Composite hydrogel as well as preparation method and application thereof

Abstract

The invention relates to composite hydrogel as well as a preparation method and application thereof. The collagen / coagulant aid composite material comprises the following components in parts by mass: 1-10 parts of collagen, 1-10 parts of a coagulant aid, 1-10 parts of a blood sample and 1-10 parts of fibrinogen. All the components are coordinated and matched through a specific matching relation, so that the composite hydrogel can form gel in situ, the gel is good in mechanical property and high in adhesion capacity, can be subjected to injection molding, can be prepared into a size matched with a part, needing to be implanted, of a patient, and can accelerate tissue repair.

Description

Technical Field

[0001] This application relates to the technical field of medical materials, and particularly to a composite hydrogel and its preparation method and application. Background Art

[0002] Hydrogels have become a promising biomaterial in the field of tissue engineering due to their unique properties and biocompatibility. This three-dimensional network composed of cross-linked polymers can absorb a large amount of water, creating a hydrated environment similar to natural tissues. This similarity to natural tissues makes hydrogels an ideal material for supporting cell growth, proliferation, and differentiation, making them a powerful tool in tissue engineering and regenerative medicine.

[0003] According to different application requirements, it is crucial to select a suitable hydrogel transition method. Currently, common methods for hydrogels to transform from sol to gel include physical cross-linking, chemical cross-linking, self-assembly, and freeze-thaw methods. However, the above traditional technologies still have limitations to a certain extent. Among them, the physical cross-linking method is easy to operate and mild, but it is usually less stable than the chemical cross-linking method. The chemical cross-linking method can provide stronger gel mechanical properties, but it may require harsh conditions, and the toxicity problems of some cross-linking agents need attention. The self-assembly method provides innovative design ideas, but it is restricted by environmental conditions and self-assembly stability. The freeze-thaw method is a relatively simple method, but its applicability is limited by low-temperature conditions.

[0004] Therefore, the traditional technologies still need to be improved. Summary of the Invention

[0005] Based on this, this application provides an injectable composite hydrogel that can form a gel in situ, and its preparation method and application.

[0006] The specific technical solutions are as follows:

[0007] In the first aspect of this application, a composite hydrogel is provided. By mass fraction, it includes the following components: 1 part to 10 parts of collagen, 1 part to 10 parts of coagulant, 1 part to 10 parts of blood sample, and 1 part to 10 parts of fibrinogen.

[0008] In some embodiments, by mass percentage, it includes the following components: 1 part to 8 parts of collagen, 1 part to 2 parts of coagulant, 1 to 5 parts of blood sample, and 1 part to 5 parts of fibrinogen.

[0009] In some embodiments, the blood sample includes bioactive factors;

[0010] Optionally, the bioactive factors include one or more of platelet-derived growth factor, transforming growth factor-β, insulin-like growth factor, epidermal growth factor, and vascular endothelial growth factor.

[0011] In some embodiments, the coagulant aid includes one or more of thrombin and calcium agent;

[0012] Optionally, the concentration of thrombin is 200U / mL~2000U / mL;

[0013] Optionally, the mass percentage of the calcium agent is 2% to 10%;

[0014] Further optionally, the calcium agent includes one or more of calcium chloride, calcium gluconate, calcium lactate and calcium citrate.

[0015] In some embodiments, the composite hydrogel further comprises one or more of stem cells and neural factors.

[0016] Another aspect of the present application also provides a method for preparing a composite hydrogel, comprising the following steps:

[0017] adding a fibrinogen solution to a blood sample to prepare a first mixture;

[0018] adding a coagulant to the collagen suspension to prepare a second mixture; and,

[0019] mixing the first mixture and the second mixture to prepare a composite hydrogel;

[0020] The composition comprises the following components by weight: 1 to 10 parts of collagen, 1 to 10 parts of coagulant, 1 to 10 parts of blood sample and 1 to 10 parts of fibrinogen.

