WO2022028244A1

WO2022028244A1 - Tissue filling material, preparation method therefor, tissue engineering scaffold and use

Info

Publication number: WO2022028244A1
Application number: PCT/CN2021/107359
Authority: WO
Inventors: 吕世文; 陈超; 李彪
Original assignee: 宁波迪创医疗科技有限公司
Priority date: 2020-08-04
Filing date: 2021-07-20
Publication date: 2022-02-10
Also published as: CN111888524A

Abstract

A tissue filling material, a preparation method therefor, a tissue engineering scaffold comprising the tissue filling material and a use of the tissue filling material. The tissue filling material comprises the following raw materials: alginate, a metal cation cross-linking agent and a water-soluble biodegradable pore-forming filling material. The content of the raw materials in the tissue filling material respectively is 0.1-20 wt%, and the balance is injection water. After the tissue filling material is implanted in body, the pore-forming filling material rapid degrades, an interconnected porous mesh structure is thereby formed, and both nutrients and cells can enter the interior; the degraded product has molecular activity, inducing myocardial cells to enter the pores and grow; therefore, the present invention can adjust cell adhesion, proliferation and differentiation, increase ventricular wall thickness, promote myocardial neovascularization, reduce myocardial infarction range, lessen post-infarction cardiac function decrease, and improve tissue regenerative and repairing capabilities.

Description

Tissue filling material and preparation method thereof, tissue engineering scaffold and application

technical field

The present application relates to the field of materials, in particular to a tissue filling material and a preparation method thereof, a tissue engineering scaffold and applications.

Background technique

Tissue engineering is a science that applies the principles and technologies of life science and engineering to develop biological substitutes for repairing, maintaining, and promoting the function and morphology of various tissues or organs in the human body after injury. Porous materials are usually used as scaffolds. materials to achieve initial support of tissue in vivo, initial adhesion of cells, and controllable tissue formation by cells. For example, tissue engineering scaffolds can be constructed using alginate hydrogels.

However, the above technical solutions have the following deficiencies in practical use: the existing alginate hydrogel has a single main body material, only sodium alginate and calcium alginate, a biopolymer material, and its therapeutic principle only uses coagulation as a The filler acts as a physical support, but does not induce the growth of cardiomyocytes, and the cells will not enter the gel; and the porosity of the gel is not easy to control, and the internal three-dimensional pores are not connected to each other, so 3D connected pores cannot be formed. Therefore, the traditional alginate-based hydrogel structure is not conducive to the growth of cardiomyocytes in the gel and the recovery of myocardial function, and cannot be used for the treatment of heart failure diseases.

SUMMARY OF THE INVENTION

The purpose of the embodiments of the present application is to provide a tissue filling material, so as to solve the problem that most of the existing tissue filling materials proposed in the above background technology can only play a physical support role, and there are problems that are not conducive to the growth of myocardial cells in the material and the recovery of myocardial function. problem.

To achieve the above purpose, the embodiments of the present application provide the following technical solutions:

A tissue filling material, comprising component A, component B, and water for injection, the component A includes alginate, the component B includes a metal cation cross-linking agent, the component A and/ Or component B further comprises a water-soluble biodegradable pore-forming filling material, the component A and component B are fully stirred to form an ion cross-linked hydrogel structure, and the pore-forming filling material is uniformly distributed in the cross-linked structure. inside the structure. The content of the alginate in the tissue filling material is 0.1-20 wt %, the content of the metal cationic cross-linking agent in the tissue filling material is 0.1-20 wt %, and the pore-forming filling material is in the The content in the tissue filling material is 0.1-20wt%. Wherein, % (g/mL) represents the content by weight in 1 mL of tissue filling material, for example, the content of the alginate in the tissue filling material is 0.1-10% (g/mL) means that 1 mL of tissue filling material contains 0.001-0.10 g of alginate, which can also be expressed as the content of the alginate in the tissue filling material is 0.1-10 wt %.

As a further solution of the present application: the tissue filling material is an injectable alginate-based composite hydrogel, the hydrogel includes a rapidly degraded pore-forming filler component, and the pore-forming filler material is degraded At the same time, its degradation products have biological activity, can promote the growth of tissue cells, and can regulate cell adhesion, proliferation and differentiation.

As a further solution of the present application: the pore-forming filling material includes any one or more of polysaccharide-based materials or protein-based materials.

As a further solution of the present application: the polysaccharide material includes any one of hyaluronic acid, dextran, chitosan or chondroitin sulfate, and the protein material includes collagen, gelatin, fiber either protein or laminin.

As a further solution of the present application: the pore filling material contains active drugs capable of promoting cell growth, and the active drugs include transforming growth factor, platelet-derived growth factor, and fibroblast growth factor.

As a further solution of the present application: the degradation period of the pore-forming filling material is less than or equal to 8 weeks.

As a further solution of the present application: the pores in the structure of the tissue filling material gradually increase as the material of the pore filling material degrades.

As a further solution of the present application: the pore-forming filling material further includes a regulator for regulating the degradation rate, and the regulator includes any one of hyaluronidase, glucanase, chitosanase, and protease .

As a further scheme of the application: the alginate is any one of sodium alginate, potassium alginate, ammonium alginate or propylene glycol alginate; the metal cationic crosslinking agent is selected from calcium alginate, glucose A combination of one or more of calcium acid, calcium carbonate, calcium sulfate or calcium chloride.

As a further scheme of the present application: the molecular weight of the alginate is 5-400kDa (kilodalton).

Preferably, the alginate is sodium alginate, and the molecular weight of the sodium alginate is 5-400kDa.

As a further solution of the present application: the metal cations in the metal cation-containing crosslinking agent are not less than divalent metal cations. Specifically, the metal cation-containing crosslinking agent is a divalent or multivalent metal cation-containing crosslinking agent.

As a further scheme of this application: the divalent metal cations include calcium cation, barium cation, zinc cation, iron cation, magnesium cation, copper cation, etc., and the multivalent metal cation includes aluminum cation, chromium cation, molybdenum cation, etc. cations, tin cations, etc.

As a further solution of the present application: the metal cation-containing crosslinking agent is selected from a combination comprising one or more of calcium alginate, calcium gluconate, calcium carbonate, calcium sulfate or calcium chloride.

As a further solution of the present application: the solid particle size of the metal cation-containing crosslinking agent is not greater than 1 mm.

As a further solution of the present application: the solid particle diameter of the metal cation-containing crosslinking agent is 10-300 μm.

