Application of tumor-derived acellular extracellular matrix in preparation of medicine for promoting angiogenesis and tissue regeneration

Abstract

The invention discloses application of a tumor-derived acellular extracellular matrix in preparation of a medicine for promoting angiogenesis and tissue regeneration. The invention provides a strategy for regulating and controlling tissue regeneration based on a tumor-derived acellular extracellular matrix microenvironment, a novel angiogenesis promoting biological material is constructed by combining the high similarity of a tumor microenvironment and a regeneration microenvironment such as angiogenesis, and high-efficiency regeneration is realized in various tissue defects. In addition, due to the characteristic of rapid proliferation of the tumor tissue, the productivity bottleneck of a traditional acellular extracellular matrix raw material source is broken through, and sustainable biological raw materials are provided for large-scale production. The method successfully solves the problem that a specific tissue-derived acellular extracellular matrix is difficult to regenerate in a complex damaged microenvironment.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the application of tumor-derived acellular extracellular matrix in the preparation of drugs for promoting angiogenesis and tissue regeneration. Background Art

[0002] Acellular extracellular matrix (dECM) constructs a functionalized bioactive scaffold by selectively removing immunogenic components in tissues and completely retaining the bioactive components and three-dimensional topological structure of the natural extracellular matrix. It shows significant advantages in effectively reducing risks such as host immune rejection, inflammatory response, and fibrosis. Currently, the raw materials for preparing dECM mainly come from single and specific tissues. Although dECM from specific tissue sources is beneficial for constructing a biomimetic tissue microenvironment, it also leads to insufficient adaptability to complex damaged microenvironments (such as the transitional region of the osteochondral interface), making it difficult to achieve multi-lineage tissue co-regeneration. Functional enhancement through the compounding of exogenous functional components or the combination with other delivery systems will also damage the integrity of the inherent signal transduction network of ECM and increase the risk of immunogenicity. Summary of the Invention

[0003] To solve at least some of the above technical problems in the prior art, the present invention provides the application of tumor-derived acellular extracellular matrix in the preparation of drugs for promoting angiogenesis and multi-directional tissue regeneration. Specifically, the present invention includes the following contents.

[0004] In the first aspect of the present invention, there is provided the application of acellular extracellular matrix derived from malignant tumor tissues in the preparation of drugs for promoting angiogenesis and / or tissue regeneration.

[0005] In certain embodiments, according to the application described in the present invention, wherein the preparation of the acellular extracellular matrix includes: obtaining the acellular extracellular matrix by treating malignant tumor tissues to remove immunogenic components.

[0006] In certain embodiments, according to the application described in the present invention, wherein the acellular extracellular matrix derived from malignant tumor tissues has a collagen network scaffold structure.

[0007] In certain embodiments, according to the application described in the present invention, wherein the malignant tumor includes epithelial or neuroectodermal malignant tumors.

[0008] In certain embodiments, according to the application described in the present invention, wherein the promotion of angiogenesis and tissue regeneration includes at least one of the following situations: (1) Promoting endothelial cell vascularization; (2) Promoting vascular regeneration in the bone defect area; (3) Promoting bone regeneration in the bone defect area; (4) Promoting tissue healing.

[0009] In certain embodiments, for the application according to the present invention, wherein the promoting effect is achieved by administering a therapeutically effective amount of the drug to a subject.

[0010] In certain embodiments, for the application according to the present invention, wherein the subject includes a mammal.

[0011] In certain embodiments, for the application according to the present invention, wherein the mammal includes a human.

[0012] In certain embodiments, for the application according to the present invention, wherein the therapeutically effective amount is 0.01 - 1000 mg / Kg.

[0013] In certain embodiments, for the application according to the present invention, wherein the drug further comprises a pharmaceutically acceptable carrier.

[0014] In certain embodiments, for the application according to the present invention, wherein the pharmaceutically acceptable carrier includes at least one of a diluent, a filler, an absorbent, a wetting agent, a binder, a lubricant, a preservative, and an antioxidant.

