WO2024045163A1 - 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用 - Google Patents

一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用 Download PDF

Info

Publication number
WO2024045163A1
WO2024045163A1 PCT/CN2022/116742 CN2022116742W WO2024045163A1 WO 2024045163 A1 WO2024045163 A1 WO 2024045163A1 CN 2022116742 W CN2022116742 W CN 2022116742W WO 2024045163 A1 WO2024045163 A1 WO 2024045163A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydrophilic
polyphenol
polymer
medical materials
functionalization
Prior art date
Application number
PCT/CN2022/116742
Other languages
English (en)
French (fr)
Inventor
徐福建
胡杨
李筱玥
杨雪
Original Assignee
北京化工大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 北京化工大学 filed Critical 北京化工大学
Priority to PCT/CN2022/116742 priority Critical patent/WO2024045163A1/zh
Publication of WO2024045163A1 publication Critical patent/WO2024045163A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
    • A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
    • C08J7/04—Coating

Definitions

  • the invention belongs to the field of medical hemostatic materials and relates to a preparation method of a polyphenol-polymer composite coating and its application in functionalization of medical materials.
  • the present invention provides a preparation method of polyphenol-polymer composite coating and its application in functionalization of medical materials.
  • the present invention specifically provides the following technical solutions:
  • a method for preparing a polyphenol-polymer composite coating and its application in functionalization of medical materials is:
  • the functionalized hemostatic function and/or protein differential adhesion function are provided.
  • the mass ratio of polyphenolic hydroxyl substances and hydrophilic uncharged or hydrophilic negatively charged polymer in step 1) is 1:2 ⁇ 8 and the soaking time in step 2) is 0.7 ⁇ 4 hours.
  • Step 3) Obtain medical materials with hemostatic function;
  • the mass ratio of polyphenolic hydroxyl substances and hydrophilic uncharged or hydrophilic negatively charged polymers in step 1) is 1:0.5 ⁇ 15 and the soaking time in step 2) is 0.5 ⁇ 20 hours, step 3) obtain medical materials with differential protein adhesion;
  • the polyphenolic hydroxyl substances described in step 1) are dopamine, tannic acid, gallic acid, catechin, epicatechin, epigallocatechin gallate, theaflavin-3-gallate, 5 -Hydroxydopamine hydrochloride or baicalein
  • the hydrophilic uncharged polymer described in step 1) is dextran, hydroxypropyl cellulose, hydroxyethyl starch, hydroxyethyl cellulose, polysaccharide, and pluronic acid Or polyethylene glycol
  • the hydrophilic and electronegative polymer described in step 1) is carboxymethylcellulose or alginic acid, sodium alginate, hyaluronic acid, sodium hyaluronate or carboxymethyl starch.
  • step 2 when the mass ratio of the polyphenolic hydroxyl material in step 1) and the hydrophilic uncharged or hydrophilic negatively charged polymer is 1:2 ⁇ 5, the soaking time in step 2) is 0.7 ⁇ 1.5 hours, step 3 ) to obtain medical materials with hemostatic function.
  • step 3 obtains It is a medical material with differential adhesion of protein, and the protein is fibrinogen, albumin, lipoprotein, glycoprotein or globulin.
  • the polyphenolic hydroxyl material in step 1) is dopamine, tannic acid or gallic acid
  • the hydrophilic uncharged polymer in step 1) is dextran, hydroxypropyl cellulose or polyethylene glycol.
  • the hydrophilic and electronegative polymer described in step 1) is carboxymethylcellulose.
  • the concentration of the polyphenolic hydroxyl substances in step 1) in the mixed solution is 0.25 to 3 mg/mL.
  • the conditions for realizing the hemostatic function are: the concentration of hydrophilic and uncharged or hydrophilic and negatively charged polymers in the mixed solution is 3 to 5 mg/mL.
  • the conditions for realizing differential protein adhesion are: the concentration of hydrophilic and uncharged or hydrophilic and negatively charged polymers in the mixed solution. The concentration in the mixed solution is 5 to 10 mg/mL.
  • the substrate in step 2) is a polymer medical material, an inorganic medical material, or a metal medical material;
  • the polymer medical material is gauze, polyvinyl alcohol sponge, polyvinyl alcohol micron-sized particles, polyethylene Alcohol microspheres, chitosan sponge, chitosan non-woven fabric, gelatin sponge, gelatin sponge micron particles, alginic acid dressing, alginic acid microspheres, alginic acid non-woven fabric or collagen sponge;
  • the inorganic medical The materials are bioglass and bioceramics;
  • the metal medical materials are nickel-titanium alloy instruments, titanium alloy medical instruments, and shape memory metal micro-spring coils.
  • the base material in step 2) is gauze, polyvinyl alcohol sponge, polyvinyl alcohol micron particles, polyvinyl alcohol microspheres, gelatin sponge, gelatin sponge micron particles, collagen sponge, biological glass or titanium alloy medical instrument.
  • the base material in step 2) is a metal medical material
  • the base material can be pretreated with polyphenol hydroxyl substances before soaking in the mixed solution to form a polyphenol coating on the surface of the base material.
  • the pretreatment method is: The base material is soaked in an alkaline aqueous solution of polyphenol hydroxyl substances with a pH of 8 to 9 for 4 to 24 hours.
  • the mixed solution described in step 1) is an alkaline aqueous solution with a pH of 8-9.
  • the first application of the present invention is to combine the oxidative self-polymerization of polyphenolic hydroxyl substances with hydrophilic uncharged or negatively charged polymers to form a coating on the surface of a substrate.
  • hydrophilic uncharged or negatively charged polymers There is a strong hydrogen bond interaction between polyphenolic hydroxyl substances and proteins.
  • the composite coating it provides a force on the key components of coagulation in the blood, such as plasma proteins (fibrinogen, etc.), causing them to adhere and aggregate; it has anti-protein properties.
  • the adhesive hydrophilic uncharged or negatively charged polymer regulates the interaction between phenolic hydroxyl groups and proteins.