Preferably, the metal cation-containing crosslinking agent is calcium alginate, and the solid particle size of the calcium alginate is 10-300 μm.

As a further solution of the present application: the water-soluble biodegradable pore-forming filling material is a water-soluble and rapidly degradable biopolymer material, namely component C. Specifically, the pore-forming filling material includes polysaccharides Any one or more of protein-like materials or protein-like materials.

It should be noted that the pore-forming filler component needs to be dissolved in an aqueous solution, has good biocompatibility, and is rapidly degraded, and the degradation product is non-toxic. It can be seen from the above description that there are many materials suitable for the pore-forming filler component, and those skilled in the art can select them according to the specific use environment and use purpose to meet clinical requirements. The content range of the above-described pore-forming filler component in the tissue-filling material of the present application is only an exemplary preferred range, and those skilled in the art can also select other pore-forming filler materials, and obtain their suitable values through limited experiments. range of use.

Preferably, the water-soluble biodegradable polymer material is hyaluronic acid.

Obviously, the addition of the pore-forming filler component endows the tissue filler with special biological properties, bringing the following advantages:

1) After the tissue filling material is injected into the myocardium, the enzymes in the body will rapidly degrade the pore-forming filler components to form an interpenetrating porous network structure, thereby solving the problem of the single pore size of the existing alginate-based hydrogels. , the inability to form 3D connected pores, and the inability of nutrients and cells to enter the interior of the scaffold. Further, after the tissue filling material is injected into the myocardium, while the pore-forming filler component is degraded in the body, the degraded product has molecular activity, can induce cells to enter the gel, and can regulate cell adhesion and proliferation. and differentiation. Further, after the tissue filling material is injected into the myocardium, while the pore-forming filler component is degraded in vivo, the degraded product can provide nutrients for cell growth and promote cell growth and blood vessel formation;

2) The tissue filling material has excellent biocompatibility, a simple manufacturing process, and the pore size and porosity of the hydrogel can be adjusted by changing the amount of the pore-forming filler component; the tissue filling material can be used either In vitro cell culture, but also in vivo injection applications.

As a further solution of the present application: in the tissue filling material, the raw material of the tissue filling material further includes an isotonicity agent and/or a regulator, specifically, the isotonicity agent is component D, the The conditioner is component E.

As a further solution of the present application: the tissue filling material further includes component D, or component E, or a combination of component D and component E. The isotonicity agent is used to adjust the osmotic pressure of the tissue filling material to meet the environmental requirements of the application field, and the adjuster is used to adjust the pH value of the tissue filling material to meet the environmental requirements of the application field. The specific amount of the isotonicity agent and/or regulator is selected according to requirements, and is not limited here.

As a further solution of the present application: the content of the isotonicity agent in the tissue filling material is 280-320 mmol/L, and the content of the adjusting agent in the tissue filling material is 0.001-25% (g/L) mL).

As a further solution of the present application: the purpose of using component D is to adjust the overall osmotic pressure of the tissue filling material so that it meets the physiological fluid requirements of the application field environment, for example, when applied to the left ventricular myocardial wall , it can avoid various high-risk damages caused by the presence of components A, B and C of the tissue filling material due to its high overall osmotic pressure, such as shrinkage of endothelial cells, loosening of intercellular connections and Broken, damaged blood-brain barrier, microcirculation disorder caused by hardening of red blood cells, increased cardiac load due to rapid increase in blood volume, cardiac electrical changes caused by atrioventricular conduction and weakened interventricular conduction and repolarization, arrhythmia and ventricular fibrillation increased incidence. For this purpose, the isotonicity agents include sodium bicarbonate, sodium dihydrogen phosphate, sodium chloride, sodium lactate, potassium chloride, calcium chloride, magnesium chloride, glucose, xylitol, mannitol, sorbitol, dextran, Any one or more of trimethylolaminomethane and the like.

As a further scheme of this application: the regulators include tromethamine, chlorobutanetriol, disodium EDTA, sodium hydroxide, calcium disodium EDTA, hydrogen chloride, meglumine, etc. any one or more of them. The use of component E has the following benefits: 1) Adjusting the overall pH value of the tissue filling material to a suitable range for human heart tissue, avoiding acidosis caused by too low pH or alkalosis caused by too high pH, to ensure that it has good 2) Improve the performance stability of the tissue filler material before the subsequently mentioned reaction stage, eg in the form of a premix during storage or transportation.

Another object of the embodiments of the present application is to provide a method for preparing a tissue filling material, and the method for preparing a tissue filling material includes the following steps:

1) premixing the pore-forming filler component with one or more of the component A or the component B in proportion to obtain a premix;

2) Weighing the remaining raw materials and the pre-mixture according to the proportion to carry out a mixing reaction to obtain the tissue filling material.

As a further solution of the present application: in the preparation method of the tissue filling material, the premixing is to uniformly mix the weighed different components in water for injection separately or together or successively.

As a further solution of the present application: in the preparation method of the tissue filling material, component A and component B do not exist in the same premix at the same time.

As a further solution of the present application: when the tissue filling material further includes an isotonicity agent and/or a regulator, the preparation method of the tissue filling material includes the following steps:

1) Premixing stage: Weigh one or more of the component C and the component A, the component B, the component D or the component E in proportion to carry out premixing, A premix is obtained; the premix is defined as uniformly mixing the required components in water for injection separately or together or successively, wherein component A and component B cannot exist in the same premix at the same time;

2) Reaction stage: mixing and reacting the premix containing component A with component B, or mixing component A with the premix containing component B, or mixing and reacting the premix containing component A The pre-mixture is mixed and reacted with the pre-mixture containing the component B, and finally a stable mixture is formed, that is, the tissue filling material is obtained.

As a further solution of the present application: in the preparation method of the tissue filling material, the component A, the component B, the component C, the component D and the component E are all sterile and pyrogen-free.

As a further solution of the present application: in the preparation method of the tissue filling material, both the premixing and the mixing reaction are performed at room temperature and under sterile conditions, and the reaction time is not more than 50 minutes. Specifically, in order to meet the actual clinical needs and ensure the safety of the formed tissue filling material, the components used in the above preparation process (including the whole preparation process in all the following examples) should be sterile Pyrogen-free, and premixing and mixing reactions are performed at room temperature under sterile conditions.

