KR102433259B1 - Medical compositions including adipose tissue-derived extracellular matrix and method of making the same - Google Patents

Abstract

The present invention is adipose tissue-derived extracellular matrix powder; And it relates to a medical composition comprising a biocompatible polymer or a cross-linked product of the biocompatible polymer and a method for preparing the same.
The medical composition according to the present invention is present in a well-aggregated state even after implantation in the body, and can maintain its volume for a certain period of time.

Description

Medical compositions including adipose tissue-derived extracellular matrix and method of making the same}

The present invention relates to a medical composition comprising an adipose tissue-derived extracellular matrix and a method for preparing the same. More specifically, allogeneic and heterogeneous adipose tissue-derived extracellular matrix powder; And it relates to a medical composition comprising a biocompatible polymer or a cross-linked product of the biocompatible polymer and a method for preparing the same.

Regenerative medicine aims to replace or regenerate human cells, tissues, and organs. Traumatic trauma that causes tissue damage and loss of function, and the emergence of new diseases in accordance with the advancement of society, provide an inevitable motive for the rapid development of the field of regenerative medicine.

Medical materials used in the field of regenerative medicine should be carefully selected according to the type of tissue and organ to be applied, the type of disease or trauma, and the patient's medical history. In general, the most frequently selected materials for research include heterogeneous-derived collagen and gelatin, microorganism-derived hyaluronic acid, chitosan, vegetable cellulose-based polymers, vegetable alginate, and the like. In addition, allogeneic substances that can be obtained from human cadavers are attracting attention as effective biomaterials that can be safely used in the field of regenerative medicine.

Among biomaterials, adipose tissue, as well as safety and effectiveness as a biomaterial, is gaining economic/industrial interest at home and abroad. Adipose tissue is a loose connective tissue composed of adipocytes, preadipocytes, fibroblasts, vascular endothelial cells, and various immune cells. Adipose tissue contains extracellular matrix such as collagen, elastin, laminin, fibronectin, glucosaminoglucan and the like. The extracellular matrix not only helps the support and proliferation of cells in the tissue in vivo, but also helps in the recovery of the damaged part of the living body by maintaining the composition of the tissue by binding to the cells.

For example, it is reported that Renuva ^® (MTF Biologics), a human adipose tissue-derived extracellular matrix product, is effective in tissue repair when applied to defect areas such as foot ulcers. As another example, it is reported that Allofill ^® (Biologica Technologies), an extracellular matrix product derived from human adipose tissue, is effective in improving wrinkles. However, medical products such as Renuva ^® and Allofill ^® have a disadvantage in that it is difficult to maintain volume in the body after use because the fine particle extracellular matrix powder derived from human adipose tissue is provided in a simple form hydrated in sterile physiological saline. have

1. Republic of Korea Patent 10-0771058 2. Republic of Korea Patent Registration 10-1628821

1. Combining decellularized human adipose tissue extracellular matrix and adipose-derived stem cells for adipose tissue engineering, Acta Biomaterialia 2013, 8921-31 2. Biocompatibility of injectable hydrogel from decellularized human adipose tissue in vitro and in vivo, Journal of Biomedical Materials Research Part B, 2018, 1684-1694 3. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells, Biomaterials, 2010, 4715-24 4. Preclinical Optimization of a Shelf-Ready, Injectable, Human-Derived, Decellularized Allograft Adipose Matrix, Tissue Engineering Part A 2019, Vol. 25, No. 3-4

Accordingly, it is an object of the present inventors to provide a medical composition and a method for preparing the same, wherein the composition remains in a well-aggregated state even after transplantation into the body and can maintain its volume for a certain period of time.

More specifically, by using an extracellular matrix powder derived from allogeneic or heterogeneous adipose tissue and a chemically cross-linked biocompatible polymer, it is possible to promote the production of autologous fat after implantation in the body and induce self-organization, and also have viscoelastic properties. An object of the present invention is to provide a medical composition having improved body volume retention and a method for manufacturing the same.

The present invention is adipose tissue-derived extracellular matrix powder; And it provides a medical composition comprising a biocompatible polymer or a cross-linked product of the biocompatible polymer.

