Composition for delaying in vivo degradation rate and reinforcing rheological properties of hyaluronic acid hydrogel for biotransplantation

Abstract

The present invention relates to a composition for delaying the in vivo degradation rate and enhancing the rheological properties of hyaluronic acid hydrogel used for biotransplantation. Specifically, the present invention pertains to a composition in which allogeneic acellular dermal particles can be mixed on-site with hyaluronic acid hydrogel having a storage modulus (G') of 300 Pa or less and used in biotransplantation for tissue restoration, whereby the composition delays the degradation rate of hyaluronic acid in the body, improves the storage modulus (G') even without crosslinkers, and offers enhanced injectability for user convenience.

Description

Composition for delaying the in vivo degradation rate and enhancing the rheological properties of hyaluronic acid hydrogels for biotransplantation

[0001] The present invention relates to a composition for delaying the rate of in vivo decomposition and enhancing the rheological properties of hyaluronic acid hydrogels for biotransplantation, and more particularly, to a composition that can be used in a medical setting by mixing homogeneous acellular dermal particles with hyaluronic acid hydrogels having a storage modulus (G') of 300 Pa or less used for biotransplantation for tissue repair.

[0002]

[0003] Hyaluronic acid (HA) is a biosynthetic natural substance that exists in various tissues in the body, including the skin. Due to its excellent biocompatibility, moisturizing properties, and physical properties, it is widely used in the form of hydrogels in biomedical applications such as tissue repair medical devices, facial fillers, and joint injections.

[0004]

[0005] However, hyaluronic acid-based hydrogels (HA-based hydrogels) are rapidly decomposed in the body, which limits their ability to maintain their function as biomaterials. To compensate for this, they are commercialized by increasing their stability in the body through chemical cross-linking using cross-linking agents such as BDDE (1,4-butanediol diglycidyl ether), PEGDE (poly ethylene glycol diglycidyl ether), and DVS (divinyl sulfone). The rheological properties of cross-linked hyaluronic acid hydrogels (cross-linked HA hydrogels) as biomaterials for tissue repair are greatly affected by the cross-linking density.

[0006]

[0007] When injecting hyaluronic acid hydrogels into the skin, different rheological properties are required for each site. For example, areas such as the forehead and bridge of the nose require compositions with relatively high storage modulus (G'). Currently, commercially available products achieve this by increasing the concentration of hyaluronic acid in the finished product or using more cross-linking agents during manufacturing. Increasing the hyaluronic acid concentration can increase injection force during the procedure or make it difficult to control the dosage after the washing process. Furthermore, increasing the amount of cross-linking agent used during manufacturing can lead to safety issues related to the biomaterial.

[0008]

[0009] Therefore, there is a need for a hyaluronic acid-based hydrogel composition and a method for manufacturing the same that can delay the in vivo decomposition of hyaluronic acid without adding more cross-linking agents during manufacturing while maintaining a relatively high storage modulus (G').

[0010]

[0011] Meanwhile, skin grafts are divided into human-derived allogeneic dermis, animal-derived xenogeneic dermis, and synthetic materials. While relatively expensive, allogeneic acellular dermal matrix-based skin grafts (HDM-based skin grafts) are considered the optimal alternative due to their decellularization, which prevents immune rejection or inflammation after transplantation. The extracellular matrix (ECM) components contained within the dermis, such as collagen, elastin, and fibronectin, are essential elements within cells and aid in cell adhesion and proliferation, leading to expected skin regeneration effects.

[0012]

[0013] "Sheet-type acellular human dermal matrix" refers to dermis that has been chemically treated to remove cells that could trigger an immune rejection response from isolated allogeneic dermis. Its main components are collagen and elastin. Sheet-type acellular dermal tissue is manufactured in various sizes depending on the target disease site and used as a graft or implant.

[0014]

[0015] "Particulated human acellular dermal matrix (phADM)" is a particle-like structure made by crushing sheet-like acellular dermis. Previously, freeze-dried particulate acellular dermis was used as a skin substitute by hydrating it with saline or distilled water. Because it can maintain a fluid state, it has the advantage of being injected using a syringe without surgery. For this reason, particulate acellular dermal tissue is used not only for the treatment of skin tissue defects caused by accidents, but also for tissue repair or reconstruction, such as the treatment of chronic diseases such as diabetic ulcers. Most particulate human acellular dermal allografts currently on the market have an average particle size of 500 μm or more, requiring a large-bore needle for injection for plastic and cosmetic purposes. Furthermore, there is a limitation in that it is difficult to homogeneously disperse it when mixed with commercialized biomaterials for tissue repair, such as hyaluronic acid hydrogel fillers (HA fillers).

[0016]

[0017] Korean Patent Publication No. 10-1523878 discloses a method for manufacturing a biotransplantable composition comprising particulate acellular dermis and cross-linked hyaluronic acid. However, the particulate allogeneic dermis used in the invention has a particle size of 300-800 μm, making it difficult to inject for cosmetic and plastic surgery purposes. Furthermore, the invention differs significantly from commercially available hyaluronic acid hydrogels, which are physically mixed with acellular dermis, in that hyaluronic acid is cross-linked to the surface of acellular dermis particles.

[0018]

[0019] To date, there have been no reports of particle-sized, allogeneic acellular dermal fillers being mixed with hyaluronic acid hydrogels to facilitate injection as a biomaterial for tissue repair. Furthermore, no studies have been reported that address the shortcomings of commercialized hyaluronic acid hydrogel products without crosslinking or other reactions, thereby improving the rheological properties of the composition.

[0020]

[0021] Although there are some literatures disclosing finished compositions comprising homogeneous acellular dermis and hyaluronic acid hydrogel, there is no known research on a method of mixing acellular dermis directly in the medical field for the purpose of delaying the in vivo degradation of commercialized hyaluronic acid hydrogel products and improving the rheological properties.