As a further solution of the present application: the preparation method of the tissue filling material includes the following steps: 1) Weighing the pore filling material according to the content of 0.1-20wt% in component A, The middle content is 0.2-40wt%, and the alginate is weighed, and the balance is water for injection, which is fully mixed and dissolved to obtain component A; and the pore-forming filling material is weighed according to the content in component B of 0.1-20wt% , according to the content of 0.2-40wt% in component B, weigh the metal cationic cross-linking agent, and the balance is water for injection, and fully mix and dissolve to obtain component B; 2) described component A and component B are carried out The same volume is fully mixed and reacted to obtain the tissue filling material.

As a further solution of the present application: the preparation method of the tissue filling material includes the following steps: 1) Weigh the pore filling material according to the content of 0.2-40wt% in component A, The alginate is weighed with a content of 0.2-40 wt%, and the balance is water for injection, which is fully mixed and dissolved to obtain component A; , and the balance is water for injection, which is fully mixed and dissolved to obtain component B; 2) the component A and component B are fully mixed and reacted in the same volume to obtain the tissue filling material.

As a further scheme of the present application: the preparation method of the tissue filling material includes the following steps: 1) Weigh the alginate according to the content of 0.2-40wt% in component A, and the balance is water for injection, and sufficient Mix and dissolve to obtain component A; weigh the pore-forming filling material according to the content in component B of 0.2-40 wt %, and weigh the metal cationic crosslinking agent according to the content of component B of 0.2-40 wt % , and the balance is water for injection, which is fully mixed and dissolved to obtain component B; 2) the component A and component B are fully mixed and reacted in the same volume to obtain the tissue filling material.

Another object of the embodiments of the present application is to provide a tissue filling material prepared by using the above-mentioned method for preparing a tissue filling material.

Another object of the embodiments of the present application is to provide a tissue engineering scaffold, which partially or wholly contains the above-mentioned tissue filling material.

As a further solution of the present application: the tissue engineering scaffold may be all of the tissue filling material, or may be the tissue filling material and the existing auxiliary materials in the preparation of the tissue engineering scaffold. Without limitation, the tissue engineering scaffolds include: bone, cartilage, blood vessels, nerves, skin and artificial organs, such as tissue scaffolds for liver, spleen, kidney, bladder and the like.

Specifically, the tissue filling material can be used as an injectable scaffold material to construct a tissue engineering scaffold, or can be used as a bone component material or a vascular tissue material to construct biological tissue. When used as an injectable scaffold material, the support strength can be adjusted in time according to the needs of the patient's heart tissue, and due to its three-dimensional porous structure, the material can achieve the initial support of the myocardial tissue in vivo, the initial adhesion of myocardial cells, and the formation of myocardial cells. Entering the material for growth, it can adapt to cell growth, transport of nutrient flow and discharge of metabolites.

Another object of the embodiments of the present application is to provide an application of the tissue filling material in the preparation of a medical material for adjuvant treatment of heart failure.

As a further solution of this application: in the application of the tissue filling material in the preparation of medical materials for adjuvant treatment of heart failure, specifically, it can be used in the preparation of injectable stents, intra-aortic balloon counterpulsation devices, artificial hearts, implantable defibrillators, pacemakers and other products.

Compared with the prior art, the beneficial effects of the present application are:

The tissue filling material provided by the present application can induce cardiomyocytes to grow in the pores of the tissue filling material. By using a skeleton component and a pore-forming filler component with a biodegradation rate greater than that of the skeleton component, when the tissue filling material is implanted into the body After that, the pore-forming filler components are rapidly degraded to form an interpenetrating porous network structure, and nutrients and cells can enter the interior. The degraded product has molecular activity, can induce cells to enter the interior of the material, and can regulate cell adhesion, proliferation and differentiation. The provided preparation method is simple, and the prepared tissue filling material is beneficial to the growth of cardiomyocytes in the material and the recovery of myocardial function, which solves the problem that most of the existing tissue filling materials can only play a physical support role, and there is a problem that is not conducive to the growth of cardiomyocytes in the material. The problem of internal growth and restoration of myocardial function has broad market prospects.

Description of drawings

FIG. 1 is a physical diagram of the tissue filling material in Example 1 of the application.

FIG. 2 is a scanning electron microscope picture of the tissue filling material without hyaluronidase added in Example 1 of the application.

3 is a scanning electron microscope picture of the tissue filling material after adding hyaluronidase for 4 hours in Example 1 of the application. The white arrows indicate the porous structure formed by the in situ degradation of the water-soluble biodegradable pore-forming filler material.

4 is a scanning electron microscope picture of the tissue filling material after adding hyaluronidase for 8 hours in Example 1 of the application. The white arrows indicate the porous structure formed by the in situ degradation of the water-soluble biodegradable pore-forming filler material.

5 is a scanning electron microscope picture of the tissue filling material after adding hyaluronidase for 18 hours in Example 1 of the application. The white arrows indicate the porous structure formed by the in situ degradation of the water-soluble biodegradable pore-forming filler material.

FIG. 6 is a graph showing the effect of the tissue filling material prepared in Example 3 of the present application on the ventricular ejection fraction of rats with myocardial infarction.

FIG. 7 is a graph showing the results of the effect of the tissue filling material prepared in Example 3 of the present application on the ventricular wall thickness of rats with myocardial infarction.

FIG. 8 is a graph showing the staining results of the tissue filling material prepared in Example 3 of the present application after 4 weeks of myocardial implantation. Black squares indicate the location of the hydrogel, black triangles indicate infarcted myocardial tissue, and black circles indicate normal myocardial tissue.

FIG. 9 is a trend diagram of myocardial infarction area change of the tissue filling material prepared in Example 3 of the present application after 4 weeks of myocardial implantation.

Fig. 10 is a picture of angiogenesis in the myocardial infarction area after 4 weeks of no treatment and intervention in rats with central infarction in Example 4 of the present application.

11 is a picture of angiogenesis after implantation of tissue fillers in the myocardial infarction area of the rat in Example 4 of the present application for 4 weeks. White triangles indicate where new blood vessels are located.

detailed description

The present application will be described in further detail below with reference to the accompanying drawings and specific embodiments. The following examples will help those skilled in the art to further understand the application, but do not limit the application in any form. It should be noted that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application. These all belong to the protection scope of the present application.

In the following examples of the present application, the methods are conventional methods unless otherwise specified, and the raw materials can be obtained from open commercial sources unless otherwise specified.

Among them, for alginate, suitable alginates include but are not limited to sodium alginate, potassium alginate, ammonium alginate, and propylene glycol alginate. In a preferred embodiment, the alginate is sodium alginate with good biocompatibility, and in a more preferred embodiment, the molecular weight of the sodium alginate is 5-400kDa.