  • Polyphenols/hydrophilic uncharged or negatively charged polymers form a procoagulant coating, which can be combined with existing hemostatic devices (such as gelatin sponges, gauze, embolism spring coils, etc.) to enhance their procoagulability without changing their original application scenarios. Coagulation properties.
  • the ratio of polyphenols to polymers and the processing time can be adjusted to form procoagulant coatings on different substrates. Through mild plasma protein adhesion, the coating changes key coagulation proteins into an activated conformation and promotes platelet adhesion, thereby achieving Optimal procoagulant properties.
  • the electropositive properties of positively charged polymers have a strong attraction to negatively charged blood cells in the blood. If positively charged polymers are used for co-deposition (regardless of the polyphenol/polymer ratio and processing time), it will Compared with the strong interaction of pure polyphenol coatings on proteins, the formed polyphenol/positively charged polymer coatings will not significantly reduce the strong interaction of the protein, and may even enhance it. Therefore, the presence of these two strong interactions will inhibit the transformation of plasma proteins into an effective conformation that promotes coagulation, making it impossible to effectively adhere to platelets, causing the surface to lose its procoagulant effect. Therefore, the present invention can only use hydrophilic uncharged or negatively charged polymer phases, but cannot use positively charged polymers.
  • the second use of the present invention is to use polyphenolic hydroxyl substances and hydrophilic uncharged or negatively charged polymers to form a coagulation-promoting coating.
  • polyphenolic hydroxyl substances Through the universal surface functionalization ability of polyphenolic hydroxyl substances, various medical materials can be surface-functionalized. Modification, by adjusting the ratio of polyphenols to polymers and the treatment time, can construct surfaces with differential protein adhesion on different types of substrates to achieve differential adhesion to different proteins.
  • the preparation and performance testing of hemostatic functional materials are Examples 1-8, Comparative Examples 1-13, and Test Examples 1-3.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5.
  • the polyphenolic hydroxyl material is tannic acid
  • the hydrophilic uncharged polymer is polyethylene glycol
  • the mass ratio is 1:3.
  • the polyphenolic hydroxyl material is gallic acid
  • the hydrophilic uncharged polymer is hydroxypropyl cellulose
  • the mass ratio is 1:2.
  • the polyphenolic hydroxyl material is gallic acid
  • the hydrophilic and electronegative polymer is carboxymethyl cellulose
  • the mass ratio is 1:5.
  • Tris buffer solution (10mM) to prepare a 1 mg/mL dopamine hydrochloride solution, adjust the pH to 8.5, soak the medical experimental titanium alloy disc in the solution for 12 hours at 25°C, and wash it three times with a large amount of deionized water;
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5
  • the soaking time is 1 hour.
  • the surface of the titanium alloy disc is not pretreated with polyphenols (that is, step 1) of Example 6 is not used, and step 2) is directly used for preparation)
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:10.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:1.
  • the polyphenolic hydroxyl substance is dopamine, and no polymer component is added.
  • the polyphenolic hydroxyl substance is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5
  • the soaking time is 5 hours.
  • the polyphenolic hydroxyl substance is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5
  • the soaking time is 0.5h.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic electropositive polymer is quaternized pullulan, and the mass ratio is 1:5.
  • the quaternized pullulan in this example is prepared as follows: weigh 1.8g sodium hydroxide and 6g 2,3-epoxypropyltrimethylammonium chloride (GTA) and dissolve it in 50mL deionized water. 6g of amylopectin was dispersed in 200 mL of deionized water to form a dispersion. Then, a mixed solution of NaOH and GTA was slowly added dropwise to the starch dispersion, and the reaction was stirred at 25°C for 24 hours. After the reaction is completed, the reaction solution is poured into a dialysis bag (MWCO, 1000Da) and dialyzed with deionized water for 2 days, changing the water at least 8 times. After the dialysis is completed, the solution in the dialysis bag is freeze-dried to obtain quaternized pullulan. .
  • GTA 2,3-epoxypropyltrimethylammonium chloride
  • the polyphenolic hydroxyl material is gallic acid
  • the hydrophilic electropositive polymer is polylysine
  • the mass ratio is 1:5.
  • Tris buffer solution (10mM) to prepare a 1 mg/mL dopamine hydrochloride solution, adjust the pH to 8.5, soak the medical experimental titanium alloy disc in the solution for 12 hours at 25°C, and wash it three times with a large amount of deionized water to remove unformed coating. layer of dopamine and dextran, and dried with nitrogen to obtain Ti3.
  • the surface of the titanium alloy disc was only pretreated with polyphenol (only step 1) of Example 6 was performed), and was not coated with a polyphenol/polymer hemostatic and procoagulant coating (Example 6 was not performed). Step 2)).
  • Test method Weigh 3mg of gelatin sponge material, cut gauze material into four layers of 0.5 ⁇ 0.5cm, weigh 5mg of gelatin sponge particle material, and take a piece of medical experimental titanium alloy disc with a diameter of 10mm. Take 100 ⁇ L of fresh anticoagulated blood and mix thoroughly with 10 ⁇ L of 0.2M CaCl2 solution, quickly add it to the material, and place the tube in a constant temperature water bath at 37°C and incubate for 1 minute. Then use 10 mL of deionized water to fully lyse the excess blood that has not formed a blood clot, and place it in a constant temperature water bath at 37°C for 3 minutes.
  • Blood coagulation index % (BCI) (Abs sample /Abs blank ) ⁇ 100%;
  • Abs sample is the absorbance at 545nm of the examples and comparative examples
  • Abs blank is the absorbance of the blank group at 545nm.
  • BCI Bood clotting index
  • Table 1 the BCI index of the materials coated with polyphenol/polymer accelerating coating obtained in Examples 1 to 6 of the present invention is only 16.3% and the highest is 44.8%, indicating that polyphenol/polymer accelerating Materials modified with coagulation coating have better procoagulant properties. It can be seen that by using the preparation method of the present invention, adding uncharged or negatively charged polymer co-deposition to polyphenols can obtain a polyphenol/polymer procoagulant coating that improves the procoagulant effect of the material.