[0022]

[0023] Accordingly, the present inventors have developed a method for mixing acellular dermal particles into a commercialized hyaluronic acid hydrogel product in a medical setting, and have found out conditions for delaying the in vivo decomposition of hyaluronic acid and improving rheological properties without the addition of a cross-linking agent, thereby completing the present invention. Specifically, by specifying the particle size of the acellular dermal particles, the relative ratio of the hyaluronic acid particle size to the acellular dermal particle size, the mixing concentration of the acellular dermal particles, and the storage modulus (G') value of the commercialized hyaluronic acid hydrogel, the conditions for delaying the in vivo decomposition of hyaluronic acid and improving the rheological properties were found.

[0024]

[0025] Object 1 of the present invention is to provide a composition for delaying the rate of in vivo degradation and enhancing the rheological properties of hyaluronic acid hydrogel used for tissue repair and biotransplantation.

[0026] Object 2 of the present invention is to provide a bio-transplant composition for on-site tissue repair.

[0027] Object 3 of the present invention is to provide a finished tissue repair biotransplant composition.

[0028] In the present invention, a method for mixing acellular dermal particles into a commercialized hyaluronic acid hydrogel product in a medical setting was specified, and conditions for delaying the in vivo degradation of hyaluronic acid and improving rheological properties without the addition of a cross-linking agent were identified. Specifically, by specifying the particle size of the acellular dermal particles, the relative ratio of the hyaluronic acid particle size to the acellular dermal particle size, the mixing concentration of the acellular dermal particles, and the storage modulus (G') value of the commercialized hyaluronic acid hydrogel, conditions for delaying the in vivo degradation of hyaluronic acid and improving the rheological properties were identified.

[0029]

[0030] The present invention can particularly increase the satisfaction of the procedure when applied to a procedure site (e.g., forehead, bridge of nose, etc.) requiring a relatively high storage modulus (G'), and can also be expected to have an effect of extending the retention period in the body of a biomaterial graft for tissue repair.

[0031]

[0032] Composition for delaying the in vivo decomposition rate of hyaluronic acid hydrogel and enhancing its rheological properties

[0033] The present invention comprises homogeneous acellular dermal particles and a medical dilution solvent.

[0034] Provided is a composition for delaying the rate of degradation in the body and enhancing the rheological properties of hyaluronic acid hydrogels having a storage modulus (G') of 300 Pa or less used for biotransplantation for tissue repair.

[0035]

[0036] In the reinforcing composition according to the present invention, the hyaluronic acid hydrogel is a hyaluronic acid hydrogel that requires delay in the rate of decomposition in the body or reinforcement of rheological properties, and may be a commercialized product already used in the medical field or a product scheduled for commercialization.

[0037] In the present invention, the rheological properties (storage modulus of elasticity, G') can be improved by optionally mixing homogeneous acellular dermal particles into a hyaluronic acid hydrogel having a storage modulus of elasticity (G') of 300 Pa or less. However, if a hyaluronic acid hydrogel having a storage modulus of elasticity (G') exceeding 300 Pa is used, there may be a problem in that the storage modulus of elasticity (G') actually decreases (see Fig. 3 and Table 4).

[0038]

[0039] In the reinforcing composition according to the present invention, the average particle size of the homogeneous acellular dermis may be 300 μm or less, specifically 200 μm or less, and more specifically 100 μm or less. If homogeneous acellular dermis having a particle size exceeding 300 μm is used, a problem may arise in which the homogeneous acellular dermis particles are not homogeneously mixed into the hyaluronic acid hydrogel (see Table 1 and Fig. 1).

[0040] In addition, the size of the homogeneous acellular dermal particles may be used at 1 / 3 or less of the average particle size of the hyaluronic acid, and preferably 1 / 5 or less. If it exceeds 1 / 3, a problem may arise in which the homogeneous acellular dermal particles are not homogeneously mixed into the hyaluronic acid hydrogel (see Table 1 and Fig. 1).

[0041]

[0042] In the reinforcing composition according to the present invention, the medical dilution solvent may be normal saline, distilled water, phosphate-buffered saline (PBS), etc., used alone or in combination of two or more thereof, and preferably, normal saline, which is most commonly used in medical settings, may be used.

[0043]

[0044] In the reinforcing composition according to the present invention, the mixed composition of the reinforcing composition and the hyaluronic acid hydrogel may contain 10 wt% or less of the homogeneous acellular dermal particles, specifically 1-10 wt%, and more specifically 1-5 wt%. If it exceeds 10 wt%, a rapid increase in injection force may occur, making the procedure difficult (see Table 2), and if it is less than 1 wt%, the effect of delaying the in vivo decomposition of hyaluronic acid and improving the rheological properties may be minimal. In addition, when the reinforcing composition according to the present invention is mixed and used with the hyaluronic acid hydrogel, there is an effect of significantly reducing the decomposition rate of hyaluronic acid by hyaluronidase in the body. However, if the mixed composition contains more than 10 wt% of the homogeneous acellular dermal particles, the effect of reducing the hyaluronic acid decomposition rate may gradually decrease (see Table 3).

[0045] For reference, when using hyaluronic acid hydrogel as a biomaterial for tissue repair for cosmetic surgery, it is generally set to a standard of 40 N or less when connected to a syringe needle due to issues such as injection convenience (Guidelines for Approval Review of Biomaterials for Tissue Repair for Cosmetic Surgery, Ministry of Food and Drug Safety, 2020).

[0046]

[0047] The above rheological properties mean that the storage modulus (G') increases. In addition, the loss factor (tan delta, tan δ) decreases and the complex viscosity improves.

[0048]

[0049] Biotransplantation composition for on-site prepared tissue repair

[0050] The present invention comprises homogeneous acellular dermal particles,

[0051] The above-mentioned homogeneous acellular dermal particles are mixed in a medical dilution solvent, and the mixed composition is mixed on-site with hyaluronic acid hydrogel.

[0052] The storage modulus (G') of the hyaluronic acid hydrogel is 300 Pa or less.

[0053] A bio-transplant composition for tissue repair is provided.

[0054]

[0055] In the biotransplant composition for tissue repair according to the present invention, the medical dilution solvent may be physiological saline solution, distilled water, phosphate buffered physiological saline solution, etc., used alone or in combination of two or more thereof, and preferably, physiological saline solution, which is most commonly used in medical settings, may be used.

