A plasma separation membrane with biomimetic self-anticoagulation function and its preparation method

Abstract

The present invention discloses a method for preparing a plasma separation membrane with a biomimetic self-anticoagulant function, comprising: (1) preparing a casting solution; (2) preparing a hollow fiber plasma separation membrane using a non-solvent phase inversion method; (3) preparing a plasma separation membrane modified with a fusion protein Hve-cad-Fc; (4) injecting an endothelial cell solution into the hollow inner cavity of the plasma separation membrane modified with the fusion protein Hve-cad-Fc, then sealing both ends of the membrane to enclose the endothelial cell solution within the membrane; then placing the membrane in an endothelial cell culture medium for incubation, allowing the endothelial cells to grow on the inner surface of the membrane, and forming a plasma separation membrane with a biomimetic self-anticoagulant function having an endothelial structure on the inner surface. The present invention forms a biomimetic endothelial functional layer on the inner surface of the membrane that is in direct contact with blood, utilizes the anticoagulant components of the endothelial cells, inhibits the adhesion and activation of platelets, significantly reduces the formation of thrombi, and simultaneously has natural advantages such as anti-inflammatory properties, thereby achieving a biomimetic function.

Description

Technical Field

[0001] The present invention belongs to the technical field of blood purification, and in particular is a plasma separation membrane with bionic self-anticoagulation function and a preparation method thereof. Background Art

[0002] Plasma separation, a rapidly developing blood purification technology, can effectively remove large pathogenic molecules and is used to treat diseases such as severe hepatitis and liver failure. The therapeutic mechanism of plasma separation membranes is the screening principle of membrane pores. After the patient's blood passes through a plasma separator, the plasma can be separated, and the pathogenic large molecules in the plasma are removed. The cellular components useful to the human body and the purified plasma are returned to the human body to achieve the purpose of treating the disease. The core element of plasma separation is a plasma separation membrane with excellent plasma screening properties and good biocompatibility. Among them, the size of the membrane pores is the main factor that determines the plasma screening properties of the membrane, and the biocompatibility of the membrane is the main factor that determines whether the biomaterial can be used in the field of plasma separation.

[0003] The sieving performance of plasma separation membranes determines whether toxins can be effectively separated from blood cells. In the preparation of plasma separation membranes by the non-solvent phase inversion method, the methods for controlling the pore size and morphology of hollow fiber membranes include polymer concentration, the types of solvents and non-solvents, the composition of additives, etc. Among them, adding polymer additives to the casting solution is considered to be the simplest method to control the thermodynamics and kinetics of phase inversion and regulate the membrane morphology and structure.

[0004] The biocompatibility of plasma separation membranes determines the safety of treatment. The biocompatibility of existing plasma separation membranes in clinical use needs to be further improved. Due to their inherent lack of anticoagulant properties, plasma separation membranes require continuous anticoagulation therapy to prevent thromboembolic events induced by coagulation factor activation during blood purification. Anticoagulation therapy requires sufficient anticoagulation to achieve the desired effect, while also avoiding the initiation or exacerbation of bleeding tendencies due to excessive anticoagulation. Modifying the anticoagulant properties of plasma separation membranes can avoid various complications caused by improper use of anticoagulants and improve treatment safety.

[0005] Currently, the main method for functionalizing the surface of blood purification materials with self-anticoagulant properties is to introduce substances or groups with anticoagulant effects, such as heparin, hirudin, and argatroban, onto the surface of the blood purification material through physical or chemical modification. However, as the blood purification time increases, the number of groups that can interact with coagulation factors decreases, and the self-anticoagulant effect of the material surface gradually decreases. Currently, the clinical use of plasma separation membranes to treat diseases still cannot get rid of the risks of continuous anticoagulation therapy, and the effectiveness of clinical application needs to be further improved. Summary of the Invention

[0006] In view of the deficiencies in the prior art, the technical problem to be solved by the present invention is to provide a plasma separation membrane with bionic self-anticoagulant function and a preparation method thereof.