[0021] In some embodiments, the volume ratio of the first mixture to the second mixture is 1:5 to 5:1;

[0022] And / or, the mixing temperature is 1°C to 37°C;

[0023] And / or, the concentration of fibrinogen in the fibrinogen solution is 10 mg / mL to 100 mg / mL;

[0024] And / or, the mass percentage of collagen in the collagen suspension is 0.5%~6%.

[0025] In some embodiments, the volume ratio of the coagulant aid to the collagen suspension is 1:1 to 1:10.

[0026] Another aspect of the present application also provides a use of the above-mentioned composite hydrogel in the preparation of a cartilage damage repair product.

[0027] Another aspect of the present application also provides a cartilage damage repair product, comprising the above-mentioned composite hydrogel.

[0028] The composite hydrogel of the present application comprises collagen, a coagulant, a blood sample, and fibrinogen with specific component ratios. Among them, collagen serves as the basic framework component of the hydrogel, providing attachment sites for cells, facilitating cell adhesion, proliferation, and differentiation, guiding tissue regeneration and repair, and at the same time endowing the hydrogel with certain physical strength and stability. The blood sample contains a large number of platelets, and after platelet activation, a variety of growth factors can be released, which can promote cell proliferation, migration, and differentiation, and accelerate the tissue repair and regeneration process. Thrombin plays a key role in the coagulation process. It can catalyze the conversion of fibrinogen into fibrin, promoting the formation of the hydrogel, so that the composite hydrogel can gel in situ, facilitating immediate molding in clinical applications. Calcium ions participate in the coagulation process. As a cofactor for thrombin activation, they enhance the activity of thrombin, promote the conversion of fibrinogen to fibrin, and play an important role in the formation and solidification of the hydrogel. Under the action of thrombin, fibrinogen is transformed into fibrin, forming a three-dimensional network structure, which is an important part of the hydrogel, endowing the hydrogel with certain mechanical strength and stability, and at the same time providing a suitable microenvironment for cell growth and tissue repair. Through the coordinated cooperation of each component with specific ratio relationships, the composite hydrogel can form a gel in situ, has good colloidal mechanical properties, strong adhesion ability, can be injection molded, made into a size matching the implantation site required by the patient, and can accelerate tissue repair.

[0029] Furthermore, the preparation process of the composite hydrogel is simple and easy to implement. All the above steps can use sterile reagents and aseptic operations to achieve the sterility of the product. It can be directly implanted as a tissue engineering scaffold without adding a sterilization process, and can maintain biological activity to the greatest extent. Specific Embodiments

[0030] To facilitate the understanding of the present application, the present application will be described more comprehensively below, and preferred embodiments of the present application are given. However, the present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure content of the present application more thorough and comprehensive.

[0031] The implementation of the present application will be described in detail below in combination with some embodiments and examples. This embodiment is implemented on the premise of the technical solution of the present application, and detailed implementation manners and specific operation processes are given, but the protection scope of the present application is not limited to the following embodiments.

[0032] 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 terms used in the specification of this application are for the purpose of describing specific embodiments only and are not intended to limit this application.

[0033] Unless otherwise stated or in case of contradiction, the terms or phrases used herein have the following meanings:

[0034] In this application, when referring to "multiple", "a variety of", etc., unless otherwise specified, it means greater than 2 or equal to 2 in quantity. For example, "one or more" means one or greater than or equal to two.

[0035] In this application, terms such as "further", "especially", etc. are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0036] In this application, among the technical features described in an open-ended manner, it includes a closed technical solution composed of the listed features, and also includes an open technical solution containing the listed features.

[0037] In this application, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values within this numerical interval is considered continuous, and includes the two numerical endpoints of this numerical interval (i.e., the minimum value and the maximum value), as well as each numerical value between these two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within this numerical interval, it includes the two endpoint integers of this numerical range, as well as each integer between the two endpoints, which is equivalent to directly listing each integer. When providing multiple numerical ranges to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all sub-ranges subsumed therein. The "numerical value" in this numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. The "numerical interval" allows for a broad inclusion of numerical interval types such as a percentage interval, a ratio interval, a ratio interval, etc.