[0056]

[0057] Specifically,

[0058] The size of the above homogeneous acellular dermal particles is 300 μm or less,

[0059] The homogeneous acellular dermal particles in the above mixed composition are 0.5-10 wt%,

[0060] The hyaluronic acid hydrogel in the above mixed composition is 0.5-2 wt%,

[0061] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0062]

[0063] Specifically,

[0064] The size of the above homogeneous acellular dermal particles is 200 μm or less,

[0065] The homogeneous acellular dermal particles in the above mixed composition are 1-10 wt%,

[0066] The hyaluronic acid hydrogel in the above mixed composition is 0.7-1.5 wt%,

[0067] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0068]

[0069] Specifically,

[0070] The size of the above homogeneous acellular dermal particles is 100 μm or less,

[0071] The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%,

[0072] The hyaluronic acid hydrogel in the above mixed composition is 0.8-1.4 wt%,

[0073] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0074]

[0075] Specifically,

[0076] The size of the above homogeneous acellular dermal particles is 100 μm or less,

[0077] The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%,

[0078] The hyaluronic acid hydrogel in the above mixed composition is 0.9-1.3 wt%,

[0079] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0080]

[0081] Specifically,

[0082] The size of the above homogeneous acellular dermal particles is 100 μm or less,

[0083] The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%,

[0084] The hyaluronic acid hydrogel in the above mixed composition is 1-1.2 wt%,

[0085] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0086]

[0087] Completed biotransplant composition for tissue repair

[0088] It is self-evident that the conditions for delaying the in vivo decomposition of hyaluronic acid and improving the rheological properties identified in the present invention can also be applied to a finished composition comprising homogeneous acellular dermis and hyaluronic acid hydrogel.

[0089]

[0090] Accordingly, the present invention provides a solution comprising a mixture of homogeneous acellular dermal particles and a medical dilution solvent; and

[0091] A bio-implantation composition for tissue repair comprising a hyaluronic acid hydrogel having a storage modulus (G') of 300 Pa or less is provided.

[0092]

[0093] In the tissue repair composition according to the present invention,

[0094] The size of the above homogeneous acellular dermal particles is 300 μm or less,

[0095] The homogeneous acellular dermal particles in the above composition are 0.5-15 wt%,

[0096] The hyaluronic acid hydrogel in the above composition is 0.5-2 wt%,

[0097] The above homogeneous acellular dermal particles may be less than 1 / 3 of the average hyaluronic acid particle size.

[0098]

[0099] The composition for delaying the rate of decomposition in vivo and reinforcing the rheological properties of a hyaluronic acid hydrogel having a storage modulus (G') of 300 Pa or less used for tissue repair and biotransplantation according to the present invention has the effect of delaying the rate of decomposition of hyaluronic acid in vivo by mixing homogeneous acellular dermal particles into the hyaluronic acid hydrogel, relatively improving the storage modulus (G') without adding a crosslinker, and providing an injection force with high usability.

[0100]

[0101] Figure 1a is an image of the properties of samples manufactured by dividing the average particle size of the acellular dermis into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermis particles and hyaluronic acid.

[0102] Figure 1b shows the results of observing the shapes of phADM particles in samples manufactured by dividing the average particle sizes of acellular dermis into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid using an optical microscope.

[0103] Figure 1c is a graph comparing the particle size distribution of a hyaluronic acid sample (sample name: HA1, manufacturer: Hugel, Korea) and four kinds of homogeneous acellular dermal powders manufactured with different average particle sizes in samples manufactured by dividing the average particle size of the acellular dermis into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid.

[0104] Figure 2a shows the results of measuring the injection force by filling a sample of a mixed composition of homogeneous acellular dermal particles (1, 5, 10, 20 wt%) and hyaluronic acid (sample name: HA2, manufacturer: Hugel, Korea) (1 wt%) into a syringe equipped with a commonly used 23G (thin-wall) needle.

[0105] Figure 2b shows the results of measuring the hyaluronic acid degradation rate in a mixed composition 24 hours after treating a sample of a mixed composition of homogeneous acellular dermal particles (0, 1, 5, 10 wt%) and hyaluronic acid (sample name: HA2, manufacturer: Hugel, Korea) (1 wt%) with hyaluronidase.

[0106] Data in Figure 2 = mean ± standard deviation; and *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0107] Figure 3a shows the results of measuring the storage modulus (G') before and after mixing phADM having an average particle size of 100 μm or less with HA3 or HA4 hyaluronic acid, respectively (sample name: HA3, manufacturer: Medytox, Korea and sample name: HA4, manufacturer: Hugel, Korea).

[0108] Figure 3b shows the results of measuring the loss modulus (G") before and after mixing phADM having an average particle size of 100 μm or less with HA3 or HA4 hyaluronic acid, respectively (sample name: HA3, manufacturer: Medytox, Republic of Korea and sample name: HA4, manufacturer: Hugel, Republic of Korea).

[0109] Figure 3c shows the results of measuring the loss factor (tan delta) before and after mixing phADM having an average particle size of 100 μm or less with HA3 or HA4 hyaluronic acid, respectively (sample name: HA3, manufacturer: Medytox, Korea and sample name: HA4, manufacturer: Hugel, Korea).

[0110] Figure 3d shows the results of measuring the complex viscosity before and after mixing phADM having an average particle size of 100 μm or less with HA3 or HA4 hyaluronic acid, respectively (sample name: HA3, manufacturer: Medytox, Republic of Korea and sample name: HA4, manufacturer: Hugel, Republic of Korea).

[0111] Data in Figure 3 = mean ± standard deviation; and *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0112] Figure 4a shows the results of measuring the storage modulus (G') before and after mixing phADM having an average particle size of 100 μm or less with each of HA5 and HA7 hyaluronic acid (sample name: HA5, manufacturer: Medytox, Korea; sample name: HA6, manufacturer: Jetema, Korea; and sample name: HA7, manufacturer: Medytox, Korea).

[0113] Figure 4b shows the results of measuring the loss modulus (G") before and after mixing phADM having an average particle size of 100 μm or less into each of HA5 and HA7 hyaluronic acid (sample name: HA5, manufacturer: Medytox, Korea; sample name: HA6, manufacturer: Jetema, Korea; and sample name: HA7, manufacturer: Medytox, Korea).