[0007] The technical solution of the present invention to solve the technical problem of the method is to provide a method for preparing a plasma separation membrane with biomimetic self-anticoagulant function, characterized in that the method comprises the following steps:

[0008] Step 1: preparing a casting solution: dissolving the film-forming material and additives in a solvent to obtain a homogeneous casting solution;

[0009] Step 2: preparing a hollow fiber plasma separation membrane using a non-solvent phase inversion method;

[0010] Step 3: Preparing a plasma separation membrane modified with the fusion protein Hve-cad-Fc: Circulating a fusion protein Hve-cad-Fc solution inside a hollow fiber plasma separation membrane for incubation, thereby coating the fusion protein on the inner surface of the membrane; after the incubation is completed, washing the inner side of the membrane to remove unreacted substances, thereby obtaining a plasma separation membrane modified with the fusion protein Hve-cad-Fc;

[0011] Step 4: Inject the endothelial cell solution into the hollow inner cavity of the plasma separation membrane modified with the fusion protein Hve-cad-Fc, and then seal both ends of the membrane to enclose the endothelial cell solution inside the membrane; then place the membrane in an endothelial cell culture medium for incubation, allowing the endothelial cells to grow on the inner surface of the membrane, forming a plasma separation membrane with an endothelial structure on the inner surface and a biomimetic self-anticoagulant function.

[0012] The technical solution of the present invention to solve the technical problem of the plasma separation membrane is to provide a plasma separation membrane prepared by the preparation method of the plasma separation membrane with bionic self-anticoagulant function, characterized in that the plasma separation membrane has an inner diameter of 100 to 400 μm, a wall thickness of 10 to 50 μm, and a packing density of 5 to 20 cm 2 / cm 3 ; The pore size of the plasma separation membrane is 0.01~2μm.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] (1) The present invention promotes the adhesion of endothelial cells to the inner surface of the plasma separation membrane that is in direct contact with blood through the fusion protein Hve-cad-Fc, thereby achieving endothelialization of the inner surface of the plasma separation membrane and forming a bionic endothelial functional layer. The anticoagulant components of endothelial cells are utilized to inhibit the adhesion and activation of platelets, significantly reducing the formation of thrombi. At the same time, it has natural advantages such as anti-inflammatory properties, thereby achieving bionic functions.

[0015] (2) The plasma separation membrane with biomimetic self-anticoagulant function prepared by the present invention not only has the plasma separation function of a traditional plasma separation membrane, but also has an inner surface covered with endothelial cells, which imparts a long-lasting self-anticoagulant function in a biomimetic manner, achieving long-lasting anticoagulation, and the anticoagulant effect is more stable, thus solving the anticoagulation problem in the clinical treatment of diseases with plasma separation membranes. This greatly reduces or even avoids the use of anticoagulants during treatment, thereby avoiding the risk of bleeding caused by continuous anticoagulant injections in clinical treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a diagram showing the adsorption of platelets by the hollow fiber plasma separation membrane prepared in step 2 of Examples 1-3 of the present invention;

[0017] Figure 2 This is a diagram showing the adsorption of platelets by the plasma separation membrane with biomimetic self-anticoagulant function prepared in Example 1 of the present invention;

[0018] Figure 3 This is a diagram showing the adsorption of platelets by the plasma separation membrane with biomimetic self-anticoagulant function prepared in Example 2 of the present invention;

[0019] Figure 4 This is a diagram showing the adsorption of platelets by the plasma separation membrane with bionic self-anticoagulant function prepared in Example 3 of the present invention. DETAILED DESCRIPTION

[0020] The specific embodiments of the present invention are given below. The specific embodiments are only used to further illustrate the present invention and do not limit the scope of protection of the claims of the present invention.