[0038] In this application, unless otherwise specified, the temperature parameter allows both constant temperature treatment and variation within a certain temperature range. It should be understood that the so-called constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within ranges such as ±5°C, ±4°C, ±3°C, ±2°C, ±1°C are allowed.

[0039] In this application, regarding the unit of a data range, if the unit is only attached after the right endpoint, it means that the units of the left endpoint and the right endpoint are the same. For example, 2~5h means that the units of the left endpoint "2" and the right endpoint "5" are both h (hours).

[0040] As shown in the background technology, traditional technologies still have certain limitations to a certain extent. For example, the transition temperature of thermosensitive hydrogels is relatively narrow, and different applications require precise temperature control, which may limit their use in specific environments. pH-sensitive hydrogels are only suitable for environments where pH changes are more significant. For systems with a smaller pH range, their reactivity may not be sensitive enough. Salt-sensitive hydrogels: changes in salt concentration may be greatly affected by environmental factors, and the effect of salt on hydrogels is relatively limited. Chemically cross-linked hydrogels may have residual toxicity of cross-linking agents. Some cross-linking agents (such as glutaraldehyde) may be toxic and not suitable for biomedical applications; chemical cross-linking reaction conditions are harsh: chemical reactions such as free radical polymerization usually require high temperatures or strong acids and bases, which may affect the stability or biocompatibility of hydrogels. Uneven cross-linking degree: In some cases, the cross-linking reaction may be uneven, resulting in unstable physical properties of the hydrogel and even affecting its function. The gelation process of self-assembled hydrogels is complex: the self-assembly process often requires specific conditions (such as pH, temperature, salt concentration, etc.), and the control is relatively complex.

[0041] Furthermore, the technical personnel of the present application have found that when the collagen suspension is mixed with the blood sample to form a gel, the gel obtained by this method will precipitate some water, shrink in volume, and have a low strength, which is not conducive to control in biomedical engineering applications. However, when the collagen suspension is mixed with the fibrinogen bioadhesive to form a gel, this method does not contain the various growth factors in the blood sample.

[0042] After a large number of creative experimental studies, the applicant found that compared with existing hydrogels, the synergistic effect of collagen, blood samples and fibrinogen makes the injectable hydrogel have good biocompatibility, high safety performance, and a wide range of uses. It can also be prepared and used clinically and has better tissue repair effects.

[0043] Based on this, one embodiment of the present application provides a composite hydrogel, which includes the following components, by mass: 1 to 10 parts of collagen, 1 to 10 parts of coagulant, 1 to 10 parts of blood sample and 1 to 10 parts of fibrinogen.

[0044] The composite hydrogel of the present application comprises collagen, a coagulant, a blood sample and fibrinogen with specific component ratios. Among them, collagen serves as the basic framework component of the hydrogel, provides attachment sites for cells, facilitates cell adhesion, proliferation and differentiation, guides tissue regeneration and repair, and at the same time imparts certain physical strength and stability to the hydrogel. The blood sample contains a large number of platelets, and after platelet activation, a variety of growth factors can be released, which can promote cell proliferation, migration and differentiation, and accelerate the tissue repair and regeneration process. Thrombin plays a key role in the blood coagulation process. It can catalyze the conversion of fibrinogen into fibrin, promoting the formation of the hydrogel, so that the composite hydrogel can gel in situ, facilitating immediate molding in clinical applications. Calcium ions participate in the blood coagulation process. As a cofactor for thrombin activation, they enhance the activity of thrombin, promote the conversion of fibrinogen to fibrin, and play an important role in the formation and solidification of the hydrogel. Under the action of thrombin, fibrinogen is transformed into fibrin, forming a three-dimensional network structure, which is an important part of the hydrogel, endows the hydrogel with certain mechanical strength and stability, and at the same time provides a suitable microenvironment for cell growth and tissue repair. Through the coordinated cooperation of each component with a specific ratio relationship, the composite hydrogel can form a gel in situ, has good colloidal mechanical properties, strong adhesion ability, can be injection molded, is made into a size matching the implantation site required by the patient, and can accelerate tissue repair.

