Tissue engineering gel scaffold and preparation method thereof, composite gel scaffold and application

Abstract

The application provides a tissue engineering gel scaffold and a preparation method, a composite gel scaffold and an application thereof, wherein the tissue engineering gel scaffold comprises a decellularized matrix of a cephalopod, and a tissue source of the decellularized matrix of the cephalopod comprises at least one of a mantle of the cephalopod and a brachialia of the cephalopod. The tissue engineering gel scaffold of the application is based on the decellularized matrix of the cephalopod, has good biocompatibility, and has a regular arrangement structure, strong water absorption performance and strong mechanical properties. The structure, water content, mechanical properties and degradation rate of the tissue engineering gel scaffold are similar to those of soft tissue, the content of high proportion of unsaturated fatty acids, in which the content of omega-3 is much greater than that of omega-6, is beneficial to the inhibition of inflammation and the repair of tissue, and the tissue engineering gel scaffold can be compounded or used in combination with various active molecules, cells and the like, is used for defect filling and tissue repair in soft tissue regeneration, and is beneficial to promoting the soft tissue regeneration process.

Description

Technical Field

[0001] This application relates to the field of bioengineering technology, specifically to a tissue engineering gel scaffold, its preparation method, a composite gel scaffold, and its applications. Background Technology

[0002] Soft tissue is an important structure in the body that connects, surrounds, or supports internal organs and bones, including muscles, tendons, ligaments, nerves, and fibrous tissue. With the rapid development of tissue engineering and regenerative medicine, the methods for repairing soft tissue injuries are becoming increasingly diverse, and related technologies are constantly advancing. Currently, while traditional soft tissue repair methods commonly used in clinical practice are widely applied, they still have significant limitations. For example, using autologous tissue flaps for filling can easily cause donor site defects; implanting artificial prostheses may trigger immune rejection reactions, leading to tissue fibrosis; and non-degradable permanent fillers used in plastic surgery may cause more serious complications due to risks such as vascular occlusion.

[0003] To overcome the aforementioned problems, tissue repair methods based on gel-filled scaffold materials have gradually gained attention. This method utilizes injectable or implantable gel scaffolds for minimally invasive filling, and can be applied to various scenarios such as soft tissue defect repair, chronic wound healing, osteoarthritis and articular cartilage treatment, tendon repair, and even organ repair. In the field of plastic and cosmetic surgery, gel-filled scaffolds can also be used alone or in combination with other techniques for wrinkle removal and restoring tissue volume loss caused by aging and fat atrophy.

[0004] Nevertheless, these gel-filled scaffold materials may still face some drawbacks in practical applications, such as a mismatch between the material's degradation rate and tissue regeneration rate, insufficient mechanical properties, uncertainty regarding long-term biocompatibility, and challenges in structural stability and functional integration under complex physiological conditions. Summary of the Invention

[0005] In view of this, this application provides a tissue engineering gel scaffold, its preparation method, a composite gel scaffold, and its application to solve the above-mentioned technical problems.

[0006] To achieve the above objectives, in a first aspect, this application provides a tissue-engineered gel scaffold comprising a decellularized matrix of cephalopods, wherein the tissue source of the decellularized matrix of cephalopods includes at least one of the mantle of cephalopods and the carpal of cephalopods.

[0007] Based on the first aspect, in some embodiments, the compressive elastic modulus of the decellularized matrix of cephalopods is from 0.2 MPa to 30 MPa.

[0008] Based on the first aspect, in some embodiments, the decellularized matrix of cephalopods absorbs water at a rate of 2 to 15 times its own weight.

[0009] Based on the first aspect, in some embodiments, the decellularized matrix of cephalopods has unsaturated fatty acids, including at least one of omega-3 and omega-6, and the unsaturated fatty acids account for more than or equal to 50% of the total fatty acids by mass.

[0010] Based on the first aspect, in some embodiments, cephalopods include at least one of squid, cuttlefish, and octopus.

[0011] Secondly, this application provides a method for preparing the above-mentioned tissue engineering gel scaffold, the method comprising: pre-treating the source tissue of the decellularized matrix of cephalopods to obtain a pre-treated sample; decellularizing the pre-treated sample to obtain a powdered decellularized matrix; and gelling the powdered decellularized matrix to prepare a tissue engineering gel scaffold.

