KR20180092348A - Composition for inducing chondrogenic differentiation or regenerating cartilage tissue comprising exosomes derived from umbilical cord stem cells and cord blood stem cells which differentiate into chondrocytes - Google Patents

Abstract

The present invention relates to a composition of regenerating cartilage tissues or inducing differentiation of chondrocytes while including exosomes isolated from stem cells derived from cord blood or umbilical cords being differentiated into the chondrocytes as an active ingredient, a medium composition for inducing the differentiation of chondrocytes including the composition, an injection agent for regenerating cartilage tissues, and a pharmaceutical composition for treating cartilage diseases. According to a specific embodiment of the present invention, the composition of regenerating cartilage tissues or inducing differentiation of chondrocytes includes the exosomes isolated from stem cells derived from cord blood or umbilical cords being differentiated into the chondrocytes as an effective factor. The exosomes secreted during the differentiation of stem cells contain growth factors related to the differentiation of chondrocytes as well as genes and proteins related to the proliferation and regeneration of stem cells, thereby having excellent properties related to regeneration of damaged cartilage tissues. The composition can accordingly minimize the side effects of an existing cell treatment agent and reliably and quickly transfer effective substances into the cells as a cell-derived carrier. The injection agent for treatment and prevention of cartilage diseases can reduce the pain of patients with a simple treatment operation and enable continuous and effective treatment of cartilage diseases after the treatment operation.

Description

TECHNICAL FIELD The present invention relates to a composition for inducing chondrocyte differentiation or regeneration of chondrogenic tissue comprising exosome isolated from umbilical cord blood-derived stem cells differentiated into cartilage cells or regenerating cartilage tissue derived from exosomes derived from umbilical cord stem cells and cord blood stem cells which differentiate into chondrocytes}

The present invention relates to a composition for inducing chondrocyte differentiation induction or cartilage tissue regeneration comprising, as an active ingredient, exosome isolated from umbilical cord stem cells and / or umbilical cord blood stem cells under differentiation into chondrocytes, A pharmaceutical composition for the treatment of cartilage diseases, and the like.

The present invention also relates to a method for inducing differentiation of chondrocytes or regenerating cartilage tissue using the exosome isolated from umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells undergoing differentiation into cartilage cells, and a method for treating and improving cartilage diseases.

Because of the nature of the cartilage tissue, it is difficult to regenerate tissue due to natural healing when the large area is damaged. Therefore, it has been treated through surgical operations such as artificial joints, articular cartilageoplasty and microperforation. However, the conventional method has scar due to the incision and has a problem in that the therapeutic effect is inferior compared to the difficult treatment method due to the regeneration of the durability fiber cartilage. Therefore, injectable solutions using hydrogels, which are easy to operate and have a rapid therapeutic effect, include injections for intraarticular administration using an alkylene diamine crosslinked hydrogel of hyaluronic acid (Korean Patent Publication No. 10-2013-0028012), collagen (Korean Patent Registration No. 10-1279812), a composition for cartilage grafting using a hydrogel containing collagen, hyaluronic acid, and fibrinogen (Korean Patent Laid-Open Publication No. 10-2012- 0046430) have been developed. However, although the above technique can temporarily reduce the pain, there is a shortage of induction of regeneration into cartilage tissue, so effective factors that help differentiate cartilage tissue are needed.

In the case of known cell-based therapies, autologous chondrocytes (Korean Patent Publication No. 10-2013-0072983), umbilical cord Derived stem cells (Korean Patent Laid-open Publication No. 10-2013-0009651), and a trophoblast-derived stem cell (Korean Patent Laid-open Publication No. 10-2016-0036031). However, in the case of autologous chondrocyte treatment, there is a limit to the treatment with only the cultured cells taken from the patient when the injured area is large, and the pain and the economic burden of the patient due to the two basic operations until the transplantation operation are great. In the case of stem cell therapy, cell therapy using stem cells such as umbilical cord and umbilical cord blood, bone marrow, adipose tissue, synovial membrane, and muscle stem have been performed, but differences in cell number and differentiation ability depending on the harvest site, , There is a problem that calcification of chondrocytes is caused by vascular permeability induced by cell apoptosis due to gene expression associated with a change in cell phenotype due to transplantation, a low differentiation rate into chondrocytes, and cell hypertrophy.