[0114] Figure 4c shows the results of measuring the loss factor (tan delta) before and after mixing phADM having an average particle size of 100 μm or less with each of HA5 and HA7 hyaluronic acids (sample name: HA5, manufacturer: Medytox, Korea; sample name: HA6, manufacturer: Jetema, Korea; and sample name: HA7, manufacturer: Medytox, Korea).

[0115] Figure 4d shows the results of measuring the complex viscosity before and after mixing phADM having an average particle size of 100 μm or less with each of HA5 and HA7 hyaluronic acid (sample name: HA5, manufacturer: Medytox, Korea; sample name: HA6, manufacturer: Jetema, Korea; and sample name: HA7, manufacturer: Medytox, Korea).

[0116] Data in Figure 4 = mean ± standard deviation; and *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0117]

[0118] Hereinafter, the present invention will be described in detail.

[0119]

[0120] The term "sheet-type human acellular dermal matrix" used in the present invention refers to dermis obtained by chemically treating isolated allogeneic dermis to remove cells that may induce immune rejection, and is mainly composed of collagen and elastin. The sheet-type acellular dermal tissue is manufactured in various areas depending on the disease site to be applied and is used in a graft or implant manner.

[0121]

[0122] The term "particulated human acellular dermal matrix (phADM)" used in the present invention refers to sheet-shaped acellular dermis pulverized into a particle structure. Previously, freeze-dried particulate acellular dermis was used as a skin substitute by hydrating it with saline or distilled water. Because it can maintain a fluid state, it has the advantage of being able to be injected using a syringe without surgery. Therefore, particulate acellular dermal tissue is used not only for the treatment of skin tissue defects caused by accidents, but also for tissue repair or reconstruction, such as the treatment of chronic diseases such as diabetic ulcers. Most particulate allografts currently on the market have an average particle size of 500 μm or more, requiring a large-bore needle for injection for plastic and cosmetic purposes. Furthermore, they present limitations in the ability to homogeneously disperse the phADM when mixed with commercialized biomaterials for tissue repair, such as hyaluronic acid hydrogel fillers (HA fillers).

[0123]

[0124] The term "hyaluronic acid (HA)" used in the present invention is a biosynthetic natural substance found in abundance in animal skin, synovial fluid, and cartilage. It is a mucopolysaccharide with many hydroxyl groups, making it hydrophilic. It combines with water to form a gel, which plays a role in lubricating joints and providing skin flexibility. Due to its high viscosity, it also plays an important role in preventing bacterial invasion and toxins from penetrating the skin. If the skin lacks hyaluronic acid, the skin becomes dry, inelastic, and wrinkled. For this reason, hyaluronic acid has been widely used as a biomaterial for tissue repair to prevent wrinkles or fill in hollow areas. Commercially available hyaluronic acid hydrogel products exhibit differences in physicochemical and biological properties depending on the method and degree of cross-linking using BDDE or other agents.

[0125]

[0126] The term "injection force" used in the present invention refers to the force applied when force is applied with a syringe, and the unit is [N]. When using hyaluronic acid hydrogel as a biomaterial for tissue repair for cosmetic surgery, it is generally set to a standard of 40 N or less when connected to an injection needle due to issues such as convenience of injection (Guidelines for Approval Review of Biomaterials for Tissue Repair for Cosmetic Surgery, Ministry of Food and Drug Safety, 2020).

[0127]

[0128] The term "storage modulus (G')" used in the present invention is a value that represents the elastic response of an object when force is applied to the object, and is an index of the characteristic of an object that changes shape when force is applied but returns to its original shape when force is removed, and its unit is [Pa]. In the properties of biomaterials for tissue repair, the higher the storage modulus, the higher the elasticity, and the harder it feels after injection into the body, and in particular, in the case of fillers for cosmetic surgery, it is good for providing volume and has the characteristic of maintaining its shape well.

[0129]

[0130] The term "viscous modulus (G)" used in the present invention refers to the degree to which energy spreads due to friction or shear force, and its unit is [Pa]. In biomaterials for tissue repair, a high loss modulus indicates high viscosity, and in particular, in the case of fillers for cosmetic surgery, it is related to the characteristic of spreading within the dermal layer after injection.

[0131]

[0132] The term "loss modulus (tan delta, tan δ)" used in the present invention represents the ratio of elasticity and viscosity, and is calculated as loss modulus / storage modulus (G" / G'), which means the degree to which it is close to a solid or liquid state. If the loss modulus is greater than 1, it is close to a liquid, if it is less than 1, it is close to a solid, and the lower it is, the more likely the biomaterial is to behave as a single mass. In biomaterials for tissue repair for cosmetic surgery, the close-to-solid characteristic means that the shape deformation after the procedure and the possibility of the contents of the implanted filler moving within the tissue are reduced. For example, a filler with a low loss modulus can better withstand the force generated when facial muscles move.

[0133]

[0134] The term "complex viscosity" used in the present invention is a resistance value to flow, which is the geometric mean of the storage modulus and the loss modulus as a function of each measurement frequency, and its unit is [Pa·S]. A high complex viscosity means that the content of the biomaterial for tissue repair is hard, and such fillers are suitable for injection into deep layers of skin tissue or for procedures to add volume. Conversely, a low complex viscosity means that they have a strong soft and flowing nature. Such fillers are intended for areas that do not have a problem with deformation after the procedure or for injection into the dermis layer. In addition, a high complex viscosity is an important factor in maintaining a clear contour after the procedure because it reduces the intra-tissue spreadability of the hyaluronic acid hydrogel filler.

[0135]

[0136] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples.

[0137]

[0138] <Manufacturing Example 1> Manufacturing of freeze-pulverized homogeneous acellular dermal particles

[0139] Skin tissue (collected from cadavers donated for non-profit purposes by tissue banks) was treated with 1.0 units / mL of dispase, a neutral proteolytic enzyme, and stirred in a shaking incubator at 37°C for 60 to 120 minutes. The tissue was then washed three times with sterile distilled water to separate the dermis and epidermis and remove the epidermis. The tissue from which the epidermis had been removed was treated with a 1% Triton X-100 solution at 30°C for 100 minutes to remove cells in the dermis. The tissue was washed three or more times with sterile distilled water to remove any processing materials used in the process, and the homologous acellular dermis was frozen at a temperature below -40°C for more than 2 hours for freeze-drying, and then dried in a freeze-dryer for more than 12 hours to remove moisture.