  • the coating can be obtained by adding uncharged Or electronegative polymers regulate the interaction between polyphenol groups and proteins, which can gently adhere to proteins and adjust the conformation of key plasma proteins, thereby promoting platelet adhesion to achieve hemostasis.
  • uncharged Or electronegative polymers regulate the interaction between polyphenol groups and proteins, which can gently adhere to proteins and adjust the conformation of key plasma proteins, thereby promoting platelet adhesion to achieve hemostasis.
  • Example 1 In Comparative Example 1, the amount of dextran added in Example 1 was increased from 5 mg/mL to 10 mg/mL.
  • the BCI of Example 1 was 43.6%, and the BCI of Comparative Example 1 was 57.6%.
  • the results show that the polyphenol/polymer composite coating material obtained by increasing the amount of dextran has a higher BCI and a poor procoagulant effect. Because the increase in the glucan content in the composite coating will enhance the shielding effect on the strong interaction between polyphenol groups and coagulation proteins, the effective interaction between the surface and the protein will be weak, making it difficult to adhere to platelets, thereby affecting the coagulation process.
  • Example 2 the amount of dextran added in Example 1 was reduced from 5 mg/mL to 1 mg/mL.
  • the BCI of Example 1 was 43.6%, and the BCI of Comparative Example 2 was 75.1%.
  • the results show that the polyphenol/polymer composite polymer material obtained by reducing the amount of dextran added has a higher BCI and a poor procoagulant effect. Because the reduction of dextran content in the composite coating will weaken the shielding effect of the strong interaction between polyphenol groups and coagulation proteins, making the protein difficult to activate, resulting in the surface being unable to effectively adhere to platelets, thereby affecting coagulation.
  • Comparative Example 3 removes the uncharged polymer in Example 1 and directly forms a polyphenol coating.
  • the BCI of Example 1 is 43.6%, and the BCI of Comparative Example 3 is 77.1%.
  • the results show that when only polyphenols are used for coating without uncharged polymer coating, the BCI of the coating material is high and the blood-promoting effect is not good. Because the phenolic hydroxyl group in the pure polyphenol coating interacts too strongly with coagulation proteins, inhibiting its activation and making it unable to transform into a conformation that effectively adheres to platelets, inhibiting platelet adhesion and thus affecting coagulation.
  • Comparative Example 4 extends the coating coating time in Example 1 from 1 hour to 5 hours.
  • the BCI of Example 1 is 43.6%, and the BCI of Comparative Example 4 is 57.7%.
  • the results show that the polyphenol/polymer composite coating material obtained by extending the coating coating time has a higher BCI and a poor procoagulant effect. Because extending the coating coating time will cause the dopamine self-polymerization coating to thicken, the polymer may be wrapped by thicker polydopamine, which will not be able to regulate the surface properties of the coating through anti-protein adhesion.
  • Example 5 the coating coating time in Example 1 was reduced from 1 h to 0.5 h.
  • the BCI of Example 1 was 43.6%, and the BCI of Comparative Example 5 was 64.7%.
  • the results show that the polyphenol/polymer composite coating material obtained by reducing the coating coating time has a higher BCI and has a poor procoagulant effect. Because reducing the coating coating time may result in the inability to form a uniform and complete coating on the surface of the material, limiting the coagulation-promoting effect of the coating.
  • Comparative Example 6 replaces the uncharged polymer (dextran) in Example 1 with a positively charged polymer (quaternized pullulan).
  • the BCI of Example 1 is 43.6%
  • the BCI of Comparative Example 6 is 59.3%.
  • the results show that when coating with electropositive polymers, the resulting material has a higher BCI and poor blood-promoting effect.
  • positively charged polymers attract negatively charged blood cells and platelets in the blood
  • the uncharged polymer component with anti-protein adhesion with positively charged polymers the polyphenols in the coating have a strong effect on proteins. Effective shielding cannot be obtained, and the addition of a large amount of positively charged polymers will also promote strong direct interactions between the material surface and blood components. In the presence of two strong interactions, it will inhibit the transformation of plasma proteins into an effective conformation that promotes coagulation, making it impossible to effectively adhere to platelets, causing the surface to lose its procoagulant effect.
  • Comparative Example 7 replaces the electronegative polymer (carboxymethylcellulose) in Example 5 with a positive polymer (polylysine).
  • the BCI of Example 5 was 42.9%, and the BCI of Comparative Example 7 was 62.7%.
  • the results show that when coating with electropositive polymers, the resulting material has a higher BCI and poor blood-promoting effect. Because positively charged polymers have a certain attraction to cells in the blood, after replacing negatively charged polymers with anti-biological adhesion with positively charged polymers, the strong force of polyphenols in the coating on proteins cannot be effectively controlled.
  • Shielding will inhibit the transformation of plasma proteins into an effective conformation that promotes coagulation, and the addition of a large number of positively charged polymers will also promote a strong direct interaction between the material surface and blood components. This strong interaction will instead inhibit the coagulation process. In the presence of two strong interactions, the surface cannot effectively adhere to platelets, thus losing its procoagulant effect.
  • Comparative Example 8 is a material corresponding to Example 6 in which the metal surface was only pretreated with polyphenol and was not coated with a polyphenol/polymer composite coating.
  • the BCI of Example 6 was 44.8%, and the BCI of Comparative Example 8 was 61.0%.
  • the BCI results show that the BCI of materials that only undergo long-term polyphenol pretreatment is higher. Since the pretreated surface is composed of polydopamine, the interaction between the phenolic hydroxyl groups and blood coagulation proteins in the pure polyphenol coating is too strong, inhibiting its Activation prevents it from transforming into a conformation that effectively adheres to platelets, inhibiting platelet adhesion, thereby affecting coagulation.
  • Comparative Examples 9 to 13 correspond to the base materials without any modification in Examples 1 to 7.
