Composition comprising fibrillated acellular dermis matrix obtained by drying, and preparation method therefor

Abstract

The present invention relates to a composition comprising a fibrillated acellular dermis matrix, and a preparation method therefor, the composition having improved physical properties and tensile strength since a biocompatible polymer is not applied thereto, thereby being more easily used than a conventional composition.

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]

[0003] 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.

[0004] 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.

[0005] 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.

[0006] 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.

[0007] Therefore, there is an urgent need to develop an acellular dermal matrix that can be fixed to the wound site.

[0008]

[0009] The purpose of the present invention is to solve the above-described problem, and 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 process of drying in a mold without applying a biocompatible polymer.

[0010]

[0011] 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, but no biocompatible polymer is added.

[0012] 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.

[0013]

[0014] 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.

[0015] 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.

[0016] Additionally, by performing the drying step in the mold, the physical properties and tensile strength are improved, making it easier for the operator to use the product during the procedure.

[0017]

[0018] 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.

[0019] Figure 2 is a photograph showing the weight before and after high-temperature drying treatment of a composition including a fibrous acellular dermal matrix according to Example 1.

[0020] FIG. 3 is a photograph showing a composition including a fibrous acellular dermal matrix produced by a dry method in Example 1 and Comparative Examples 1 to 3.

[0021] Figure 4 is a graph showing the results of Experimental Example 2.

[0022] Figure 5 is a photograph showing the results of Experimental Example 3.

[0023] Figure 6 is a graph showing the results of Experimental Example 4.

[0024]

[0025] 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.

[0026] 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.

[0027] 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.

[0028]

[0029] Hereinafter, a composition including a fibrous acellular dermal matrix according to one embodiment of the present invention will be described in detail.

[0030] The composition comprising the fibrous acellular dermal matrix of the present embodiment comprises a fibrous acellular dermal matrix and water.

[0031] 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%.

[0032] 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 acellular dermal matrix, and if it exceeds 52 wt%, it may be difficult to implement the desired formulation due to excessive use.

[0033] 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.

[0034] 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.

[0035] In this embodiment, stem cells, growth factors or mixtures thereof may be further included.

[0036] Additionally, the present embodiment may further include one or more selected from among pharmaceutically used antibacterial agents, excipients, and additives.

[0037] Antibacterial agents include short-chain alcohols, benzoalkonium chloride (BAC), didecyl dimethyl ammonium chloride (DDAC), zeolites (CWT-A), isothiazolones, alkyl dimethyl ammonium chlorides, triazines, 2-thiocyanomethylthiobenzothiazole, methylene bis thiocyanate, acrolein, dodecyl guanidine hydrogen chloride, chlorophenols, quaternary ammonium salts, gluteraldehyde, dithiocarbamate, 2-mercaptobenzothiazole, para-chloro-meta-xylenol, silver, chlorhexidine, polyhexamethylene biguanide, n-halamines, triclosan, phospholipids, alpha hydroxy acids, 2,2-dibromo-3-nitrilopropionamide, One or more selected from 2-bromo-2-nitro-1,3-propanediol, farnesol, iodine, bromine, hydrogen peroxide, chlorine dioxide, vegetable oils, plant extracts, benzalkonium chloride, chlorine, and sodium hypochlorite may be used.

[0038] 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.

[0039] 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.

[0040] The composition may further comprise one or more of stem cells and growth factors.

[0041] In this embodiment, the composition comprising the fibrous acellular dermal matrix may 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 may be preferably in the form of a sheet, and may 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 grafting material.

[0042]

[0043] 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.

[0044] 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.

[0045] Referring to Fig. 1, first, an acellular dermal matrix is ​​pulverized to produce a fibrous acellular dermal matrix (S10).

[0046] The acellular dermal matrix can be ground 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 to produce a fibrous acellular dermal matrix. 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.

[0047] 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.

[0048]

[0049] Next, water is added to the fibrous acellular dermal matrix to rehydrate it (S20).

[0050] 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. If the water content is less than 300 parts by weight, it may be difficult to hydrate the fibrotic acellular dermal matrix, and if it exceeds 2,000 parts by weight, the heat drying time described below may be prolonged due to unnecessary use, which is not preferable. In this embodiment, the water may be sterilized distilled water.

[0051]

[0052] Then, the rehydrated fibrous acellular dermal matrix is ​​heat-dried (S30).

[0053] In the above step S20, the rehydrated fibrous acellular dermal matrix can be heat-dried in a mold, preferably a lattice-shaped mold, 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, until the moisture content becomes 0.5 to 52 wt%. If the temperature is less than 30°C, drying may take an excessive amount of time, and if it exceeds 50°C, the human tissue may become deformed, which may result in a decrease in amino acid content as well as a decrease in tensile strength and hydration degree.

[0054] The average tensile strength of the fibrous acellular dermal matrix under the above-described heat-drying conditions was 0.2 N / mm. 2 By satisfying the above, the practitioner can use it more easily than before during the procedure.

[0055]

[0056] After the above step S30, a step of processing the heat-dried fibrous acellular dermal matrix into a sheet type may be further included.

[0057] 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, and of course, it can be manufactured in various sizes according to the width, length, and thickness required by the wound site or surgical characteristics.

[0058] 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.).

[0059]

[0060] 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.

[0061]

[0062] Manufacturing Example 1. Manufacturing of acellular dermal matrix

[0063] 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.