[0140] Using a surgical knife, cut the above homogeneous acellular dermis into pieces of approximately 1 x 1 cm in length and width. 2 After cutting into the size of , 100 g of the allogeneic acellular dermis was placed in a micro grinder and ground for 3 minutes at 5,000 rpm, which is the optimal condition for preventing the denaturation of collagen and elastin, to produce allogeneic acellular dermis particles that passed through sieves of 500 and 300 μm in a sterile environment. In order to produce allogeneic acellular dermis particles with smaller particles, the particles were ground using a freeze grinder and then allogeneic acellular dermis particles that passed through a sieve of 100 μm were produced. In summary, through the above manufacturing process, particles with an average size of less than 100 μm, particles with an average size of 100-300 μm, particles with an average size of 300-500 μm, and phADM particles larger than an average size of 500 μm were obtained, respectively.

[0141]

[0142] <Manufacturing Example 2> On-site manufacturing of a mixed composition of homogeneous acellular dermal particles and hyaluronic acid hydrogel

[0143] (1) Preparation of a solution containing homogeneous acellular dermal particles

[0144] 5-10 mL of normal saline was placed in a syringe, and the homogeneous acellular dermal particles prepared in Manufacturing Example 1 were divided into portions according to the desired concentration in another syringe. After attaching a Luer-lock connector, the plungers on both sides were reciprocated 5 to 10 times to prepare an homogeneous acellular dermal solution containing the homogeneous acellular dermal particles and normal saline.

[0145] (2) Preparation of a mixed composition of homogeneous acellular dermal solution and hyaluronic acid hydrogel

[0146] Depending on the concentration and volume to be produced, the above-mentioned homogeneous acellular dermal solution and the commercialized hyaluronic acid hydrogel were each placed in different syringes, and after attaching a Luer lock connector, the plunger was reciprocated 5 to 10 times to produce a mixed composition of the hyaluronic acid hydrogel and the homogeneous acellular dermal solution.

[0147]

[0148] Injection power evaluation

[0149] Injection force was evaluated using a ZwickiLine (Zwick / Roell, Germany) device. A pre-filled syringe containing the sample to be measured was inserted into a jig, and the syringe plunger was adjusted to be centered on the fixture plate. Injection force was measured at a speed of 12 mm / min.

[0150]

[0151] Particle size measurement

[0152] The particle size measurement and analysis of the present invention were performed using a particle size analyzer (LS13320XR, Beckman Coulter, USA). The sample to be measured was diluted in physiological saline solution to 0.1 to 1 wt% and then measured using wet mode.

[0153]

[0154] Measurement of enzymatic degradation of hyaluronic acid

[0155] The decomposition test of the present invention was conducted according to the method of JL Reissig et al. (A modified colorimetric method for the estimation of N-acetylamino sugar, J. Biol. Chem. 1955, 217:959-966). After putting the same mass of hyaluronic acid hydrogel in each test tube, PBS (pH 7.4) containing 10 IU / mL of hyaluronidase (hyaluronidase, Sigma-Aldrich, USA) was added. The mixture was reacted at 37°C for 24 hours. The enzymatic reaction was stopped with 0.1 N HCl (40 vol%), and only the supernatant was obtained after centrifugation, and the amount of N-acetylglucosamine (NAG) decomposed was measured using the carbazole assay.

[0156] For the carbazole assay, 0.5 mL of the sample was added to a H2SO4 reagent containing 0.025 M sodium tetraborate·10H2O and heated at 99°C for 10 minutes. After gradual cooling, 0.1 mL of a 0.125% carbazole reagent was added, and the reaction was terminated after heating for another 15 minutes. The absorbance of the sample after the carbazole colorimetric reaction was measured at a wavelength of 530 nm using a UV-vis spectrophotometer. The degree of degradation of the crosslinker was set to 100% based on the content indicated on the product, and the degradation rates of the samples were analyzed.

[0157]

[0158] Viscoelastic property analysis

[0159] The viscoelastic property analysis of the present invention was performed using an oscillating rotational rheometer equipped with parallel plates. The sample to be measured was placed between the plates, and the viscoelasticity was measured while applying a constant horizontal shear stress by rotating one of the plates. The testing equipment was a rheometer (MCR302e, Anton Paar Ltd., Austria), and the test items were expressed as storage elastic modulus (G', Pa·s), loss elastic modulus (G", Pa·s), loss modulus (G" / G', dimensionless), and complex viscosity (Pa·s). The frequency, temperature, shear strain, and measurement interval were set to 0.1 Hz, 25 ℃, 1%, and 1.0 mm, respectively, and the sample to be measured was filled, and the parallel plates were 25 mm in diameter and had a circular shape.

[0160]

[0161] <Experimental Example 1> Evaluation of physical properties of mixed compositions according to the average particle size of homogeneous acellular dermal particles

[0162] After manufacturing the homogeneous acellular dermal powder according to the method of Manufacturing Example 1, it was filtered using a sieve according to the average particle size. The average particle size ranges to be filtered were divided into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more. Each of the above four types of powders was mixed in physiological saline at a concentration of 10%, and then 1 mL of the solution and 1 mL of a commercially available hyaluronic acid hydrogel (sample name: HA1, manufacturer: Hugel, Korea) were each mixed according to Manufacturing Example 2. The content of the particulate homogeneous acellular dermal powder (phADM) in the resulting mixture composition was 5 wt%. The results comparing the properties of each mixture composition are shown in Fig. 1 and Table 1.

[0163]

[0164] Figure 1a is an image observing the properties of samples manufactured by dividing the size of the acellular dermal particles into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid.

[0165] Figure 1b shows the results of observing the shapes of phADM particles in samples manufactured by dividing the sizes of acellular dermal particles into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid using an optical microscope.