[0012] Based on the second aspect, in some embodiments, gelling the powdered decellularized matrix includes mixing the powdered decellularized matrix with tissue exudate and / or blood to form a gel.

[0013] Based on the second aspect, in some embodiments, gelling the powdered decellularized matrix includes mixing the powdered decellularized matrix with a polymer solution, the polymer solution including at least one of hyaluronic acid, chitosan, gelatin, dextran and collagen.

[0014] Based on the second aspect, in some embodiments, the mass ratio of the powdered decellularized matrix to the polymer solution is 1:(1~5).

[0015] Based on the second aspect, in some embodiments, the mass concentration of the polymeric solute in the polymeric solution is 1% to 30%.

[0016] Thirdly, this application provides a composite gel scaffold comprising: i) at least one of the tissue engineering gel scaffold described above and the tissue engineering gel scaffold prepared by the above preparation method; and ii) at least one of a biomacromolecule and an active factor, wherein the biomacromolecule includes collagen and the active factor includes at least one of growth factors and cytokines.

[0017] Fourthly, this application also provides the use of at least one of the above-mentioned tissue-engineered gel scaffolds, the tissue-engineered gel scaffolds prepared by the above-mentioned preparation method, and the composite gel scaffolds in the preparation of soft tissue defect filling agents.

[0018] In addition, this application provides the use of at least one of the above-mentioned tissue-engineered gel scaffold, the tissue-engineered gel scaffold prepared by the above-mentioned preparation method, and the composite gel scaffold in the preparation of soft tissue injury repair agents.

[0019] The tissue-engineered gel scaffold of this application is based on decellularized cephalopod matrix, exhibiting good biocompatibility and a regular arrangement structure, strong water absorption, and strong mechanical properties. Its structure, water content, mechanical properties, and degradation rate are similar to soft tissue. It contains a high proportion of unsaturated fatty acids, with omega-3 content significantly exceeding omega-6 content, which is beneficial for inhibiting inflammation and repairing tissues. It can also be used in combination or in combination with various active molecules and cells for filling defects and repairing tissues during soft tissue regeneration, thus promoting the soft tissue regeneration process. Attached Figure Description

[0020] Figure 1 The microstructure test results of the squid mantle membrane provided in Example 1 of this application after decellularization and gelation treatment.

[0021] Figure 2 The microstructure test results of the squid mantle membrane provided in Example 2 of this application after decellularization and gelation treatment.

[0022] Figure 3 The results of microscopic morphology testing of the octopus tentacles after decellularization and gelation treatment provided in Example 3 of this application.

[0023] Figure 4 The results of macroscopic morphology tests on the tissues provided in Example 1 and Comparative Example 1 of this application after decellularization and powdering.

[0024] Figure 5 The morphological test results of the tissue-engineered gel scaffolds provided in Example 4 and Comparative Example 2 of this application.

[0025] Figure 6 The results of the performance test of squid mantle powder after decellularization and powdering in the application of Example 1 of this application on the repair of subcutaneous soft tissue defects in dogs. Detailed Implementation

[0026] To facilitate understanding of the technical solutions of this application, a more comprehensive description of the technical solutions of this application will be provided below with reference to the accompanying drawings. Preferred embodiments of the technical solutions of this application are shown in the drawings. However, the technical solutions of this application can be implemented in many different forms and are not limited to the embodiments described herein. Rather, these embodiments are provided to enable a more thorough and comprehensive understanding of the disclosure of the technical solutions of this application.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0028] To address the shortcomings of gel-filled scaffold materials in practical applications, this application proposes a novel tissue engineering gel scaffold based on decellularized matrix. While removing cells, the decellularized matrix retains intact collagen and bioactive components (such as growth factors), exhibiting excellent biocompatibility and biodegradability.

[0029] One embodiment of this application provides a tissue-engineered gel scaffold comprising a decellularized matrix of cephalopods, wherein the tissue source of the decellularized matrix of cephalopods includes at least one of the mantle of cephalopods and the carpal of cephalopods.