[0045] In some of these embodiments, by mass percentage, it comprises the following components: 1 part to 8 parts of collagen, 1 part to 2 parts of coagulant, 1 to 5 parts of blood sample and 1 part to 5 parts of fibrinogen.

[0046] By further regulating the component ratio relationship of collagen, coagulant, blood sample and fibrinogen, the colloidal mechanical properties of the composite hydrogel can be better, the adhesion ability can be stronger, it can be injection molded, is made into a size matching the implantation site required by the patient, and can accelerate tissue repair.

[0047] In some of these embodiments, the above-mentioned blood sample contains bioactive factors.

[0048] Optionally, the above-mentioned bioactive factors include one or more of platelet-derived growth factor, transforming growth factor-β, insulin-like growth factor, epidermal growth factor and vascular endothelial growth factor.

[0049] It can be understood that the blood sample contains a variety of bioactive factors and nutrients, can recruit stem cells, provide adhesion conditions for stem cells and induce directional differentiation, and at the same time does not cause an immune response in the body. It has fluidity and cannot be directly used for cartilage repair, so it needs to be combined with a scaffold material, and is fixed and adhered to the cartilage defect by the scaffold material.

[0050] In some of these embodiments, the above-mentioned blood sample includes at least one of peripheral blood and whole blood. Optionally, the peripheral blood may be, but is not limited to, autologous peripheral blood, and the whole blood may be, but is not limited to, autologous whole blood.

[0051] Optionally, the above-mentioned blood sample includes one or more of platelet-rich plasma and platelet-poor plasma.

[0052] Platelet-rich plasma is a platelet concentrate obtained by centrifuging whole blood, which contains a large number of bioactive factors, such as platelet-derived growth factor (PDGF), transforming growth factor (TGF-β), insulin-like growth factor (IGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and can promote tissue growth and repair.

[0053] In some of these embodiments, the above-mentioned coagulant includes at least one of thrombin and calcium agent.

[0054] Optionally, the concentration of the above-mentioned thrombin is 200 U / mL to 2000 U / mL, and the mass percentage of the above-mentioned calcium agent is 2% to 10%.

[0055] It should be noted that the value range of the concentration of thrombin is "200 U / mL to 2000 U / mL", that is, the minimum value and the maximum value of the range of 200 U / mL to 2000 U / mL can be taken, as well as each value between this minimum value and the maximum value. Specific examples include, but are not limited to, the point values in the embodiments and the following point values: 200 U / mL, 300 U / mL, 400 U / mL, 500 U / mL, 600 U / mL, 700 U / mL, 800 U / mL, 900 U / mL, 1000 U / mL, 1100 U / mL, 1200 U / mL, 1300 U / mL, 1400 U / mL, 1500 U / mL, 1600 U / mL, 1700 U / mL, 1800 U / mL, 1900 U / mL or 2000 U / mL; or the ranges composed of any two of these values. As an example, it includes: 500 U / mL to 1000 U / mL.

[0056] The value range of the mass percentage of the calcium agent is "2% to 10%", that is, the minimum value and the maximum value of the range of 2% to 10% can be taken, as well as each value between this minimum value and the maximum value. Specific examples include, but are not limited to, the point values in the embodiments and the following point values: 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%; or the ranges composed of any two of these values. As an example, it includes: 3% to 10%.

[0057] Further optionally, the above calcium agent includes at least one of calcium chloride, calcium gluconate, calcium lactate or calcium citrate.

[0058] It can be understood that thrombin not only plays a key role in the conversion of fibrinogen, but also works synergistically with calcium agents and other components (such as collagen suspension and platelet-rich plasma). In the entire gel formation system, the activity of thrombin cooperates with the presence of calcium agents to ensure the efficiency of the coagulation process.

[0059] In some of these embodiments, the above composite hydrogel further includes at least one of stem cells and nerve factors.

[0060] Other growth factors and nerve factors can also be added externally to the above composite hydrogel to form a gel. During cartilage repair, the additionally added growth factors can further supplement and cooperate with the growth factors in PRP, enhancing the stimulatory effect on chondrocytes. The addition of nerve factors helps to improve local nerve innervation, which may have a positive significance for reducing pain and promoting tissue repair. For example, in some patients with cartilage damage accompanied by pain, nerve factors can help regulate nerve conduction, relieve pain symptoms, and promote the repair of cartilage tissue at the same time.

