Composition comprising fibrous acellular dermal matrix prepared by drying, and method for preparing same

Abstract

The present invention relates to: a composition comprising a fibrous acellular dermal matrix, which does not employ a biocompatible polymer and is subjected to a drying process using a sieve, thereby exhibiting improved physical properties, tensile strength, and suture strength, and thus providing enhanced usability compared to conventional compositions; and a method for preparing same.

Description

Composition comprising a fibrous acellular dermal matrix of a dry type and method for preparing the same

[0001] The present invention relates to a composition comprising a fibrous acellular dermal matrix and a method for producing the same, and more particularly, to a composition comprising a fibrous acellular dermal matrix to which a drying method is applied and a method for producing the same.

[0002] A wound is a condition in which the continuity of the skin or other tissue is disrupted or damaged by external pressure. It typically refers to damage to the dermal layer, resulting in an opening of the skin. Wound surgery fundamentally involves suturing and dressing the wound site to prevent exposure to the external environment, thereby preventing infection and suppressing inflammation. Wound dressing materials can be broadly categorized into allogeneic and xenogeneic dermis. Wound dressings can be manufactured by extracting specific polymers from the material or by utilizing a dermal matrix.

[0003] Acellular dermal matrix (ADM) is a dermal matrix obtained by removing the epidermis and dermal cells from donated cadaveric skin to eliminate immune rejection. It is widely used in soft tissue reconstructive surgery and as an allograft for burn treatment. Dermal tissue is composed of 80-90% collagen, elastin, and glycosaminoglycans.

[0004] Meanwhile, in 1994, LifeCell developed an acellular dermal matrix (ADM) product (Alloderm (allograft)) made by decellularizing and freeze-drying skin tissue harvested from cadavers, and used it for burn treatment and skin reconstruction. Alloderm (allograft) was safer than other products and showed a significantly better survival rate and healing effect. Similar products were developed, such as AlloMax from Bard Dabol and FlexHD from Ethicon. However, these products had the problem of poor flexibility, making them difficult to use on curved or deep wound areas.

[0005] To solve these problems, LifeCell developed a technology to granulate acellular dermal matrix in 1999 and launched AlloDerm (Cymetra) in an injectable micronized form. Wright Medical Group also developed the Graftjacket product in an injectable form and launched the Graftjacket Xpress product. In 2014, L&C Bio developed a composition that granulated acellular dermal matrix and cross-linked it with hyaluronic acid. However, the composition that cross-links acellular dermal matrix particles and a biocompatible polymer as described above has high structural stability, but it also has high cross-linking degree, viscoelasticity, hardness, and extrusion force, making it suitable for use as a filler or implant. However, it is still inadequate for use as a wound dressing. In the case of the micronized form, there was a problem that it could not be fixed to the wound site in liquid form and flowed out of the dressing, preventing it from being evenly distributed on the wound.

[0006] Therefore, there is an urgent need to develop an acellular dermal matrix that can be fixed to the wound site.

[0007] The purpose of the present invention is to provide a composition including a fibrous acellular dermal matrix with enhanced physical properties and a method for manufacturing the same, which is easy to apply to curved wound areas by applying a drying process in a sieve without applying a biocompatible polymer.

[0008] In order to achieve the above object, a composition comprising a fibrous acellular dermal matrix in a dry manner according to one embodiment of the present invention comprises 48 to 95.5 wt% of a fibrous acellular dermal matrix and 0.5 to 52 wt% of water, wherein no biocompatible polymer is added, and the fibrous acellular dermal matrix comprises an antimicrobial agent at a concentration of 1 to 1,000,000 U / mL.

[0009] A method for producing a composition comprising a fibrous acellular dermal matrix in a dry manner according to another embodiment of the present invention comprises the steps of (a) producing a fibrous acellular dermal matrix by pulverizing an acellular dermal matrix, (b) rehydrating the fibrous acellular dermal matrix by adding water, and (c) drying the rehydrated fibrous acellular dermal matrix.