In addition, the present invention is adipose tissue-derived extracellular matrix powder; And it provides a method for producing a medical composition comprising the step of mixing a biocompatible polymer or a cross-linked product of the biocompatible polymer.

The present invention is adipose tissue-derived extracellular matrix powder; And it provides a biocompatible polymer or a medical composition comprising a cross-linked product of the biocompatible polymer and a method for producing the same.

The medical composition according to the present invention is present in a well-aggregated state even after implantation in the body, and can maintain its volume for a certain period of time. Specifically, in the present invention, by using an extracellular matrix powder derived from allogeneic or heterogeneous adipose tissue and a biocompatible polymer or a chemically cross-linked biocompatible polymer, it is possible to promote the production of autologous fat after implantation in the body and induce self-organization. , In addition, the viscoelastic properties are improved and the body volume retention is excellent.

1 is a photograph confirming the physical properties according to the mixing ratio of the cross-linked product of extracellular matrix powder and biocompatible polymer during gamma sterilization in the manufacture of a medical composition according to an example of the present invention.
Figure 2 is a graph measuring the viscosity modulus, elastic modulus, and complex viscosity of the medical composition according to an example of the present invention.
3 is a photograph showing a composition extracted from a nude mouse after 6 weeks of injection of a medical composition according to an example of the present invention, and a graph showing the volume of the composition.
Figure 4 is a graph quantifying the influx of cells and photographs performed with H & E (haematoxylin and eosin) staining to histologically analyze the composition extracted from nude mice after 6 weeks of injection of the medical composition according to an example of the present invention;
5 is a photograph and quantitative graph showing Oil Red O staining performed to analyze fat production in a composition extracted from nude mice after 6 weeks of injection of a medical composition according to an example of the present invention.

The present invention is adipose tissue-derived extracellular matrix powder; And it relates to a medical composition comprising a biocompatible polymer or a cross-linked product of the biocompatible polymer.

In an embodiment of the present invention, by preparing a medical composition comprising a cross-linked product of adipose tissue-derived extracellular matrix powder and a biocompatible polymer, the medical composition can maintain its formulation even during radiation sterilization, and has excellent viscoelastic properties Confirmed. In addition, by conducting an in vivo experiment on the medical composition, it was confirmed that the ability to maintain volume in the body, self-organization ability, and autologous fat production ability were superior to those of the case of using the HA-CMC carrier.

Hereinafter, the medical composition according to the present invention will be described in more detail.

The medical composition of the present invention includes adipose tissue-derived extracellular matrix powder; and a biocompatible polymer or a cross-linked product of a biocompatible polymer.

In the present invention, adipose tissue-derived extracellular matrix powder (hereinafter referred to as extracellular matrix powder) is used as a medical material, and can promote autologous fat production and induce autologous tissue after transplantation in the body.

In one embodiment, the extracellular matrix (ECM) refers to a complex aggregate of biopolymers filling the tissue or extracellular space. The components of the extracellular matrix may vary depending on the type of cell or the degree of cell differentiation, and fibrous proteins such as collagen and elastin, complex proteins such as proteoglycans and glycosaminoglycans, and fibronectin, laminin, etc. of cell-adherent glycoproteins.

In one embodiment, the adipose tissue may be allogeneic or xenogeneic adipose tissue. The same species refers to humans, and heterogeneous refers to animals other than humans, ie, mammals such as pigs, cattle, and horses, as well as fish.

In one embodiment, the average particle diameter of the adipose tissue-derived extracellular matrix powder may be 100 to 800 μm. It is suitable for in vivo injection in the above particle size range, and injection is possible with a syringe.

In one embodiment, the content of the adipose tissue-derived extracellular matrix powder may be 1 to 30 parts by weight, 5 to 15 parts by weight, or 3 to 8 parts by weight relative to the total weight of the composition. In the above range, it is possible to inject with a syringe.

In the present invention, the biocompatible polymer or a cross-linked product of the biocompatible polymer can improve the viscoelastic properties of the medical composition, and can improve body volume retention. In this case, the crosslinked product of the biocompatible polymer refers to one or more chemically crosslinked biocompatible polymers.