[0021] The present invention provides a method for preparing a plasma separation membrane with biomimetic self-anticoagulant function (hereinafter referred to as the method), characterized in that the method comprises the following steps:

[0022] Step 1: preparing a casting solution: dissolving the film-forming material and additives in a solvent to obtain a homogeneous casting solution;

[0023] Preferably, in step 1, the membrane-forming material is polyethersulfone, polyvinylidene fluoride, polysulfone, ethylene-vinyl alcohol polymer or polypropylene (preferably polyvinylidene fluoride); the additive is at least one of polyvinyl alcohol, polyethylene glycol or polyvinyl pyrrolidone, which is used to form pores and enhance blood compatibility; and the solvent is dimethylformamide, dimethylacetamide or dimethyl sulfoxide.

[0024] Preferably, in step 1, the film-forming material accounts for 10 to 30 wt.% of the total mass of the casting solution, and the additives account for 1 to 10 wt.% of the total mass of the casting solution.

[0025] Preferably, in step 1, the dissolution process is: stirring at 60-120° C. for 6-24 hours.

[0026] Step 2: preparing a hollow fiber plasma separation membrane using a non-solvent phase inversion method;

[0027] Preferably, step 2 specifically comprises: placing the casting liquid in a reaction kettle and allowing it to stand for degassing; placing the core liquid in the core liquid kettle; then starting spinning, under the action of pressure, the casting liquid and the core liquid flow into the spinneret together, are extruded into the air through the spinneret, and are immersed in a coagulation bath after passing through an air bath to solidify, thereby forming a hollow fiber membrane; and then removing additives and solvents with deionized water to obtain a hollow fiber plasma separation membrane;

[0028] Preferably, in step 2, the temperature of the reactor of the hollow fiber spinning machine is 60 to 120° C., the pressure is 0.1 to 1 MPa, and the degassing time is 1 to 24 hours.

[0029] Preferably, in step 2, the core liquid is a 40-90 wt.% aqueous solution of dimethylformamide, dimethylacetamide, or dimethyl sulfoxide. The core liquid kettle temperature is 60-120° C. The core liquid flow rate is 25-250 mL / min.

[0030] Preferably, in step 2, the length of the air bath is 1 to 10 cm. The temperature of the coagulation bath is 20 to 60° C., and the coagulation bath is an aqueous solution of dimethylformamide, dimethylacetamide or dimethyl sulfoxide with a mass fraction of 0 to 30 wt.%.

[0031] Preferably, in step 2, the pore size of the hollow fiber plasma separation membrane is 0.01 to 2 μm.

[0032] Step 3: Preparing a plasma separation membrane modified with the fusion protein Hve-cad-Fc: Circulating a fusion protein Hve-cad-Fc solution inside a hollow fiber plasma separation membrane for incubation, thereby coating the fusion protein on the inner surface of the membrane; after the incubation is completed, washing the inner side of the membrane to remove unreacted substances, thereby obtaining a plasma separation membrane modified with the fusion protein Hve-cad-Fc;

[0033] Preferably, in step 3, the concentration of the fusion protein Hve-cad-Fc solution is 10 to 800 ng / mL, and the circulating flow rate is 1 to 10 mL / min.

[0034] Preferably, in step 3, the incubation temperature is 25-40° C. (preferably 37° C.) and the incubation time is 1-24 h (preferably 1-4 h).

[0035] Preferably, in step 3, the washing is performed by circulating physiological saline or PBS buffer at a flow rate of 1 to 10 mL / min on the inner side of the membrane for 4 to 24 hours.

[0036] Preferably, in step 3, before the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane, a polyethyleneimine grafting modification process is added to construct active functional groups on the inner surface of the hollow fiber plasma separation membrane, and then the fusion protein Hve-cad-Fc is grafted. Then, step 3 specifically comprises: first, the polyethyleneimine solution is circulated on the inner side of the hollow fiber plasma separation membrane to perform a Michael addition reaction; after the reaction is completed, the inner side of the membrane is washed to remove unreacted substances; then, the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane for incubation, and then the fusion protein is grafted on the inner surface of the membrane through an amidation reaction; after the incubation is completed, the inner side of the membrane is washed to remove unreacted substances to obtain a plasma separation membrane modified with the fusion protein Hve-cad-Fc.