[0064]

[0065] Example 1. Preparation of a composition comprising a fibrous acellular dermal matrix

[0066] The human-derived acellular dermal matrix manufactured in the above Manufacturing Example 1 was ground with a cutting mill and fiberized to manufacture a fibrous acellular dermal matrix. 10 times the amount of sterile distilled water was added to rehydrate, placed in a grid-shaped mold, heat-dried at 30°C to 50°C for 6 hours or more, and then applied to the mold to shape it and processed into a sheet type to manufacture a composition including a fibrous acellular dermal matrix.

[0067] Meanwhile, the weight of the composition including the fibrous acellular dermal matrix according to Example 1 before and after heat drying treatment at a high temperature of 126°C for 10 minutes or more is shown in Figure 2.

[0068] When checking Fig. 2(c), it can be confirmed that the composition including the fibrous acellular dermal matrix contains 48.32 wt% of the fibrous acellular dermal matrix and 51.68 wt% of water, from the pre-drying weight of 0.387 g shown in Fig. 2(a) and the post-drying weight of 0.187 g shown in Fig. 2(b).

[0069] ※ In Example 1, drying did not occur overnight at RT, a temperature below 30℃.

[0070]

[0071] Comparative Example 1. Preparation of a composition including a fibrous acellular dermal matrix

[0072] It was manufactured in the same manner as Example 1, except that the heat treatment was performed at 60°C for more than 6 hours.

[0073]

[0074] Comparative Example 2. Preparation of a composition including a fibrous acellular dermal matrix

[0075] It was manufactured in the same manner as Example 1, except that the heat treatment was performed at 70°C for more than 6 hours.

[0076]

[0077] Comparative Example 3. Preparation of a composition including a fibrous acellular dermal matrix

[0078] It was manufactured in the same manner as Example 1, except that the heat treatment was performed at 100°C for more than 6 hours.

[0079]

[0080] Experimental Example 1. Appearance Evaluation

[0081] The appearance of the compositions including the fibrous acellular dermal matrix produced in Example 1 and Comparative Examples 1 to 3 was visually confirmed and evaluated.

[0082] As a result, as shown in Fig. 3, it was confirmed that Example 1 had a structure very similar to the dermal matrix of a wound site of a human body in appearance, Comparative Examples 1 and 2 showed some differences in appearance from the dermal matrix of a wound site of a human body, and it was confirmed that the tissue of Comparative Example 3 was charred.

[0083] In this way, it can be confirmed that the composition including the fibrous acellular dermal matrix of the present invention does not require heating to a high temperature because no biocompatible polymer is added, so there is no change in physical properties due to heat, and since a step for heat-treating the biocompatible polymer is not required, the manufacturing process can be simplified and the manufacturing cost can be reduced.

[0084]

[0085] Experimental Example 2. Evaluation of Amino Acid Content According to Heat Treatment Temperature

[0086] To confirm the change in amino acid content according to heat treatment temperature, the amino acid content of Example 1 and Comparative Examples 1 and 2 was measured using an amino acid auto-analyzer (S433D, Sykam GmbH Co., Germany), and the results are shown in Fig. 4.

[0087] Referring to Figure 4, it was confirmed that the amino acid content decreased from Example 1 to Comparative Examples 1 and 2, and it was expected that this was because human tissue deformation occurred in Comparative Examples 1 and 2.

[0088] As described above, it can be seen that the composition including the fibrous acellular dermal matrix manufactured according to an embodiment of the present invention has a structure very similar to the dermal matrix of a wound site in a human body, and thus has excellent bioadhesiveness and body preservation properties, thereby improving the therapeutic effect and making it easy to apply to a curved wound site.

[0089]

[0090] Experimental Example 3. Evaluation of Hydration Degree According to Heat Treatment Temperature

[0091] In order to check the degree of hydration according to the heat treatment temperature, Example 1, Comparative Examples 1 and 2 were placed in a tube containing water and observed, and the results are shown in Fig. 5.

[0092] Referring to Figure 5, it was confirmed that Example 1 sank within a few seconds, but Comparative Examples 1 and 2 did not sink but continued to float, which was expected to be due to the effects of drying and shrinkage caused by high temperature.

[0093] As such, it can be seen that a composition including a fibrous acellular dermal matrix manufactured according to an embodiment of the present invention has an excellent therapeutic effect due to good maintenance of a moist environment and is easy to apply to a curved wound area.

[0094]

[0095] Experimental Example 4. Tensile strength evaluation according to heat treatment temperature

[0096] To confirm the tensile strength according to the heat treatment temperature, the results are shown in Fig. 5.

[0097] Referring to Figure 6, it was confirmed that the tensile strength of Comparative Examples 1 and 2 decreased compared to Example 1.

[0098] As such, the composition including the fibrous acellular dermal matrix manufactured according to the embodiment of the present invention does not require heating to a high temperature because no biocompatible polymer is added, so there is no change in physical properties due to heat, and by performing a heat-drying step, particularly, a heat-drying step in a mold, the physical properties and tensile strength are improved, making it easy for a practitioner to use during a procedure.

[0099]

[0100] 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.

[0101] 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. 48-95.5 wt% of fibrous acellular dermal matrix; and A composition comprising a fibrous acellular dermal matrix, comprising 0.5 to 52 wt% of water; but no biocompatible polymer is added.

2. In paragraph 1, The average tensile strength is 0.2 N / 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 producing a fibrous acellular dermal matrix by crushing an 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 mold.

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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