[0166] Figure 1c is a graph comparing the particle size distribution of a hyaluronic acid sample (sample name: HA1, manufacturer: Hugel, Korea) and four kinds of homogeneous acellular dermal powders manufactured with different average particle sizes in samples manufactured by dividing the acellular dermal particle sizes into 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid.

[0167] The results of Figure 1c are summarized and shown in Table 1 below.

[0168] HA1phADM(<100 μm)phADM(100-300 μm)phADM(300-500 μm)phADM(>500 μm)Average particle size (μm, mean±standard deviation)541±6.6245.1±1.25152±0.569663±34.6829±43.6Moderate particle size (μm, mean±standard deviation)534±29.240.4±2.14140±0.066434±80.7605±32.0

[0169]

[0170] As shown in Figs. 1a and 1b, the mixture prepared with acellular dermal particles having an average size of 100 μm or less showed a homogeneous distribution of acellular dermal particles within the mixture, while the mixture prepared with particles having an average size of 100 to 300 μm showed a slightly clumped appearance of particles. The solutions prepared with particles having an average size of 300 to 500 μm and particles larger than an average size of 500 μm showed a greater degree of particle clumping within the mixture.

[0171] From this, it can be seen that when the average size of the phADM particles is 300 μm or less, they are homogeneously mixed with the existing hyaluronic acid hydrogel, but when the average size is 300 μm or more, the particles are relatively widely distributed and do not form a homogeneous mixed composition.

[0172]

[0173] As shown in Fig. 1c and Table 1, the mean particle size (mean) of HA1 was 541±6.62 μm (mean±standard deviation, n=3) and the mode particle size (mode) was 534±29.2 μm (mean±standard deviation, n=3). As a result of measurement according to the sieve pore size range, the four powders of 100 μm or less, 100 to 300 μm, 300 to 500 μm, and 500 μm or more had average particle sizes of 45.1±1.25, 152±0.569, 663±34.6, and 829±43.6 μm (mean±standard deviation, n=3), respectively, and the most frequent particle sizes were 40.4±2.14, 140±0.0666, 434±80.7, and 605±32.0 μm (mean±standard deviation, n=3), respectively. From this, it was found that when the average size of the mixed phADM particles was approximately 1 / 3 smaller than the average size of the hyaluronic acid hydrogel particles, a more homogeneous mixed composition was formed.

[0174]

[0175] <Experimental Example 2> Changes in the physical properties of a mixed composition according to the concentration of a homogeneous acellular dermal particle solution

[0176] After manufacturing the homogeneous acellular dermal powder according to the method of Manufacturing Example 1, the particles were filtered using a sieve with a 100 μm diameter to obtain only the particles that passed through the sieve. The powder was mixed with physiological saline solution at concentrations of 2, 10, 20, and 40 wt%, respectively, and then 1 mL of the solution according to Manufacturing Example 2 and 1 mL of a commercially available hyaluronic acid hydrogel (sample name: HA2, manufacturer: Hugel, Korea) were mixed, respectively, so that the final concentrations of the homogeneous acellular dermal powder in the mixed composition were 1, 5, 10, and 20 wt%, and the final concentration of the hyaluronic acid (HA2) was 1 wt%, thereby manufacturing samples.

[0177] The above mixtures were filled into a syringe equipped with a commonly used 23G (thin-wall) needle, and the injection force was measured, and the results are shown in Fig. 2a and Table 2.

[0178]

[0179] Figure 2a shows the results of measuring the injection force by filling a sample of a mixed composition of homogeneous acellular dermal particles (1, 5, 10, 20 wt%) and hyaluronic acid (HA2) (1 wt%) into a syringe equipped with a commonly used 23G (thin-wall) needle (n=3).

[0180] The results of Figure 2a are summarized and shown in Table 2 below.

[0181]

[0182] HA2 (wt%) 1.0 phADM (wt%) 1.0 5.0 10.0 20.0 Main input (N, mean ± standard deviation) 19.5 ± 0.5 3 2 4.9 ± 0.7 3 3 0.3 ± 1.26 1 15 ± 0.236

[0183]

[0184] As shown in Fig. 2a and Table 2, the mixed composition prepared at a concentration of 1 wt% of phADM showed the lowest injection force of 19.5 N, and the mixed compositions prepared at concentrations of 5 and 10 wt% obtained injection force values ​​of 24.9 and 30.3 N, respectively. On the other hand, the mixed composition prepared at a concentration of 20 wt% was measured to have a required injection force of 115.1 N, exceeding 40 N, confirming that the injection procedure was difficult.

[0185]

[0186] Next, the results of measuring the enzymatic degradation of hyaluronic acid in a mixed composition of homogeneous acellular dermal particles and hyaluronic acid (HA2) are shown in Fig. 2b and Table 3.

[0187]

[0188] Figure 2b shows the results of measuring the hyaluronic acid degradation rate in a mixed composition 24 hours after treating a sample of a mixed composition of homogeneous acellular dermal particles (0, 1, 5, 10 wt%) and hyaluronic acid (HA2) (1 wt%) with hyaluronidase, a hyaluronic acid decomposing enzyme (n=3).

[0189] The results of Figure 2b are summarized and shown in Table 3 below.

[0190]

[0191] HA2 (wt%) 2.0 1.0 phADM (wt%) 0.0 1.0 5.0 10.0 Relative degradation rate of hyaluronic acid (%, mean ± standard deviation) 93.3 ± 0.84 156.5 ± 2.9 159.4 ± 5.8 364.2 ± 1.68

[0192]

[0193] As shown in Fig. 2b and Table 3, the existing hyaluronic acid hydrogel (HA2) without phADM showed a degradation rate of about 93% after 24 hours of hyaluronidase treatment, but in the composition containing 1 to 10 wt% of phADM, the degradation rate significantly decreased to 56 to 64%, confirming an increase in enzyme resistance. From this, it was confirmed that mixing granulated phADM into the hyaluronic acid hydrogel increases the resistance to hyaluronidase, thereby delaying the degradation rate when the developed tissue repair mixture composition is administered in the body, and significantly improving the duration of treatment efficacy.