[0037] Preferably, in step 3, the flow rate of the polyethyleneimine solution circulating inside the hollow fiber plasma separation membrane is 1 to 10 mL / min; the concentration of the polyethyleneimine solution is 0.5 to 30 wt.%; the amidation reaction environment is an alkaline environment (preferably pH = 9 to 13), the temperature is 30 to 80°C, and the time is 1 to 4 hours.

[0038] Preferably, in step 3, before the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane, a dopamine adhesion modification process is added to construct active functional groups on the inner surface of the hollow fiber plasma separation membrane, and then the fusion protein Hve-cad-Fc is grafted. Then, step 3 specifically comprises: firstly circulating the dopamine solution on the inner side of the hollow fiber plasma separation membrane so as to physically adhere to the inner surface of the membrane, and then washing the inner side of the membrane to remove unreacted substances; then circulating the fusion protein Hve-cad-Fc solution on the inner side of the hollow fiber plasma separation membrane for incubation, and then grafting the fusion protein on the inner surface of the membrane through an esterification reaction; after the incubation is completed, washing the inner side of the membrane to remove unreacted substances to obtain a plasma separation membrane modified with the fusion protein Hve-cad-Fc.

[0039] Preferably, in step 3, the dopamine solution circulates inside the hollow fiber plasma separation membrane at a flow rate of 1 to 10 mL / min; the concentration of the dopamine solution is 0.5 to 15 wt.%. The esterification reaction is carried out in an alkaline environment (preferably pH = 8.5), at a temperature of 30 to 80° C., and for 1 to 4 hours.

[0040] Preferably, in step 3, the hollow fiber plasma separation membrane obtained in step 2 is made into an internal pressure assembly, the plasma separation membrane assembly and related pipelines and instruments are sterilized and placed in a biosafety cabinet, and the plasma separation membrane assembly is connected to the polypropylene pipeline to establish a circulation path.

[0041] Preferably, in step 3, the fusion protein Hve-cad-Fc can be an existing product, which is formed by combining human E-cadherin protein, human vascular endothelial growth factor and the Fc domain of immunoglobulin IgG.

[0042] Step 4: Inject the endothelial cell solution into the hollow inner cavity of the plasma separation membrane modified with the fusion protein Hve-cad-Fc, and then seal both ends of the membrane to enclose the endothelial cell solution in the membrane to ensure that the endothelial cells grow only on the inner surface of the membrane; then place the membrane in an endothelial cell culture medium for incubation to allow the endothelial cells to grow on the inner surface of the membrane, forming a plasma separation membrane with an endothelial structure (i.e., a bionic functional layer) on the inner surface and having a bionic self-anticoagulant function.

[0043] Preferably, in step 4, the endothelial cell solution fills the hollow lumen of the plasma separation membrane modified with the fusion protein Hve-cad-Fc.

[0044] Preferably, in step 4, the concentration of the endothelial cell solution is 1×10 3 ~1×10 4 / mL.

[0045] Preferably, in step 4, the incubation temperature is 36-38° C. (preferably 37° C.), the incubation time is 8-48 h, and the incubation environment is a 5 v / v% CO 2 incubator (ie, the volume fraction of CO 2 is 5%).

[0046] The present invention also provides a plasma separation membrane prepared by the preparation method of the plasma separation membrane with bionic self-anticoagulant function, characterized in that the plasma separation membrane has an inner diameter of 100 to 400 μm (preferably 200 to 300 μm), a wall thickness of 10 to 50 μm (preferably 20 to 30 μm), and a packing density of 5 to 20 cm 2 / cm 3 ; The pore size of the plasma separation membrane is 0.01 to 2 μm (preferably 0.3 to 1 μm).