  • the BCI of the gelatin sponge is 83.7%, and the BCI after modification is 43.6%; the BCI of the gelatin sponge particles is 41.9%, and the BCI after modification is 16.6%, the BCI of commercially available gauze is 70.4%, the modified BCI is 34.2%, the BCI of medical experimental titanium alloy discs is 57.3%, the modified BCI is 44.8%, the BCI of medical glass sheets is 71.7%, modified Post-sex BCI was 37.5%. It can be seen that after coating the substrate with the polyphenol/polymer coating of the present invention, the BCI index is significantly reduced and the procoagulant effect is significantly enhanced.
  • Test method Weigh 3 mg of gelatin sponge material and 5 mg of gelatin sponge particle material, and put them into 2 mL plastic centrifuge tubes respectively. Centrifuge the fresh sodium citrate anticoagulated blood taken from the rat heart at 150g for 10 minutes, collect the supernatant PRP, and dilute it with PBS to the volume of the original blood. Add 100 ⁇ L of diluted PRP to the material and place it in a constant temperature water bath at 37°C and incubate for 5 minutes. After the incubation is completed, wash the material with PBS for a total of three times, using 100 ⁇ L PBS each time to remove unadhered platelets on the surface. Collect the washing solution and dilute it to 1 mL with PBS.
  • the LDH content sample is the LDH content in the lysate measured by the LDH kit in the examples and comparative examples
  • the LDH content blank is the LDH content in the lysate measured by the LDH kit in the blank group.
  • the platelet adhesion test can characterize the material's adhesion effect on platelets. Generally, the higher the material's adhesion rate to platelets, the better the material's procoagulant effect.
  • Example 1 In Comparative Example 1, the amount of dextran added in Example 1 was increased from 5 mg/mL to 10 mg/mL. The platelet adhesion rate of Example 1 was 44.1%, and the platelet adhesion rate of Comparative Example 1 was 39.5%. The results show that the platelet adhesion rate of the polyphenol/polymer composite coating obtained by increasing the amount of dextran is relatively low, which corresponds to a higher BCI and poor procoagulation effect. Because the increase in the glucan content in the composite coating will enhance the shielding effect on the strong interaction between polyphenol groups and coagulation proteins, the effective interaction between the surface and the protein will be weaker, and fewer platelets will adhere.
  • Example 2 the amount of dextran added in Example 1 was reduced from 5 mg/mL to 1 mg/mL.
  • the platelet adhesion rate of Example 1 was 44.1%, and the platelet adhesion rate of Comparative Example 2 was 33.9%.
  • the results show that the polyphenol/polymer composite coating obtained by reducing the amount of dextran has a lower platelet adhesion rate, which corresponds to a higher BCI and poor procoagulation effect. Because the reduction of the glucan content in the composite coating will weaken the shielding effect of the strong interaction between the polyphenol group and the coagulation protein, making it difficult for the protein to activate and form a protein conformation that is conducive to platelet adhesion, making it difficult to adhere to platelets.
  • Example 3 the uncharged polyhydroxy polymer (dextran) in Example 1 was removed.
  • the platelet adhesion rate of Example 1 was 44.1%, and the platelet adhesion rate of Comparative Example 3 was 24.3%.
  • the results show that when only polyphenols are used for coating without adding the uncharged polymer (dextran) component, the platelet adhesion rate of the coated material is low, corresponding to a high BCI and poor procoagulation effect. Because the polyphenol coating is directly formed, the interaction between the phenolic hydroxyl groups in the pure polyphenol coating and the coagulation protein is too strong, inhibiting its activation and preventing the formation of a protein conformation that is conducive to platelet adhesion.
  • Comparative Examples 9 and 10 correspond to the base materials without any modification in Examples 1 and 2.
  • the platelet adhesion rate of gelatin sponge is 23.1%, and the platelet adhesion rate after modification is 44.1%; the platelet adhesion of gelatin sponge particles The rate was 20.4%, and the platelet adhesion rate after modification was 37.1%.
  • the results show that the present invention can increase the platelet adhesion rate by coating commercially available hemostatic materials with a polyphenol/polymer procoagulant coating, thereby improving the procoagulant performance.
  • sample GS1 (Implementation GS6 (right GS comparison
  • the rat femoral artery hemostasis test simulates and evaluates the hemostatic effect of the material in real application scenarios by measuring the hemostatic time and blood loss when the material is used to treat the injured location in the rat femoral artery injury model.
  • the better the procoagulant effect of the material the better the hemostatic effect.
  • Example 3 the hydrophilic uncharged polymer (dextran) in Example 1 was removed and a polyphenol coating was directly formed.
  • the blood loss in Example 1 was (33.9 mg) which was less than that in Comparative Example 3 ( 146.2 mg), the hemostatic time of Example 1 (2 min) was shorter than the hemostatic time of Comparative Example 3 (13.3 min).
  • Comparative Example 10 corresponds to the base material without any modification in Example 1.
  • the blood loss of the commercial gelatin sponge 353.1 mg
  • the blood loss of Example 1 (33.9 mg)
  • the hemostasis time of the commercial gelatin sponge is 7.5 minutes.
  • the hemostatic time is longer than that of Example 1 (2 minutes).
  • results show that after the medical substrate is coated according to the present invention, it has excellent and sufficient procoagulant ability, has small blood loss and short hemostasis time in the rat femoral artery injury model, and can play a better role in injury. Hemostatic effect.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:10.
  • the polyphenolic hydroxyl material is dopamine
  • the hydrophilic uncharged polymer is dextran
  • the mass ratio is 1:5.
  • the polyphenolic hydroxyl substance is dopamine, and no polymer component is added.
  • Unmodified glass sheet GL Unmodified glass sheet GL.
  • Test method Cut the glass sheet material into 1 ⁇ 1cm size and place it in a 24-well plate for testing. Weigh 3mg of the gelatin sponge material and place it into a 2mL centrifuge tube for testing. Taking human fibrin as an example, prepare human fibrinogen into a 10 mg/mL physiological saline solution, add 50 ⁇ L of fibrinogen solution to the material, place the material in a constant temperature water bath at 37°C and incubate it with a glass slide. Incubate the materials for 30 minutes and the gelfoam materials for 5 minutes.