For the crosslinking agent, the divalent metal cations it contains include but are not limited to calcium cations, barium cations, zinc cations, iron cations, magnesium cations, and copper cations, and the multivalent metal cations it contains include but are not limited to aluminum cations, Chromium cation, molybdenum cation, tin cation. In a preferred embodiment, the cross-linking agent includes a combination of one or more of calcium alginate, calcium gluconate, calcium carbonate, calcium sulfate or calcium chloride; further, considering the cost performance, it is preferred to use Calcium alginate; further, the solid particle diameter of calcium alginate is not greater than 1mm, so that it has better dispersibility in water; further, the solid particle diameter of calcium alginate is preferably available in the market. The conventional specification is 1-300μm.

For the in vitro enzymatic hydrolysis experiment of tissue filling materials: according to the specific selection of component C, configure the corresponding enzyme solution, if component C is hyaluronic acid, configure hyaluronidase solution, such as component C is chitosan, Then configure chitosanase solution, if component C is collagen, then configure collagenase solution, and so on. Put the tissue filling material fully mixed according to the preparation method provided in this application in a petri dish, all the materials used have been sterilized by irradiation, and placed in the same volume of enzyme solution under aseptic operation conditions. In a 37°C cell incubator, samples were taken regularly, and the samples were weighed after being freeze-dried in a vacuum freeze dryer, and the structural characteristics such as the distribution of the internal network structure of the hydrogel were observed by scanning electron microscopy.

For in vivo histocompatibility testing of tissue fillers: The tissue fillers must be prepared under sterile conditions, and each raw material must be sterile. Therefore, after the fully mixed tissue filling material according to the preparation method provided by the present application was injected into the myocardial wall of the experimental rat during thoracotomy, the experimental rats were reared for 4 weeks, and the tissue filling material in the left ventricular wall was dissected and observed. The shape of the tissue was fixed in formalin solution, and the fixed tissue material was dehydrated, embedded, sliced, analyzed by H&E staining (hematoxylin-eosin staining) and Masson staining, and observed under an optical microscope. Microscopic morphology of myocardial tissue within the ventricular wall.

For the test of the support strength (ie: mechanical support performance) of the tissue filling material: the tissue filling material fully mixed according to the preparation method provided in this application is placed in a cylindrical cavity mold, and the hydrogel biomaterial sample is fully coagulated. After gluing, make a cylindrical sample with a diameter of 20mm and a height of 20mm. Use a universal material testing machine to test the support strength of the sample. Set the pressing speed to 2mm/min and the clamping distance to 20mm, and output directly through the universal material testing machine. data to obtain the support strength of the specimen.

The various advantages brought by different component formulations and preparation methods of the tissue filling material provided by the present application are described below with reference to specific examples, including: the convenience of the preparation process, the in vitro enzymatic hydrolysis test of the tissue filling material, the formation of The control ability of the mechanical support of tissue filling materials, the in vivo histocompatibility, and the test results of animal experiments will be further elaborated.

Example 1

A tissue filling material, the preparation method of which specifically comprises the following steps:

1) Component B (select calcium alginate, the solid particle size of calcium alginate is 75 μm), component C (select hyaluronic acid, the molecular weight of hyaluronic acid is 200kDa) and component D ( The three powders of mannitol) are dissolved together in water for injection, and fully dissolved to obtain the first premix (i.e., cross-linking agent+pore-forming filler system), so that component B, component C and component D are here The content in the premix is 1.5% (g/mL), 1% (g/mL) and 302mmol/L, respectively. This premix is formed into a water-soluble solution, and then the component E (sodium hydroxide solution) is added to adjust pH is 7.0;

2) take by weighing the calculated amount of component A (select sodium alginate, molecular weight is 180kDa) and these two powders of component D (mannitol) are dissolved in water for injection together, fully dissolve, obtain the second premix (that is, seaweed). sodium system), so that the content of component A and component D in this premix is 2% (g/mL) and 302 mmol/L, respectively, this premix is formed into a water-soluble solution, the pH value of this solution is 7.0;

3) The sodium alginate system prepared above and the cross-linking agent+pore-forming filler system are respectively filled into a sterile syringe in a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube Or the injection system mentioned in this application, the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the tissue filling material contains component A, component B, and component C. The content of component D is 1% (g/mL), 0.75% (g/mL), 0.5% (g/mL) and 302mmol/L, respectively.

In this example, the pH value of the obtained tissue filling material was 7.0, and the mechanical support strength was 7±0.8 kPa. The pore-forming filling material is evenly distributed inside the ion-crosslinked hydrogel structure, and the hydrogel is in a colorless and transparent state, as shown in FIG. 1 .

Example 2

The tissue filling material prepared in Example 1 was subjected to an in vitro enzymatic hydrolysis experiment, specifically taking a volume of 1 cm ^of the tissue filling material and placing it in 20 mL of hyaluronidase (150 μ/mL) solution under sterile conditions, Placed in a 37°C cell incubator, sampled regularly at 4, 8, 12, and 18 h, respectively, and weighed after being freeze-dried in a vacuum freeze dryer. Scanning electron microscopy was used to observe the distribution of the internal network structure of the hydrogel formed by the tissue filling material. and other structural features. The specific distribution results of the internal network structure of the tissue filling material are shown in Figures 2, 3, 4, and 5, respectively, wherein Figure 2 is a scanning electron microscope picture of the tissue filling material without hyaluronidase, and Figure 3 is a The scanning electron microscope picture of the tissue filling material after adding hyaluronidase for 4 hours, Figure 4 is the scanning electron microscope picture of the tissue filling material after adding hyaluronidase for 8 hours, and Figure 5 is the tissue filling material after adding hyaluronidase for 18 hours Scanning electron microscope picture.

As can be seen from Figures 2-4, as the reaction time between the tissue filling material and hyaluronidase increases, the pores inside the tissue filling material become larger, and an interpenetrating porous network structure is gradually formed, so that the The material has a three-dimensional porous structure inside, and the three-dimensional pore size is interconnected, which realizes the initial support of the material to the myocardial tissue in the body, the initial adhesion of the myocardial cells, and the high porosity porous structure that allows the myocardial cells to enter the material, and the interior of the material is connected. It can adapt to cell growth, nutrient flow transport and metabolite excretion.