[0061] In some of these embodiments, the addition amount of the above stem cells is 1 part to 10 parts.

[0062] In some of these embodiments, the addition amount of the above nerve factors is 1 part to 10 parts.

[0063] An embodiment of the present application also provides a preparation method of a composite hydrogel, including step S10 to step S30.

[0064] Step S10: Add a fibrinogen solution to a blood sample to prepare a first mixture.

[0065] It can be understood that after fibrinogen forms a gel, it can slowly release various growth factors such as platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and vascular endothelial growth factor (VEGF) in the blood sample, prolonging the action time of the growth factors at the repair site and continuously stimulating the regeneration and repair process of chondrocytes.

[0066] Step S20: Add a coagulant to the collagen suspension to prepare a second mixture.

[0067] It is understandable that collagen itself is an important component of the extracellular matrix of chondrocytes. Controlling the gel formed by mixing it with other components at a specific concentration can provide a supporting structure close to the physiological environment for chondrocytes. Chondrocytes can better maintain their phenotypes in this gel scaffold and carry out normal metabolic and functional activities, such as synthesizing and secreting the extracellular matrix, and promoting the reconstruction of cartilage tissue.

[0068] Step S30: Mix the above first mixture and the above second mixture at 1°C to 37°C to prepare a composite hydrogel; by mass, it includes the following components: 1 to 10 parts of collagen, 1 to 10 parts of a coagulant aid, 1 to 10 parts of a blood sample, and 1 to 10 parts of fibrinogen.

[0069] After the above coagulant aid (calcium agent and thrombin) is mixed with the collagen suspension, it is then uniformly mixed with PRP containing fibrinogen to form a gel in situ at the injection site. The three-dimensional structure of this gel is similar to the extracellular matrix of cartilage tissue, providing good attachment sites for chondrocytes.

[0070] In some of these embodiments, the above mixing time is 3 min to 60 min, for example, it can be 3 min, 5 min, 7 min, 9 min, 11 min, 13 min, 15 min, 17 min, 19 min, 21 min, 23 min, 25 min, 27 min, 29 min, 31 min, 33 min, 35 min, 37 min, 39 min, 41 min, 43 min, 45 min, 47 min, 49 min, 51 min, 53 min, 55 min, 57 min, 59 min or 60 min. In some examples, it can be within the range formed by any two of these point values as the end values. The same applies hereinafter.

[0071] It is understandable that in some cartilage repair surgeries that require precise shaping, the gel formation time can be extended to have enough time to inject the mixture into the cartilage defect site and perform shaping, so that it better fits the defect shape and improves the repair effect. In some emergency situations or scenarios with high requirements for operation time, the gel formation can be accelerated to quickly provide a stable scaffold for cartilage repair.

[0072] In some of these embodiments, the volume ratio of the above first mixture and the above second mixture is 1:5 to 5:1, for example, it can be 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1 or 5:1.

[0073] In some of these embodiments, the temperature of the mixture is from 1°C to 37°C; for example, it can be 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C or 37°C.

[0074] In some of these embodiments, the concentration of fibrinogen in the fibrinogen solution is from 10 mg / mL to 100 mg / mL; for example, it can be 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL or 100 mg / mL.

[0075] In some of these embodiments, the mass percentage of collagen in the collagen suspension is from 0.5% to 6%; for example, it can be 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5% or 6%.

[0076] In some of these embodiments, the volume ratio of the above-mentioned coagulant aid to the above-mentioned collagen suspension is from 1:1 to 1:10; for example, it can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0077] Furthermore, the preparation process of this composite hydrogel is simple and easy to implement. All of the above steps can use sterile reagents and aseptic operations to achieve the sterility of the product, and it can be directly implanted as a tissue engineering scaffold without adding a sterilization process, which can maximize the preservation of biological activity.