[0047] Example 1

[0048] Step 1: dissolve 200 g of polyvinylidene fluoride, 70 g of polyvinyl pyrrolidone, and 30 g of polyethylene glycol in 700 mL of dimethylacetamide, and stir at 90° C. for 8 h to prepare a uniformly dissolved casting solution;

[0049] Step 2: preparing a hollow fiber plasma separation membrane with excellent blood compatibility by using a non-solvent phase inversion method;

[0050] Step 3: A 100 ng / mL fusion protein Hve-cad-Fc solution was circulated through the inner side of the membrane at a flow rate of 10 mL / min and incubated at 40°C for 2 h. Then, the fusion protein Hve-cad-Fc solution was replaced with sterile saline at a flow rate of 10 mL / min, and the inner surface of the membrane was circulated and washed for 24 h to obtain a plasma separation membrane modified with the fusion protein Hve-cad-Fc.

[0051] Step 4: Add 1×10 4 / mL endothelial cell solution replaces physiological saline and flows into a plasma separation membrane modified with the fusion protein Hve-cad-Fc; then the two ends of the membrane are sealed and the membrane is placed in a culture dish with endothelial cell culture medium; then the membrane and the culture dish are moved to a CO2 incubator with a volume fraction of 5% and incubated at 37°C for 24 hours to allow endothelial cells to grow on the inner surface of the membrane, forming a plasma separation membrane with an endothelial structure on the inner surface and a biomimetic self-anticoagulant function.

[0052] Example 2

[0053] Step 1 is the same as in Example 1;

[0054] Step 2 is the same as in Example 1;

[0055] Step 3, a 0.5 wt.% polyethyleneimine solution with a pH of 12 was circulated through the inside of the membrane at a flow rate of 2 mL / min, and a Michael addition reaction was performed at 60°C for 2 hours; after the reaction was completed, the inner surface of the membrane was circulated and washed with sterile saline for 24 hours at a flow rate of 10 mL / min; then a 100 ng / mL fusion protein Hve-cad-Fc solution was circulated through the inside of the membrane at a flow rate of 10 mL / min and incubated at 40°C for 4 hours; then the fusion protein Hve-cad-Fc solution was replaced with sterile saline, and the inner surface of the membrane was washed for 24 hours at a flow rate of 10 mL / min to obtain a plasma separation membrane modified with the fusion protein Hve-cad-Fc;

[0056] Step 4 is the same as in Example 1.

[0057] Example 3

[0058] Step 1 is the same as in Example 1;

[0059] Step 2 is the same as in Example 1;

[0060] Step 3, a dopamine solution with a concentration of 0.5 wt.%, pH = 8.5, a flow rate of 5 mL / min, and a flow rate of 40°C for 4 hours was circulated on the inside of the membrane; after the reaction was completed, the inner surface of the membrane was circulated and washed with sterile saline for 24 hours at a flow rate of 10 mL / min; then a fusion protein Hve-cad-Fc solution with a concentration of 100 ng / mL was circulated on the inside of the membrane at a flow rate of 10 mL / min and incubated at 40°C for 12 hours; then the fusion protein Hve-cad-Fc solution was replaced with sterile saline, and the inner surface of the membrane was washed for 24 hours at a flow rate of 10 mL / min to obtain a plasma separation membrane modified with the fusion protein Hve-cad-Fc;

[0061] Step 4 is the same as in Example 1.

