A nanofiber membrane for adsorbing inflammatory factors and a preparation method thereof

Abstract

The invention discloses a nanofiber membrane for adsorbing inflammatory factors and a preparation method thereof, belonging to the technical field of biomedical materials. The nanofiber membrane structure comprises a positively charged nanofiber membrane and a negatively charged nanofiber membrane arranged in sequence; the positively charged nanofiber membrane is obtained by in-situ crosslinking after electrospinning with a main polymer, a positively charged ligand and a crosslinking agent as raw materials; the positively charged ligand is at least one of polylysine, polyethyleneimine, polyarginine or protamine; the negatively charged nanofiber membrane is obtained by in-situ crosslinking after electrospinning with a main polymer, a negatively charged ligand and a crosslinking agent as raw materials; the negatively charged ligand is at least one of heparin, chondroitin sulfate or heparan sulfate. The nanofiber membrane has a simple preparation process, has abundant charged binding sites on the membrane surface, has a large charge density, high mechanical strength, and has a good adsorption effect on inflammatory factors.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical materials, and specifically relates to a nanofiber membrane for adsorbing inflammatory factors and a preparation method thereof. Background Art

[0002] Inflammatory factors are an important group of proteins that play a role in information transmission in the human immune system. There are many types of them, including interleukins, tumor necrosis factors, interferons, and other major categories, each of which contains many groups. When the body is infected or tissues are damaged, inflammatory factors produced by inflammatory cells or other cells regulate immune responses and inflammatory reactions through synergistic and antagonistic effects to maintain the physiological health of the body, but their excessive or unregulated expression may lead to inflammatory diseases and cause damage to the body itself.

[0003] Symptoms such as death due to cerebral ischemia, tissue damage caused by organ separation, infection, etc., lead to excessive secretion of inflammatory factors in the separated organs, causing inflammation, ischemia-reperfusion injury and other symptoms, which affect the quality of donor organs. Among the large number of donor organs provided by the transplant team, only a small number of organs meet the transplantation standards, and most of the donor organs become marginal donor organs because they do not meet the transplantation standards. In order to expand the donor pool, medical personnel mechanically perfuse marginal donor organs, and perform functional repair and quality assessment of damaged organs by adjusting cell metabolism, adding antibiotics and immunomodulatory drugs, and testing physiological indicators.

[0004] However, mechanical perfusion itself may also present a negative pro-inflammatory environment. Donor organs may be subjected to reperfusion injury or mechanical ventilation injury, which may trigger inflammatory responses and oxidative stress, leading to further increases in the concentration of inflammatory factors and causing tissue damage. Therefore, removing over-secreted inflammatory factors is a potential strategy to help repair damaged organs. In view of this situation, the development of adsorbent materials that can effectively remove inflammatory factors in perfusion fluids has significant economic and social benefits.

[0005] The Chinese patent document with publication number CN117942792A discloses a nanofiber filtration adsorption membrane for adsorbing interleukin, and the adsorption performance of the nanofiber membrane is improved by introducing functional groups that can react with interleukin on the surface of the nanofiber filtration membrane. The chemical reaction of introducing functional groups that can react with interleukin includes nucleophilic substitution reaction, electrophilic substitution reaction, nucleophilic addition reaction or electrophilic addition reaction. This process requires the use of a large amount of chemical reagents, which may have potential biosafety issues.

[0006] A Chinese patent document with publication number CN116966756A discloses a method for preparing a blood purification membrane for removing pro-inflammatory cytokines. The invention constructs a pro-inflammatory cytokine adsorption functional layer on the outer surface of a hollow fiber plasma separation membrane, which effectively improves the blood compatibility of the material and the adsorption performance of pro-inflammatory cytokines. A Chinese patent document with publication number CN118718758A discloses a plasma filtration adsorption membrane, which includes a three-layer structure-a cellulose-based adsorption layer, a fiber membrane-based filtration layer, and a nanofiber membrane layer, wherein the nanofiber membrane layer is a nanofiber filtration membrane modified with nitrogen-containing organic functional groups, which can adsorb at least one interleukin. The above invention has cumbersome steps and involves many chemical raw materials. Summary of the invention

[0007] The present invention provides a nanofiber membrane for adsorbing inflammatory factors, which has a simple preparation process, has abundant charged binding sites on the membrane surface, has a large charge density, and has high mechanical strength, and has a good adsorption effect on inflammatory factors.