[0010] According to one embodiment of the present invention, a composition including a fibrous acellular dermal matrix in a dry manner and a method for manufacturing the same do not require a biocompatible polymer to be added, so there is no change in physical properties due to heat, and a step for heat-treating a biocompatible polymer is not required, so that the manufacturing process can be simplified and the manufacturing cost can be reduced.

[0011] In addition, since it has a structure very similar to the dermal matrix of the wound site of the human body, it has excellent bioadhesiveness and body preservation, and it is good at maintaining a moist environment, so it has an excellent treatment effect and can be easily applied to curved wound sites.

[0012] In addition, by performing the drying step on a sieve, the physical properties, tensile strength and sealing strength are improved, so that the practitioner can use it more easily during the procedure, and it can also act as a physical barrier and be used as an anti-adhesion agent.

[0013] FIG. 1 is a process diagram illustrating a method for manufacturing a composition including a fibrous acellular dermal matrix in a dry manner according to another embodiment of the present invention.

[0014] Figures 2 and 3 are graphs showing the results according to Experimental Example 1.

[0015] Figure 4 is a photograph showing the results according to Experimental Example 2.

[0016] Hereinafter, the present invention will be described in detail with reference to the attached drawings, using exemplary embodiments. However, the following exemplary embodiments are provided as illustrative examples of the present invention. If a detailed description of a technology or configuration well known to those skilled in the art is judged to unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the following claims and equivalents interpreted therefrom.

[0017] In addition, the terms used in this specification are terms used to appropriately express preferred embodiments of the present invention, and may vary depending on the intention of the user or operator, or the customs of the field to which the present invention belongs. Therefore, the definitions of these terms should be determined based on the contents throughout this specification. Throughout the specification, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components may be included, unless specifically stated otherwise.

[0018] 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 stated.

[0019]

[0020] Hereinafter, a composition including a fibrous acellular dermal matrix according to one embodiment of the present invention will be described in detail.

[0021] The composition comprising the fibrous acellular dermal matrix of the present embodiment comprises a fibrous acellular dermal matrix and water, wherein the fibrous acellular dermal matrix comprises an antimicrobial agent.

[0022] Acellular dermal matrix (ADM) is a component widely used in plastic surgery and orthopedics for the purpose of reconstruction, regeneration, and strengthening of skin, tendons, and ligaments due to its excellent biocompatibility with high cell adhesion ability and low immune response compared to existing animal-derived products. The acellular dermal matrix used in this example may contain 90% or more of the long axis length of 10 to 3,000 ㎛, preferably 100 to 2,000 ㎛, and more preferably 50 to 90% of the long axis length of 200 to 800 ㎛. However, if a large portion is contained less than 100 ㎛, the shape may not be properly formed when processed into a sheet type, or it may be formed only in a film form, making it difficult to flexibly adhere to a wound site. The acellular dermal matrix may be contained in an amount of 48 to 95.5 wt%.

[0023] Meanwhile, the fibrous acellular dermal matrix may contain an antimicrobial agent, which may be present at a concentration of 1 to 1,000,000 U / mL, for example, 300,000 to 700,000 U / mL. If the concentration of the antimicrobial agent is less than 1 U / mL, it may be difficult to achieve an antimicrobial effect with a value of Log 5 or higher, and if the concentration exceeds 1,000,000 U / mL, the risk of toxicity may increase, and the effect due to excessive use may not be that great.

[0024] In this example, the antibacterial agent is a component added to provide antibacterial properties, and includes Polymyxin B, Vancomycin, Amphotericin B, short-chain alcohol, benzoalkonium chloride (BAC), didecyl dimethyl ammonium chloride (DDAC), zeolite (CWT-A), isothiazolone, alkyl dimethyl ammonium chloride, triazine, 2-thiocyanomethylthio benzothiazole, methylene bis thiocyanate, acrolein, dodecyl guanidine hydrogen chloride, chlorophenol, quaternary ammonium salt, gluteraldehyde, dithiocarbamate, 2-mercaptobenzothiazole, para-chloro-meta-xylenol, silver, chlorhexidine, polyhexamethylene biguanide, At least one selected from among n-halamine, triclosan, phospholipid, alpha hydroxy acid, 2,2-dibromo-3-nitrilopropionamide, 2-bromo-2-nitro-1,3-propanediol, farnesol, iodine, bromine, hydrogen peroxide, chlorine dioxide, vegetable oil, plant extract, benzalkonium chloride, chlorine and sodium hypochlorite, for example, at least one selected from among Polymyxin B, vancomycin and amphotericin B, can be used, and when the above conditions are satisfied, a high antibacterial effect of Log 5 or higher can be exhibited.