Thus, in the prior art, chondrocytes or adult stem cells obtained from a patient were cultured in vitro, dispersed in a hydrogel, and then transplanted into a defect site. Although it is possible to regenerate tissues close to normal cartilage by injecting cells directly, it is difficult to perform the inevitable surgical procedure to obtain cells, difficulties of in vitro culture process, limitation of cell number according to the size of damaged tissue site, I have a problem. In addition, once administration, it is possible to temporarily reduce the pain and increase the motility of the joint, but ultimately it has been difficult to regenerate damaged cartilage tissue.

Accordingly, the present inventors have conducted intensive studies to solve the conventional problems. As a result, the present inventors have found that when exosome isolated from umbilical cord-derived stem cells and / or cord blood-derived stem cells undergoing differentiation into chondrocytes is used, induction of differentiation into chondrocytes and / A composition for inducing chondrocyte differentiation induction or cartilage tissue regeneration comprising an exosome isolated from umbilical cord stem cells and / or umbilical cord blood stem cells, which is confirmed to be excellent in cartilage tissue regeneration, and which is differentiated into cartilage cells, A medium composition for inducing differentiation of cartilage cells, an injectable preparation for cartilage tissue regeneration, and a pharmaceutical composition for treating cartilage disease.

Korean Patent Publication No. 10-2013-0028012 (March 31, 2013) Korean Registered Patent No. 10-1279812 (2013.06.28) Korean Patent Publication No. 10-2012-0046430 (2012.05.10) Korean Patent Publication No. 10-2013-0072983 (2013.07.02) Korean Patent Publication No. 10-2013-0009651 (Feb. 23, 2013) Korean Patent Publication No. 10-2016-0036031 (2014.04.01)

It is an object of the present invention to provide a composition for inducing differentiation of chondrocytes or regenerating cartilage tissue, which comprises, as an active ingredient, exosome isolated from umbilical cord stem cells and / or cord blood-derived stem cells undergoing differentiation into cartilage cells.

Another object of the present invention is to provide a medium composition for inducing chondrocyte differentiation induction or composition for regenerating cartilage tissue.

It is still another object of the present invention to provide an injection for regeneration of cartilage tissue comprising the composition for inducing differentiation of cartilage cells or regenerating cartilage tissue.

It is still another object of the present invention to provide a pharmaceutical composition for treating cartilage diseases, comprising the composition for inducing differentiation of cartilage cells or regenerating cartilage tissue.

Another object of the present invention is to provide a method of inducing differentiation of cartilage cells, a method of regenerating cartilage tissue, a method of regenerating cartilage disease, and a method of regenerating cartilage by using exosome isolated from umbilical cord stem cells and / And to provide a method of treatment and improvement.

In order to achieve the above object, one embodiment of the present invention provides a method for inducing chondrocyte differentiation or cartilage differentiation comprising chondrocyte-derived stem cells and / or umbilical cord blood-derived stem cells differentiated into chondrocytes as an active ingredient A composition for tissue regeneration is provided.

As used herein, the term "cord-derived stem cell and / or cord blood-derived stem cell under differentiation into chondrocytes" refers to stem cells derived from umbilical cord tissue and / or cord blood derived stem cells, It refers to stem cells that are differentiating into cartilage cells. From this, it is possible to separate exosomes containing chondrocyte genetic information, proteins, and growth factors.

Specifically, when stem cells derived from cord tissue and / or stem cells derived from cord blood are differentiated into chondrocytes, the shape and characteristics of cells are changed, and then exosome is extracted. Therefore, it is different from extracting exosome from normal stem cells.

The term "exosome " as used herein in the specification of the present invention refers to a pseudocyst of a membrane structure secreted from various kinds of cells. It is a protein that binds to other cells and tissues and transmits membrane components, proteins, It is known to play a role.

The exosome isolated from umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells differentiated into chondrocytes may have the basic characteristics of stem cells, and may contain important growth factors, various bioactive proteins, and gene information at the time of cartilage differentiation ≪ / RTI >

The exosome can be prepared by using the exosome separation method known in the art, for example,

1) proliferation of umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells;

2) differentiating the proliferated umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells into chondrocytes; And

3) Separation and purification of exosome from umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells differentiated into cartilage cells can be used.

The expression "inducing differentiation of chondrocytes" may mean inducing stem cells to differentiate into chondrocytes.