[0008] The specific technical solutions adopted are as follows:

[0009] A nanofiber membrane for adsorbing inflammatory factors, the structure of which includes a positively charged nanofiber membrane and a negatively charged nanofiber membrane arranged in sequence;

[0010] The positively charged nanofiber membrane is obtained by electrospinning a main polymer, a positively charged ligand and a cross-linking agent as raw materials and then in-situ cross-linking; the positively charged ligand is at least one of polylysine, polyethyleneimine, polyarginine or protamine;

[0011] The negatively charged nanofiber membrane is obtained by in-situ crosslinking after the main polymer, negatively charged ligand and crosslinking agent are mixed and electrospun as raw materials; the negatively charged ligand is at least one of heparin, chondroitin sulfate or heparan sulfate.

[0012] Both the positively charged nanofiber membrane and the negatively charged nanofiber membrane are prepared by co-blending electrospinning. The charged ligands (positively charged ligands / negatively charged ligands) are evenly dispersed on the surface of the nanofibers, with good dispersibility and a large grafting amount. In situ crosslinking can achieve covalent bonding between the charged ligands and the main polymer, with strong binding force and good fixation. The membrane material having a double-layer structure of positively charged nanofiber membrane and negatively charged nanofiber membrane can adsorb a variety of inflammatory factors through electrostatic-electrostatic interactions, and has broad application prospects in the adsorption of inflammatory factors. Specifically, it can be used to remove pathogenic inflammatory factors from the perfusion solution during mechanical perfusion of isolated organs.

[0013] The inflammatory factors include interleukin-8, interleukin-10, interleukin-1β, interleukin-6, interleukin-4, interferon, etc. The experimental results show that the nanofiber membrane for adsorption of inflammatory factors has a clearance rate of 1000 pg / mL interleukin-8, interleukin-10, and interleukin-1β of more than 95%.

[0014] Preferably, the thickness of the positively charged nanofiber membrane is 110-300 μm, and the thickness of the negatively charged nanofiber membrane is 150-280 μm; further preferably, the thickness of the positively charged nanofiber membrane is 150-270 μm, and the thickness of the negatively charged nanofiber membrane is 150-250 μm.

[0015] Preferably, the zeta potential of the positively charged nanofiber membrane is 3~60 mV, and the zeta potential of the negatively charged nanofiber membrane is -23~-50 mV; further preferably, the zeta potential of the positively charged nanofiber membrane is 20~60 mV, and the zeta potential of the negatively charged nanofiber membrane is -32~-50 mV.

[0016] The main polymer includes but is not limited to at least one of nylon 66, silk protein, gelatin, polyvinyl alcohol, zein or cellulose acetate.

[0017] The cross-linking agent is at least one of genipin and glutaraldehyde.

[0018] Furthermore, the mass ratio of the main polymer and the charged ligand (positively charged ligand / negatively charged ligand) is 1~40:1, and the mass ratio of the charged ligand (positively charged ligand / negatively charged ligand) and the cross-linking agent is 5~80:1; further preferably, the mass ratio of the main polymer and the charged ligand (positively charged ligand / negatively charged ligand) is 1.5~20:1, and the mass ratio of the charged ligand (positively charged ligand / negatively charged ligand) and the cross-linking agent is 5~40:1.

[0019] The present invention also provides a method for preparing the nanofiber membrane for adsorbing inflammatory factors, comprising the following steps:

[0020] A first casting solution comprising components including a main polymer, a positively charged ligand and a cross-linking agent is prepared, and a second casting solution comprising components including a main polymer, a negatively charged ligand and a cross-linking agent is prepared; the first casting solution and the second casting solution are formed into a double-layer membrane structure using electrostatic spinning technology, and the nanofiber membrane for adsorbing inflammatory factors is obtained after cross-linking, washing and drying.