[0015] A second aspect of the present invention provides a pharmaceutical composition for promoting angiogenesis and / or tissue regeneration, which comprises a decellularized extracellular matrix derived from malignant tumor tissue.

[0016] A third aspect of the present invention provides a method for promoting tissue regeneration, which includes the step of contacting a tumor-derived decellularized extracellular matrix with endothelial cells in vitro.

[0017] A fourth aspect of the present invention provides a method for promoting endothelial cell vascularization, which includes the step of contacting a tumor-derived decellularized extracellular matrix with endothelial cells in vitro.

[0018] The present invention breaks through the traditional tissue source limitation and innovatively proposes a strategy for regulating tissue regeneration based on the tumor-derived dECM microenvironment. Combining the high similarity between the tumor microenvironment and the regeneration microenvironments such as angiogenesis, a novel angiogenesis-promoting biomaterial is constructed, achieving high-efficiency regeneration in various tissue defects. In addition, the characteristic of rapid proliferation of tumor tissue breaks through the production capacity bottleneck of the traditional dECM raw material source, providing a sustainable biological raw material for large-scale production. The present invention successfully solves the problem of difficult regeneration of specific tissue source dECM for complex damaged microenvironments. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Shows the characterization of the decellularized extracellular matrix derived from tumor tissue.

[0020] Figure 2 The proteomic analysis results of acellular extracellular matrix from different tumor tissue sources are shown.

[0021] Figure 3 The immunofluorescence and flow cytometry results after mixing the acellular extracellular matrix from tumor tissue sources with endothelial cells are shown.

[0022] Figure 4 It is shown that the dECM from liver cancer tissue can promote angiogenesis and bone regeneration in the skull defect area of rats.

[0023] Figure 5 It is shown that the dECM from liver cancer tissue can promote angiogenesis and osteochondral regeneration in the osteochondral defect area of the rat knee joint.

[0024] Figure 6 It is shown that the dECM from liver cancer tissue can promote angiogenesis and skin healing in the skin defect area of mice.

[0025] Figure 7 They are HE and Masson tissue sections of the skin defect area after being treated with acellular extracellular matrix from liver cancer. Detailed implementation manners

[0026] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0027] It should be understood that the terms described in the present invention are only for describing specific implementation manners and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that the upper and lower limits of the range and each intermediate value therebetween are specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0028] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0029] Application One aspect of the present invention provides the use of tumor-derived acellular extracellular matrix in the preparation of a medicament for promoting angiogenesis and / or tissue regeneration. In the present invention, considering the high similarity between the tumor microenvironment and the regeneration microenvironments such as angiogenesis and the ability to retain more active ingredients, thus, in a preferred embodiment, the acellular extracellular matrix of the present invention is derived from tumor tissue, rather than normal tissue or cell sources, so as to increase the adaptability to the complex damaged microenvironment and achieve multi-lineage tissue collaborative regeneration.

[0030] In the present invention, the tumor includes solid tumors with high malignancy and fast growth, and its examples include but are not limited to: breast cancer, melanoma, osteosarcoma, neuroblastoma, pancreatic cancer, lung cancer, rhabdomyosarcoma, Ewing's sarcoma, bladder cancer, colon cancer, liver cancer, ovarian cancer, cervical cancer, nasopharyngeal cancer, laryngeal cancer, gastric cancer, kidney cancer, head and neck tumors, esophageal cancer, thyroid cancer or brain cancer. The malignant tumor of the present invention can be a primary tumor or a secondary tumor. In a preferred embodiment, the tumor includes epithelial malignant tumors, such as liver cancer, colon cancer, lung cancer, etc. In another preferred embodiment, the tumor includes neuroectodermal malignant tumors, such as melanoma. In yet another preferred embodiment, the tumor is a non-germ cell tumor. In a most preferred embodiment, the acellular extracellular matrix is derived from liver cancer tissue.