The expression "cartilage tissue regeneration" may mean regeneration of damaged cartilage tissue or induction of insufficient cartilage tissue regeneration.

The composition for regenerating cartilage according to the present invention is different from the conventional techniques in that it uses exosomes isolated from umbilical cord-derived stem cells and / or umbilical cord blood-derived stem cells that are differentiated into cartilage cells as an effective substance for cartilage regeneration . In particular, stem cells derived from human umbilical cord and / or stem cells derived from umbilical cord blood are superior to bone marrow or adipose derived stem cells in terms of cell proliferation and ability to differentiate into cartilage cells, thereby effectively inducing regeneration of cartilage tissue. The composition for regenerating cartilage according to the present invention is capable of effectively regenerating cartilage tissue that is sustainable by various growth factors related to the proliferation and differentiation of cells carried on the separated and purified exosome, The problem of in vitro cell culture of fibroblast, regeneration of fibrocartilage, or calcification of tissue due to cell death can be solved.

The exosomes derived from the umbilical cord stem cells derived from the umbilical cord stem cells and the umbilical cord blood stem cell-derived exosomes during the differentiation into chondrocytes according to the present invention not only have the characteristics of the stem cells but also effectively transmit the differentiation-related active factors to the chondrocytes , The side effects can be minimized and the same effect as the treatment using existing stem cells can be expected.

The exosome-derived stem cell-derived exosomes and / or cord blood stem cell-derived exosomes during the differentiation into cartilage cells according to the present invention are biomembrane endoplasmic reticulum secreted from cells. In addition, since it has a lipid structure similar to a cell membrane, it is excellent in absorption rate into peripheral cells when injected into the body, and it is possible to induce effective regeneration of cartilage tissue by rapid delivery of an effective substance.

Another embodiment of the present invention provides a medium composition for inducing differentiation of chondrocytes comprising the composition for inducing differentiation of cartilage cells or regenerating cartilage tissue.

The media composition is prepared by mixing the exosomes with 1 to 10 x 10 ⁷ particles / mL concentration, specifically 1 to 5 x 10 ⁷ particles / mL concentration, for example 3 x 10 ⁷ particles / mL Concentration, but is not limited thereto.

The medium composition for inducing differentiation of cartilage cells is characterized in that dexamethasone, insulin, ascorbate, chondrogenic growth factor IGF (Insulin) are used to differentiate umbilical cord stem cells and / or cord blood derived stem cells into cartilage cells. -like Growth Factor) and / or TGF-? 1 (Transforming Growth Factor? 1), but the present invention is not limited thereto.

Another embodiment of the present invention provides an injection for regeneration of cartilage tissue comprising the composition for inducing differentiation of cartilage cells or regenerating cartilage tissue.

The injection may further include phosphate-buffered saline (PBS). That is, the injectable preparation can be used by supporting a composition for inducing chondrocyte differentiation or regenerating cartilage tissue in phosphate buffered saline.

The injection may contain a hydrogel instead of phosphate buffered saline.

The hydrogel may be any one or more selected from the group consisting of hyaluronic acid, gelatin, alginate, chitosan, fibrin, elastin, collagen and methylcellulose, and may be hyaluronic acid hydrogel, but is not limited thereto.

The injectable formulation may contain exosomes at a concentration of 1 to 10 x 10 ⁹ particles / mL, specifically 1 to 5 x 10 ⁹ particles / mL, for example, 3 x 10 ⁹ particles / mL concentration But is not limited thereto.

The injection may be injected into a damaged area of cartilage of a mammal such as a mouse, a mouse, a livestock, or a human.

The composition for cartilage regeneration according to the present invention is economical in terms of operation time and cost because it is easy to inject into the body as an injectable composition, thereby reducing patient suffering, sequelae and economic burden. In addition, when the stem cells are differentiated into chondrocytes, the secreted exo-somes contain various extracellular matrix derivatives and various growth factors related to cell proliferation and differentiation, thus enabling effective regeneration of injured cartilage tissue. Therefore, long-term effects can be expected with a single procedure, which can overcome the problems that had to be performed periodically in order to obtain a lasting effect.

Another embodiment of the present invention provides a pharmaceutical composition for the treatment of cartilage diseases comprising the composition for inducing differentiation of cartilage cells or regenerating cartilage tissue.