[0021] Preferably, the mass concentration of the first casting solution and the second casting solution (including the total mass concentration of each solute) are both 15-22wt%.

[0022] The process parameters of electrospinning are: casting liquid advancement rate 0.1~1 mL / h, receiving distance 8~15 cm, collector speed 10~300 rpm, voltage 15~30 kV, and electrospinning time 2~20 h.

[0023] Preferably, the cross-linking temperature is 30-80° C., and the cross-linking time is 12-48 h. Under the above preferred parameters, it is helpful to achieve the covalent bonding of the charged ligand and the main polymer, and the bonding force is strong and the fixation is good.

[0024] Preferably, the washing method is oscillating washing, the washing temperature is 25-80° C., and the washing time is 3-48 h.

[0025] Preferably, the drying method is vacuum drying, the drying temperature is 30-60°C, and the drying time is 12-48 h.

[0026] The present invention also provides an inflammatory factor adsorption method, using the nanofiber membrane for inflammatory factor adsorption. Specific application scenarios include removing pathogenic inflammatory factors from the perfusion solution during the mechanical perfusion of an isolated organ.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] (1) The present invention uniformly disperses positively charged ligands or negatively charged ligands on the surface of the main polymer by means of co-blending electrospinning, thereby providing abundant charged binding sites and effectively improving the binding performance of inflammatory factors.

[0029] (2) The present invention achieves the fixation of charged ligands by in-situ cross-linking, which greatly simplifies the preparation process, has mild conditions, and can reduce the use of organic solvents.

[0030] (3) The membrane material prepared by the present invention comprises a double-layer structure of positively charged nanofiber membrane and negatively charged nanofiber membrane. By enriching charged binding sites on the membrane surface, it regulates the electrostatic interaction and hydrogen bond interaction between the charged ligand and the inflammatory factor carrying the opposite charge, thereby causing the inflammatory factor to undergo structural changes and produce different conformations, thereby achieving adsorption of the inflammatory factor on the membrane surface.

[0031] (4) The preparation process of the nanofiber membrane for adsorbing inflammatory factors provided by the present invention is simple, the raw materials are easy to obtain, the equipment requirements are low, the amount of solvent required is small, and it is easy to achieve large-scale production. The prepared nanofiber membrane contains mixed charges, has a large charge density, high mechanical strength, and good adsorption effect, and can simultaneously adsorb inflammatory factors with different charges.

[0032] (5) Adsorption tests show that the nanofiber membrane for adsorption of inflammatory factors provided by the present invention has a clearance rate of >95% for interleukin-8, interleukin-10, and interleukin-1β at a concentration of 1000 pg / mL, and can be used to repair damaged organs during mechanical perfusion. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is a microscopic morphology of the positively charged nanofiber membrane in the nanofiber membrane for adsorption of inflammatory factors in Example 3.

[0034] Figure 2 This is a microscopic morphology of the negatively charged nanofiber membrane in the nanofiber membrane for adsorption of inflammatory factors in Example 3.

[0035] Figure 3 It is a schematic diagram of the structure of the nanofiber membrane for adsorbing inflammatory factors. DETAILED DESCRIPTION

[0036] In order to make the purpose, features and advantages of the present invention more obvious and easy to understand, the following is a detailed description through specific embodiments. In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below. The technical features in each embodiment of the present invention can be combined accordingly without conflicting with each other.

[0037] The operating methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. The contents not described in detail in this specification belong to the prior art known to professionals in the field. The experimental materials used in the following examples, unless otherwise specified, can be purchased from conventional biochemical reagent companies.

[0038] Example 1

[0039] (1) Nylon 66, polyethyleneimine and genipin were dissolved in formic acid to prepare the first casting solution, and magnetic stirring was performed for 12 h to fully dissolve and mix them; in the first casting solution, the mass ratio of nylon 66 to polyethyleneimine was 40:1, the mass ratio of polyethyleneimine to genipin was 80:1, and the mass concentration of the first casting solution was 22wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 1 mL / h, the receiving distance was controlled to 8 cm, the collector drum speed was controlled to 20 rpm, a 20 kV DC high voltage was applied, and electrospinning was performed for 2 h to prepare the first nanofiber membrane.