In one embodiment, the molecular weight of the biocompatible polymer or the crosslinked product of the biocompatible polymer may be 10 kDa to 2 MDa.

In one embodiment, one or more selected from the group consisting of collagen, hyaluronic acid, chitosan, carboxymethylcellulose, alginate and gelatin may be used as the biocompatible polymer.

In one embodiment, the cross-linked product of the biocompatible polymer may be a cross-linked product of one or more biocompatible polymers selected from the group consisting of collagen, hyaluronic acid, chitosan, carboxymethyl cellulose, alginate and gelatin.

In one embodiment, the biocompatible polymer is crosslinked by a crosslinking agent, and the crosslinking agent is 1,4-butandiol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE). ), hexanediol diglycidyl ether (1,6-hexanediol diglycidyl ether), propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetra Methylene glycol diglycidyl ether (polytetramethylene glycol diglycidyl ether), neopentyl glycol diglycidyl ether (neopentyl glycol diglycidyl ether), polyglycerol polyglycidyl ether (polyglycerol polyglycidyl ether), diglycerol polyglycidyl ether ( diglycerol polyglycidyl ether, glycerol polyglycidyl ether, tri-methylpropane polyglycidyl ether, bisepoxypropoxyethylene (1,2-(bis(2,3-epoxypropoxy) ) ethylene), pentaerythritol polyglycidyl ether (pentaerythritol polyglycidyl ether), and sorbitol polyglycidyl ether (sorbitol polyglycidyl ether) may be at least one selected from the group consisting of.

In one embodiment, the content of the biocompatible polymer may be 0.1 to 20 parts by weight, 1 to 15 parts by weight, 1 to 11 parts by weight, or 9 to 11 parts by weight based on the total weight of the composition. Within the above range, the physical properties of the biocompatible polymer may be improved, and the body volume retention may be improved.

In the present invention, the viscosity modulus of the medical composition may be 3,000 to 20,000 Pa, the elastic modulus may be 1,000 to 10,000 Pa, and the complex viscosity may be 1,000 to 10,000 Pa·s. The viscous modulus, elastic modulus, and complex viscosity refer to results measured by a rotary rheometer analyzer (frequency: 0.1 to 10 Hz, temperature: 25°C, strain: 1%).

Viscoelasticity refers to a phenomenon in which the properties of a liquid and properties of a solid appear at the same time when a force is applied to an object. In the present invention, the viscous modulus, the elastic modulus and the complex viscosity can be measured by measuring the force that resists the force applied to the composition and the force that is lost.

The viscous modulus (loss modulus, viscous modulus, G'') is a measure of lost energy and means the viscous component of the material. In the present invention, the viscosity coefficient of the medical composition may be 5,000 to 10,000 Pa or 6,000 to 8,000 Pa. The elastic modulus (storage modulus, elastic modulus, G') means the ratio of stress and strain that an elastic body has within the elastic limit. The greater the modulus of elasticity, the harder the composition and the higher the ability to resist deformation. In the present invention, the elastic modulus of the medical composition may be 1,000 to 5,000 Pa Ehsms 1,000 to 3000 pa. The complex viscosity is a frequency-dependent viscosity calculated by the vibration measurement method, and the above numerical values reflect G'' and G' and the frequency values to be measured. In the present invention, the complex viscosity of the medical composition may be 1,000 to 3,000 Pa·s or 1,500 to 2,500 Pa·s.

In addition, the extrusion force of the medical composition, that is, the injection pressure may be 110 N or less. In the present invention, the extrusion force is a value measured using a universal testing machine. Specifically, the cannula is fastened to a syringe (20 G) containing the contents, and the contents in the syringe are removed by pressing the syringe at a test speed of 12 mm/min. It represents the maximum load value (N) when discharged to the outside of the cannula.

The extrusion force means the extrusion force at an injection rate comfortable for the patient. "Patient-friendly" is used to define an injection rate that does not cause injury or undue pain to the patient when injected into the skin. As used herein, "comfort" includes not only the comfort of the patient, but also the comfort or ability of a physician or healthcare professional to inject the composition. In general, those having a low extrusion force do not have tenderness during injection of the composition and are easy to control. In the present invention, the extrusion force of the medical composition may be 100 N or less, 70 N or less, 60 N or less, 40 N or less, 35 N or less, or 30 N or less.

