KR-102961974-B1 - Manufacturing method of animal cartilage-derived injectable composition and uses thereof

Abstract

The present invention relates to a method for preparing an injectable animal cartilage-derived injectable composition, an injectable composition prepared by said method, and the use thereof. The injectable formulation according to the present invention comprises a collagen-containing biomaterial in a formulation suitable for intra-articular injection, and can induce cartilage tissue regeneration by promoting tissue repair through direct injection into the application site without surgical incision. Furthermore, the animal cartilage-derived extracellular matrix contained in the injectable formulation not only serves to protect articular cartilage tissue but also stimulates an environment capable of regenerating damaged joint tissue by inducing intra-articular stem cells to differentiate into chondrocytes, thereby treating osteoarthritis; thus, it can be applied as a treatment for arthritis.

Inventors

• 강경선
• 이승희
• 안종찬
• 김미진

Assignees

• 주식회사 강스템바이오텍
• 에이템즈 주식회사

Dates

Publication Date

20260511

Application Date

20230410

Priority Date

20201127

Claims (9)

1. a) a step of solubilizing a composition comprising a sterilized animal cartilage-derived cell-free extracellular matrix (cartilage acellular marix; CAM) sponge by stirring at a temperature of 33°C to 37°C and performing the process for less than 24 hours; and b) a step of centrifuging the aqueous solution of a) above and collecting the solution to prepare an injectable composition; comprising a method for preparing an injectable composition for the prevention or treatment of osteoarthritis, wherein the injectable composition has tissue repair and cartilage tissue regeneration effects.
2. delete
3. delete
4. The method of claim 1, wherein the method further comprises the step of sterilizing the manufactured injectable composition.
5. A method according to claim 1, wherein the animal is a pig.
6. An injectable composition for the prevention or treatment of osteoarthritis prepared by the method of claim 1.
7. In claim 6, the injectable composition is one that contains collagen at a concentration of 120 μg/mg or more based on the total weight of the composition.
8. In claim 6, the injectable composition is one having tissue repair and cartilage tissue regeneration effects.
9. A method for the prevention or treatment of osteoarthritis comprising the step of administering the injectable composition of claim 6 into the joint cavity of an individual other than a human.

Description

Manufacturing method of animal cartilage-derived injectable composition and uses thereof The present invention relates to a method for preparing an injectable animal cartilage-derived injectable composition, an injectable composition prepared by said method, and the use thereof. Conventional methods for treating damaged cartilage include debridement, bone marrow stimulating techniques, osteochondrocyte transplantation, and autologous chondrocyte transplantation. These are invasive procedures primarily performed when cartilage damage has progressed significantly, whereas the injection of hyaluronic acid products into the joint cavity is commonly used as a treatment applicable in the early stages of cartilage damage. Most invasive treatment methods present problems such as surgical resection, periosteum harvesting, complexity of use, high treatment costs, leakage of adult stem cells induced during bone marrow stimulation, and the formation of abnormal fibrotic cartilage due to hemostasis issues. Furthermore, the injection of hyaluronic acid products serves only the simple role of alleviating pain through lubrication (Frizziero L, et al., Clinical and Experimental Rheumatology, 01 Jul 1998, 16(4):441-449). Therefore, to address the aforementioned problems, there is an urgent need to develop a therapeutic agent that enables minimally invasive procedures and possesses therapeutic efficacy beyond simple lubrication. Meanwhile, the extracellular matrix (ECM) is the part of a tissue excluding cells, and it is an aggregate of biomolecules that creates an environment capable of maintaining the natural structure of biological tissues by filling the gaps between cells and physically supporting the tissue. In particular, collagen, a major component of the extracellular matrix of cartilage tissue, is known to be a key component that creates a microenvironment for cell growth and differentiation, as well as tissue composition, in cartilage. Against this backdrop, the inventors, through diligent efforts to treat damaged cartilage, have completed the present invention by developing an animal cartilage-derived injectable agent capable of inducing cartilage tissue regeneration by promoting tissue repair through the direct injection of an animal cartilage-derived collagen and extracellular matrix-containing biomaterial in an injectable formulation into the application site without surgical incision. Figure 1 is a figure confirming the injectable raw material and water-soluble conditions. Figure 2 is a photograph of a CAM solution prepared for 6 hours at three different water-soluble temperature conditions (4℃, 25℃, 37℃). Figure 3 is a figure confirming the injectable sterilization conditions. FIG. 4 is a schematic diagram showing a method for preparing an injectable porcine cartilage-derived injectable (CAM solution) according to the present invention. Figure 5 is a figure showing the protein content of an injectable porcine cartilage-derived injectable before and after sterilization, confirmed by SDS-PAGE. Figure 6 shows the results of confirming the collagen content according to the water-soluble process conditions. Figure 7 shows the results of cartilage differentiation ability according to the water-soluble process conditions. Figure 8 shows the results of confirming the cartilage tissue regenerative capacity in a rat anterior cruciate ligament amputation-induced osteoarthritis model when umbilical cord blood-derived mesenchymal stem cells were administered for 8 weeks (stem cell administration group) or when stem cells and a CAM solution prepared by the process of Figure 5 were administered together for 8 weeks (stem cell + CAM solution administration group). Scale bar = 100 µm. The present invention will be explained in more detail below by way of examples. However, these examples are intended to illustrate the present invention and the scope of the present invention is not limited by these examples, and it will be obvious to those skilled in the art to which the present invention pertains. Example 1. Verification of injectable raw materials and water solubilization conditions To secure a process for manufacturing an injectable form of CAM solution, three raw materials (sterile CAM-WS powder, non-sterile CAM-WS sponge, sterile CAM-WS sponge) (ATEMS, Korea) were eluted in a stirred incubator under three water solubilization temperature conditions (4℃, 25℃, 37℃) and three water solubilization time conditions (3 hours, 6 hours, 18 hours), and then centrifuged at 300g for 5 minutes to prepare CAM solutions, and the injectability of each solution was analyzed. Sterilization was performed by irradiating with gamma rays of VDmax25, which exhibited 25–40 kGy. As a result, as shown in Figure 1 and Figure 2, which illustrates CAM solutions prepared for 6 hours at three different water-soluble temperature conditions (4℃, 25℃, 37℃), it was confirmed that an injectable CAM solution could not be prepared using the raw material of the