[0030] The tissue-engineered gel scaffold of this application is based on decellularized cephalopod matrix, exhibiting good biocompatibility and a regular arrangement structure, strong water absorption, and strong mechanical properties. Its structure, water content, mechanical properties, and degradation rate are similar to soft tissue. It contains a high proportion of unsaturated fatty acids, with omega-3 content significantly exceeding omega-6 content, which is beneficial for inhibiting inflammation and repairing tissues. It can also be used in combination or in combination with various active molecules and cells for filling defects and repairing tissues during soft tissue regeneration, thus promoting the soft tissue regeneration process.

[0031] In some embodiments, the compressive elastic modulus of the decellularized cephalopod matrix is ​​between 0.2 MPa and 30 MPa. For example, the compressive elastic modulus of the decellularized matrix can be 0.2 MPa, 1 MPa, 4 MPa, 7 MPa, 10 MPa, 13 MPa, 16 MPa, 19 MPa, 22 MPa, 25 MPa, 28 MPa, 30 MPa, or any value within the range of any two of the above values. Controlling the compressive elastic modulus of the decellularized matrix within the above range is beneficial for optimizing the mechanical properties of the resulting tissue-engineered gel scaffold, making it similar to soft tissue.

[0032] In some embodiments, the water absorption capacity of the decellularized matrix from cephalopods is 2 to 15 times its own weight. For example, the water absorption capacity of the decellularized matrix can be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times its own weight, or any value within the range of any two of the above values. Controlling the water absorption capacity of the decellularized matrix within the above range is beneficial for optimizing the water retention properties of the resulting tissue-engineered gel scaffold, making it similar to soft tissue.

[0033] In some embodiments, the decellularized matrix of cephalopods contains unsaturated fatty acids, including at least one of omega-3 and omega-6, with the unsaturated fatty acids accounting for more than or equal to 50% of the total fatty acids by mass. Controlling the mass percentage of the unsaturated fatty acids in the total fatty acids within the aforementioned range helps to improve the inflammation relief and tissue repair performance of the resulting tissue-engineered gel scaffold.

[0034] In some embodiments, cephalopods include at least one of squid, cuttlefish, and octopus.

[0035] An embodiment of this application also provides a method for preparing the above-mentioned tissue engineering gel scaffold, the method comprising:

[0036] Step 1: Pre-treat the tissue from which the decellularized matrix of cephalopods is derived to obtain a pre-treated sample.

[0037] In related technologies, animal tissues are easily contaminated or degraded after being separated from the host, so they usually need to be cryopreserved. To facilitate subsequent preparation, the removed animal tissues need to be pretreated.

[0038] In some embodiments, the pretreatment includes a thawing process. Preferably, the thawing process is performed in water or a buffer solution.

[0039] In some embodiments, the pretreatment further includes treating the thawed animal tissue with a buffer solution. For example, the buffer solution may include, but is not limited to, antioxidants, antibiotics, proteases, protease inhibitors, etc., to promote the disinfection and stabilization of the animal tissue.

[0040] In some embodiments, antioxidants include, but are not limited to, ascorbic acid, vitamin A, vitamin C, vitamin E, beta-carotene, etc.

[0041] In some embodiments, antibiotics include, but are not limited to, streptomycin, penicillin, etc.

[0042] In some embodiments, the protease includes, but is not limited to, dispersase II.

[0043] In some embodiments, protease inhibitors include, but are not limited to, analgin, aprotinin, benzoamide, phenbutastatin, DFP, EDTA, EGTA, leucinogen, pepsin, phosphonyl dipeptide, PMSF, etc.

[0044] Step 2: Decellularize the pretreated sample to obtain a powdered decellularized matrix.

[0045] Decellularization helps remove immunogenic cellular components (cell membrane, nucleus, cytoplasm, etc.) while preserving the natural extracellular matrix.

[0046] In some embodiments, decellularization may be performed by physical treatment, chemical treatment, or a combination of physical and chemical treatment.

[0047] In some embodiments, physical processing includes mechanical methods such as repeated freeze-thaw cycles or high-speed centrifugation of the pre-treated sample.