In one embodiment, the medical composition of the present invention may be injected or inserted into a living body through injection into a syringe or the like. Such a medical composition can be used as a general medical material, and can be used as a tissue repair agent, a filler, an anti-adhesion agent, a cosmetic aid, an arthritis agent, a wound coating agent, a hemostatic agent, or a lipodystrophy agent. At this time, lipodystrophy has a symptom in which adipose tissue is lost, and the production of autologous fat can be promoted by the medical composition of the present invention.

The present invention also relates to a method for preparing the aforementioned medical composition.

The method for preparing the medical composition includes adipose tissue-derived extracellular matrix powder; and mixing a biocompatible polymer or a cross-linked product of the biocompatible polymer.

In the present invention, the adipose tissue-derived extracellular matrix powder may be a commercially available product, or may be prepared and used in a laboratory.

The adipose tissue-derived extracellular matrix powder is a delipidation step of removing the lipid component from the adipose tissue;

a decellularization step of removing cells from the adipose tissue from which the lipid component has been removed;

a freeze-drying step of freeze-drying the adipose tissue from which the cells have been removed; and

It may be prepared through a powdering step of pulverizing the freeze-dried lyophilisate.

The present invention may perform a washing step prior to performing the defatting step. In the washing step, the adipose tissue may be washed with sterile distilled water. Through the above step, impurities in the adipose tissue can be removed.

In the present invention, the delipidation step is a step of removing the lipid component from the adipose tissue.

In one embodiment, delipidation refers to the removal of a lipid component from a tissue.

In one embodiment, the removal of the lipid component may be performed by physical treatment or chemical treatment, and the physical treatment and chemical treatment may be performed together. When performed together, the order of execution is not limited.

In one embodiment, the type of physical treatment is not particularly limited and may be performed using pulverization. The pulverization may be performed using a pulverizing means known in the art, for example, a mixer, a homogenizer, a frozen pulverizer, an ultrasonic pulverizer, a hand blender, a plunger mill, and the like.

When performing the pulverization, the particle size of the pulverized product, that is, the pulverized adipose tissue may be 0.01 to 1 mm.

In one embodiment, the type of chemical treatment is not particularly limited, and may be performed using a delipidation solution. The delipidation solution may include a polar solvent, a non-polar solvent, or a mixed solvent thereof. Water, alcohol, or a mixed solution thereof may be used as the polar solvent, and methanol, ethanol or isopropyl alcohol may be used as the alcohol. As the non-polar solvent, hexane, heptane, octane, or a mixed solution thereof may be used. Specifically, in the present invention, a mixed solution of isopropyl alcohol and hexane may be used as the delipidation solution. In this case, the mixing ratio of isopropyl alcohol and hexane may be 40:60 to 60:40.

The treatment time of the delipidation solution may be 4 to 30 hours, or 10 to 20 hours.

In one embodiment, the defatting step may be performed by sequentially applying a physical treatment and a chemical treatment. The lipid component may be primarily removed from the adipose tissue by physical treatment, and the lipid component not removed by the physical treatment may be removed by chemical treatment.

In the present invention, the decellularization step is a step of removing cells from the adipose tissue from which the lipid component has been removed by the delipidation step.

In one embodiment, decellularization refers to the removal of other cellular components other than the extracellular matrix from a tissue, for example, a nucleus, a cell membrane, a nucleic acid, and the like.

In one embodiment, decellularization may be performed using a basic solution, and one or more selected from the group consisting of sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium carbonate, magnesium hydroxide, calcium hydroxide and ammonia may be used as the basic solution. have. In the present invention, sodium hydroxide (NaOH) may be used as the basic solution. The present invention has the advantage that there is no cytotoxicity by using a basic solution during decellularization.

In one embodiment, the concentration of the basic solution may be 0.01 to 1 N, 0.06 to 0.45 N, 0.06 to 0.2 N, or 0.08 to 1.02 N. It is easy to remove the cells in the above concentration range.