[0031] In the present invention, "promote" means to improve this condition after the occurrence of an injury. Compared with the untreated reference group under the same conditions, measured by any standard technique, this degree of promotion or improvement is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95% or 100%. "Injury" refers to the destruction of the structure of tissues or organs such as skin, bone, cartilage, viscera, etc. caused by various traumatic factors acting on the body or the local and systemic reactions brought about by it. Injuries include physical injuries, chemical injuries, biological injuries, etc. In the present invention, the medicament is particularly used for bone injuries (or bone defects), cartilage injuries (or cartilage defects) or skin injuries (or skin defects).

[0032] In the present invention, beneficial or desired clinical outcomes include but are not limited to the following detectable or undetectable outcomes, such as promoting angiogenesis and / or tissue regeneration. The promotion of angiogenesis and tissue regeneration includes at least one of the following situations: (1) promoting endothelial cell vascularization; (2) promoting angiogenesis in the bone defect area; (3) promoting bone regeneration in the bone defect area; (4) promoting the healing of tissues (such as defective skin).

[0033] In the present invention, promoting angiogenesis and tissue regeneration is achieved by administering a therapeutically effective amount of the said medicament to a subject. Subjects include but are not limited to mammals, and examples of mammals include but are not limited to humans, mice, rabbits, cats, dogs, cows, sheep, pigs, etc.

[0034] In the present invention, the drug contains a therapeutically effective amount of tumor-derived acellular extracellular matrix, and optionally, a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, and such pharmaceutically acceptable material, composition or vehicle participates in transporting or delivering the medicament from one organ or part of the body to another organ or another part of the body. Each carrier is "acceptable" in that it is compatible with the other ingredients of the formulation and does not harm the patient.

[0035] In the present invention, the pharmaceutically acceptable carrier includes, but is not limited to, at least one of diluents, fillers, absorbents, wetting agents, binders, lubricants, preservatives and antioxidants. Among them, examples of diluents include, but are not limited to, normal saline, aqueous buffer solutions, solvents, dispersion media, etc.; fillers include, but are not limited to, starch, lactose, mannitol, microcrystalline cellulose, etc.; absorbents include, but are not limited to, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents include, but are not limited to, water, ethanol, etc.; binders include, but are not limited to, synthetic materials or natural synthetic materials, etc.; lubricants include, but are not limited to, magnesium stearate, talc, polyethylene glycol, sodium dodecyl sulfate, colloidal silica, talc, etc.; preservatives include, but are not limited to, parabens, chlorobutanol, phenol, sorbic acid, etc.; antioxidants include, but are not limited to, ascorbic acid, methionine, etc.

[0036] In certain embodiments, the synthetic biomaterials include, but are not limited to: polyethylene glycol, polyethylene glycol derivatives, polylactic acid, polylactic acid-alcohol copolymers, polyanhydrides, polyesters, polyamino acids, polyethylene oxide, polyesters, polymethyl methacrylate, polycarbonates, polyurethanes, polycaprolactone, polyhydroxyalkanoates, polysiloxanes, polyethylene, polyvinyl chloride, polytetrafluoroethylene, polystyrene, polypropylene, N-(3-aminopropyl) methacrylamide hydrochloride (APMA), N-[tris(hydroxymethyl)methyl]acrylamide (THMA), maleic anhydride graft copolymers, polyacrylamides, polyacetals, polypyrrole or any combination thereof. In certain embodiments, the natural biomaterials include, but are not limited to: natural proteins, collagen and collagen derivatives, gelatin and gelatin derivatives, agar and agar derivatives, proteoglycans, alginates and alginate derivatives, Matrigel, propolis, cellulose and cellulose derivatives, chitin and chitin derivatives, silk fibroin and its derivatives, laminin and its derivatives, fibronectin and its derivatives, sodium hyaluronate and hyaluronic acid derivatives, agarose and its derivatives, dextran and its derivatives, sucrose and sucrose derivatives, starch, chitosan and chitosan derivatives or any combination thereof, preferably gelatin and gelatin derivatives, chitosan and chitosan derivatives.