[0048] In some embodiments, chemical treatment includes treating the pretreated sample with acids, bases, ionic salts, detergents (ionic and nonionic detergents), oxidants, ascorbic acid, etc.

[0049] In some embodiments, the decellularization process includes the following steps:

[0050] S1: The cleaned tissue is placed in a surfactant for treatment to promote decellularization.

[0051] In some embodiments, the surfactants include, but are not limited to, SDS (sodium dodecyl sulfate), Triton X-100, sodium deoxycholate, etc.

[0052] In some embodiments, the concentration of the surfactant is 0.5% to 5%. For example, the concentration of the surfactant can be 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, or any value within the range of any two of the above values.

[0053] In some embodiments, the surfactant treatment time is from 1 h to 5 h. For example, the surfactant treatment time can be 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, or any value within the range of any two of the above values.

[0054] S2: Place the surfactant-treated tissue in a weak acid or weak alkali to assist and enhance the decellularization effect.

[0055] In some embodiments, the weak acid includes, but is not limited to, acetic acid, citric acid, hypochlorous acid, phosphoric acid, etc.

[0056] In some embodiments, the concentration of the weak acid is 0.5% to 5%. For example, the concentration of the weak acid can be 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, or any value within the range of any two of the above values.

[0057] In some embodiments, the weak acid treatment time is from 1 h to 5 h. For example, the weak acid treatment time is 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, or any value within the range of any two of the above values.

[0058] In some embodiments, the weak base includes, but is not limited to, sodium bicarbonate, sodium carbonate, ammonia, etc.

[0059] In some embodiments, the concentration of the weak base is 0.5% to 5%. For example, the concentration of the weak base can be 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, or any value within the range of any two of the above values.

[0060] In some embodiments, the weak alkali treatment time is from 1 h to 5 h. For example, the weak alkali treatment time can be 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, or any value within the range of any two of the above values.

[0061] S3: Place the tissue treated with a weak acid or weak alkali into an antibacterial agent to inactivate the virus in the tissue.

[0062] In some embodiments, the antibacterial agent includes hydrogen peroxide.

[0063] In some embodiments, the concentration of hydrogen peroxide is 2% to 6%. For example, the concentration of hydrogen peroxide can be 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, or any value within the range of any two of the above values.

[0064] In some embodiments, the hydrogen peroxide treatment time is from 1 h to 4 h. For example, the hydrogen peroxide treatment time is 1 h, 1.3 h, 1.6 h, 1.9 h, 2.2 h, 2.5 h, 2.8 h, 3.1 h, 3.4 h, 3.7 h, 4 h, or any value within the range of any two of the above values.

[0065] In some embodiments, the decellularized matrix obtained after decellularization is prepared into a powdered decellularized matrix by freeze-drying. For example, the decellularized matrix powder is obtained by crushing after freeze-drying.

[0066] Step 3: Gel the powdered decellularized matrix to prepare a tissue engineering gel scaffold.

[0067] In some embodiments, gelling the powdered decellularized matrix includes mixing the powdered decellularized matrix with tissue exudate and / or blood to form a gel. The powdered decellularized matrix of this application can absorb 6-10 times its own weight in solution, and therefore can directly combine with tissue exudate, blood, etc., to form a gel.

[0068] In other embodiments, gelling the powdered decellularized matrix includes mixing the powdered decellularized matrix with a polymer solution, the polymer solution including at least one of hyaluronic acid, chitosan, gelatin, dextran, and collagen. The powdered decellularized matrix of this application can also form a gel after mixing with a polymer solution. Taking gelatin as an example, after mixing the powdered decellularized matrix of this application with gelatin, a stable gel can be formed through non-covalent interactions such as hydrogen bonds and van der Waals forces.

[0069] In some embodiments, the mass ratio of the powdered decellularized matrix to the polymer solution is 1:(1~5). For example, the mass ratio of the powdered decellularized matrix to the polymer solution can be 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, or any value within the range of any two of the above values. Controlling the mass ratio of the powdered decellularized matrix to the polymer solution within the above range helps to improve the injectability of the resulting tissue-engineered gel scaffold while promoting stable binding gel formation.