[0040] (2) Nylon 66, heparin and genipin were dissolved in formic acid to prepare a second casting solution, and magnetic stirring was performed for 6 h to fully dissolve and mix them. In the second casting solution, the mass ratio of nylon 66 to heparin was 40:1, and the mass ratio of heparin to genipin was 80:1; the mass concentration of the second casting solution was 22wt%. The second casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 1 mL / h, the receiving distance was controlled to 8 cm, the collector drum speed was controlled to 10 rpm, and a 15 kV DC high voltage was applied for electrospinning for 2 h. On the basis of the first nanofiber membrane, electrospinning was performed to obtain a second nanofiber membrane; a double-layer structure of a mixed charge nanofiber membrane with different charges on both sides was formed.

[0041] (3) The nanofiber membrane obtained in step (2) was placed in an oven at a temperature of 30°C for cross-linking for 48 hours; after being taken out, it was shaken and washed at 25°C for 48 hours, and then vacuum dried for 48 hours at a drying temperature of 30°C. The nanofiber membrane for adsorbing inflammatory factors was obtained.

[0042] The nanofiber membrane for adsorbing inflammatory factors includes a positively charged nanofiber membrane with a thickness of 300 μm and a negatively charged nanofiber membrane with a thickness of 280 μm. The zeta potential of the positively charged nanofiber membrane is 3 mV, and the zeta potential of the negatively charged nanofiber membrane is -23 mV.

[0043] Example 2

[0044] (1) Nylon 66, polyethyleneimine and genipin were dissolved in formic acid to prepare the first casting solution, and magnetic stirring was performed for 12 h to fully dissolve and mix them. In the first casting solution, the mass ratio of nylon 66 to polyethyleneimine was 10:1, the mass ratio of polyethyleneimine to genipin was 40:1, and the mass concentration of the first casting solution was 22wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.2 mL / h, the receiving distance was controlled to 12 cm, the collector drum speed was controlled to 50 rpm, a DC high voltage of 22 kV was applied, and electrospinning was performed for 10 h to prepare the first nanofiber membrane.

[0045] (2) Nylon 66, heparin and genipin were dissolved in formic acid to prepare the second casting solution, and magnetic stirring was performed for 6 h to fully dissolve and mix them. In the second casting solution, the mass ratio of nylon 66 to heparin was 9:1, and the mass ratio of heparin to genipin was 20:1; the mass concentration of the second casting solution was 18wt%. The second casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.5 mL / h, the receiving distance was controlled to 9 cm, the collector drum speed was controlled to 50 rpm, and a 29 kV DC high voltage was applied for 4 h. The second nanofiber membrane was electrospun on the basis of the first nanofiber membrane to obtain a second nanofiber membrane; a double-layer structure of mixed charge nanofiber membranes with different charges on both sides was formed.

[0046] (3) The nanofiber membrane obtained in step (2) was placed in an oven at a temperature of 40°C for cross-linking for 36 hours; after being taken out, it was shaken and washed at 37°C for 12 hours, and then vacuum dried for 36 hours at a drying temperature of 45°C. The nanofiber membrane for adsorbing inflammatory factors was obtained.

[0047] The nanofiber membrane for adsorbing inflammatory factors includes a positively charged nanofiber membrane with a thickness of 280 μm and a negatively charged nanofiber membrane with a thickness of 250 μm. The zeta potential of the positively charged nanofiber membrane is 28 mV, and the zeta potential of the negatively charged nanofiber membrane is -42 mV.

[0048] Example 3

[0049] (1) Nylon 66, polyethyleneimine and genipin were dissolved in formic acid to prepare the first casting solution, and magnetic stirring was performed for 12 h to fully dissolve and mix them. In the first casting solution, the mass ratio of nylon 66 to polyethyleneimine was 3:2, the mass ratio of polyethyleneimine to genipin was 20:1, and the mass concentration of the first casting solution was 20wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.4 mL / h, the receiving distance was controlled to 10 cm, the collector drum speed was controlled to 200 rpm, a DC high voltage of 25 kV was applied, and electrospinning was performed for 5 h to prepare the first nanofiber membrane.