[0025] Water is an ingredient added for rehydration and can be included in an amount of 0.5 to 52 wt%. If the amount of water is less than 0.5 wt%, it may be difficult to hydrate the fibrous acellular dermal matrix, and if it exceeds 52 wt%, it may be difficult to implement the desired formulation due to excessive use.

[0026] In this embodiment, a biocompatible polymer is not added, which eliminates heat-induced changes in physical properties and eliminates the need for heat-treating the biocompatible polymer, thereby simplifying the manufacturing process and reducing manufacturing costs. Furthermore, because the composition has a structure very similar to the dermal matrix of human wound sites, it exhibits excellent bioadhesiveness and body preservation properties, maintains a moist environment well, and thus exhibits excellent therapeutic effects and can be easily applied to curved wound sites.

[0027] In this embodiment, the term "fibrous acellular dermal matrix" may mean a particle-shaped, individual acellular dermal matrix having a thin and long fiber shape like a thread rather than a spherical or streamlined shape.

[0028] In this embodiment, stem cells, growth factors or mixtures thereof may be further included.

[0029] Additionally, the present embodiment may further include one or more excipients and additives selected from pharmaceutically used excipients and additives.

[0030] As excipients, one or more selected from stabilizers, antioxidants, osmotic pressure-adjusting agents, buffers, and pH-adjusting agents may be used, 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 may be used.

[0031] As an additive, one or more selected from physiologically biocompatible buffers (tromethamine hydrochloride), chelating agents (DTPA or DTPA-bisamide), and calcium chelating complexes (calcium DTPA, CaNaDTPA-bisamide) may be used, and optionally, calcium or sodium salts (calcium chloride, calcium ascorbate, calcium gluconate, or calcium lactate) may be used.

[0032] The composition may further comprise one or more of stem cells and growth factors.

[0033] In this embodiment, the composition including the fibrous acellular dermal matrix can be manufactured in the form of an aqueous solution, suspension, emulsion, paste, cream, balm, ointment, foam, sheet, gel, gum, spray, slurry, film, granule, patch, powder, etc., but is preferably in the form of a sheet, and can be used as a covering material, an adhesive, a surgical and medical device, an artificial skin, a bandage, a foaming agent, an anti-adsorption agent, or a graft material.

[0034]

[0035] Hereinafter, a method for manufacturing a composition including a fibrous acellular dermal matrix according to another embodiment of the present invention will be described in detail with reference to the drawings.

[0036] FIG. 1 is a process diagram illustrating a method for manufacturing a composition including a fibrous acellular dermal matrix according to another embodiment of the present invention.

[0037] Referring to Figure 1, first, an antimicrobial agent is added to an acellular dermal matrix, allowed to penetrate, freeze-dry, and then pulverized to produce a fibrotic acellular dermal matrix (S10).

[0038] A fibrous acellular dermal matrix can be manufactured by adding an antimicrobial agent at a concentration of 1 to 1,000,000 U / mL, for example, 300,000 to 700,000 U / mL, to an acellular dermal matrix, infiltrating the matrix in a low-temperature reactor, freeze-drying for 20 to 80 hours, and then grinding the matrix using one or more grinders selected from a cutting mill, a food processor, an agate grinder, a freeze grinder, a micronizer, a vibrating micro mill, a jaw crusher, a mortar grinder, a planetary mill, a disk mill, a ball mill, a knife mill, and a variable speed rotor mill. At this time, it is preferable to set the grinder to a rotation speed of 500 to 2000 rpm to facilitate grinding in a short period of time.