  • the protein content sample is the protein content in the washing liquid measured by the BCA kit in the examples and comparative examples
  • the protein content blank is the protein content in the dilution liquid measured by the BCA kit in the blank group.
  • the protein adhesion rate is an important indicator to characterize the protein adhesion on the material surface. The greater the adhesion percentage, the better the protein adhesion performance of the material.
  • Example 9 of the present invention has high adhesion to fibrinogen and medium adhesion to albumin;
  • Example 10 has Fibrinogen has ultra-high adhesion and globulin has moderate adhesion.
  • Comparative Example 14 removes the uncharged polymer in Example 9 and directly forms a polyphenol coating. Compared with Comparative Example 15 (unmodified glass sheet), the adhesion rate to both proteins is increased (from 6.2 to 6.2). % to 8.1%, 3.1% to 6.6%), indicating that the pure polyphenol coating can improve adhesion to proteins broadly and has no selectivity.
  • Example 9 when dextran is added to form a polyphenol/polymer composite coating, the adhesion rate of Example 9 to albumin remains basically unchanged (the adhesion rate ranges from 3.1% to 3.2% ), the adhesion rate to fibrinogen became higher (the adhesion rate increased from 6.2% to 8.2%). It can be seen that the coating-modified glass sheet prepared in the present invention can achieve differential adhesion to proteins. Differential adhesion.
  • Example 10 and Comparative Example 16 unmodified gelatin sponge
  • the modified gelatin sponge has a higher adhesion rate to fibrinogen.
  • the adhesion rate became higher (the adhesion rate increased from 6.4% to 17.2%), and the adhesion rate to albumin decreased (the adhesion rate decreased from 6.6% to 3.1%).
  • the coated modified sponge prepared in the present invention can achieve differential adhesion to proteins.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Organic Chemistry (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Surgery (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Materials For Medical Uses (AREA)

Abstract

一种多酚-聚合物复合涂层的制备方法，步骤为：1)将多酚羟基类物质和亲水不带电或亲水带负电聚合物配制成混合溶液；2)将基材在所述的混合溶液中浸泡；3)洗涤，干燥。该多酚-聚合物复合涂层能够在医用材料功能化中应用；所述的功能化为止血功能和/或蛋白差异化黏附功能。实现止血功能条件为：多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:2～8和浸泡时间为0.7～4小时，得到具备止血功能的医用材料。实现蛋白差异化黏附条件为：多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:0.5～15和浸泡时间为0.5～20小时，得到具备蛋白差异化黏附的医用材料。

Description

一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用 技术领域
本发明属于医用止血材料领域，涉及一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用。
背景技术
不受控制的出血是交通事故、外科手术和自然灾害中造成伤亡的重要原因。目前常用的止血材料/医疗器械存在性能不足的缺陷，因此开发高效安全的新型止血材料十分重要。通过构建新型促凝血涂层对现有医用止血材料/器械进行表面改性，可以在不改变现有止血材料/器械的应用场景基础上，提高其止血性能、以更好地满足临床需求。在目前表面材料抗凝血性相关研究中(Acta Biomaterialia,2017,64:187-199)发现，当材料表面对蛋白的黏附能力过强时，会和蛋白产生较为强烈的相互作用，在表面与血液接触的过程中，这种较强的黏附作用不利于纤维蛋白原向其有效构象发生转变，进而抑制血小板的黏附与激活，导致抗凝。因此，有必要开发具有可控/温和的蛋白黏附能力的促凝涂层，通过促进关键凝血蛋白转化为有效构象，从而促进血小板的黏附与激活，最终促进凝血。将多酚类物质与多羟基聚合物共沉积制备复合止血涂层，这一技术国内未见报道。