[0078] One embodiment of this application also provides an application of the above-mentioned composite hydrogel in the preparation of a product for repairing cartilage damage.

[0079] One embodiment of this application also provides a product for repairing cartilage damage, including the above-mentioned composite hydrogel.

[0080] It is understandable that the above composite hydrogel is used in products for cartilage injury repair and has better clinical operability. When used for cartilage defect repair, through the microfracture technique, bone marrow mesenchymal stem cells in the bone under the injury site flow in and proliferate and differentiate into new cartilage, which can better cover the cartilage defect area, adhere more firmly to the cartilage defect site, and the tissue repair effect is better.

[0081] In order to make the purpose, technical solutions and advantages of the present application more concise and clear, the present application is described by the following specific embodiments, but the present application is by no means limited to these embodiments. The embodiments described below are only the preferred embodiments of the present application and can be used to describe the present application, and should not be construed as a limitation on the scope of the present application. It should be noted that any modifications, equivalent replacements and improvements made within the spirit and principle of the present application should be included within the protection scope of the present application.

[0082] To better illustrate the present application, the content of the present application will be further described below in conjunction with embodiments.

[0083] Example 1

[0084] 1. At room temperature, centrifugation is used to prepare platelet-rich plasma (PRP) from whole blood for standby. The amount of the obtained PRP can be controlled at 1 mL to 5 mL. Human fibrinogen (commercially available HuGulaiShi, the specification can be 1 - 5 ml) is added to the prepared PRP, and the added fibrinogen concentration is 50 mg / mL and completely dissolved. It is prepared and used immediately to prepare the first mixture.

[0085] 2. Prepare a coagulant aid at room temperature: calcium agent + thrombin, where the mass concentration of the calcium agent is 5%, specifically calcium chloride; the thrombin concentration is 500 U / mL.

[0086] 3. Mix the coagulant aid with the collagen suspension at room temperature. The concentration of collagen is 3%, and the volume ratio of the coagulant aid to the collagen suspension is 1:4 to prepare the second mixture.

[0087] 4. Mix the first mixture and the second mixture at 25°C through a dual syringe, and the volume ratio of the two is 1:1. By mass fraction, the composite hydrogel includes 8 parts of collagen, 2 parts of coagulant aid, 5 parts of blood sample, and 5 parts of fibrinogen; it is injected into a mold to be shaped into a gel, and uniform mixing of PRP and the collagen suspension containing the coagulant aid is achieved during the injection process. The gel time is 10 min, and it can form a gel in situ.

[0088] Example 2

[0089] The preparation method of the composite hydrogel in Example 2 is basically the same as that in Example 1, except that the ratios of collagen, coagulant, blood sample and fibrinogen are different. Specifically, by mass fraction, the composite hydrogel includes 1 part of collagen, 1 part of coagulant, 1 part of blood sample and 1 part of fibrinogen.

[0090] Example 3

[0091] The preparation method of the composite hydrogel in Example 3 is basically the same as that in Example 1, except that the ratios of collagen, coagulant, blood sample and fibrinogen are different. Specifically, by mass fraction, the composite hydrogel includes 6 parts of collagen, 2 parts of coagulant, 10 parts of blood sample and 2 parts of fibrinogen.

[0092] Example 4

[0093] The preparation method of the composite hydrogel in Example 4 is basically the same as that in Example 1, except that the ratios of collagen, coagulant, blood sample and fibrinogen are different. Specifically, by mass fraction, the composite hydrogel includes 10 parts of collagen, 3 parts of coagulant, 7 parts of blood sample and 2 parts of fibrinogen.

[0094] Example 5

[0095] The preparation method of the composite hydrogel in Example 5 is basically the same as that in Example 1, except that the temperature at which the first mixture and the second mixture are mixed is different, specifically 37°C.

[0096] Other steps and conditions are the same as those in Example 1.

[0097] Comparative Example 1

[0098] The preparation method of the composite hydrogel in Comparative Example 1 is basically the same as that in Example 1, except that fibrinogen is not added.

[0099] Other steps and conditions are the same as those in Example 1.

