellular dermal matrix in which the individual loose fibers are not spherical or streamlined but have a thin, thread-like fiber shape.

[0024] This embodiment may further comprise stem cells, growth factors, or a mixture thereof.

[0025] In this embodiment, the composition may further contain one or more selected from antimicrobial agents, excipients, and additives used pharmaceutically.

[0026] Antibacterial agents include short-chain alcohols, benzalkonium chloride (BAC), and didecyl dimethyl ammonium chloride. chloride, DDAC), zeolite (CWT-A), isothiazolones, alkyldimethylammonium chlorides, triazines, 2-thiocyanomethylthiobenzothiazole, methylene bisthiocyanate, acrolein, dodecylguanidine hydrochloride, chlorophenols, quaternary ammonium salts, glutaraldehyde, dithiocarbamates, 2-mercaptobenzothiazole, para-chloro-meta-xylenol, silver, chlorhexidine, polyhexamethylene biguanide, n-halamines, triclosan, phospholipids, alpha-hydroxy acids, 2,2-dibromo-3-nitrilopropionamide, 2-bromo-2-nitro-1,3-propanediol, panesol, iodine, bromine, hydrogen peroxide, chlorine dioxide, vegetable oils, vegetable extracts, benzalkonium chloride, chlorine, and sodium hypochlorite can be used.

[0027] The excipient may be one or more selected from stabilizers, antioxidants, osmolality adjusters, buffers, and pH adjusters, and specifically, one or more selected from starch, cellulose, glucose, lactose, sucrose, gelatin, corn, rice, wheat flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, glycerol, propylene glycol, water, and ethanol.

[0028] The additives may be one or more selected from physiologically biocompatible buffers (tromethamine hydrochloride), chelating agents (DTPA or DTPA-bisamide), and calcium chelate complexes (calcium DTPA, CaNaDTPA-bisamide), and in some cases, calcium or sodium salts (calcium chloride, calcium ascorbate, calcium gluconate, or calcium lactate).

[0029] The composition may further comprise one or more of stem cells and growth factors.

[0030] In this embodiment, the composition containing the fibrous acellular dermal matrix can be prepared in the form of an aqueous solution, suspension, emulsion, paste, cream, balm, ointment, foam, sheet, gel, gum, spray, slurry, film, granules, patch, powder, etc., preferably in the form of a sheet, and can be used as a dressing, adhesive, surgical and medical device, artificial skin, bandage, foaming agent, anti-adsorption agent, or graft material.

[0031] Hereinafter, a method for producing a composition containing a fiberized acellular dermal matrix according to another embodiment of the present invention will be described in detail with reference to the drawings.

[0032] FIG. 1 is a flow chart illustrating a method for producing a composition containing a fiberized acellular dermal matrix according to another embodiment of the present invention.

[0033] Referring to FIG. 1, first, an acellular dermal matrix is ​​pulverized to prepare a fibrous acellular dermal matrix (S10).

[0034] The acellular dermal matrix can be pulverized using one or more mills selected from the group consisting of a cutting mill, food processor, agate mill, freeze mill, micronizer, vibratory micro mill, jaw crusher, mortar grinder, planetary mill, disk mill, ball mill, knife mill, and variable speed rotor mill to produce a fibrous acellular dermal matrix. In this case, the mill is preferably set to a rotation speed of 500 to 2000 rpm to facilitate pulverization in a short period of time.

[0035] Meanwhile, in this embodiment, the acellular dermal matrix may be prepared by the steps of preparing human-derived skin tissue that has not been subjected to a separate epidermal and fat removal process, treating the skin tissue with a hypotonic solution containing a surfactant, and washing the treated skin tissue with an isotonic solution. Here, it is preferable to use an acellular dermal matrix having a thickness of 0.1 mm to 3 mm.

[0036] Next, water is added to the fiberized acellular dermal matrix to rehydrate it (S20).

[0037] The acellular dermal matrix prepared in step S10 can be hydrated by adding 300 to 2,000 parts by weight of water to 100 parts by weight of the acellular dermal matrix. If the water content is less than 300 parts by weight, it becomes difficult to hydrate the fiberized acellular dermal matrix, and if it exceeds 2,000 parts by weight, excessive use may result in a longer heat drying time, as described below, which is not preferable. In this embodiment, the water may be sterilized distilled water.

[0038] The rehydrated fibrous acellular dermal matrix is ​​then heat-dried (S30).