[0062] The hollow fiber plasma separation membrane obtained in step 2 of the above Examples 1-3 and the plasma separation membrane with biomimetic self-anticoagulant function obtained in step 4 were characterized by the following method, specifically:

[0063] 1. Platelet adhesion test:

[0064] Fresh venous blood was centrifuged at 1000 rpm for 15 minutes to obtain platelet-rich plasma, which was then injected into the hollow fiber plasma separation membrane and the plasma separation membrane with biomimetic self-anticoagulant function described in Examples 1-3. The membranes were incubated at 37°C for 90 minutes, and the inner side of the membranes was rinsed with PBS and fixed with 2.5% glutaraldehyde / PBS buffer at 4°C for 2 hours. The membranes were then dehydrated stepwise with ethanol, dried under vacuum, and then sprayed with gold. The platelet adhesion on the inner surface of the membranes was observed using a field emission scanning electron microscope. Specific platelet adsorption results are shown in [ 1 ]. Figure 1-Figure 4 .

[0065] Depend on Figure 1-Figure 4 It can be seen that compared with the hollow fiber plasma separation membrane, the inner surface of the plasma separation membrane with bionic self-anticoagulant function obtained in Examples 1-3 has reduced adsorption of platelets, and no pseudopodia appear on the platelets adsorbed on the membrane surface, indicating that the inner surface of the plasma separation membrane with bionic self-anticoagulant function will further reduce the adhesion of platelets on the membrane surface, avoid platelet activation-induced coagulation response, and avoid coagulation phenomenon.

[0066] 2. Coagulation test:

[0067] The hollow fiber plasma separation membranes and the plasma separation membranes with biomimetic self-anticoagulant function described in Examples 1-3 were immersed in fresh human blood. After incubation at 37°C for 2 hours, the blood was transferred to a centrifuge tube and centrifuged at 3000 rpm for 15 minutes at 4°C. Fresh plasma was collected and measured for prothrombin time, activated partial thromboplastin time, thrombin time, and fibrinogen using an automated coagulometer. Specific coagulation test results are shown in Table 1.

[0068] Table 1

[0069]

[0070] As shown in Table 1, compared with the hollow fiber plasma separation membrane, the inner surface of the plasma separation membrane with biomimetic self-anticoagulant function obtained in Examples 1-3 can significantly prolong the activated partial thromboplastin time while reducing the fibrinogen content. The hollow fiber plasma separation membrane and the inner surface of the plasma separation membrane with biomimetic self-anticoagulant function had no significant effect on the prothrombin time and thrombin time. Therefore, the inner surface of the plasma separation membrane with biomimetic self-anticoagulant function obtained in Examples 1-3 has enhanced self-anticoagulability, especially the inner surface of the plasma separation membrane with biomimetic self-anticoagulant function prepared in Example 3. After blood comes into contact with it, its activated partial thromboplastin time is extended to 3 times that of the inner surface of the hollow fiber plasma separation membrane.

[0071] 3. Plasma screening properties:

[0072] Fresh plasma was circulated through the inner side of the hollow fiber plasma separation membrane and the plasma separation membrane with biomimetic self-anticoagulant function in Examples 1-3. The blood pump flow rate was set to 200 ml / min. Plasma at the membrane inlet, plasma at the outlet, and plasma on the filtrate side were collected over a period of 60 minutes. The plasma protein concentration was measured using a fully automatic biochemical analyzer, and the membrane sieving rate SC for plasma components was calculated according to formula (1).

[0073]

[0074] In formula (1), C f is the protein concentration measured by filtration, C in and C out is the protein concentration at the inlet and outlet sides.

[0075] The specific results of the plasma screening experiment are shown in Table 2.

[0076] Table 2

[0077]

[0078]

[0079] As can be seen from Table 2, compared with the hollow fiber plasma separation membrane, the plasma separation membrane with bionic self-anticoagulant function obtained in Examples 1-3 has no obvious change in the plasma total protein screening rate and albumin screening rate, indicating that endothelial modification of the inner surface of the hollow fiber plasma separation membrane will not affect the screening performance of the membrane, and toxin-containing plasma can be effectively separated from the plasma separation membrane with bionic self-anticoagulant function obtained in Examples 1-3 to the outside of the membrane, thereby achieving toxin removal.