[0039] Meanwhile, in the present embodiment, the acellular dermal matrix may be prepared by the steps of preparing human-derived skin tissue that has not undergone separate de-epidermization and de-fat processes, treating the skin tissue with a storage solution containing a surfactant, and washing the treated skin tissue with an isotonic solution. Here, it is preferable to apply the acellular dermal matrix having a thickness of 0.1 mm to 3 mm.

[0040]

[0041] Next, water is added to the fibrous acellular dermal matrix to rehydrate it (S20).

[0042] The fibrous 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. If the water content is less than 300 parts by weight, it may be difficult to hydrate the fibrous acellular dermal matrix, and if it exceeds 2,000 parts by weight, the heat drying time described below may be prolonged due to excessive use, which is not preferable. In this embodiment, the water may be sterilized distilled water.

[0043]

[0044] Then, the rehydrated fibrous acellular dermal matrix is ​​heat-dried (S30).

[0045] In the above step S20, the rehydrated fibrous acellular dermal matrix can be heat-dried at 30 to 50°C for at least 6 hours, for example, at least 10 hours, for example, 10 to 24 hours, preferably 16 to 18 hours, through a sieve, preferably a sieve having a mesh size of 0.0001 to 5 mm, until the moisture content becomes 0.5 to 52 wt%. If the temperature is less than 30°C, excessive drying time may be required, and if it exceeds 50°C, the human tissue may be deformed, which may result in a decrease in amino acid content as well as a decrease in tensile strength and hydration degree.

[0046] The maximum tensile strength of the fibrous acellular dermal matrix under the above-described heat-drying conditions was 9.7 N / mm. 2 The maximum ideal and sealing strength is 7.3 N / mm 2 By satisfying these requirements, practitioners can utilize the material more easily than before during procedures. Furthermore, drying through a sieve not only provides a dense structure but also enables it to function as a physical barrier.

[0047]

[0048] After the above step S30, a step of processing the heat-dried fibrous acellular dermal matrix into a sheet type may be further included.

[0049] In the above S30 step, the heat-dried fibrous acellular dermal matrix can be applied to a mold or frame and then shaped and processed into a sheet type. It goes without saying that it can be manufactured in various sizes according to the width, length, and thickness required by the wound site or surgical characteristics.

[0050] Meanwhile, prior to processing into a sheet type, non-chemical crosslinking using radiation irradiation, reduced pressure, and heat, or chemical crosslinking using a crosslinking agent may be additionally performed depending on the physical properties (crosslinking degree, degree of curing, etc.).

[0051]

[0052] Hereinafter, the present invention will be described in more detail using examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0053]

[0054] Manufacturing Example 1. Manufacturing of acellular dermal matrix

[0055] Skin tissues were purchased from EURO skin bank, Allosource, and CTS, and tissues with a thickness of 0.5 mm or more were selected. The adipose tissue attached to the skin tissues was removed using forceps, and the tissues were washed three times with sterile water. Then, the tissues were immersed in a hypotonic solution (Tris-HCl, EDTA, NaOH, and SDS) and treated for 6 hours. Then, the skin tissues were washed with PBS at 4°C to remove the remaining fat, epidermis, cells, and hypotonic solution, and left overnight. After washing with an isotonic solution (Tris-HCl, EDTA, NaCl, and NaOH) for 6 hours, a deepidermal, defatted, and decellularized human-derived acellular dermal matrix was prepared.

[0056]

[0057] Example 1. Preparation of a composition comprising a fibrous acellular dermal matrix

[0058] In the human-derived acellular dermal matrix manufactured in the above Manufacturing Example 1, a 14.3% maltitol solution and an antimicrobial agent at a concentration of 1,000,000 U / mL were added and infiltrated in a low-temperature reactor at 4°C. Then, after placing it in a Tyvek (Korea Advanced Materials, Korea), it was dried in a freeze-dryer with a vacuum of 5 torr for 24 hours and pulverized with a cutting mill to fiberize it, thereby producing a fibrous acellular dermal matrix. To this, 10 times the amount of sterilized distilled water was added to rehydrate it, and it was placed on a sieve with a size of 100 μm and heat-dried at 30°C to 50°C for more than 6 hours, then applied to a mold to shape it and processed into a sheet type to produce a composition including a fibrous acellular dermal matrix.