Also, in one embodiment, the decellularization step may be performed for 60 to 48 minutes, 70 to 200 minutes, or 90 to 150 minutes. Removal of cells in this time range is easy.

In the present invention, after performing the decellularization step, a centrifugation step may be additionally performed before performing the freeze-drying step. Through the centrifugation step, impurities in the delipidation step and the decellularization step can be removed, and a high-purity extracellular matrix material (precipitate) can be obtained.

In one embodiment, centrifugation may be performed at 4,000 to 10,000 rpm, or 8,000 rpm for 5 to 30 minutes, 5 to 20 minutes, or 10 minutes.

In addition, before and/or after centrifugation, a washing step may be additionally performed, and sterile distilled water may be used for washing.

In the present invention, the freeze-drying step is a step of freeze-drying the obtained product after the aforementioned step, that is, the decellularization step or the centrifugation step. The freeze-drying is a method of absorbing moisture in a vacuum after rapid cooling of the tissue in a frozen state. can

In one embodiment, freeze-drying may be performed at -50 to -80°C for 24-96 hours.

In the present invention, the powdering step is a step of powdering the lyophilized lyophilisate, that is, the extracellular matrix.

The particle diameter of the powdered extracellular matrix powder may be 100 to 800 μm.

In addition, the adipose tissue-derived extracellular matrix powder of the present invention includes a washing step of washing the adipose tissue;

a delipidation step of removing lipid components from the washed adipose tissue;

a decellularization step of removing cells from the adipose tissue from which the lipid component has been removed;

a centrifugation step of centrifuging the decellularized adipose tissue;

a freeze-drying step of freeze-drying the precipitate after the centrifugation; and

It can be prepared through; a powdering step of pulverizing the freeze-dried lyophilisate.

In the present invention, a commercially available product may be used as the biocompatible polymer or a cross-linked product of the biocompatible polymer. In addition, the cross-linked product may be prepared and used in a laboratory using a biocompatible polymer.

The cross-linking step of cross-linking the biocompatible polymer using a cross-linking agent; and

It can be prepared through a freeze-drying step of freeze-drying the cross-linked cross-linked product.

In the present invention, the crosslinking step is a step of crosslinking the biocompatible polymer using a crosslinking agent. In the above step, the biocompatible polymer and the crosslinking agent may be of the types described above.

In one embodiment, the biocompatible polymers may be bound through an amide bond.

In one embodiment, the content of the crosslinking agent may be 0.5 to 10 parts by weight based on the biocompatible polymer.

In the present invention, the freeze-drying step is a step of freeze-drying the biocompatible polymer crosslinked in the above step.

In one embodiment, lyophilization may be performed at -50 to -80°C for 24 to 96 hours.

Adipose tissue-derived extracellular matrix powder in the present invention; and a biocompatible polymer or a cross-linked product of the biocompatible polymer may be mixed by physical mixing.

In one embodiment, the content of the adipose tissue-derived cell matrix powder in the mixture may be 1 to 30 parts by weight, 5 to 15 parts by weight, or 3 to 8 parts by weight.

In addition, the content of the crosslinked material of the biocompatible polymer in the mixture may be 0.1 to 20 parts by weight, 1 to 15 parts by weight, 1 to 11 parts by weight, or 9 to 11 parts by weight.

In one embodiment, the mixture may be prepared by dissolving a cross-linked product of a freeze-dried biocompatible polymer in a solvent and mixing it with the extracellular matrix powder. In this case, physiological saline may be used as the solvent.

The present invention may further comprise the step of sterilizing the mixture.

Through the sterilization step, it is possible to remove the immunity in the medical composition, and it is possible to effectively destroy bacteria and the like.

In one embodiment, the sterilization step may be performed by irradiating radiation, and the irradiation range of radiation may be 10 to 30 kGy.

The invention also relates to the use of the aforementioned medical composition.

The medical composition according to the present invention can promote autologous fat production and induce self-organization after transplantation, and also has an excellent effect of maintaining body volume due to improved viscoelastic properties.

Accordingly, in one embodiment, the medical composition of the present invention may be injected or inserted into a living body through injection or the like with a syringe, and a tissue repair agent, a filler, an anti-adhesion agent, a molding aid, an arthritis treatment agent, a wound coating agent, a hemostatic agent, Or it may be used as a therapeutic agent for lipodystrophy.