[0080] The operating principle and workflow of the present invention are as follows: a patient's blood is drawn from the body through one lumen of a double-lumen cannula and connected via tubing to a plasma separator constructed from a plasma separation membrane with biomimetic self-anticoagulant function. Under the action of a blood pump, the blood is drawn from the body into the inner surface of the plasma separation membrane with biomimetic self-anticoagulant function. Contact between the blood and the inner surface of the membrane prolongs the clotting time, thereby avoiding the need for continuous anticoagulant injections. The blood pump creates a pressure differential between the inner and outer sides of the membrane. This pressure differential, through the sieving mechanism of the membrane pores, allows toxin-containing plasma to pass through the membrane pores and be separated to the outer side of the plasma separation membrane with biomimetic self-anticoagulant function, effectively separating the toxin-containing plasma from blood cells. The toxins are then removed in one of two ways: The toxins are directly discarded, while fresh plasma is replenished and then re-infused with the beneficial cellular components on the inner side of the membrane, returning to the body through the other lumen of the double-lumen cannula to complete the treatment process. Secondly, the separated toxin-containing plasma flows into the plasma perfusion device, and the toxins are adsorbed on the plasma perfusion device. After the purified plasma merges with the cellular components on the inner side of the membrane that are useful to the human body, it is returned to the human body through the other cavity of the double-lumen cannula to complete the treatment process.

[0081] Any matters not described in the present invention are applicable to the prior art.

Claims

1. A method for preparing a plasma separation membrane with biomimetic self-anticoagulant function, characterized in that: The method comprises the following steps: Step 1: preparing a casting solution: dissolving the film-forming material and additives in a solvent to obtain a homogeneous casting solution; The film-forming material is polyethersulfone, polyvinylidene fluoride, polysulfone, ethylene-vinyl alcohol polymer or polypropylene; the additive is at least one of polyvinyl alcohol, polyethylene glycol or polyvinyl pyrrolidone; Step 2: preparing a hollow fiber plasma separation membrane using a non-solvent phase inversion method; Step 3: Preparing a plasma separation membrane modified with the fusion protein Hve-cad-Fc: Circulating a fusion protein Hve-cad-Fc solution inside a hollow fiber plasma separation membrane for incubation, thereby coating the fusion protein on the inner surface of the membrane; after the incubation is completed, washing the inner side of the membrane to remove unreacted substances, thereby obtaining a plasma separation membrane modified with the fusion protein Hve-cad-Fc; Step 4: Inject the endothelial cell solution into the hollow inner cavity of the plasma separation membrane modified with the fusion protein Hve-cad-Fc, and then seal both ends of the membrane to enclose the endothelial cell solution inside the membrane; then place the membrane in an endothelial cell culture medium for incubation, allowing the endothelial cells to grow on the inner surface of the membrane, forming a plasma separation membrane with an endothelial structure on the inner surface and a biomimetic self-anticoagulant function.

2. The method for preparing a plasma separation membrane having biomimetic self-anticoagulant function according to claim 1, characterized in that: In step 1, the film-forming material accounts for 10-30 wt% of the total mass of the casting solution, and the additive accounts for 1-10 wt% of the total mass of the casting solution; The solvent is dimethylformamide, dimethylacetamide or dimethyl sulfoxide.

3. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 1, characterized in that: Step 2 is specifically as follows: placing the casting liquid in a reactor and letting it stand for degassing; placing the core liquid in a core liquid reactor; then starting spinning. Under the action of pressure, the casting liquid and the core liquid flow into the spinneret together, are extruded into the air through the spinneret, and after passing through an air bath, are immersed in a coagulation bath for solidification to form a hollow fiber membrane; and then deionized water is used to remove additives and solvents to obtain a hollow fiber plasma separation membrane.

4. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 3, characterized in that: In step 2, the temperature of the reactor is 60~120°C, the pressure is 0.1~1Mpa; the standing degassing time is 1~24h; the core liquid adopts an aqueous solution of dimethylformamide, dimethylacetamide or dimethyl sulfoxide with a mass fraction of 40~90wt%; the temperature of the core liquid reactor is 60~120°C; the core liquid flow rate is 25~250mL / min; the length of the air bath is 1~10cm; the temperature of the coagulation bath is 20~60°C, and the coagulation bath adopts an aqueous solution of dimethylformamide, dimethylacetamide or dimethyl sulfoxide with a mass fraction of 0~30wt%.

5. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 1, characterized in that: In step 3, the concentration of the fusion protein Hve-cad-Fc solution is 10-800 ng / mL, and the circulating flow rate is 1-10 mL / min; the incubation temperature is 25-40°C, and the time is 1-24 h; and washing is performed by circulating physiological saline or PBS buffer on the inner side of the membrane at a flow rate of 1-10 mL / min for 4-24 h.

6. The method for preparing a plasma separation membrane having biomimetic self-anticoagulant function according to claim 1 or 5, characterized in that: In step 3, before the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane, a polyethyleneimine grafting modification process is added to construct active functional groups on the inner surface of the hollow fiber plasma separation membrane, and then the fusion protein Hve-cad-Fc is grafted. Step 3 specifically comprises: first, the polyethyleneimine solution is circulated on the inner side of the hollow fiber plasma separation membrane to perform a Michael addition reaction; after the reaction is completed, the inner side of the membrane is washed to remove unreacted substances; then, the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane for incubation, and the fusion protein is grafted on the inner surface of the membrane through an amidation reaction; after the incubation is completed, the inner side of the membrane is washed to remove unreacted substances, thereby obtaining a plasma separation membrane modified with the fusion protein Hve-cad-Fc; The polyethyleneimine solution circulates inside the hollow fiber plasma separation membrane at a flow rate of 1-10 mL / min; the concentration of the polyethyleneimine solution is 0.5-30 wt%; the amidation reaction is carried out in an alkaline environment at a temperature of 30-80° C. for 1-4 hours.

7. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 1 or 5, characterized in that: In step 3, before the fusion protein Hve-cad-Fc solution is circulated on the inner side of the hollow fiber plasma separation membrane, a dopamine adhesion modification process is added to construct active functional groups on the inner surface of the hollow fiber plasma separation membrane, and then the fusion protein Hve-cad-Fc is grafted. Step 3 specifically comprises: firstly circulating the dopamine solution on the inner side of the hollow fiber plasma separation membrane so as to physically adhere to the inner surface of the membrane, and then washing the inner side of the membrane to remove unreacted substances; then circulating the fusion protein Hve-cad-Fc solution on the inner side of the hollow fiber plasma separation membrane for incubation, and then grafting the fusion protein on the inner surface of the membrane through an esterification reaction; after the incubation is completed, washing the inner side of the membrane to remove unreacted substances, thereby obtaining a plasma separation membrane modified with the fusion protein Hve-cad-Fc; The dopamine solution circulates inside the hollow fiber plasma separation membrane at a flow rate of 1-10 mL / min; the concentration of the dopamine solution is 0.5-15 wt%; the esterification reaction is carried out in an alkaline environment at a temperature of 30-80° C. for 1-4 hours.

8. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 1, characterized in that: In step 4, the concentration of the endothelial cell solution was 1×10 3 ~1×10 4 pieces / mL.

9. The method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to claim 1, characterized in that: In step 4, the incubation temperature is 36-38°C, the incubation time is 8-48 hours, and the incubation environment is 5 v / v% CO2.

10. A plasma separation membrane prepared by the method for preparing a plasma separation membrane with biomimetic self-anticoagulant function according to any one of claims 1 to 9, characterized in that: The inner diameter of the plasma separation membrane is 100~400μm, the wall thickness is 10~50μm, and the packing density is 5~20cm 2 / cm 3 ;The pore size of the plasma separation membrane is 0.01~2μm.

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