[0070] In some embodiments, the mass concentration of the polymeric solute in the polymer solution is from 1% to 30%. For example, the mass concentration of the polymeric solute in the polymer solution can be 1%, 3%, 5%, 7%, 9%, 11%, 13%, 15%, 17%, 19%, 21%, 23%, 25%, 28%, 30%, or any value within the range of any two of the above values. Controlling the mass concentration of the polymeric solute in the polymer solution within the above range helps to improve the injectability of the resulting tissue-engineered gel scaffold while promoting stable binding gel formation.

[0071] One embodiment of this application also provides a composite gel scaffold, the composite gel scaffold comprising:

[0072] i) at least one of the tissue-engineered gel scaffolds described above and the tissue-engineered gel scaffolds prepared by the above preparation method; and ii) at least one of biomacromolecules and active factors.

[0073] The tissue-engineered gel scaffold of this application can also be used in combination with various active molecules or cells.

[0074] In some embodiments, the biomacromolecules include collagen.

[0075] In some embodiments, the active factor includes at least one of growth factors and cytokines.

[0076] One embodiment of this application also provides the use of at least one of the above-described tissue-engineered gel scaffolds, the tissue-engineered gel scaffolds prepared by the above-described preparation method, and the composite gel scaffolds in the preparation of soft tissue defect filling formulations.

[0077] The tissue-engineered gel scaffold of this application is based on decellularized cephalopod matrix, exhibiting good biocompatibility and a regular arrangement structure, strong water absorption, and strong mechanical properties. Its structure, water content, mechanical properties, and degradation rate are similar to soft tissue. It contains a high proportion of unsaturated fatty acids, with omega-3 content significantly exceeding omega-6, which is beneficial for inhibiting inflammation and repairing tissues. It can also be used in combination or in combination with various active molecules and cells for filling defects and repairing tissues during soft tissue regeneration, thus promoting the soft tissue regeneration process. Therefore, the tissue-engineered gel scaffold of this application possesses good biocompatibility, facilitating good integration with surrounding tissues when used for filling soft tissue defects. Simultaneously, the tissue-engineered gel scaffold of this application exhibits excellent mechanical properties, providing durable mechanical support after integration with surrounding tissues.

[0078] One embodiment of this application also provides the use of at least one of the above-described tissue-engineered gel scaffolds, the tissue-engineered gel scaffolds prepared by the above-described preparation method, and the composite gel scaffolds in the preparation of soft tissue injury repair agents.

[0079] The tissue-engineered gel scaffold of this application possesses mechanical properties and degradation rates similar to soft tissue, which is beneficial for providing continuous and suitable mechanical and biological support to soft tissue defect areas. Simultaneously, its excellent biocompatibility, mechanical properties, and active integration with tissues, along with its high proportion of unsaturated fatty acids (with omega-3 content significantly higher than omega-6), facilitate the synergistic construction of a microenvironment conducive to cell growth and functional reconstruction, thereby improving the effectiveness of soft tissue injury repair. Therefore, the tissue-engineered gel scaffold of this application can be used for filling soft tissue defects and synergistically promoting tissue repair.

[0080] The present application will be described below through specific embodiments and comparative examples. Those skilled in the art should understand that the preparation methods described in this application are merely examples, and any other suitable preparation methods are within the scope of this application.

[0081] Example 1:

[0082] A tissue-engineered gel scaffold, the preparation method of which includes:

[0083] Step 1: Clean the squid's outer membrane with pure water.

[0084] Step 2: Place the cleaned tissue in a 3% Triton X-100 solution for decellularization for 3 hours, then treat it with a 2% phosphoric acid solution for 1 hour to enhance the decellularization process. After the above treatment, place it in a 2% hydrogen peroxide solution for virus inactivation for 1 hour.

[0085] Step 3: The tissue is cleaned to remove residual reagents, and the decellularized squid mantle (i.e., decellularized matrix) is obtained. The decellularized matrix is ​​freeze-dried and pulverized to obtain powdered decellularized matrix.

[0086] Step 4: The powdered decellularized matrix is ​​directly combined with blood to form a gel, thus completing the gelation process and obtaining a tissue-engineered gel scaffold.