[0037] In a preferred embodiment, the carrier is selected from at least one of collagen, gelatin, polyvinyl alcohol, and hydroxyapatite.

[0038] In the present invention, the administration mode of the drug is not particularly limited, and representative administration modes include, but are not limited to: parenteral (intravenous, intramuscular or subcutaneous) and topical administration. Accordingly, the drugs of the present invention can be formulated into various clinically acceptable dosage forms, including injection dosage forms, topical dosage forms or external use dosage forms, etc. In certain embodiments, the dosage form of the drug containing tumor-derived acellular extracellular matrix can be a composite film, patch, gel, microneedle, porous sponge, fiber scaffold, 3D printing structure, injection, etc.

[0039] The therapeutically effective amount described in the present invention refers to the pharmaceutically recognized effective dosage, that is, the amount of tumor-derived acellular extracellular matrix is sufficient to significantly improve the condition without causing serious side effects. The daily dosage of tumor-derived acellular extracellular matrix is usually 0.01 - 1000 mg / Kg, preferably 0.01 - 500 mg / Kg, or 0.01 - 400 mg / Kg, or 0.01 - 300 mg / Kg, or 0.01 - 200 mg / Kg, or 0.01 - 150 mg / Kg, or 0.01 - 100 mg / Kg, or 0.01 - 50 mg / Kg, or 0.01 - 40 mg / Kg, or 0.01 - 30 mg / Kg, and most preferably 0.01 - 20 mg / Kg. Exemplary effective dosages are, for example, 0.01 mg / Kg, 0.05 mg / Kg, 0.1 mg / Kg, 0.2 mg / Kg, 0.3 mg / Kg, 0.4 mg / Kg, 0.5mg / Kg, 0.75 mg / Kg, 0.95 mg / Kg, 1 mg / Kg, 1.25 mg / Kg, 1.5 mg / Kg, 1.75 mg / Kg, 2 mg / Kg, 2.5 mg / Kg, 2.75 mg / Kg, 3 mg / Kg, 3.25 mg / Kg, 3.5 mg / Kg, 3.75 mg / Kg, 4 mg / Kg, 4.25 mg / Kg, 4.5 mg / Kg, 4.75 mg / Kg, 5 mg / Kg, 5.25 mg / Kg, 5.5 mg / Kg, 5.75 mg / Kg, 6 mg / Kg, 6.25mg / Kg, 6.5 mg / Kg, 6.75 mg / Kg, 7 mg / Kg, 7.25 mg / Kg, 7.5 mg / Kg, 7.75 mg / Kg, 8 mg / Kg, 8.25 mg / Kg, 8.5 mg / Kg, 8.75 mg / Kg, 9 mg / Kg, 9.25 mg / Kg, 9.5 mg / Kg, 9.75 mg / Kg, 10mg / Kg, 11 mg / Kg, 12 mg / Kg, 13 mg / Kg, 14 mg / Kg, 15 mg / Kg, 16 mg / Kg, 17 mg / Kg, 18 mg / Kg, 19 mg / Kg, 20 mg / Kg. It can be administered as a single dose once a day, administered in multiple doses per day, or used at intervals.

[0040] Those skilled in the art will understand that the tumor-derived acellular extracellular matrix can also be used in combination with other drugs to prepare drugs for jointly promoting angiogenesis and / or tissue regeneration. The other drugs are not specifically limited and can be any suitable drugs for promoting angiogenesis and / or tissue regeneration.

[0041] In the present invention, the method for preparing the tumor-derived acellular extracellular matrix is not particularly limited, and the methods for preparing acellular matrix known in the art can be used. In a preferred embodiment, the acellular extracellular matrix is obtained by treating malignant tumor tissues to remove immunogenic components (including tumor cells and various cytokines). The malignant tumor tissues can be from tumor-bearing animal models or from organoids obtained by primary three-dimensional culture of tumor cells.