Example 3

1) Component B (select calcium alginate, the solid particle size of calcium alginate is 75 μm), component C (select hyaluronic acid, the molecular weight of hyaluronic acid is 200kDa) and component D ( Select mannitol) to be dissolved in water for injection together, fully dissolve, obtain the first premix (i.e. cross-linking agent+pore-forming filler system), so that component B, component C and component D are in the first premix The content of 1.5% (g/mL), 1% (g/mL) and 302mmol/L respectively, this first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH is 7.0;

2) Component A (select sodium alginate, molecular weight is 180kDa), component C (select hyaluronic acid, the molecular weight of hyaluronic acid is 200kDa) of calculated amount and component D (mannitol) are dissolved in injection together. In water, fully dissolve to obtain the second premix (that is, sodium alginate system+pore-forming filler system), so that the content of component A, component C and component D in the second premix is respectively 2% ( g/mL), 1% (g/mL), and 302 mmol/L, the second premix was formed into a water-soluble solution, the pH of which was 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. It can be injected into the injection system using the same injection system, and the stable tissue filling material can be formed after equilibration for 8 minutes (ie: gelation time) after injection. The contents were 1% (g/mL), 0.75% (g/mL), 1% (g/mL) and 302mmol/L, respectively.

In this example, the obtained tissue filling material has a pH value of 7.0 and a mechanical support strength of 9±0.5kPa.

Example 4

The tissue filling material prepared in Example 3 was subjected to a small animal treatment experiment for myocardial infarction, specifically, the tissue filling material was injected into the junction between the myocardial infarction area and the normal myocardium of the experimental rat heart, 40uL was injected at a single point, and 3 points were injected. , Ejection fraction (EF, ejection fraction, calculated as %), ventricular wall thickness, and cardiac output were tested and recorded before and after operation, and the rats were reared for 4 weeks after operation. The ventricular wall thickness was tested and recorded again, wherein the effect of the tissue filling material on the ventricular ejection fraction of the myocardial infarction rats is shown in Figure 6, and the effect of the tissue filling material on the ventricular wall thickness of the myocardial infarction rats is as follows shown in Figure 7.

The tissue filling material prepared in Example 3 was subjected to an in vivo histocompatibility test: after the fully mixed tissue filling material was injected into the myocardial wall of experimental rats during thoracotomy, the experimental rats were reared for 4 weeks and dissected. The morphology of the tissue filling material in the left ventricular wall was observed, the material was fixed in formalin solution, and the fixed tissue material was dehydrated, embedded, sliced, stained with H&E and Masson, and observed under an optical microscope. Its microscopic morphology with myocardial tissue within the wall of the left ventricle. The specific staining results of the tissue filling material after 4 weeks of myocardial implantation are shown in Figure 8. The pore filling material can be degraded and absorbed in a short period of time, and a porous cavity is formed in the alginate hydrogel. At the same time, it guides the growth of cells in the lumen, induces the growth of cardiomyocytes, and restores myocardial function.

FIG. 9 is a graph showing the change trend of myocardial infarction area 4 weeks after myocardial implantation of the tissue filling material prepared in Example 3 of the present application. We found that the injected tissue filler material significantly delayed the deterioration of cardiac function after myocardial infarction in rats, reduced the size of myocardial infarction, maintained the level of left ventricular EF, and improved cardiac function with the passage of time after injection. Injection of hydrogel can significantly improve cardiac function. The thickness of the ventricular wall in the infarct area; without the injection of this material, the myocardial infarction area gradually increased with time, and the cardiac function continued to deteriorate.

In particular, we further carried out batch animal experimental studies, including materials incorporated into the prior art, for statistically significant comparison of results, and found that: for rats with multiple myocardial infarction, no treatment was given. 4 weeks after the injection, the myocardial infarction area was 8.00±1.08 mm ² , and the injection of prior art materials, such as alginic acid-based hydrogel without adding pore filling material, the myocardial infarction area 4 weeks after the injection Compared with the injection of the tissue filling material provided by the present application, the myocardial infarction area after 4 weeks of myocardial implantation was 4.37± ^{1.72mm 2} ^. After statistical analysis, it was concluded that the experiment without any treatment There was no significant difference in myocardial infarction area after 4 weeks between the group and the experimental group injected with the materials of the prior art, while the experimental group injected with the tissue filling material provided by the present application and the experimental group injected with the materials of the prior art, the myocardial infarction area was obtained after 4 weeks. Significant reduction, and the reduction range is 41.7%, it can be seen that the tissue filling material provided by the present application can effectively restrain or even reverse the degree of deterioration of cardiac function after myocardial infarction, greatly reduce the size of myocardial infarction, and significantly improve cardiac function.

Figure 10 is a picture of angiogenesis in the myocardial infarction area after 4 weeks of no treatment intervention in myocardial infarction rats. It was found that the new blood vessels in the myocardial infarction area and the myocardial infarction junction area of the myocardial infarction rats were rare, and the lumen structure was small. Figure 11 is a picture of angiogenesis after 4 weeks of implantation of the tissue filling material prepared in Example 3 in the myocardial infarction area of the rat with myocardial infarction. The myocardial infarction area and the myocardial infarction junction area of the rat showed more neovascularization, and the lumen structure was relatively simple The new blood vessels in the myocardial infarction group were enlarged. The above results suggest that injection of the hydrogel provided by the present application can promote the regeneration of intramyocardial blood vessels, which is beneficial to the recovery of cardiac function after myocardial infarction.

In summary, the hydrogel in Example 3 can be injected into the myocardium to increase the thickness of the ventricular wall and promote myocardial neovascularization, which can promote myocardial angiogenesis, reduce the scope of myocardial infarction, improve the decline of cardiac function after infarction, and improve the ability of tissue regeneration and repair. , the treatment is safe and effective, and provides a new method for clinical myocardial revascularization.

Example 5

1) Component B (select calcium alginate, its solid particle diameter is 75 μm), component C (select carboxylated chitosan, and molecular weight is 100kDa) and component D (select mannitol) of calculated amount Dissolve together in water for injection, fully dissolve to obtain a first premix, so that the content of component B, component C and component D in the first premix is 1.5% (g/mL), 1.5% (g/mL), respectively. /mL) and 302mmol/L, the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH to 7.0;

2) Component A (selecting sodium alginate, molecular weight is 180kDa), component C (selecting carboxylated chitosan, and molecular weight is 100kDa) and component D (mannitol) of calculated amount are dissolved in water for injection together , fully dissolve to obtain a second premix, so that the contents of component A, component C and component D in the second premix are 2% (g/mL), 1.5% (g/mL) and 302 mmol, respectively /L, the second premix is formed into a water-soluble solution, and the pH of this solution is 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. It can be injected into the injection system using the same injection system, and the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the composition of component A, component B, component C and component D The contents were 1% (g/mL), 0.75% (g/mL), 1.5% (g/mL) and 302 mmol/L, respectively.