[0100] Comparative Example 2

[0101] The preparation method of the composite hydrogel in Comparative Example 2 is basically the same as that in Example 1, except that the blood sample is not added.

[0102] Other steps and conditions are the same as those in Example 1.

[0103] Comparative Example 3

[0104] The preparation method of the composite hydrogel in Comparative Example 3 is basically the same as that in Example 1, except that the ratios of collagen, coagulant, blood sample and fibrinogen are different. Specifically, by mass fraction, the composite hydrogel comprises 12 parts of collagen, 2 parts of coagulant, 4 parts of blood sample and 2 parts of fibrinogen.

[0105] Test:

[0106] 1. Test the colloidal mechanical properties of the composite hydrogels prepared in the examples and comparative examples. Specifically, the compression method is used to detect the elastic modulus.

[0107] 2. Test the tissue repair effect of the composite hydrogels prepared in the examples and comparative examples. Specifically, L929 fibroblasts are used to conduct a cell scratch experiment for detection and verification. The results are shown in Table 1.

[0108] Table 1

[0109]

[0110] In summary, compared with the comparative examples, the composite hydrogel prepared by the technical solution of the present application has good colloidal mechanical properties, strong adhesion ability, and better tissue repair effect.

[0111] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of concise description, 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, it should be considered as the scope described in this specification.

[0112] The above-described embodiments merely represent several implementation manners of the present application. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can still be made, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the appended claims.

Claims

1. A composite hydrogel, characterized in that, By mass parts, it includes the following components: 1 part to 10 parts of collagen, 1 part to 10 parts of coagulant aid, 1 part to 10 parts of blood sample, and 1 part to 10 parts of fibrinogen.

2. The composite hydrogel according to claim 1, wherein By mass percentage, it includes the following components: 1 part to 8 parts of collagen, 1 part to 2 parts of coagulant aid, 1 part to 5 parts of blood sample, and 1 part to 5 parts of fibrinogen.

3. The composite hydrogel according to claim 1, wherein The blood sample includes bioactive factors; Optionally, the bioactive factors include one or more of platelet-derived growth factor, transforming growth factor-β, insulin-like growth factor, epidermal growth factor, and vascular endothelial growth factor.

4. The composite hydrogel according to any one of claims 1 to 3, characterized in that, The coagulant aid includes one or more of thrombin and calcium agent; Optionally, the concentration of the thrombin is 200 U / mL to 2000 U / mL; Optionally, the mass percentage of the calcium agent is 2% to 10%; Further optionally, the calcium agent includes one or more of calcium chloride, calcium gluconate, calcium lactate, and calcium citrate.

5. The composite hydrogel according to any one of claims 1 to 3, characterized in that, The composite hydrogel further includes one or more of stem cells and nerve factors.

6. A preparation method of a composite hydrogel, characterized in that, It includes the following steps: Adding a fibrinogen solution to the blood sample to prepare a first mixture; Adding a coagulant aid to the collagen suspension to prepare a second mixture; and, Mixing the first mixture and the second mixture to prepare a composite hydrogel; By mass parts, it includes the following components: 1 part to 10 parts of collagen, 1 part to 10 parts of coagulant aid, 1 part to 10 parts of blood sample, and 1 part to 10 parts of fibrinogen.

7. The preparation method of the composite hydrogel according to claim 6, characterized in that, The volume ratio of the first mixture to the second mixture is 1:5 to 5:1; And / or, the temperature of the mixing is 1°C to 37°C; And / or, the concentration of fibrinogen in the fibrinogen solution is 10 mg / mL to 100 mg / mL; And / or, the mass percentage of collagen in the collagen suspension is 0.5% to 6%.

8. The preparation method of the composite hydrogel according to any one of claims 6 to 7, characterized in that, The volume ratio of the coagulant aid to the collagen suspension is 1:1 to 1:

10.

9. Use of the composite hydrogel according to any one of claims 1 to 5 in the preparation of a cartilage injury repair product.

10. A cartilage injury repair product, characterized in that, It includes the composite hydrogel according to any one of claims 1 to 5.

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