[0042] Exemplary methods for preparing the acellular extracellular matrix include the following steps: (1) obtaining tumor tissues; (2) chopping the tumor tissues, immersing them in double-distilled water, and oscillating at 0-10°C and 50-300 rpm for 5-20 hours to obtain tissue samples; (3) sequentially placing the tissue samples in the following buffers for continuous stirring treatment: First, reacting overnight in a hypotonic Tris buffer (1-50 mM Tris, 1-10 mM EDTA) at 50-300 rpm; second, washing with ddH2O at 50-300 rpm for 0.5-5 hours; then, reacting in a hypertonic Tris buffer (20-80 mM Tris, 0.5-2 M NaCl, 1-20 mM EDTA) at 30-40°C and 50-300 rpm for 20-30 hours; then, washing with ddH2O at 100-300 rpm for 0.5-5 hours; subsequently, treating with 0.5-5% (v / v) Triton X-100 at 30-40°C and 50-300 rpm for 12-24 hours; washing with ddH2O at 50-300 rpm for 0.5-5 hours; treating with DNase I at 30-40°C and 50-300 rpm for 3-6 hours, and finally, washing with ddH2O at 50-300 rpm overnight; (4) performing supercritical carbon dioxide extraction, with the operating conditions being: pressure 200-400 bar, temperature 30-40°C, ethanol as an adjuvant (concentration 1-10%), total extraction flow rate 20-40 g / L, extraction time 1-5 hours. After extraction, the obtained acellular extracellular matrix is ground into powder for standby.

[0043] Method for promoting tissue regeneration or endothelial cell vascularization On the other hand, the present invention also provides a method for promoting tissue regeneration or endothelial cell vascularization, including the step of contacting the tumor-derived acellular extracellular matrix with endothelial cells in vitro. The method of the present invention can be used for non-diagnostic and non-therapeutic purposes (such as but not limited to disease mechanism research, drug screening and evaluation, organoid model construction, organ-on-a-chip construction, artificial blood vessel development, etc.), and it can be used to promote the expression of tip cell markers by endothelial cells or to promote the transformation of endothelial cells into tip cells.

[0044] Example 1 1. Preparation and characterization of acellular extracellular matrix from different tumor tissues. The preparation method is as follows: (1) After negative tumor-bearing nude mice and Balb / c mice were euthanized in a carbon dioxide anesthesia chamber, necropsy was performed to remove tumors and livers.

[0045] (2) The above tissues were chopped and immersed in double-distilled water (ddH2O), and shaken in a shaker at 4°C and 150 rpm for 12 hours to achieve fat separation.

[0046] (3) Subsequently, the tissue samples were successively placed in the following designated buffers for continuous stirring treatment: First, react overnight in hypotonic Tris buffer (10 mM Tris, 5 mM EDTA, pH 8.0) at 150 rpm; second, wash with ddH2O for 1 hour at 150 rpm; then, react in hypertonic Tris buffer (50 mM Tris, 1 M NaCl, 10 mM EDTA, pH 8.0) at 37°C and 150 rpm for 24 hours; then, wash with ddH2O for 1 hour at 150 rpm; subsequently, treat with 1% (v / v) Triton X-100 at 37°C and 150 rpm for 12 - 24 hours; then wash with ddH2O for 1 hour at 150 rpm; finally, treat with 20 U / ml DNase I at 37°C and 150 rpm for 3 - 6 hours. Finally, wash with ddH2O overnight at 150 rpm. Unless otherwise specified, all steps were carried out at 4°C.

[0047] (4) In the final treatment stage, supercritical carbon dioxide (scCO2) extraction was used, and its operating conditions were: pressure 300 bar, temperature 37°C, ethanol as an adjuvant (concentration 3%), total extraction flow rate 30 g / L, and extraction time 3 hours. After extraction, the obtained acellular extracellular matrix (dECM) was ground into powder.