[0050] (2) Nylon 66, heparin and genipin were dissolved in formic acid to prepare a second casting solution, and magnetic stirring was performed for 6 h to fully dissolve and mix them. In the second casting solution, the mass ratio of nylon 66 to heparin was 17:3, and the mass ratio of heparin to genipin was 10:1; the mass concentration of the second casting solution was 20wt%. The second casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.4 mL / h, the receiving distance was controlled to 10 cm, the collector drum speed was controlled to 150 rpm, and 30 kV DC high voltage was applied for electrospinning for 5 h. The second nanofiber membrane was electrospun on the basis of the first nanofiber membrane to form a double-layer structure of mixed charge nanofiber membranes with different charges on both sides.

[0051] (3) The nanofiber membrane obtained in step (2) was placed in an oven at a temperature of 60°C for cross-linking for 24 h; after being taken out, it was shaken and washed at 50°C for 24 h, and then vacuum dried for 12 h at a drying temperature of 60°C. The nanofiber membrane for adsorbing inflammatory factors was obtained.

[0052] The nanofiber membrane for adsorbing inflammatory factors includes a positively charged nanofiber membrane with a thickness of 160 μm and a negatively charged nanofiber membrane with a thickness of 200 μm. The zeta potential of the positively charged nanofiber membrane is 60 mV, and the zeta potential of the negatively charged nanofiber membrane is -50 mV.

[0053] Example 4

[0054] (1) Nylon 66, polylysine and genipin were dissolved in formic acid to prepare the first casting solution, and magnetic stirring was performed for 12 h to fully dissolve and mix them. In the first casting solution, the mass ratio of nylon 66 to polylysine was 10:1, the mass ratio of polylysine to genipin was 10:1, and the mass concentration of the first casting solution was 20wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.5 mL / h, the receiving distance was controlled to 11 cm, the collector drum speed was controlled to 300 rpm, 27 kV DC high voltage electricity was applied, and electrospinning was performed for 4 h to prepare the first nanofiber membrane;

[0055] (2) Nylon 66, heparin and glutaraldehyde were dissolved in formic acid to prepare the second casting solution, and magnetic stirring was performed for 6 h to fully dissolve and mix them. In the second casting solution, the mass ratio of nylon 66 to heparin was 9:1, and the mass ratio of heparin to glutaraldehyde was 20:1; the mass concentration of the second casting solution was 15wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.25 mL / h, the receiving distance was controlled to 10 cm, the collector drum speed was controlled to 300 rpm, and 25 kV DC high voltage was applied for electrospinning for 8 h. On the basis of the first nanofiber membrane, electrospinning was performed to obtain the second nanofiber membrane; a double-layer structure of mixed charge nanofiber membranes with different charges on both sides was formed.

[0056] (3) The nanofiber membrane obtained in step (2) was placed in an oven at a temperature of 50°C for cross-linking for 24 hours; after being taken out, it was shaken and washed at 45°C for 20 hours, and then vacuum dried for 36 hours at a drying temperature of 45°C. The nanofiber membrane for adsorbing inflammatory factors was obtained.

[0057] The nanofiber membrane for adsorbing inflammatory factors includes a positively charged nanofiber membrane with a thickness of 200 μm and a negatively charged nanofiber membrane with a thickness of 220 μm. The zeta potential of the positively charged nanofiber membrane is 16 mV, and the zeta potential of the negatively charged nanofiber membrane is -38 mV.

[0058] Example 5

[0059] (1) Nylon 66, polyethyleneimine and genipin were dissolved in formic acid to prepare the first casting solution, and magnetic stirring was performed for 12 h to fully dissolve and mix them. In the first casting solution, the mass ratio of nylon 66 to polyethyleneimine was 1:1, the mass ratio of polyethyleneimine to genipin was 10:1, and the mass concentration of the first casting solution was 19wt%. The first casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.5 mL / h, the receiving distance was controlled to 14 cm, the collector drum speed was controlled to 100 rpm, a DC high voltage of 28 kV was applied, and electrospinning was performed for 4 h to prepare the first nanofiber membrane.