[0059]

[0060] Comparative Example 1. Preparation of a composition including a fibrous acellular dermal matrix

[0061] It was manufactured in the same manner as Example 1, except that freeze-drying was performed instead of heat drying.

[0062]

[0063] Comparative Example 2. Preparation of a composition including a fibrous acellular dermal matrix

[0064] It was manufactured in the same manner as Example 1, except that heat drying was performed in a mold instead of a sieve.

[0065]

[0066] Experimental Example 1: Physical Characteristics Evaluation

[0067] In order to determine the physical properties of the present invention, the tensile strength and sealing strength of Example 1 and Comparative Examples 1 and 2 were measured, and the results are shown in FIGS. 2 and 3.

[0068] Referring to FIGS. 2 and 3, it was confirmed that the tensile strength and sealing strength of Comparative Examples 1 and 2 were far below those of Example 1.

[0069] It was found that the composition including the fibrous acellular dermal matrix manufactured according to Example 1 of the present invention has a dense structure and improved physical properties, tensile strength and suture strength by performing a heat drying step, particularly, a heat drying step in a sieve, so that it is easy for a practitioner to use during a procedure.

[0070]

[0071] Experimental Example 2: Antibacterial Performance Evaluation

[0072] In order to confirm the antibacterial properties of the present invention, an experiment was conducted in accordance with JIS Z 2801:2012, and the results are shown in Fig. 4.

[0073] The bacterial strains used for the antibacterial performance evaluation were Escherichia coli ATCC 8739 and Staphylococcus aureus subsp. Aureus ATCC 33591, provided by ATCC.

[0074] Referring to Fig. 4, Example 1 showed antibacterial performance of Log 5 or higher.

[0075] Therefore, it was confirmed that the composition including the fibrous acellular dermal matrix manufactured according to Example 1 of the present invention has excellent antibacterial and antifungal performance.

[0076]

[0077] 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.

[0078] All technical terms used in this invention, unless otherwise defined, have the same meaning as commonly understood by those skilled in the art. The contents of all publications cited herein as references are incorporated herein by reference.

Claims

1. Contains 48 to 95.5 wt% of acellular fibrous dermal matrix and 0.5 to 52 wt% of water, but no biocompatible polymer is added. A composition comprising a fibrous acellular dermal matrix, wherein the fibrous acellular dermal matrix comprises an antimicrobial agent at a concentration of 1 to 1,000,000 U / mL.

2. In paragraph 1, Maximum tensile strength 9.7 N / mm 2 Ideal and maximum bond strength 7.3N / mm 2 A composition comprising a fibrous acellular dermal matrix satisfying the above.

3. In paragraph 1, A composition comprising a fibrous acellular dermal matrix, further comprising stem cells, growth factors or a mixture thereof.

4. In paragraph 1, The composition above is a composition comprising a fibrous acellular dermal matrix in sheet form. 5.(a) A step of manufacturing a fibrotic acellular dermal matrix by adding an antibacterial agent to the acellular dermal matrix, permeating the acellular dermal matrix, freeze-drying the acellular dermal matrix, and then pulverizing the acellular dermal matrix; (b) a step of rehydrating the fibrous acellular dermal matrix by adding water; and (c) a step of heat-drying the rehydrated fibrotic acellular dermal matrix; a method for producing a composition comprising a fibrotic acellular dermal matrix.

6. In paragraph 5, A method for producing a composition including a fibrous acellular dermal matrix, wherein in the step (c) above, heat drying is performed at 30 to 50°C for 6 hours or longer.

7. In paragraph 5, A method for producing a composition including a fibrous acellular dermal matrix, wherein in the step (c) above, heat drying is performed in a sieve.

8. In paragraph 5, A method for producing a composition including a fibrous acellular dermal matrix, further comprising the step of (d) processing the heat-dried fibrous acellular dermal matrix into a sheet type after the step (c).

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