材料在与生理环境接触时总是立即被蛋白质覆盖，这是生物学中的普遍规则。在生物医用材料植入体内后，生物材料表面会在几秒内迅速与蛋白质发生吸附，然后再与生物体内其他物质接触，并且材料表面对蛋白黏附作用也会影响其对体内其他物质的作用。通过构建蛋白差异化吸附涂层对现有生物医用材料/器械进行表面改性，可以调控其对蛋白的黏附作用，通过简便的制备方法，满足不同的临床需求。因此，有必要开发具有可控的蛋白黏附能力的涂层，通过不同蛋白质的差异化黏附，从而更进一步指导调控生物医用表面与细胞、组织与器官层面上的相互作用。
发明内容
有鉴于此，本发明提供一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用。本发明具体提供了如下的技术方案：
一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，步骤为：
1)将多酚羟基类物质和亲水不带电或亲水带负电聚合物配制成混合溶液；
2)将基材在所述的混合溶液中浸泡；
3)洗涤，干燥，得到功能化医用材料；
所述的功能化为止血功能和/或蛋白差异化黏附功能；
实现止血功能条件：步骤1)中的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:2～8和步骤2)中的浸泡时间为0.7～4小时，步骤3)得到具备止血功能的医用材料；
实现蛋白差异化黏附条件：步骤1)中的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:0.5～15和步骤2)中的浸泡时间为0.5～20小时，步骤3)得到具备蛋白差异化黏附的医用材料；
步骤1)所述的多酚羟基类物质为多巴胺、单宁酸、没食子酸、儿茶素、表儿茶素、表没食子儿茶素没食子酸酯、茶黄素-3-没食子酸酯、5-羟基多巴胺盐酸盐或黄芩素，步骤1)所述的亲水不带电聚合物为葡聚糖、羟丙基纤维素、羟乙基淀粉、羟乙基纤维素、普罗兰多糖、普兰尼克或聚乙二醇，步骤1)所述的亲水负电聚合物为羧甲基纤维素或海藻酸、海藻酸钠、透明质酸、透明质酸钠或羧甲基淀粉。
进一步，当步骤1)的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:2～5，步骤2)所述的浸泡时间为0.7～1.5小时，步骤3)得到具备止血功能的医用材料。
进一步，当步骤1)的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:5～10，步骤2)的浸泡时间为1～5小时，步骤3)得到的是具备蛋白差异化黏附的医用材料，所述的蛋白为纤维蛋白原、白蛋白、脂蛋白、糖蛋白或球蛋白。
进一步，步骤1)所述的多酚羟基类物质为多巴胺、单宁酸或没食子酸，步骤1)所述的亲水不带电聚合物为葡聚糖、羟丙基纤维素或聚乙二醇，步骤1)所述的亲水负电聚合物为羧甲基纤维素。
进一步，步骤1)所述的多酚羟基类物质在混合溶液中的浓度为0.25～3mg/mL。
进一步，止血功能的实现条件为：亲水不带电或亲水负电聚合在混合溶液中的浓度为3～5mg/mL，蛋白差异化黏附的实现条件为：亲水不带电或亲水负电聚合在混合溶液中的浓度为5～10mg/mL。
进一步，步骤2)所述的基材为高分子医用材料、无机类医用材料、金属类医用材料；所述的高分子医用材料为纱布、聚乙烯醇海绵、聚乙烯醇微米级颗粒、聚乙烯醇微球、壳聚糖海绵、壳聚糖无纺布、明胶海绵、明胶海绵微米级颗粒、海藻酸敷料、海藻酸微球、海藻酸无纺布或胶原蛋白海绵；所述的无机类医用材料是生物玻璃、生物陶瓷；所述的金属类医用材料是镍钛合金器械、钛合金医用器械、形状记忆金属微弹簧圈。
进一步，步骤2)所述的基材为纱布、聚乙烯醇海绵、聚乙烯醇微米级颗粒、聚乙烯醇微球、明胶海绵、明胶海绵微米级颗粒、胶原蛋白海绵、生物玻璃或钛合金医用器械。
进一步，步骤2)所述基材为金属类医用材料时，基材在浸泡混合溶液前可以用多酚羟基类物质进行预处理，在基材表面形成多酚涂层，预处理方法为：将基材在多酚羟基类物质的pH为8～9的碱性水溶液中浸泡4～24小时。
进一步，步骤1)所述的混合溶液为碱性水溶液，pH为8～9。
本发明的有益效果在于：
本发明的第一种用途是利用多酚羟基物质的氧化自聚与亲水不带电或负电的聚合物相结合，在基材表面形成涂层。多酚羟基物质与蛋白之间具有强的氢键相互作用，在复合涂层中它提供对血液中凝血关键成分如血浆蛋白(纤维蛋白原等)的作用力，使之黏附聚集；具有抗蛋白黏附作用的亲水不带电或负电聚合物在与多酚羟基物质共沉积的过程中，调控了酚羟基与蛋白间的作用，通过削弱酚羟基和血浆中关键凝血蛋白的氢键等作用力，实现“温和”/中等强度的蛋白黏附，使蛋白转变为易黏附血小板的构象，促进血小板黏附，进而达到促凝血的效果。
多酚/亲水不带电或负电聚合物形成促凝涂层，可与现有止血器械(如明胶海绵、纱布、栓塞弹簧圈等)复合，在不改变其原有应用场景的同时增强其促凝血性能。可调节多酚与聚合物的比例与处理时间，在不同基材上形成促凝涂层，该涂层通过温和的血浆蛋白黏附，使关键凝血蛋白转变为激活构象，促进血小板的黏附，从而达到最佳的促凝血性能。
而正电性聚合物的正电性对血液中的负电的血细胞具有很强的吸引作用，若使用正电性聚合物进行共沉积(无论何种多酚/聚合物比例与处理时间)，相比于纯多酚涂层对蛋白的强作用力，形成的多酚/正电性聚合物涂层都不会明显降低该蛋白强相互作用、甚至有可能增强。因此，此两种强相互作用存在的情况下，反而会抑制血浆蛋白向促进凝血的有效构象转变，无法有效黏附血小板，使表面失去促凝血作用。因此本发明只能采 用亲水不带电或负电的聚合物相，而不能使用正电性聚合物。
本发明的第二种用途是利用多酚羟基物质与亲水性不带电或负电性聚合物形成促凝涂层，通过多酚羟基物质通用的表面功能化能力，可对多种医用材进行表面改性，通过调节多酚与聚合物的比例与处理时间，可以在不同类型的基材上构建具有差异化蛋白黏附的表面，实现对不同蛋白的差异化黏附。
具体实施方式
下面对本发明的优选实施例进行详细的描述。
止血功能材料的制备和性能检测是实施例1-8，对比例1-13，测试例1-3。
蛋白差异化黏附功能材料的制备和性能检测是实施例9-10，对比例14-16，测试例4。
实施例1
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS1。
本实施例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5。
实施例2
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵微米级颗粒浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GSP1。
本实施例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5。
实施例3
将单宁酸50mg，聚乙二醇150mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵加入该溶液中，25℃浸泡1h，用大量去离子水洗涤三次，除去未形成涂层的单宁酸与聚乙二醇，干燥得到GS2。