[0048] HE DAPI results showed that after treatment with the above decellularization method, the cells in liver, hepatocellular carcinoma, colon cancer, lung cancer, and melanoma dECM were completely removed, and the collagen network scaffold was relatively intact. Safranin staining results indicated that the proteoglycans in dECM were also retained. In summary, this preparation method removed the cellular immunogenic components in the tissue while completely retaining the bioactive components and three-dimensional topological structure of the natural extracellular matrix. Therefore, in this example, a functionalized acellular extracellular matrix ( Figure 1 ) was successfully constructed.

[0049] 2. Proteomics study of dECM Proteomic analysis was performed on the acellular extracellular matrices of four tumors obtained by decellularizing liver cancer, colon cancer, lung cancer, and melanoma tissues respectively. The results are as Figure 2 shown Figure 2 The GSVA results in the left figure show that compared with the other three tumors, the liver cancer dECM has a closer connection with stem cell vital activities, angiogenesis, immune response, and histogenesis. Figure 2 The results of the regeneration potential score in the right figure further show that the liver cancer dECM is more related to regeneration.

[0050] Example 2 This example shows that the dECM derived from liver cancer tissue can promote the expression of tip cell markers by endothelial cells.

[0051] 1 ml of gelatin was mixed with 10 mg of liver cancer dECM and human umbilical vein endothelial cells. After photo-curing for 30 s, endothelial cell medium was added. As Figure 3 shown, after culturing for 24 h, immunofluorescence and flow cytometry results showed that adding gelatin with liver cancer dECM (GelMA+H-dECM) could significantly promote the expression of tip cell markers (CD34) by endothelial cells compared with adding liver dECM (GelMA+L-dECM). Figure 3 Among them, the quantitative results of the fluorescence intensity of CD34-positive endothelial cells are as follows: liver 6.293±1.464; liver cancer 22.713±3.651( P <0.05). The percentage of CD34-positive endothelial cells in endothelial cells: liver 39.875%±0.386, liver cancer 48.1%±0.917( P <0.05).

[0052] Example 3 This example shows that the dECM derived from liver cancer tissue can promote angiogenesis and bone regeneration in the skull defect area of rats.

[0053] A rat animal model with a skull defect area was constructed. At 2 weeks and 4 weeks after applying collagen-hydroxyapatite loaded with liver cancer dECM (COL+HA+H) at the defect, angiogenesis and bone regeneration in the defect area were measured.

[0054] Figure 4 Shows the real-time blood perfusion and CT photos at 2 weeks and 4 weeks after applying collagen loaded with liver cancer dECM at the rat skull defect. The control is collagen-hydroxyapatite loaded with liver dECM (COL+HA+L). The results of the vascular area coefficient at 4 weeks show: liver 0.213±0.028; liver cancer 0.281±0.041( P<0.05). The results of bone mass / tissue mass at 4 weeks showed: liver 32.607% ± 6.589; liver cancer 49.175% ± 1.082( P <0.05). The results indicated that the dECM derived from liver cancer tissues could significantly promote angiogenesis and bone regeneration in the skull defect area of rats.

[0055] Example 4 This example demonstrated that the dECM derived from liver cancer tissues could promote angiogenesis and osteochondral regeneration in the osteochondral defect area of rat knee joints.

[0056] A rat animal model with an osteochondral defect area in the knee joint was constructed. At 2 weeks and 4 weeks after applying gelatin-loaded liver cancer dECM (GelMA+H) to the defect, angiogenesis and osteochondral regeneration in the osteochondral defect area were measured.