The present invention will be described in more detail through the following examples. However, the scope of the present invention is not limited to the following examples, and it will be understood by those skilled in the art that various modifications, modifications or applications are possible within the scope without departing from the technical matters derived from the matters described in the appended claims. will be.

Example

Example 1. Human adipose tissue-derived extracellular matrix particles and chemically crosslinked biocompatibility Medical composition in which a polymer is physically mixed

(1) Preparation of extracellular matrix derived from human adipose tissue

Human adipose tissue was pulverized with a grinder to remove fat. In order to remove the fat that did not fall off, it was subjected to a de-fat process for 16 hours using 40% to 60% isopropyl alcohol and 40% to 60% hexane. Cells were removed by treatment with 0.1N sodium hydroxide (NaOH) in the tissue from which the fat was removed.

The supernatant was removed by centrifugation at 8,000 rpm for 10 minutes to wash the extracellular matrix from which fat and cells were removed, and the washing process was repeated 5 to 10 times. The support was freeze-dried so that the moisture content of the human adipose tissue-derived extracellular matrix was 10% or less, preferably 1% to 8%.

The lyophilized human adipose tissue-derived extracellular matrix was finely divided using a micro grinder.

(2) Preparation of chemically crosslinked biocompatible polymers

HA-CMC carrier was prepared by mixing hyaluronic acid (HA) and carboxymethylcellulose (CMC) with 1,4-butanediol diglycidyl ether (BDDE).

Specifically, a reaction solvent was prepared by adding 1 to 10 ml of BDDE per 100 ml of 0.1 to 1 N aqueous sodium hydroxide solution. To the reaction solvent, 1 to 10 g of CMC and 1 to 20 g of HA were added and then homogeneously mixed to prepare a mixed solution. Crosslinking was performed by heating the mixed solution at 50° C. for 3 hours.

The cross-linked reactant was placed in a dialysis membrane and dialyzed at room temperature with 5 L of phosphate-buffered saline. After 2 hours, it was replaced with 5 L of 50% EtOH and dialyzed at room temperature for 1 hour. After dialyzing with sterile distilled water at room temperature for 72 hours, the reaction product was lyophilized to finally obtain an HA-CMC carrier.

(3) Preparation of medical composition

The human adipose tissue-derived extracellular matrix (5% to 15% by weight) prepared in (1) and the HA-CMC carrier (1% to 10% by weight) prepared in (2) were mixed with sterile physiological saline.

The mixed final product was sterilized with 25 kGy gamma rays to prepare a medical composition.

Experimental Example 1. Verification of formulation retention ability during radiation sterilization of medical compositions

(1) Method

The formulation retention ability of the medical composition prepared in Example 1 was verified.

After preparing a sample with the contents of Table 1 below (the rest: sterile physiological saline), sterilization was performed with 25 kGy gamma rays. During gamma sterilization, the physical properties according to the content ratio of each component were confirmed.

On the other hand, the extrusion force was measured for each sample. The extrusion force is measured by fastening the cannula to the syringe containing the contents using a universal testing machine, and measuring the maximum load value (N) when the contents in the syringe are discharged to the outside of the cannula by pressing the syringe at a test speed of 12 mm/min. did.

(2) Results

The results of measurement of physical properties according to the content ratio of each component of the sample during gamma sterilization are shown in FIG. 1 .

In addition, the results of measuring the extrusion force of the sample are shown in Table 2.

sample number Human adipose tissue-derived extracellular matrix content (%) Carrier content (%) Extrusion force (injection pressure, N) 17G 20G One 5 2 26N 28N 2 10 2 23N 20N 3 15 2 32N 37N 4 5 4 26N 20N 5 10 4 19N 30N 6 15 4 35N 41N 7 5 6 22N 27N 8 10 6 23N 30N 9 15 6 36N 65N 10 5 8 28N 27N 11 10 8 23N 41N 12 15 8 38N 72N 13 5 10 19N 28N 14 10 10 23N 68N 15 15 10 48N 100N

As shown in Figure 1 and Table 2, a medical composition containing 5 wt% of human adipose tissue-derived extracellular matrix (ECM) and 10 wt% of an HA-CMC carrier has the best physical properties, and the pressure of 40 N or less at 20G It can be confirmed that it is easy to inject in vivo through a syringe by having an output.