[0060] (2) Nylon 66, heparin and genipin were dissolved in formic acid to prepare the second casting solution, and magnetic stirring was performed for 6 h to fully dissolve and mix them. In the second casting solution, the mass ratio of nylon 66 to heparin was 20:1, and the mass ratio of heparin to genipin was 5:1; the mass concentration of the first casting solution was 22wt%. The second casting solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.1 mL / h, the receiving distance was controlled to 15 cm, the collector drum speed was controlled to 250 rpm, and 28 kV DC high voltage was applied for electrospinning for 20 h. On the basis of the first nanofiber membrane, electrospinning was performed to obtain a second nanofiber membrane; a double-layer structure of mixed charge nanofiber membranes with different charges on both sides was formed.

[0061] (3) The nanofiber membrane obtained in step (2) was placed in an oven at 80°C for crosslinking for 12 h; after being taken out, it was shaken and washed at 80°C for 3 h, and then vacuum dried for 24 h at a drying temperature of 50°C. The nanofiber membrane for adsorbing inflammatory factors was obtained.

[0062] The nanofiber membrane for adsorbing inflammatory factors includes a positively charged nanofiber membrane with a thickness of 110 μm and a negatively charged nanofiber membrane with a thickness of 150 μm. The zeta potential of the positively charged nanofiber membrane is 53 mV, and the zeta potential of the negatively charged nanofiber membrane is -30 mV.

[0063] Comparative Example 1

[0064] Nylon 66 was dissolved in formic acid and magnetically stirred for 6 h to obtain a nylon casting solution with a mass concentration of 20wt%. The solution was transferred to a 10 mL syringe, the solution propulsion rate was controlled to 0.2 mL / h, the receiving distance was controlled to 10 cm, the drum speed was 200 rpm, a 20 kV DC high voltage was applied, and electrospinning was performed for 10 h to prepare a nanofiber-based membrane.

[0065] Comparative Example 2

[0066] The first nanofiber membrane after crosslinking, washing and drying in Example 3 is used as the sample of Comparative Example 2, and the conditions of crosslinking, washing and drying are the same as those in Example 3.

[0067] Comparative Example 3

[0068] The second nanofiber membrane after crosslinking, washing and drying in Example 3 was used as the sample of Comparative Example 3 (electrospinning was performed directly on the collector instead of electrospinning on the basis of the first nanofiber membrane), and the conditions of crosslinking, washing and drying were the same as those in Example 3.

[0069] Sample analysis

[0070] (1) Surface morphology

[0071] The microscopic morphology of the nanofiber membrane for adsorption of inflammatory factors in Example 3 was analyzed, and the SEM images of the positively charged nanofiber membrane and the negatively charged nanofiber membrane were shown as follows: Figure 1 and Figure 2 As shown in the figure, the fibers in the positively charged nanofiber membrane and the negatively charged nanofiber membrane are all nanometer-sized, have good pore characteristics, and can provide abundant binding sites; the structural schematic diagram of the nanofiber membrane for adsorption of inflammatory factors is shown in the figure. Figure 3 shown.

[0072] (2) Adsorption performance

[0073] Interleukin-8 with a net positive charge, interleukin-10 with a near-neutral slightly net positive charge, and interleukin-1β with a net negative charge were added to a PBS buffer solution with a pH of 7.35-7.45, and the concentration was controlled to be about 1000 pg / mL. 20 mg of the nanofiber membrane for adsorption of inflammatory factors in the embodiments or comparative examples was cut into a glass test tube, 1 mL of inflammatory factor aqueous solution was added, the sealing film was sealed, and oscillated in a metal bath for 6 h, the oscillation temperature was 37°C, and the oscillation rate was 400 rpm. Take a small amount of aqueous solution for detection.

[0074] According to the instruction manual of the ELISA kit, a standard curve was established for the detection of sample concentration. The concentrations of interleukin-8, interleukin-10 and interleukin-1β in the solution before and after adsorption were obtained using the four-parameter regression method. Each group of experiments was repeated three times, and the clearance rate of inflammatory factors was calculated according to the following formula.