The cartilage disease may be a cartilage disease derived from damage of cartilage tissue and is selected from the group consisting of osteoarthritis, degenerative arthropathy, cartilage dyssynergia, degenerative arthritis, rheumatoid arthritis, osteomalacia, But is not limited thereto.

The treatment of the cartilage disease may mean that cartilage tissue damage is treated by regenerating the damaged cartilage by intra-articular injection of the pharmaceutical composition for treating cartilage disease.

The pharmaceutical composition according to the above embodiments may be various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.

Examples of the pharmaceutical composition according to the present invention include non-aqueous solutions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

Dosage forms of the pharmaceutical compositions according to the above embodiments may also be used in the form of their pharmaceutically acceptable salts and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable set. The salt is not particularly limited as long as it is pharmaceutically acceptable so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid.

The pharmaceutical composition according to the above embodiments may be administered parenterally or orally according to the purpose, and may be administered once to several times to be administered in an amount of 0.1 to 500 mg and 1 to 100 mg per kg of body weight per day can do. The dosage for a particular patient may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease, and the like.

The pharmaceutical compositions according to the above embodiments may be formulated into oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories and sterilized injection solutions And may be formulated in any form suitable for pharmaceutical preparations.

The pharmaceutical compositions according to the above embodiments may be administered to mammals such as rats, mice, livestock, and humans in various routes such as parenteral, oral, etc., and all manner of administration may be expected, Or by intravenous, intramuscular, subcutaneous, intramural or intracerebroventricular injection.

The pharmaceutical composition according to the above embodiment may be used for differentiating stem cells derived from umbilical cord and / or stem cells derived from cord blood into chondrocytes such as dexamethasone, insulin, ascorbate, chondrogenic growth factor IGF -like Growth Factor) and / or TGF-? 1 (Transforming Growth Factor? 1), but the present invention is not limited thereto.

In another aspect of the present invention, the pharmaceutical composition for the treatment of cartilage diseases is characterized in that the exosome is at least one of a surface, an inside, or a surface selected from the group consisting of a solid or a semi-scaffold scaffold and a membrane, Or may be contained in a liquid formulation. For example, the support and the membrane may be formed of at least one biodegradable synthetic polymer selected from the group consisting of polylactic acid (PLA), polyglutamic acid (PGA) and poly lactic-co-glycolic acid (PLGA), collagen, matrix.

Another aspect of the present invention relates to a method for producing a chondrocyte-derived stem cell, comprising the steps of: (a) removing at least one selected from the group consisting of exosome isolated from umbilical cord stem cells differentiated into cartilage cells and exosome isolated from cord blood- (B) preparing a composition for inducing chondrocyte differentiation or cartilage tissue regeneration prepared in the step (a) to a cartilage tissue or its surroundings; Thereby inducing chondrocyte differentiation or cartilage tissue regeneration. In the chondrocyte differentiation induction or cartilage tissue regeneration method of the present invention, the composition for inducing chondrocyte differentiation or cartilage tissue regeneration is at least one surface selected from the group consisting of solid or semi-solid scaffolds and membranes, , Or both on the surface and inside, or contained in a liquid dosage form.

Yet another further aspect of the present invention relates to a method for the treatment of osteoblast-derived stem cells comprising the steps of: (a) culturing at least 1 < RTI ID = 0.0 > Comprising the steps of: (a) preparing a pharmaceutical composition for treating a cartilage disorder, comprising the step of: (a) applying a pharmaceutical composition for treating cartilage disease prepared in step (a) Treatment and improvement. In the cartilage disease treating and improving method of the present invention, the pharmaceutical composition for treating cartilage disease is at least one kind selected from the group consisting of solid or semi-solid scaffolds and membranes, , Or may be contained in a liquid formulation.

The composition for inducing chondrocyte differentiation or cartilage regeneration according to an embodiment of the present invention contains exosomes isolated from cord-derived stem cells and / or cord blood-derived stem cells differentiating into chondrocytes as an effective factor. The exosomes secreted during the differentiation of stem cells contain genes and proteins related to cell proliferation and regeneration of stem cells, as well as growth factors related to chondrocyte differentiation, and thus induce regeneration of injured cartilage tissue. In addition, the adverse effect on the existing cell treatment agent can be minimized, and the cell can be stably and rapidly delivered as a cell-derived carrier. Therefore, it is an injectable composition for prevention and treatment of cartilage diseases, which not only reduces the suffering of patients by simple procedures, but also enables continuous and effective treatment of cartilage disease after the treatment.