[0039] The fiberized acellular dermal matrix rehydrated in step S20 can be heat-dried in a mold, preferably a lattice-shaped mold, at 30 to 50°C for 6 hours or more, for example, 10 hours or more, for example, 10 to 24 hours, preferably 16 to 18 hours, until the moisture content reaches 0.5 to 52% by weight. If the temperature is lower than 30°C, drying may take an excessively long time, while if the temperature exceeds 50°C, the human tissue may be deformed, resulting in a decrease in amino acid content, as well as a decrease in tensile strength and degree of hydration.

[0040] The average tensile strength of the fiberized acellular dermal matrix under the above-mentioned heat drying conditions was 0.2 N / mm 2 By satisfying the above requirements, it may be easier for the practitioner to use the device during treatment than before.

[0041] After step S30, the method may further include processing the heat-dried fiberized acellular dermal matrix into a sheet.

[0042] The heat-dried fiberized acellular dermal matrix in step S30 can be applied to a mold or frame, shaped, and processed into a sheet type, and it can be manufactured in various width, length, and thickness sizes required depending on the wound site or surgical characteristics.

[0043] Meanwhile, before processing into a sheet type, a non-chemical crosslinking process using radiation, reduced pressure, and heat or a chemical crosslinking process using a crosslinking agent may be further carried out depending on the physical properties (degree of crosslinking, degree of hardening, etc.).

[0044] The present invention will be described in more detail below using examples. These examples are merely for the purpose of more specifically explaining the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples.

[0045] Manufacturing Example 1. Manufacturing of acellular dermal matrix Skin tissues were purchased from EURO Skin Bank, Allosource, and CTS. Tissues with a thickness of 0.5 mm or greater were selected. Adherent adipose tissue was removed using forceps and then washed three times in a sterile environment. The tissues were then immersed in a hypotonic solution (Tris-HCl, EDTA, NaOH, and SDS) for six hours. The skin tissues were then washed with PBS at 4°C to remove any remaining adipose tissue, epidermis, cells, and hypotonic solution. The tissues were then left overnight and then washed in an isotonic solution (Tris-HCl, EDTA, NaCl, and NaOH) for six hours to prepare de-epidermized, de-lipidated, and decellularized human-derived acellular dermal matrices.

[0046] Example 1. Preparation of a composition containing a fiberized acellular dermal matrix The human-derived acellular dermal matrix prepared in Preparation Example 1 was pulverized using a cutting mill to produce a fibrous acellular dermal matrix. The matrix was rehydrated by adding 10 times sterile distilled water, placed in a lattice-shaped mold, and dried by heating at 30°C to 50°C for at least 6 hours. The matrix was then applied to a metal mold, shaped, and processed into a sheet to produce a composition containing a fibrous acellular dermal matrix.

[0047] Meanwhile, the weight of the composition containing the fibrous acellular dermal matrix of Example 1 before and after heat drying treatment at a high temperature of 126° C. for 10 minutes or more is shown in FIG.

[0048] Referring to Figure 2(c), it can be seen that the composition containing fibrous acellular dermal matrix contains 48.32 wt% fibrous acellular dermal matrix and 51.68 wt% water, based on the pre-drying weight of 0.387 g shown in Figure 2(a) and the post-drying weight of 0.187 g shown in Figure 2(b).

[0049] *In Example 1, drying did not progress overnight at room temperature, which is a temperature below 30°C.

[0050] Comparative Example 1. Preparation of a composition containing a fibrous acellular dermal matrix The same method as in Example 1 was used, except that the heat treatment was carried out at 60° C. for 6 hours or more.

[0051] Comparative Example 2. Preparation of a composition containing a fibrous acellular dermal matrix The same method as in Example 1 was used except that the heat treatment was carried out at 70° C. for 6 hours or more.

[0052] Comparative Example 3. Preparation of a composition containing a fibrous acellular dermal matrix The same method as in Example 1 was used, except that the heat treatment was carried out at 100° C. for 6 hours or more.

[0053] Experimental example 1. Appearance evaluation The appearance of the compositions containing the fibrous acellular dermal matrices prepared in Example 1 and Comparative Examples 1 to 3 was evaluated by visual inspection.

[0054] As a result, as shown in Figure 3, it was confirmed that Example 1 had a structure that was very similar in appearance to the dermal matrix of a wound site in a human body, while Comparative Examples 1 and 2 showed slight differences in appearance from the dermal matrix of a wound site in a human body, and it was confirmed that the tissue in Comparative Example 3 was burned.