Experimental Example 2. Analysis of viscoelastic properties of medical compositions

(1) Method

A medical composition (Sample 13) comprising 5% by weight of human adipose tissue-derived extracellular matrix (ECM) and 10% by weight of HA-CMC carrier selected in Experimental Example 1 as an optimal mixing ratio was used as an experimental group (medical composition), and the extracellular matrix A composition containing 10% by weight of HA-CMC without including HA-CMC was controlled (HA-CMC), and viscoelastic properties were compared.

Specifically, the elastic modulus, the viscous modulus, and the complex viscosity were measured using a rotary rheometer analyzer with a frequency of 0.1 to 10 Hz, a temperature of 25° C., and a strain of 1%.

(2) Results

The measurement results of the elastic modulus, the viscous modulus and the complex viscosity are shown in FIG. 2 .

As shown in FIG. 2, the medical composition according to the present invention exhibited about 7 times higher values in the modulus of elasticity and viscous modulus than HA-CMC, a control, and 6 times higher values in complex viscosity.

Experimental Example 3. In vivo performance verification of medical composition derived from human adipose tissue

(1) Method

An animal experiment was conducted to verify the performance of the medical composition.

The extracellular matrix powder (extracellular matrix) prepared in (1) of Example 1 and the HA-CMC carrier (HA-CMC) prepared in Example 1 (2) were used as controls, respectively, and the optimum in Experimental Example 1 A medical composition (Sample 13) comprising 5% by weight of human adipose tissue-derived extracellular matrix (ECM) powder and 10% by weight of HA-CMC carrier selected at a mixing ratio was used as an experimental group.

0.2cc of each of the composition of the control group and the experimental group was implanted under the dorsal subcutaneous tissue of BALB/c nude mice, and the results were analyzed by sacrificing the experimental animals 6 weeks after transplantation.

(2) Results

(A) Verification of body volume maintenance ability

Six weeks after the injection of the composition in the control group and the experimental group, samples extracted from nude mice were photographed, and the volume was measured using a digital caliper.

3 is a graph showing a photograph and a volume (Residual vol.) showing a composition extracted from a nude mouse.

As shown in FIG. 3 , it can be confirmed that the experimental group medical composition has superior volume retention in the body compared to the control group HA-CMC and the extracellular matrix.

From the graph, it can be confirmed that the medical composition maintains volume 4 times or more than the extracellular matrix after 6 weeks.

(B) Self-organization verification

For the samples extracted in (A), it was checked whether autologous tissue was formed by tissue staining. H&E (haematoxylin and eosin) staining was performed to perform tissue analysis, and cell influx was quantified.

FIG. 4 is a graph quantifying cell influx and photographs obtained by performing H&E (haematoxylin and eosin) staining for histological analysis of compositions extracted from nude mice.

As shown in FIG. 4 , as a result of tissue staining, it can be confirmed that cells are introduced into the medical composition, which is the experimental group compared to the control group HA-CMC and the extracellular matrix, and blood vessels are formed.

In addition, through the graph, it can be confirmed that the cell influx of the medical composition is increased 8 times more than that of HA-CMC after 6 weeks.

(C) Verification of the effect of autologous fat production

Adipogenesis was verified for the samples extracted in (A).

To analyze the adipogenesis in the extracted samples, Oil Red O staining was performed, and the adipogenesis was quantified.

5 is a photograph and quantified graph of Oil Red O staining to analyze fat production in a composition extracted from nude mice.

As shown in FIG. 5, as a result of staining with Oil Red O, it can be confirmed that lipogenesis in the medical composition of the experimental group is induced more than that of HA-CMC and the extracellular matrix of the control group.

In addition, through the graph, it can be confirmed that the medical composition increased lipogenesis by 8% or more compared to the extracellular matrix after 6 weeks.