[0087] Example 2:

[0088] The difference from Example 1 is that in the first step, the mantle of the squid is replaced with the mantle of the cuttlefish.

[0089] Example 3:

[0090] The difference from Example 1 is that in the first step, the squid's mantle is replaced with the octopus's tentacles.

[0091] Example 4:

[0092] The difference from Example 1 is that the gelation step in the fourth step is replaced by mixing the powdered decellularized matrix with a 20% gelatin solution and stirring at 37 °C to complete the gelation.

[0093] Comparative Example 1:

[0094] A tissue-engineered gel scaffold is prepared by decellularizing pig skin using the method of Example 1, and then processed into powder form to combine with blood to form a gel, thereby completing the gelation process and obtaining the tissue-engineered gel scaffold.

[0095] Comparative Example 2:

[0096] The difference from Comparative Example 1 is that the fourth step of gelation is replaced by mixing the powdered decellularized matrix with a 20% gelatin solution and stirring at 37 °C to complete the gelation.

[0097] The following morphological analysis was performed in this application:

[0098] Morphological analysis of the decellularized mantle of squid, cuttlefish, and octopus in Examples 1-3 was performed using a Thermo Fisher Scientific Prisma E scanning electron microscope. Please refer to [link to relevant documentation]. Figure 1 , Figure 2 and Figure 3 After decellularization and powdering, the internal structures of the three cephalopods are all porous mesh structures, with the squid tissue being more loose and more suitable for cell growth.

[0099] Please see Figure 4 Taking Example 1 as an example, the mantle of squid, cuttlefish and octopus after decellularization treatment in this application example are processed into a uniform powder; while in Comparative Example 1, the powder made from decellularized pigskin has coarser particles.

[0100] This application also provides a morphological analysis of the tissue-engineered gel scaffolds obtained in Example 4 and Comparative Example 2. Please refer to [link to relevant documentation]. Figure 5 Both can form a stable gel and can be injected, but due to the higher fat content of pigskin, the resulting gel is more viscous. In contrast, Example 4 exhibits better fluidity during tissue injection and is more capable of filling any type of wound, such as burn wounds.

[0101] Furthermore, this application conducted the following performance analysis:

[0102] The fat content of the tissue-engineered gel scaffolds obtained in Example 1 and Comparative Example 1 was tested using the GB5009.6 standard method. Neither Example 1 nor Comparative Example 1 involved a defatting process in their preparation methods (see Table 1). The results show that Example 1 retained the original fat content, while under this mild decellularization process, the pig skin in Comparative Example 1 still contained a large amount of fat, which would increase the risk of infection and inflammation, affecting tissue healing.

[0103] Table 1. Test results of fat content of the tissue-engineered gel scaffolds provided in Example 1 and Comparative Example 1 of this application.

[0104]

[0105] This application also tested the water absorption and compression modulus of the tissue-engineered gel scaffolds obtained in Examples 1-3 and Comparative Example 1. The compression modulus test was performed using the material before powdering. The test results are shown in Table 2.

[0106] Table 2. Performance test results of tissue-engineered gel scaffolds of Examples 1-3 and Comparative Example 1 of this application

[0107]

[0108] This application also tested the fatty acid content of tissue-engineered gel scaffolds based on decellularized cephalopod matrix according to the GC-MS method of GB5009.168. Taking Example 1 as an example, please refer to Table 3. The results show that the treatment by this mild preparation process has little effect on the fatty acid content of the tissue itself, and can effectively retain the high-quality fatty acids in marine fish, especially unsaturated fatty acids mainly composed of ω-3 (DHA / EPA), which can effectively reduce inflammation at the defect site and promote tissue repair.

[0109] Table 3. Fatty acid content test results of the tissue-engineered gel scaffold in Example 1 of this application.