Hereinafter, the present invention will be described in more detail in the following Examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Example 1 Isolation of exosomes from stem cells differentiated into chondrocytes

Human umbilical cord-derived stem cells were cultured in a culture medium (10% fetal bovine serum, Minimum Essential Medium Eagle-Alpha Modification (Alpha MEM) containing 50 mg / mL gentamicin) to separate exosomes from chondrocyte- ], 10 [mu] g / mL TGF [gamma]), 10 [mu] g / mL streptomycin, 100 nM dexamethasone, 0.15 mM ascorbic acid, (Alpha-MEM containing [beta] 1 (Transforming Growth Factor [beta] 1), and the cells were cultured for 5 weeks to induce differentiation into chondrocytes.

After replacing the differentiation medium, the stem cells were washed with phosphate buffered saline (PBS) before the exosomes were separated, and the differentiation medium was replaced with DMEM medium without phenol red in serum-free medium After 24 hours of holding, the cell culture supernatant was recovered. The recovered cell culture supernatant was centrifuged at 300 × g for 10 minutes to remove the cells, and the remaining cell secretion was removed using a 0.22 μm filter. Thereafter, a process of removing mycoplasma and micro-vesicles using a 0.1 mu m filter was performed. From the solution containing exosomes and other proteins, other protein impurities except exosomes were removed using Tangential Flow Filtration (TFF, cut off 500 kDa) system, the exosomes were concentrated and the concentrated exo The cells were washed with phosphate buffered saline. Then, the exosomal solution obtained in the first step is sonicated to disperse the nanosized exosomes in the solution evenly, and again, through the TFF system, from the stem-derived stem cells which are finally differentiated into chondrocytes A high-purity exosome (chondrogenic differentiating umbilical cord stem cell-derived exosome (UC-DEXO)) was isolated.

≪ Comparative Example 1 > Isolation of exosome from proliferating umbilical cord-derived stem cells

Umbilical cord-derived exosomes (UCB-PEXO) were isolated from proliferating umbilical cord stem cells to compare the efficacy of exosomes isolated from umbilical cord-derived stem cells differentiated into chondrocytes. Specifically, exosomes were isolated from human umbilical cord-derived stem cells by performing the same method as in Example 1, except that the differentiation medium was not used.

<Example 2> Characterization of exosome

The exosomes isolated from the stem cells differentiated into chondrocytes of Example 1 and the exosomes isolated from the proliferating stem cells of Comparative Example 1 were analyzed by a transmission electron microscope and a dynamic light scattering And the exosomal membrane surface proteins were identified using Exo-Check ^™ exosome antibody arrays showing the presence or absence of specific protein expression.

Example 3 Evaluation of cartilage regeneration in vivo using exosomes isolated from stem cells differentiated into cartilage cells

Actual in vivo cartilage regeneration effect of exosomes isolated from umbilical cord-derived stem cells differentiated into cartilage cells using a mouse model of destabilization of the medial meniscus (DMM) arthritis commonly used as an osteoarthritis model Respectively.

Specifically, the periphery of the knee of the mouse was cleaned, and the knee joint part was cut about 1 cm along the side of the patella using a surgical tool. After exposing the joint capsule, the medial meniscotibial ligament connected to the medial meniscus was cut, the joint capsule and skin were closed in order, and the suture was closed. Respectively.

From 5 to 10 weeks of osteoarthritis, the composition containing the exosomes was injected into the joints once a week. The exosome composition was prepared by loading exosomes (UC-DEXO) isolated from umbilical cord-derived stem cells differentiated into cartilage cells of Example 1 in phosphate buffered saline. The exosomal composition was injected with 30 μL (1 × 10 ⁸ particles / 30 μL per mouse) in the joints of the mice at a final concentration of 3.3 × 10 ⁹ particles / mL. As a negative control, phosphate buffered saline was used. As a comparative control group, Umbilical cord-derived exosome (UCB-PEXO) extracted from umbilical cord-derived stem cells and human fat-derived stem cells differentiated into chondrocytes Chondrogenic differentiating adipose stem cell-derived exosome (ASC-DEXO) was used. After 11 weeks, cartilage tissue regeneration was confirmed using safranin-o staining.