Claims (17)

adipose tissue-derived extracellular matrix powder; and a cross-linked product of a biocompatible polymer,
The content of the crosslinked product of the biocompatible polymer is 0.1 to 20% by weight based on the total weight of the composition,
The cross-linked product of the biocompatible polymer is a medical composition that is a cross-linked product of hyaluronic acid and carboxymethyl cellulose.
The method of claim 1,
The average particle diameter of the adipose tissue-derived extracellular matrix powder is 100 to 800 μm medical composition.
The method of claim 1,
The content of the adipose tissue-derived extracellular matrix powder is 1 to 30% by weight based on the total weight of the composition for a medical composition.
delete The method of claim 1,
The cross-linked product of the biocompatible polymer is produced by cross-linking with a cross-linking agent,
The crosslinking agent is butanediol diglycidyl ether (1,4-butandiol diglycidyl ether, BDDE), ethylene glycol diglycidyl ether (EGDGE), hexanediol diglycidyl ether (1,6-hexanediol) diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, polytetramethylene glycol diglycidyl ether, Neo Pentyl glycol diglycidyl ether (neopentyl glycol diglycidyl ether), polyglycerol polyglycidyl ether (polyglycerol polyglycidyl ether), diglycerol polyglycidyl ether (diglycerol polyglycidyl ether), glycerol polyglycidyl ether ), tri-methylpropane polyglycidyl ether, bisepoxypropoxyethylene (1,2-(bis(2,3-epoxypropoxy)ethylene), pentaerythritol polyglycidyl ether Polyglycidyl ether), and sorbitol polyglycidyl ether (sorbitol polyglycidyl ether) at least one selected from the group consisting of a medical composition.
delete The method of claim 1,
The viscosity coefficient of the medical composition is 3,000 to 20,000 Pa,
The modulus of elasticity is 1,000 to 10,000 Pa,
The complex viscosity is 1,000 to 10,000 Pa s,
The extrusion force is 110 N or less medical composition.
The method of claim 1,
The medical composition is a medical composition that is used as a tissue repair agent, filler, anti-adhesion agent, cosmetic aid, arthritis treatment agent, wound coating agent, hemostatic agent, or lipodystrophy treatment agent.
adipose tissue-derived extracellular matrix powder; and mixing a cross-linked product of a biocompatible polymer,
The content of the crosslinked product of the biocompatible polymer is 0.1 to 20% by weight based on the total weight of the composition,
The cross-linked product of the biocompatible polymer is a method for producing a medical composition that is a cross-linked product of hyaluronic acid and carboxymethyl cellulose.
10. The method of claim 9,
Adipose tissue-derived extracellular matrix powder is a delipidation step of removing the lipid component from the adipose tissue;
a decellularization step of removing cells from the adipose tissue from which the lipid component has been removed;
a freeze-drying step of freeze-drying the adipose tissue from which the cells have been removed; and
A method of manufacturing a medical composition that is prepared through; a powdering step of powdering the freeze-dried lyophilisate.
11. The method of claim 10,
Removal of the lipid component is carried out by physical treatment and/or chemical treatment,
The physical treatment is grinding,
A method for producing a medical composition wherein the chemical treatment is performed using a delipidation solution.
12. The method of claim 11,
The delipidation solution is a method for producing a medical composition comprising a polar solvent, a non-polar solvent, or a mixed solvent thereof.
11. The method of claim 10,
Decellularization is performed using a basic solution,
The basic solution is a method of manufacturing a medical composition comprising at least one selected from the group consisting of sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium carbonate, magnesium hydroxide, calcium hydroxide and ammonia.
11. The method of claim 10,
After performing the decellularization step, the method for producing a medical composition to further perform the step of centrifugation.
10. The method of claim 9,
A crosslinking step of crosslinking the biocompatible polymer using a crosslinking agent; and
A method for preparing a medical composition that is prepared through a freeze-drying step of freeze-drying the cross-linked cross-linked product.
16. The method of claim 15,
The content of the crosslinking agent is 0.5 to 10% by weight relative to the biocompatible polymer method for producing a medical composition.
10. The method of claim 9,
The method for preparing a medical composition further comprising the step of sterilizing the mixed mixture.

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