In this example, the pH value of the obtained tissue filling material was 7.0, and the mechanical support strength was 7.54±0.5 kPa.

Example 6

1) Component B (for selection of calcium alginate, whose solid particle diameter is 75 μm) and component D (for selection of mannitol) of calculated amount are dissolved in water for injection together, and fully dissolved to obtain the first premix, so that The content of component B and component D in the first premix is 1.5% (g/mL) and 302 mmol/L, respectively, and the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide) is added. solution) to adjust the pH to 7.0;

2) Component A (select sodium alginate, molecular weight is 180kDa) of calculated amount, component C (select chondroitin sulfate sodium salt, molecular weight 50kDa) and component D (mannitol) are dissolved in water for injection together, Fully dissolve to obtain the second premix, so that the content of component A, component C and component D in the second premix is 2% (g/mL), 2% (g/mL) and 302mmol/L respectively , the second premix is formed into a water-soluble solution, the pH of this solution is 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. It can be injected into the injection system using the same injection system, and the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the composition of component A, component B, component C and component D The contents were 1% (g/mL), 0.75% (g/mL), 1% (g/mL) and 302mmol/L, respectively.

In this example, the pH value of the obtained tissue filling material was 7.0, and the mechanical support strength was 9.67±0.5 kPa.

Example 7

1) Component B (select calcium alginate, its solid particle size is 75 μm), component C (select collagen, molecular weight 70-200kDa) and component D (select mannitol) of calculated amount are dissolved in together. In water for injection, fully dissolve to obtain a first premix, so that the contents of component B, component C and component D in the first premix are 1.5% (g/mL) and 2% (g/mL) respectively and 302mmol/L, the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH to be 7.0;

2) take the calculated amount of component A (select sodium alginate, molecular weight is 180kDa) and be dissolved in water for injection together with component D (mannitol), fully dissolve, obtain the second premix, make component A and group The content of Part D in the second premix is 2% (g/mL) and 302 mmol/L respectively, the second premix is formed into a water-soluble solution, and the pH of this solution is 7.0;

In this example, the obtained tissue filling material has a pH value of 7.0 and a mechanical support strength of 7.33±0.5kPa.

Example 8

1) Component B (select calcium alginate, its solid particle diameter is 75 μm), component C (select fibrinogen) and component D (select mannitol) of calculated amount are dissolved in water for injection together, Fully dissolve to obtain a first premix, so that the contents of component B, component C and component D in the first premix are 1.5% (g/mL), 0.2% (g/mL) and 302mmol/L respectively , the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH to 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. It can be injected into the injection system using the same injection system, and the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the composition of component A, component B, component C and component D The contents were 1% (g/mL), 0.75% (g/mL), 0.1% (g/mL) and 302mmol/L, respectively.

In this example, the obtained tissue filling material has a pH value of 7.0 and a mechanical support strength of 8.45±0.7kPa.

Example 9

1) Component B (select calcium alginate for use, its solid particle diameter is 75 μm), component C (select dextran) and component D (select mannitol) of calculated amount are dissolved in water for injection together, Fully dissolve to obtain the first premix, so that the content of component B, component C and component D in the first premix is 1.5% (g/mL), 20% (g/mL) and 302mmol/L respectively , the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH to 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. It can be injected into the injection system using the same injection system, and the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the composition of component A, component B, component C and component D The contents were 1% (g/mL), 0.75% (g/mL), 10% (g/mL) and 302 mmol/L, respectively.

In this example, the obtained tissue filling material has a pH value of 7.0 and a mechanical support strength of 6.67±0.5kPa.

Example 10

1) Component B (select calcium alginate, its solid particle diameter is 75 μm), component C (select laminin, 805kD) and component D (select mannitol) of calculated amount are dissolved in water for injection together , fully dissolve to obtain a first premix, so that the contents of component B, component C and component D in the first premix are 1.5% (g/mL), 0.2% (g/mL) and 302 mmol, respectively /L, the first premix is formed into a water-soluble solution, and then component E (sodium hydroxide solution) is added to adjust the pH to 7.0;

In this example, the obtained tissue filling material has a pH value of 7.0 and a mechanical support strength of 6.00±0.5kPa.

Example 11

Compared with Example 10, except that the contents of component A, component B, component C and component D in the tissue filling material are 0.1% (g/mL), 0.1% (g/mL), 0.1%, respectively % (g/mL) was the same as in Example 10 except that it was 280 mmol/L.

Example 12

Compared with Example 10, except that the contents of component A, component B, component C and component D in the tissue filling material are 20% (g/mL), 20% (g/mL), 20% % (g/mL) is the same as Example 10 except that it is 320 mmol/L.

Example 13

Compared with Example 10, except that the contents of component A, component B, component C and component D in the tissue filling material are 0.1% (g/mL), 15% (g/mL), 10 % (g/mL) was the same as in Example 10 except that it was 290 mmol/L.

Example 14

Compared with Example 10, except that the contents of component A, component B, component C and component D in the tissue filling material are 5% (g/mL), 10% (g/mL), 15% % (g/mL) was the same as in Example 10 except that it was 310 mmol/L.

Example 15

Compared with Example 10, except replacing mannitol with sorbitol, the rest is the same as Example 10.

Example 16

Compared with Example 10, except that mannitol was replaced with glucose, other parts were the same as Example 10.

Example 17

Compared with Example 10, the rest is the same as Example 10 except that laminin is replaced by chitosan.

Example 18

Compared with Example 1, the rest is the same as Example 1, except that calcium alginate is replaced by calcium gluconate with a solid particle diameter of 1 μm.

Example 19

Compared with Example 1, the rest is the same as Example 1, except that calcium alginate is replaced by calcium carbonate with a solid particle size of 100 μm.

Example 20

Compared with Example 1, the rest is the same as Example 1, except that calcium alginate is replaced by calcium sulfate with a solid particle diameter of 300 μm.

Example 21

Compared with Example 1, the rest is the same as Example 1, except that calcium alginate is replaced by calcium chloride with a solid particle size of 500 μm.

Example 22

Compared with Example 1, the rest is the same as Example 1, except that calcium alginate is replaced by calcium gluconate with a solid particle diameter of 1000 μm.

Example 23

Compared with Example 1, except that sodium alginate is replaced by potassium alginate with a molecular weight of 5 kDa, the rest is the same as Example 1.