[0075] AP = (C0-C1) / C0*100%

[0076] Where AP is the clearance rate (%), C0 is the concentration of interleukin-8, interleukin-10, and interleukin-1β in the solution before adsorption (pg / mL), and C1 is the concentration of interleukin-8, interleukin-10, and interleukin-1β in the solution after adsorption (pg / mL).

[0077] The adsorption results are shown in Table 1.

[0078] Table 1 Test results of adsorption performance of nanofiber membranes prepared in Examples 1-5 and Comparative Examples 1-3.

[0079]

[0080] As shown in Table 1, compared with the comparative example, the embodiment proves that the method of the present invention achieves a significant improvement in the adsorption performance of inflammatory factors by fixing charged ligands with different charges. This is because the different charged ligands are evenly distributed on the surface, providing abundant positive and negative charge binding sites, so that the charged nanofiber membrane can achieve the adsorption of inflammatory factors with different charges through electrostatic interactions.

[0081] The embodiments described above provide a detailed description of the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, supplements or similar substitutions made within the scope of the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A nanofiber membrane for adsorbing inflammatory factors, characterized in that: The structure includes a positively charged nanofiber membrane and a negatively charged nanofiber membrane arranged in sequence; the thickness of the positively charged nanofiber membrane is 110-300 μm, and the thickness of the negatively charged nanofiber membrane is 150-280 μm; the zeta potential of the positively charged nanofiber membrane is 20-60 mV, and the zeta potential of the negatively charged nanofiber membrane is -32--50 mV; The positively charged nanofiber membrane is obtained by electrospinning a main polymer, a positively charged ligand and a cross-linking agent as raw materials and then in-situ cross-linking; the positively charged ligand is at least one of polylysine, polyethyleneimine, polyarginine or protamine; The negatively charged nanofiber membrane is obtained by in-situ cross-linking after the main polymer, negatively charged ligand and cross-linking agent are mixed and electrospun as raw materials; the negatively charged ligand is at least one of heparin, chondroitin sulfate or heparan sulfate; The inflammatory factor is interleukin-8, interleukin-10, interleukin-1β, interleukin-6, interleukin-4 or interferon; The cross-linking agent is at least one of genipin or glutaraldehyde; The mass ratio of the main polymer to the positively charged ligand or the negatively charged ligand is 1.5-20:1, and the mass ratio of the positively charged ligand or the negatively charged ligand to the cross-linking agent is 5-40:

1.

2. The nanofiber membrane for adsorbing inflammatory factors according to claim 1, characterized in that: The main polymer includes at least one of nylon 66, silk protein, gelatin, polyvinyl alcohol, zein or cellulose acetate.

3. The method for preparing a nanofiber membrane for adsorbing inflammatory factors according to claim 1 or 2, characterized in that: The following steps are involved: A first casting solution comprising components including a main polymer, a positively charged ligand and a cross-linking agent is prepared, and a second casting solution comprising components including a main polymer, a negatively charged ligand and a cross-linking agent is prepared; the first casting solution and the second casting solution are formed into a double-layer membrane structure using electrostatic spinning technology, and the nanofiber membrane for adsorbing inflammatory factors is obtained after cross-linking, washing and drying.

4. The method for preparing a nanofiber membrane for adsorbing inflammatory factors according to claim 3, characterized in that: The mass concentrations of the first casting solution and the second casting solution were both 15~22wt%; the process parameters of electrospinning were: casting solution advancement rate 0.1~1 mL / h, receiving distance 8~15 cm, collector speed 10~300 rpm, voltage 15~30 kV, and electrospinning time 2~20 h.

5. The method for preparing a nanofiber membrane for adsorbing inflammatory factors according to claim 3, characterized in that: The cross-linking temperature is 30~80℃, and the cross-linking time is 12~48 h.

6. A method for adsorbing inflammatory factors, characterized in that: Utilize the nanofiber membrane for adsorbing inflammatory factors as described in claim 1 or 2.

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