本实施例中，多酚羟基类物质为单宁酸，亲水不带电聚合物为聚乙二醇，质量比为1:3。
实施例4
将没食子酸75mg，羟丙基纤维素150mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售纱布浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的没食子酸与羟丙基纤维素，冷冻干燥得到G1。
本实施例中，多酚羟基类物质为没食子酸，亲水不带电聚合物为羟丙基纤维素，质量比为1:2。
实施例5
将没食子酸50mg，羧甲基纤维素250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售纱布浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的没食子酸与羧甲基纤维素，冷冻干燥得到G2。
本实施例中，多酚羟基类物质为没食子酸，亲水负电性聚合物为羧甲基纤维素，质量比为1:5。
实施例6
1)用Tris缓冲溶液(10mM)配制1mg/mL的盐酸多巴胺溶液，调节pH为8.5，将医用实验钛合金圆片在25℃下在溶液中浸泡12h，使用大量去离子水洗涤三次；
2)将盐酸多巴胺5mg，葡聚糖25mg溶于5mL Tris缓冲溶液(10mM)中，将1)中处理后的钛合金圆片浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，氮气吹干得到Ti1。
本实施例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5。
实施例7
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将医用玻璃片浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，氮气吹干得到GL1。
本实施例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5，浸泡时间1h。
实施例8
将盐酸多巴胺5mg，葡聚糖25mg溶于5mL Tris缓冲溶液(10mM)中，将医用实验钛合金圆片浸泡在该混合溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，氮气吹干得到Ti2。
本对比例中，钛合金圆片表面不进行多酚预处理(即不采用实施例6的步骤1)，直接采用步骤2)进行制备)
对比例1
将盐酸多巴胺50mg，葡聚糖500mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS3。
本对比例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:10。
对比例2
将盐酸多巴胺50mg，葡聚糖50mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS4。
本对比例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:1。
对比例3
将盐酸多巴胺50mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS5。
本对比例中，多酚羟基类物质为多巴胺，不添加聚合物组分。
对比例4
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡5h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS6。
本对比例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5，浸泡时间为5h。
对比例5
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡0.5h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS7。
本对比例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5，浸泡时间为0.5h。
对比例6
将盐酸多巴胺50mg，季铵化支链淀粉250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡1h后取出，使用大量 去离子水洗涤三次，除去未形成涂层的多巴胺与季铵化支链淀粉，冷冻干燥得到GS8。
本对比例中，多酚羟基类物质为多巴胺，亲水正电性聚合物为季铵化支链淀粉，质量比为1:5。
本实施例子中的季铵化支链淀粉按照如下方法制备得到：称取1.8g氢氧化钠和6g2,3-环氧丙基三甲基氯化铵(GTA)溶于50mL去离子水中，将6g支链淀粉分散在200mL去离子水中形成分散液，随后，将NaOH与GTA的混合溶液缓缓滴加到淀粉分散液中，25℃下搅拌反应24h。反应完成后，将反应溶液倒入透析袋(MWCO,1000Da)中使用去离子水透析2天，至少换水8次，透析完成后，将透析袋内溶液冻干，得到季铵化支链淀粉。
对比例7
将没食子酸50mg，聚赖氨酸250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售纱布浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的没食子酸与聚赖氨酸，冷冻干燥得到G3。
本对比例中，多酚羟基类物质为没食子酸，亲水正电性聚合物为聚赖氨酸，质量比为1:5。
对比例8
用Tris缓冲溶液(10mM)配制1mg/mL的盐酸多巴胺溶液，调节pH为8.5，将医用实验钛合金圆片在25℃下在溶液中浸泡12h，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，氮气吹干得到Ti3。
本对比例中，钛合金圆片表面仅用多酚进行预处理(只做了实施例6的步骤1))，不用多酚/聚合物止血促凝涂层进行涂覆(不做实施例6的步骤2))。
对比例9
市售明胶海绵GS(未作任何改性)。
对比例10
市售明胶海绵颗粒GSP(未作任何改性)。
对比例11
市售纱布G(未作任何改性)。
对比例12
医用实验钛合金圆片Ti(未作任何改性)。
对比例13
医用玻璃片(未作任何改性)。
测试例1体外凝血效果测试
检测条件：将各实验例、对比例得到的材料进行凝血效果对比实验，检测用血液为取自大鼠心脏的新鲜柠檬酸钠抗凝血液。
测试方法：将明胶海绵材料称取3mg，纱布材料裁剪四层0.5×0.5cm，明胶海绵颗粒材料称取5mg，医用实验钛合金圆片取直径为10mm的一片。取100μL的新鲜抗凝血与10μL的0.2M CaCl 2溶液充分混合后，迅速加在材料上，将其管置于37℃的恒温水浴锅中孵育1分钟。接着用10mL去离子水将未形成血凝块的多余血液充分裂解，置于37℃的恒温水浴锅中孵育3分钟。吸取1mL裂解后的液体离心(2500rpm，3分钟)。取100μL离心后的上清液加入96孔板，用酶联免疫检测仪测试545nm处的吸光度(Abs)。空白组是将100μL新鲜抗凝血加入10mL去离子水，37℃的恒温水浴锅中孵育3分钟，取100μL加入96孔板，测试545nm处的吸光度Abs。最后通过以下公式来计算凝血指数(BCI)。