Example 24

Compared with Example 1, except that sodium alginate is replaced by ammonium alginate with a molecular weight of 80 kDa, the rest is the same as Example 1.

Example 25

Compared with Example 1, the rest is the same as Example 1, except that sodium alginate is replaced by propylene glycol alginate with a molecular weight of 200 kDa.

Example 26

Compared with Example 1, except that sodium alginate is replaced by propylene glycol alginate with a molecular weight of 400 kDa, the rest is the same as Example 1.

Example 27

Compared with Example 1, except that hyaluronic acid was replaced with dextran, the rest was the same as Example 1.

Example 28

Compared with Example 1, except that the hyaluronic acid is replaced by chitosan, other parts are the same as Example 1.

Example 29

Compared with Example 1, except replacing hyaluronic acid with collagen, the rest is the same as Example 1.

Example 30

Compared with Example 1, except replacing hyaluronic acid with gelatin, other parts are the same as Example 1.

Example 31

Compared with Example 10, except that the contents of component A, component B, component C and component D in the tissue filling material are 10% (g/mL), 10% (g/mL), 10% % (g/mL) was the same as in Example 10 except that it was 310 mmol/L.

Example 32

2) Component A (selecting sodium alginate for use, molecular weight is 180kDa), component D (mannitol) and hyaluronidase of calculated amount are dissolved in water for injection together, fully dissolve, obtain the second premix (i.e. Sodium alginate + enzyme system), so that the content of component A, component D and hyaluronidase in this premix is 2% (g/mL), 302mmol/L, 150U/mL, respectively, this premix forms It is a water-soluble solution, and the pH value of this solution is 7.0;

3) The sodium alginate+enzyme system and the cross-linking agent+pore-forming filler system prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, and then mixed with a three-way syringe and connected to the injection. Needle tube or injection system mentioned in this application, equilibrated for 7 minutes after injection (ie: gelation time) can form the stable tissue filling material, the tissue filling material in component A, component B, group The contents of fraction C, fraction D and hyaluronidase were 1% (g/mL), 0.75% (g/mL), 0.5% (g/mL), 302 mmol/L and 75 U/mL, respectively.

The tissue filling material in this embodiment can accelerate the formation of porous structure due to the additional addition of hyaluronidase.

Example 33

Compared with Example 32, it is the same as Example 32 except that hyaluronic acid is replaced by glucan and hyaluronidase is replaced by glucanase.

Example 34

Compared with Example 32, the rest is the same as Example 32 except that hyaluronic acid is replaced by chitosan and hyaluronidase is replaced by chitosanase.

Example 35

Compared with Example 32, it is the same as Example 32 except that hyaluronic acid is replaced by fibrin and hyaluronidase is replaced by protease.

Example 36

1) Component B (select calcium alginate, the solid particle diameter of calcium alginate is 75 μm), component C (select hyaluronic acid, the molecular weight of hyaluronic acid is 200kDa) of calculated amount is dissolved in water for injection together , fully dissolve to obtain a first premix, so that the contents of component B and component C in this premix are 1.5% (g/mL) and 1% (g/mL) respectively, and this premix is formed to have Water-soluble solution, while adjusting pH to 7.0;

2) take the calculated amount of component A (select sodium alginate, molecular weight is 180kDa) and dissolve in water for injection, fully dissolve, obtain the second premix, so that the content of component A in this premix is respectively 2% (g/mL), the premix is formed into a water-soluble solution with a pH of 7.0;

3) The first premix and the second premix prepared above are respectively filled into a sterile syringe at a volume ratio of 1:1, then mixed with a three-way syringe and connected to the injection needle tube or mentioned in this application. The injection system is injected, and the stable tissue filling material can be formed after equilibration for 7 minutes after injection (ie: gelation time), and the content of component A, component B, and component C in the tissue filling material is 1 % (g/mL), 0.75% (g/mL), 0.5% (g/mL).

According to the above results, it can be seen that the addition of component C endows the tissue filler with special biological properties, bringing the following advantages:

1) After the tissue filling material is injected into the myocardium, the enzymes in the body will rapidly degrade the pore-forming filler to form an interpenetrating porous network structure, thereby solving the problem that the existing alginate-based hydrogel has a single pore size and cannot be used. The disadvantage of forming 3D connected pores, nutrients and cells cannot enter the inside of the scaffold; after the tissue filling material is injected into the myocardium, while the pore filling agent is degraded in the body, the degraded products have molecular activity and can Inducing cells into the gel can regulate cell adhesion, proliferation and differentiation, and at the same time, the degraded products can provide nutrients for cell growth and promote cell growth and blood vessel formation;

2) The tissue filling material has excellent biocompatibility, and animal experiments show that the gel has no adverse and serious reactions with cells around the tissue, can improve the cardiac function index of animals with heart failure, effectively treat heart failure, and prevent the deterioration of heart failure, and Tissue can grow within the gel and have angiogenesis;

3) The preparation process of the tissue filling material is simple, and the preparation can be realized by mixing, and the size and porosity of the pore size of the hydrogel can be adjusted by changing the amount of the pore-forming filler; the tissue filling material can be used for in vitro cells. In culture, in vivo injection applications can also be achieved.

In conclusion, the tissue filling material provided by the present application can meet the performance requirements of injectable tissue engineering and cell culture, etc., and can treat heart failure diseases and restore heart function at the same time.

The beneficial effects of the present application are as follows. The tissue filling material prepared by the present application is composed of two natural biopolymer materials with very different degradation rates in vivo. One is alginate that degrades slowly as a skeleton, and the other is fast degrading alginate. The water-soluble biodegradable polymer material, when the tissue filling material is implanted into the body, the rapidly degraded water-soluble biodegradable polymer material rapidly degrades to form an interpenetrating porous network structure, thus solving the problem of existing alginate-based water. The pore size of the gel is single, the 3D connected pores cannot be formed, and the nutrients and cells cannot enter the scaffold. At the same time, the rapidly degraded biopolymer materials are degraded in vivo, and the degraded products have molecular activity and can induce cells. Entering the inside of the gel can regulate cell adhesion, proliferation and differentiation. Animal experiments have good results and can effectively treat heart failure diseases. However, the provided preparation method is simple, adopts a simple and effective way to construct a porous structure with interpenetrating pore shape, and can induce the growth of cardiomyocytes in the pores, which solves the problem that most of the existing tissue filling materials can only play a physical supporting role, and there are inconveniences. It is beneficial to the growth of cardiomyocytes in the material and the recovery of myocardial function, and it can also solve the problem of the prior art due to the limitation of the formulation of the component materials. It will enter the inside of the gel, and the porosity of the gel is not easy to control, and the internal three-dimensional pore size is not connected to each other, so 3D connected pores cannot be formed, and nutrients and cells cannot enter the inside of the scaffold, which is not conducive to the growth of cardiomyocytes inside the gel and myocardial cells. Function recovery and other issues have broad market prospects.