[0194] In a composition containing 1 to 10 wt% of phADM, the decomposition rate tends to decrease significantly, which is desirable. However, as the content of phADM increases, the decomposition rate of hyaluronic acid also tends to gradually increase, so it is expected that containing more than 10 wt% of phADM in the mixed composition is not desirable.

[0195]

[0196] <Experimental Example 3> Evaluation of physical properties of mixed compositions according to elastic modulus (G') of hyaluronic acid hydrogel

[0197] The phADM having an average particle size of 100 μm or less prepared in Manufacturing Example 1 was mixed with physiological saline at a concentration of 10% to prepare a mixture. In order to measure the physical properties of the mixture composition, two types of hyaluronic acid hydrogels having a storage modulus (G') of 300 Pa or less and more than 300 Pa before mixing, which are commonly available on the market, were prepared (G'≤300 Pa sample name: HA3, manufacturer: Medytox, Korea, USA, n=3; G'>300 Pa sample name: HA4, manufacturer: Hugel, Korea). According to Manufacturing Example 2, 1 mL of the phADM solution and 1 mL of the hyaluronic acid hydrogel were mixed, respectively, and viscoelasticity characteristics were analyzed after preparation (n=3). At this time, the final concentration of phADM was 5.0 wt%, and the concentration of hyaluronic acid was 1.0 wt%.

[0198]

[0199] Figure 3a shows the results of measuring the storage modulus (G') before and after mixing phADM of 100 μm or less into HA3 or HA4 hyaluronic acid, respectively (n=3).

[0200] Figure 3b shows the results of measuring the loss modulus (G") before and after mixing phADM of 100 μm or less into HA3 or HA4 hyaluronic acid, respectively (n=3).

[0201] Figure 3c shows the results of measuring the loss factor (tan delta) before and after mixing phADM of 100 μm or less into HA3 or HA4 hyaluronic acid, respectively (n=3).

[0202] Figure 3d shows the results of measuring the complex viscosity before and after mixing phADM of 100 μm or less into HA3 or HA4 hyaluronic acid, respectively (n=3).

[0203] The results of Figures 3a to 3d are summarized and shown in Table 4 below.

[0204]

[0205] HA3HA4phADM Mixed or not XOXO Storage modulus (Pa, mean ± standard deviation) 289 ± 6.30 338 ± 18.8 396 ± 12.2 371 ± 7.36 Loss modulus (Pa, mean ± standard deviation) 73.1 ± 2.6 366.7 ± 5.6 060.3 ± 2.00 45.7 ± 2.73 Tangent delta (mean ± standard deviation) 0.25 ± 0.00 420.20 ± 0.00 580.15 ± 0.00 950.12 ± 0.00 54 Complex viscosity (Pa s, mean ± standard deviation) 469 ± 17.3 548 ± 31.1 615 ± 13.4 595 ± 11.7

[0206]

[0207] As shown in Figure 3 and Table 4,

[0208] The mixture of hyaluronic acid (HA3) hydrogel and acellular dermal powder solution, whose storage modulus (G') before mixing was less than 300 Pa, was measured to have an average increase in storage modulus from 289 Pa to 338 Pa, an increase of 17.0% after mixing. On the other hand, the mixture using hyaluronic acid (HA4) hydrogel, whose G' before mixing exceeded 300 Pa, was measured to have an average decrease in storage modulus from 396 Pa to 371 Pa, an increase of 6.3% after mixing.

[0209] Regardless of the physical properties of the hyaluronic acid hydrogels before mixing, the loss modulus decreased from 73 Pa to 67 Pa (when G'≤300 Pa of the hyaluronic acid hydrogel before mixing) and from 60 Pa to 36 Pa (when G'>300 Pa of the hyaluronic acid hydrogel before mixing) after mixing.

[0210] The loss factor (tan delta) decreased from 0.25 to 0.20 (when G'≤300 Pa of hyaluronic acid hydrogel before mixing) and from 0.15 to 0.12 (when G'>300 Pa of hyaluronic acid hydrogel before mixing) when mixing with phADM solution. Therefore, it was confirmed that mixing with phADM increased the relative elasticity ratio of the existing hyaluronic acid hydrogel.

[0211] The complex viscosity increased from 469 Pa·S to 548 Pa·S on average when the G' of the hyaluronic acid hydrogel before mixing was less than 300 Pa, and decreased from 615 Pa·S to 595 Pa·S when the G' of the hyaluronic acid hydrogel before mixing was more than 300 Pa.

[0212]

[0213] <Experimental Example 4> Changes in the physical properties of mixed compositions using various hyaluronic acid hydrogels with a storage modulus (G') of 300 Pa or less

[0214] The acellular dermal powder of 100 μm or less prepared in Manufacturing Example 1 was mixed with physiological saline at a concentration of 10 wt%. In order to confirm the measurement results of FIG. 3 and Table 4, three types of other hyaluronic acid hydrogels having a storage modulus (G') of 300 Pa or less before mixing were prepared (n=3), and then 1 mL of phADM solution and 1 mL of hyaluronic acid hydrogel were mixed according to Manufacturing Example 2, and post-manufacture analysis was performed (G'≤300 Pa sample name: HA5, manufacturer: Medytox, Korea, USA; G'≤300 Pa sample name: HA6, manufacturer: Jetema, Korea; G'≤300 Pa sample name: HA7, manufacturer: Medytox, Korea). The final concentration of phADM was 5 wt%, and the concentration of hyaluronic acid was 1.0 wt%.

[0215]

[0216] Figure 4a shows the results of measuring the storage modulus (G') before and after mixing phADM with an average thickness of 100 μm or less into each of HA5 to HA7 hyaluronic acid (n=3).

[0217] Figure 4b shows the results of measuring the loss modulus (G") before and after mixing phADM with an average thickness of 100 μm or less into each of HA5 to HA7 hyaluronic acid (n=3).

[0218] Figure 4c shows the results of measuring the loss factor (tan delta) before and after mixing phADM with an average thickness of 100 μm or less into each of HA5 to HA7 hyaluronic acid (n=3).

[0219] Figure 4d shows the results of measuring the complex viscosity before and after mixing phADM with an average thickness of 100 μm or less into each of HA5 to HA7 hyaluronic acid (n=3).