[0057] Figure 5 The histological sections and gross views of angiogenesis at 2 weeks and 4 weeks after applying gelatin-loaded liver cancer dECM to the osteochondral defect of rat knee joints were shown. The quantitative results of the fluorescence intensity of CD31-positive endothelial cells showed: liver 66.368 ± 19.198; 134.165 ± 20.696( P <0.05). The results of the gross view score of ICRS cartilage at 4 weeks (full score 12 points) showed: liver 5.6 ± 0.894; liver cancer 9.4 ± 0.548( P <0.05). The results indicated that, compared with the dECM from the liver (GelMA+L), the dECM derived from liver cancer tissues could significantly promote angiogenesis and osteochondral regeneration in the osteochondral defect area of rat knee joints.

[0058] Example 5 This example demonstrated that the dECM derived from liver cancer tissues could promote angiogenesis and skin healing in the skin defect area of mice.

[0059] A mouse animal model with a skin defect area was constructed. At 10 days after applying PVA-loaded liver cancer dECM (PVA+H) to the defect, angiogenesis and skin healing in the skin defect area of mice were measured.

[0060] Figure 6 The histological sections of angiogenesis and the healing rate at 10 days after applying PVA-loaded liver cancer dECM to the skin defect of mice were shown. The blood vessel density results ( / mm 2 ) showed: liver 103.242 ± 11.510; liver cancer 140.987 ± 9.288( P <0.05). The results of the skin wound healing rate (%) at day 10 showed: liver 89.892 ± 3.245; liver cancer 96.533 ± 0.501( P(<0.05). The results showed that compared with the liver dECM (PVA+L), the dECM derived from HCC tissues could significantly promote angiogenesis and skin healing in the skin defect area of mice.

[0061] Example 6 This example shows the safety evaluation of the HCC-derived decellularized extracellular matrix. After treatment with the HCC-derived decellularized extracellular matrix, HE and Masson staining were performed on the skin defect area for observation, specifically as Figure 7 shown. Figure 7 The HE and Masson tissue section results of the skin defect area after treatment with the HCC-derived decellularized extracellular matrix are shown. The section results showed that compared with the liver decellularized extracellular matrix group, the HCC-derived decellularized extracellular matrix treatment group formed an epidermal layer, a dermal layer, and skin appendage structures that were closer to healthy tissues, accelerated the healing of skin defects, and no abnormal tumor cells were generated.

[0062] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features. These modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. Use of acellular extracellular matrix derived from malignant tumor tissues in the preparation of drugs for promoting angiogenesis and / or tissue regeneration.

2. The application according to claim 1, characterized in that, The preparation of the acellular extracellular matrix includes: obtaining the acellular extracellular matrix by treating malignant tumor tissues to remove immunogenic components.

3. The application according to claim 1, characterized in that The acellular extracellular matrix derived from malignant tumor tissues has a collagen network scaffold structure.

4. The application according to claim 1, characterized in that The malignant tumors include epithelial and / or neuroectodermal malignant tumors.

5. A pharmaceutical composition for promoting angiogenesis and / or tissue regeneration, characterized in that, It contains acellular extracellular matrix derived from malignant tumor tissues.

6. The pharmaceutical composition for promoting angiogenesis and / or tissue regeneration according to claim 5, characterized in that, The amount of the acellular extracellular matrix derived from malignant tumor tissues can be administered at a therapeutically effective amount of 0.01 - 1000 mg / Kg.

7. The pharmaceutical composition for promoting angiogenesis and / or tissue regeneration according to claim 5, characterized in that, The drug further includes a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, characterized in that, The pharmaceutically acceptable carrier includes at least one of diluents, fillers, absorbents, wetting agents, binders, lubricants, preservatives, and antioxidants.

9. A method for promoting tissue regeneration, characterized in that, It includes the step of contacting the tumor-derived acellular extracellular matrix with endothelial cells in vitro.

10. The method for promoting tissue regeneration according to claim 9, wherein, The tissue regeneration includes at least one of the following situations: (1) Promoting endothelial cell vascularization; (2) Promoting blood vessel regeneration in the bone defect area; (3) Promoting bone regeneration in the bone defect area; (4) Promoting tissue healing.

Images