Moreover, the tissue filling material can be used as an injectable stent material, with a support strength closer to that of human heart tissue, or the support strength can be adjusted in time according to the needs of the patient's heart tissue, and the material has a three-dimensional porous structure inside, and the three-dimensional pores penetrate each other. Connected to realize the initial support of the material to the myocardial tissue in the body, the initial adhesion of the cardiomyocytes, and the entry of the cardiomyocytes into the material. The high porosity porous structure of the material can be adapted to cell growth, nutrient flow transport and metabolites of discharge.

It should be noted that, compared with the prior art, the present application has at least the following advantages:

1) Due to the introduction of the pore-forming filler component in the tissue filling material provided in this application, after the tissue filling material is implanted into the body, the rapidly degraded pore-forming filler component is rapidly degraded to form an interpenetrating porous network structure, thereby It solves the shortcomings of the existing alginate-based hydrogels with single pore size, unable to form 3D connected pores, and inability of nutrients and cells to enter the interior of the scaffold.

2) While the pore-forming filler component in the tissue filling material provided by this application is degraded in vivo, the degraded product has molecular activity, can induce cells to enter the gel, and can regulate cell adhesion, proliferation and differentiation, which The degraded products can provide nutrients for cell growth and promote cell growth and the formation of blood vessels.

3) The tissue filling material provided by the present application has excellent biocompatibility, animal experiments show that the gel has no serious adverse reaction with the surrounding cells of the tissue, and the tissue can grow in the gel and have angiogenesis.

4) The tissue filling material provided by the present application has a simple manufacturing process and can be mixed. By changing the amount of the pore-forming filler component, the pore size and porosity of the hydrogel can be adjusted.

5) The tissue filling material provided in this application can be used for both in vitro cell culture and in vivo injection applications.

The preferred embodiments of the present application have been described in detail above, but the present application is not limited to the above-mentioned embodiments. Within the scope of knowledge possessed by those of ordinary skill in the art, various aspects can be made without departing from the purpose of the present application. kind of change. There is no need and cannot be exhaustive of all implementations here. However, the obvious changes or changes derived from this are still within the protection scope of the present application.

Claims

A tissue filling material, the tissue filling material includes component A, component B, and water for injection, the component A includes alginate, the component B includes a metal cation cross-linking agent, the Component A and/or Component B further comprise a water-soluble biodegradable pore-forming filling material, and after the component A and component B are fully stirred to form an ionically cross-linked hydrogel structure, the pore-forming filling material uniformly distributed inside the ion cross-linked hydrogel structure; the content of the alginate in the tissue filling material is 0.1-20wt%, and the content of the metal cationic cross-linking agent in the tissue filling material is 0.1-20 wt %, and the content of the pore filling material in the tissue filling material is 0.1-20 wt %.
The tissue filling material according to claim 1, wherein when the pore filling material is degraded, the degraded product has biological activity, which can promote the growth of tissue cells and regulate cell adhesion, proliferation and differentiation.
The tissue filling material according to claim 1, wherein the pore filling material comprises any one or more of polysaccharide-based materials or protein-based materials.
The tissue filling material according to claim 3, wherein the polysaccharide material comprises any one of hyaluronic acid, dextran, chitosan or chondroitin sulfate; the protein material comprises Any of collagen, gelatin, fibrin or laminin.
The tissue filling material according to claim 1, wherein the pore filling material contains active drugs capable of promoting cell growth, and the active drugs include transforming growth factor, platelet-derived growth factor, and fibroblast growth factor.
The tissue filling material according to claim 1, wherein the degradation period of the pore filling material is less than or equal to 8 weeks.
The tissue filler material of claim 1, wherein the pores within the tissue filler material structure gradually increase as the pore filler material degrades.
The tissue filling material according to claim 1, wherein the pore-forming filling material further comprises a regulator for regulating the degradation rate, the regulator comprises hyaluronidase, glucanase, chitosanase, protease any of the .
The tissue filling material according to claim 1, wherein the alginate is any one of sodium alginate, potassium alginate, ammonium alginate or propylene glycol alginate; the metal cationic cross-linking agent is selected from A combination of one or more of calcium alginate, calcium gluconate, calcium carbonate, calcium sulfate or calcium chloride.
A method for preparing a tissue filling material according to any one of claims 1-9, comprising the following steps: 1) weighing the pore-forming filling material according to the content in component A of 0.1-20wt% , according to the content of 0.2-40wt% in component A, the alginate is weighed, and the balance is water for injection, and fully mixed and dissolved to obtain component A; For the pore-forming filling material, the metal cationic cross-linking agent is weighed according to the content of 0.2-40wt% in component B, and the balance is water for injection, which is fully mixed and dissolved to obtain component B; 2) the component A and component B are thoroughly mixed and reacted in the same volume to obtain the tissue filling material.
A preparation method of tissue filling material according to any one of claims 1-9, the preparation method of tissue filling material comprises the following steps: 1) according to the content in component A of 0.2-40wt% Take the pore-forming filling material, weigh the alginate according to the content in component A of 0.2-40wt%, and the balance is water for injection, fully mix and dissolve to obtain component A; according to the content in component B Weigh the metal cationic cross-linking agent at 0.2-40 wt%, the balance is water for injection, and fully mix and dissolve to obtain component B; 2) the component A and component B are fully mixed and reacted in the same volume to obtain the Tissue filling material.
A method for preparing a tissue filling material according to any one of claims 1 to 9, wherein the method for preparing a tissue filling material comprises the following steps: 1) according to the content of component A in the range of 0.2-40wt % Weigh the alginate, the balance is water for injection, fully mix and dissolve to obtain component A; weigh the pore-forming filling material according to the content in component B of 0.2-40wt%, and press The middle content is 0.2-40wt%, and the metal cationic crosslinking agent is weighed, and the balance is water for injection, which is fully mixed and dissolved to obtain component B; 2) the component A and component B are fully mixed and reacted in the same volume, The tissue filler material is obtained.
A tissue engineering scaffold, wherein part or all of the tissue filling material according to any one of claims 1-9 is contained.
An application of the tissue filling material according to any one of claims 1 to 9 in the preparation of a medical material for adjuvant treatment of heart failure.