[0055] As described above, the composition containing the fiberized acellular dermal matrix of the present invention does not need to be heated to a high temperature because a biocompatible polymer is not added, and there is no change in physical properties due to heat. Furthermore, since a step for heat treatment of the biocompatible polymer is not required, it has been confirmed that the manufacturing process can be simplified and manufacturing costs can be reduced.

[0056] Experimental Example 2: Evaluation of amino acid content by heat treatment temperature To confirm the change in amino acid content depending on the heat treatment temperature, the amino acid content of Example 1 and Comparative Examples 1 and 2 was measured using an automatic amino acid analyzer (S433D, Sykam GmbH Co., Germany), and the results are shown in FIG.

[0057] Referring to FIG. 4, it can be seen that the amino acid content decreases from Example 1 to Comparative Examples 1 and 2, which is presumed to be due to the occurrence of human tissue deformation in Comparative Examples 1 and 2.

[0058] As described above, the composition containing the fibrous acellular dermal matrix prepared according to the examples of the present invention has a structure very similar to that of the dermal matrix at the wound site of the human body, and therefore has excellent bioadhesion and in vivo preservation properties, thereby improving the therapeutic effect and facilitating application to curved wound sites.

[0059] Experimental Example 3. Evaluation of the degree of hydration depending on the heat treatment temperature In order to confirm the degree of hydration depending on the heat treatment temperature, Example 1 and Comparative Examples 1 and 2 were placed in a tube containing water and observed, and the results are shown in FIG.

[0060] Referring to FIG. 5, it can be seen that Example 1 sank within a few seconds, while Comparative Examples 1 and 2 continued to float without sinking, which is expected to be due to the effects of drying and shrinkage caused by high temperature.

[0061] As described above, it can be seen that the composition containing the fiberized acellular dermal matrix prepared according to the examples of the present invention maintains a moist environment well, has excellent therapeutic effects, and is easily applied to curved wound sites.

[0062] Experimental Example 4. Evaluation of tensile strength depending on heat treatment temperature The results are shown in Figure 5 to confirm the effect of heat treatment temperature on tensile strength.

[0063] Referring to FIG. 6, it was confirmed that the tensile strength of Comparative Examples 1 and 2 was reduced compared to that of Example 1.

[0064] As described above, the composition containing the fiberized acellular dermal matrix prepared according to the embodiment of the present invention does not need to be heated to a high temperature because a biocompatible polymer is not added, and therefore the physical properties are not changed by heat. Furthermore, the physical properties and tensile strength are improved by performing a step of thermal drying, particularly thermal drying in a mold, making it easy for practitioners to use during treatment.

[0065] Although exemplary 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.

[0066] Unless otherwise defined, all technical terms used in this specification are used in the same sense as commonly understood by those of ordinary skill in the art in the relevant field of this invention. The contents of all publications referenced in this specification are incorporated herein by reference.

Claims

1. 48 to 95.5% by weight of a fibrous acellular dermal matrix; 0.5 to 52% by weight of water; A composition comprising a fiberized acellular dermal matrix, comprising: The composition does not contain an added biocompatible polymer, The fiberized acellular dermal matrix is ​​heat-dried at 30 to 50°C for 6 hours or more, The average tensile strength of the fiberized acellular dermal matrix was 0.2 N / mm ２ The above conditions are met, The composition is used as a wound dressing. A composition comprising a fibrous acellular dermal matrix.

2. 10. The composition comprising the fiberized acellular dermal matrix of claim 1, further comprising stem cells, growth factors, or a mixture thereof.

3. The composition comprising a fiberized acellular dermal matrix according to claim 1 , wherein the composition is in a sheet form.

4. (a) pulverizing an acellular dermal matrix to produce a fibrous acellular dermal matrix; (b) adding water to rehydrate the fiberized acellular dermal matrix; (c) heat-drying the rehydrated fiberized acellular dermal matrix; A method for producing a composition comprising a fiberized acellular dermal matrix, comprising: In step (c), the heat drying is carried out at 30 to 50°C for 6 hours or more; The average tensile strength of the fiberized acellular dermal matrix was 0.2 N / mm ２ The above conditions are met, The composition is used as a wound dressing. A method for producing a composition comprising a fiberized acellular dermal matrix.

5. The method for producing a composition comprising a fiberized acellular dermal matrix according to claim 4 , wherein the heat drying in step (c) is carried out in a mold.

6. 5. The method for producing a composition comprising a fiberized acellular dermal matrix according to claim 4, further comprising, after step (c), (d) processing the heat-dried fiberized acellular dermal matrix into a sheet type.

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