[0220] The results of Figures 4a to 4d are summarized and shown in Table 5 below.

[0221]

[0222] HA5HA6HA7phADM Mixed or notXOXOXOStorage modulus (Pa, mean±standard deviation)138±15.6216±10.5167±2.27191±11.2289±6.30338±18.8Loss modulus (Pa, mean±standard deviation)40.3±1.5241.3±4.1640.5±0.9326.9±2.8673.1±2.6366.7±5.60Tan delta (mean±standard deviation)0.29±0.0240.19±0.0100.24±0.00280.14±0.00700.25±0.00420.20±0.0058Complex viscosity (Pa·s, (Mean ± standard deviation)243 ± 2.65350 ± 17.7271 ± 6.18307 ± 18.2469 ± 17.3548 ± 31.1

[0223]

[0224] As shown in Figure 4 and Table 5,

[0225] The storage moduli of HA5, HA6, and HA7 before mixing were measured to be 138, 167, and 289 Pa on average, respectively, and the samples mixed with phADM solution showed an average increase of 56.5, 14.4, and 17.0% to 216, 191, and 338 Pa on average, respectively.

[0226] The loss modulus of hyaluronic acid hydrogels before mixing was measured to be 40, 40, and 73 Pa, respectively, for HA5, HA6, and HA7, but when the phADM solution was mixed, it was measured to be 41, 27, and 67 Pa, respectively.

[0227] The loss coefficients before mixing were measured to be 0.29, 0.24, and 0.25 for HA5, HA6, and HA7, respectively, on average, but when the phADM solution was mixed, they all decreased to 0.19, 0.14, and 0.20, respectively.

[0228] Before mixing, the complex viscosities were measured to be 243, 271, and 469 Pa·s for HA5, HA6, and HA7, respectively, on average, but when the phADM solution was mixed, they all increased to 350, 307, and 548 Pa·s, respectively.

[0229] From these results, it was confirmed that when the storage elastic modulus of the hyaluronic acid hydrogel before mixing is 300 Pa or less, mixing particles of homogeneous acellular dermis with an average size of 300 μm or less at a concentration of 10 wt% or less increases the elasticity and complex viscosity of the tissue repair mixture composition and reduces the loss factor.

[0230]

[0231] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. Containing homogeneous acellular dermal particles and a medical dilution solvent, A composition for delaying the in vivo degradation rate and enhancing the rheological properties of a hyaluronic acid hydrogel having a storage modulus (G') of 300 Pa or less used in biotransplantation for tissue repair.

2. In paragraph 1, A reinforcing composition, wherein the average particle size of the homogeneous acellular dermis is 300 μm or less.

3. In paragraph 2, A reinforcing composition, wherein the average particle size of the homogeneous acellular dermis is 100 μm or less.

4. In paragraph 1, A reinforcing composition, wherein the above homogeneous acellular dermal particles have a size of 1 / 3 or less of the average hyaluronic acid particle size.

5. In paragraph 1, A reinforcing composition, wherein the medical dilution solvent is at least one selected from the group consisting of saline solution, distilled water, and phosphate buffered saline solution.

6. In paragraph 1, A reinforcing composition, wherein the homogeneous acellular dermal particles are contained in an amount of 10 wt% or less in the mixed composition of the reinforcing composition and the hyaluronic acid hydrogel.

7. In paragraph 1, A reinforcing composition having the above rheological properties, wherein the storage modulus (G') increases.

8. In paragraph 1, A reinforcing composition, wherein the decomposition rate of hyaluronic acid hydrogel in the body is reduced by using the reinforcing composition.

9. Contains homogeneous acellular dermal particles, The above-mentioned homogeneous acellular dermal particles are mixed in a medical dilution solvent, and the mixed composition is administered on-site with hyaluronic acid hydrogel. The storage elastic modulus (G') of the hyaluronic acid hydrogel is 300 Pa or less. A bio-graft composition for tissue repair.

10. In paragraph 9, A bio-transplant composition for tissue repair, wherein the medical dilution solvent is at least one selected from the group consisting of physiological saline solution, distilled water, and phosphate buffered saline solution.

11. In paragraph 9, The size of the above homogeneous acellular dermal particles is 300 μm or less, The homogeneous acellular dermal particles in the above mixed composition are 0.5-10 wt%, The hyaluronic acid hydrogel in the above mixed composition is 0.5-2 wt%, The above homogeneous acellular dermal particles are less than 1 / 3 of the average hyaluronic acid particle size. A bio-graft composition for tissue repair.

12. In paragraph 11, The size of the above homogeneous acellular dermal particles is 200 μm or less, The homogeneous acellular dermal particles in the above mixed composition are 1-10 wt%, The hyaluronic acid hydrogel in the above mixed composition is 0.7-1.5 wt%, The above homogeneous acellular dermal particles are less than 1 / 3 of the average hyaluronic acid particle size. A bio-graft composition for tissue repair.

13. In paragraph 12, The size of the above homogeneous acellular dermal particles is 100 μm or less, The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%, The hyaluronic acid hydrogel in the above mixed composition is 0.8-1.4 wt%, The above homogeneous acellular dermal particles are less than 1 / 3 of the average hyaluronic acid particle size. A bio-graft composition for tissue repair.

14. In paragraph 13, The size of the above homogeneous acellular dermal particles is 100 μm or less, The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%, The hyaluronic acid hydrogel in the above mixed composition is 0.9-1.3 wt%, The above homogeneous acellular dermal particles are less than 1 / 3 of the average hyaluronic acid particle size. A bio-graft composition for tissue repair.

15. In paragraph 14, The size of the above homogeneous acellular dermal particles is 100 μm or less, The homogeneous acellular dermal particles in the above mixed composition are 1-5 wt%, The hyaluronic acid hydrogel in the above mixed composition is 1-1.2 wt%, The above homogeneous acellular dermal particles are less than 1 / 3 of the average hyaluronic acid particle size. A bio-graft composition for tissue repair.

16. A solution containing homogeneous acellular dermal particles and a medical dilution solvent; and A bio-implantation composition for tissue repair, comprising a hyaluronic acid hydrogel having a storage modulus (G') of 300 Pa or less.

Images