Although the present invention has been described with reference to the above embodiments, the present invention is not limited thereto. It will be understood by those skilled in the art that modifications and variations may be made without departing from the spirit and scope of the invention, and that such modifications and variations are also contemplated by the present invention.

Claims (14)

Inducing chondrocyte differentiation comprising at least one member selected from the group consisting of exosome isolated from umbilical cord-derived stem cells differentiated into cartilage cells and exosome isolated from cord blood-derived stem cells differentiated into cartilage cells as an active ingredient Or cartilage tissue. A medium composition for inducing differentiation of chondrocytes comprising at least one selected from the group consisting of exosome isolated from umbilical cord-derived stem cells differentiated into cartilage cells and exosome isolated from cord blood-derived stem cells differentiated into cartilage cells . 3. The method of claim 2,
Wherein the medium composition comprises the exosome at a concentration of 1 to 10 x 10 ⁷ particles / mL. For regenerating cartilage tissue containing at least one selected from the group consisting of exosome isolated from umbilical cord-derived stem cells differentiated into cartilage cells and exosome isolated from cord blood-derived stem cells differentiated into cartilage cells as an active ingredient Injection. 5. The method of claim 4,
Wherein the injection comprises exosomes at a concentration of 1 to 10 x 10 ⁹ particles / mL. A method for treating cartilage disease comprising at least one selected from the group consisting of exosome isolated from umbilical cord-derived stem cells differentiated into cartilage cells and exosome isolated from cord blood-derived stem cells differentiated into cartilage cells as an active ingredient A pharmaceutical composition. The method according to claim 6,
Characterized in that the exosome is carried on at least one surface, inside, or both on the surface and inside, selected from the group consisting of a solid or semi-solid scaffold and a membrane, or contained in a liquid formulation A pharmaceutical composition for treating diseases. 8. The method of claim 7,
The support and the membrane may be formed of at least one biodegradable synthetic polymer selected from the group consisting of polylactic acid (PLA), polyglutamic acid (PGA) and poly lactic-co-glycolic acid (PLGA), collagen, or an extracellular matrix &Lt; / RTI &gt; wherein the pharmaceutical composition for treating cartilage disease is a pharmaceutical composition for treating cartilage disease. 9. The method according to any one of claims 6 to 8,
Wherein the cartilage disease is selected from the group consisting of osteoarthritis, deformed arthritis, cartilage dysplasia, degenerative arthritis, rheumatoid arthritis, osteomalacia, fibrous osteitis, and osteoarthritis. (a) inducing chondrocyte differentiation comprising at least one selected from the group consisting of exosome isolated from umbilical cord-derived stem cells differentiated into cartilage cells and exosome isolated from cord blood-derived stem cells differentiated into cartilage cells Or a composition for regenerating cartilage tissue,
(b) applying a composition for inducing chondrocyte differentiation or regeneration of cartilage tissue prepared in the step (a) to cartilage tissue or its periphery, or inducing cartilage differentiation or cartilage tissue regeneration. 11. The method of claim 10,
The composition for inducing differentiation of cartilage cells or regenerating cartilage tissue is supported on at least one surface, inside, or both on the surface and inside of at least one selected from the group consisting of solid or semi-solid scaffolds and membranes, Wherein the method further comprises the step of introducing into the formulation a cartilage cell differentiation inducing or cartilage tissue regeneration method. (a) at least one selected from the group consisting of exosomes isolated from umbilical cord-derived stem cells differentiated into chondrocytes and exosomes isolated from cord blood-derived stem cells differentiated into chondrocytes; Preparing a pharmaceutical composition,
(b) applying the pharmaceutical composition for treating cartilage disease prepared in step (a) to a cartilage disease site or its periphery. 13. The method of claim 12,
The pharmaceutical composition for treating cartilage disease is coated on at least one surface selected from the group consisting of solid or semispherical scaffolds and membranes, inside, or both on the surface and inside, or contained in the liquid formulation &Lt; / RTI &gt; further comprising the steps of: The method according to claim 12 or 13,
Wherein the cartilage disease is selected from the group consisting of osteoarthritis, degenerative arthritis, cartilage dysplasia, degenerative arthritis, rheumatoid arthritis, osteomalacia, fibrous osteitis, and osteoarthritis.