[0110]

[0111] This application also tested the compressive modulus of the tissue-engineered gel scaffolds obtained in Examples 1-3. The test results were as follows: Example 1: 21.7 MPa, Example 2: 30 MPa, and Example 3: 23 MPa, indicating that the tissue-engineered gel scaffolds of this application have good mechanical properties. Furthermore, the tissue-engineered gel scaffolds obtained in Examples 1-3 of this application, when lyophilized, contained a high proportion of unsaturated fatty acids, with omega-3 content exceeding omega-6 content, which is beneficial for inhibiting inflammation and repairing tissues. It was verified that the powdered decellularized matrix of Example 4 of this application, when mixed with a polymer solution of a predetermined mass concentration, completes gelation, which can promote stable binding gel formation while improving the injectability of the resulting tissue-engineered gel scaffold.

[0112] This application also verifies that the tissue powder obtained in Example 1 was directly applied to subcutaneous soft tissue defects in dogs. Please refer to [link to relevant documentation]. Figure 6 , Figure 6 (a) in the text represents the area before filling. Figure 6 (b) shows that after filling the decellularized tissue powder obtained in Example 1 for two days, the results indicate that the tissue healing effect is accelerated by rapidly absorbing blood and tissue exudate from the tissue defect to form a gel.

[0113] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0114] The embodiments described above are merely illustrative of several implementations of the technical solution of this application, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the technical solution of this application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the inventive concept of this application, and these modifications and improvements all fall within the protection scope of this application. Therefore, the protection scope of this application should be determined by the appended claims.

Claims

1. A tissue-engineered gel scaffold, characterized in that, The tissue-engineered gel scaffold comprises a decellularized matrix of cephalopods, wherein the tissue source of the decellularized matrix of the cephalopods includes at least one of the mantle of the cephalopods and the brachiopods of the cephalopods.

2. The tissue-engineered gel scaffold as described in claim 1, characterized in that, The compressive elastic modulus of the decellularized matrix of the cephalopods is from 0.2 MPa to 30 MPa.

3. The tissue engineering gel scaffold as described in claim 1, characterized in that, The decellularized matrix of the cephalopods absorbs water at a rate of 2 to 15 times its own weight.

4. The tissue-engineered gel scaffold as described in claim 1, characterized in that, The decellularized matrix of the cephalopod contains unsaturated fatty acids, including at least one of omega-3 and omega-6, and the unsaturated fatty acids account for more than or equal to 50% of the total fatty acids by mass.

5. The tissue-engineered gel scaffold as described in claim 1, characterized in that, The cephalopods include at least one of squid, cuttlefish, and octopus.

6. A method for preparing a tissue-engineered gel scaffold as described in any one of claims 1-5, characterized in that, The preparation method includes: The decellularized matrix-derived tissue of the cephalopod was pretreated to obtain a pretreated sample; The pretreated sample was subjected to decellularization to obtain a powdered decellularized matrix; The powdered decellularized matrix is ​​gelled to prepare the tissue-engineered gel scaffold.

7. The preparation method according to claim 6, characterized in that, Gelming the powdered decellularized matrix includes one of the following steps: (1) The powdered decellularized matrix is ​​mixed with tissue exudate and / or blood to form a gel; (2) The powdered decellularized matrix is ​​mixed with a polymer solution, wherein the polymer solution includes at least one of hyaluronic acid, chitosan, gelatin, dextran and collagen.

8. The preparation method according to claim 7, characterized in that, The preparation method also satisfies at least one of the following conditions: (1) The mass concentration of the polymer solute in the polymer solution is 1% to 30%; (2) The mass ratio of the powdered decellularized matrix to the polymer solution is 1:(1~5).

9. A composite gel scaffold, characterized in that, The composite gel scaffold includes: i) at least one of the tissue-engineered gel scaffolds as described in any one of claims 1-5, and the tissue-engineered gel scaffolds prepared by the preparation method as described in any one of claims 6-8; and ii) At least one of a biomacromolecule and an active factor, wherein the biomacromolecule includes collagen and the active factor includes at least one of a growth factor and a cytokine.

10. The use of at least one of the tissue-engineered gel scaffolds according to any one of claims 1-5, the tissue-engineered gel scaffolds prepared by the preparation method according to any one of claims 6-8, and the composite gel scaffolds according to claim 9 in the preparation of at least one of the following formulations: (1) Soft tissue defect filling preparations; (2) Soft tissue injury repair agents.

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