凝血指数％(BCI)＝(Abs 样品/Abs 空白)×100％...................式1
式中：Abs 样品是实施例与对比例在545nm处的吸光度；Abs 空白是空白组在545nm处的吸光度。
表1体外凝血效果测试
Figure PCTCN2022116742-appb-000001
BCI(Blood clotting index，凝血指数)，可以表征材料的凝血效果，通常BCI的数值越小，表示材料的凝血效果越好。从表1可以看出，本发明实施例1～6得到的包覆多酚/聚合物促凝涂层的材料的BCI指数最低的只有16.3％，最高的44.8％，说明多酚/聚合物促凝涂层改性的材料具有更优异的促凝血性能。由此可见，采用本发明的制备方法，向多酚中加入不带电或负电性聚合物共沉积可以得到提升材料促凝血效果的多酚/聚合 物促凝涂层，该涂层通过加入不带电或负电性聚合物调节多酚基团与蛋白的相互作用，可以温和地黏附蛋白并调节关键血浆蛋白的构象，从而促进血小板黏附以达到止血的目的。而对比例止血效果不好的原因如下：
对比例1是将实施例1中的葡聚糖添加量由5mg/mL提高为10mg/mL，实施例1的BCI为43.6％，对比例1的BCI为57.6％。结果表明，提高葡聚糖添加量得到的多酚/聚合物复合涂层的材料BCI较高，促凝血效果不好。因为复合涂层中葡聚糖含量的提高会增强对于多酚基团与凝血蛋白强作用力的屏蔽作用，使表面与蛋白产生有效相互作用偏弱，难以黏附血小板，从而影响凝血过程。
对比例2是将实施例1中的葡聚糖添加量由5mg/mL降低为1mg/mL，实施例1的BCI为43.6％，对比例2的BCI为75.1％。结果表明，降低葡聚糖添加量得到的多酚/聚合物复合聚合物的材料BCI较高，促凝血效果不好。因为复合涂层中葡聚糖的含量的降低会减弱对于多酚基团与凝血蛋白强作用力的屏蔽作用，使蛋白不易活化，导致表面无法有效黏附血小板，进而影响凝血。
对比例3是将实施例1中的不带电聚合物去掉，直接形成多酚涂层，实施例1的BCI为43.6％，对比例3的BCI为77.1％。结果表明，仅使用多酚进行包覆，不用不带电聚合物包覆，得到涂层的材料BCI较高，促凝血效果不好。因为纯多酚涂层中的酚羟基和凝血蛋白相互作用力过强，抑制其活化，使其无法向有效黏附血小板的构象转变，抑制了血小板黏附，进而影响凝血。
对比例4是将实施例1中的涂层包覆时间由1h延长为5h，实施例1的BCI为43.6％，对比例4的BCI为57.7％。结果表明，延长涂层包覆时间得到的多酚/聚合物复合涂层的材料BCI较高，促凝血效果不好。因为延长涂层包覆的时间会导致多巴胺自聚的涂层增厚，聚合物可能会被更厚的聚多巴胺包裹起来，无法起到通过抗蛋白黏附作用对涂层表面性能进行调控。
对比例5是将实施例1中的涂层包覆时间由1h降低为0.5h，实施例1的BCI为43.6％，对比例5的BCI为64.7％。结果表明，降低涂层包覆时间得到的多酚/聚合物复合涂层的材料BCI较高，促凝血效果不好。因为降低涂层包覆的时间可能会导致材料表面不能形成均匀完整的涂层，限制了该涂层促凝效果的发挥。
对比例6是将实施例1中的不带电聚合物(葡聚糖)替换为正电性聚合物(季铵化支链淀粉)，实施例1的BCI为43.6％，对比例6的BCI为59.3％。结果表明，采用正 电性聚合物进行涂覆，得到的材料BCI较高，促凝血效果不好。因为正电性聚合物对血液中的负电的血细胞与血小板具有吸引作用，将具有抗蛋白黏附的不带电聚合物组分替换成正电性聚合物后，涂层中多酚对蛋白的强作用力无法得到有效的屏蔽，且大量正电性聚合物的加入也会促使材料表面与血液组分直接产生强相互作用。两种强相互作用存在的情况下，反而会抑制血浆蛋白向促进凝血的有效构象转变，无法有效黏附血小板，使表面失去促凝血作用。
对比例7是将实施例5中的负电性聚合物(羧甲基纤维素)替换为正电性聚合物(聚赖氨酸)。实施例5的BCI为42.9％，对比例7的BCI为62.7％。结果表明，采用正电性聚合物进行涂覆，得到的材料BCI较高，促凝血效果不好。因为正电性聚合物对血液中的细胞具有一定的吸引作用，将具有抗生物黏附的负电性聚合物替换成正电性聚合物后，涂层中多酚对蛋白的强作用力无法得到有效的屏蔽，会抑制血浆蛋白向促进凝血的有效构象转变，且大量正电性聚合物的加入也会促使材料表面与血液组分直接产生强相互作用，这种强相互作用反而会抑制凝血过程。两种强相互作用存在的情况下，该表面无法有效黏附血小板，从而失去促凝血作用。
对比例8是对应实施例6中对金属表面仅进行多酚预处理，不进行多酚/聚合物复合涂层的包覆的材料。实施例6的BCI为44.8％，对比例8的BCI为61.0％。BCI结果表明，仅进行长时间多酚预处理的材料的BCI较高，由于预处理后的表面由聚多巴胺组成，纯多酚涂层中的酚羟基和凝血蛋白相互作用力过强，抑制其活化，使其无法向有效黏附血小板的构象转变，抑制了血小板黏附，进而影响凝血。
对比例9～13对应实施例1-7中不做任何改性的基材，明胶海绵的BCI为83.7％，改性后为43.6％；明胶海绵颗粒的BCI为41.9％，改性后BCI为16.6％，市售纱布的BCI为70.4％，改性后BCI为34.2％，医用实验钛合金圆片的BCI为57.3％，改性后BCI为44.8％，医用玻璃片的BCI为71.7％，改性后BCI为37.5％。由此可见，采用本发明的多酚/聚合物涂层对基材进行包覆后，BCI指数显著降低，促凝血效果明显增强。
测试例2体外血小板黏附效果测试
检测条件：将各实验例、对比例得到的材料进行体外血小板黏附效果测试，检测用血液为取自大鼠心脏的新鲜柠檬酸钠抗凝血液，检测使用乳酸脱氢酶(LDH)试剂盒。
测试方法：将明胶海绵材料称取3mg，明胶海绵颗粒材料称取5mg，分别放入2mL塑料离心管。将取自大鼠心脏的新鲜柠檬酸钠抗凝血液150g离心10min，收集上清液 PRP，用PBS将其稀释至原血液的体积。向材料中加入100μL稀释PRP，置于37℃的恒温水浴锅中孵育5分钟。孵育完成后，用PBS洗涤材料，共洗涤三遍，每遍使用100μL PBS，去除表面未黏附的血小板。将洗涤液收集，并用PBS稀释至1mL，向其中1mL加入2％的Triton X-100溶液(用来裂解血小板，使其释放出LDH)置于37℃的恒温水浴锅中孵育60分钟。使用乳酸脱氢酶试剂盒测定裂解液中的LDH含量。空白组是将100μL稀释PRP用PBS稀释至1mL后，向其中1mL加入2％的Triton X-100溶液裂解全部血小板，使用乳酸脱氢酶试剂盒测定裂解液中的LDH含量。最后通过以下公式计算血小板黏附率。
血小板黏附率η％＝(1－LDH含量 样品/LDH含量 空白)×100％.........式2
式中：LDH含量 样品是实施例与对比例通过LDH试剂盒测定的裂解液中的LDH含量；LDH含量 空白是空白组通过LDH试剂盒测定的裂解液中的LDH含量。
表2体外血小板黏附效果测试
Figure PCTCN2022116742-appb-000002
血小板黏附测试可以表征材料对于血小板的黏附效果，通常材料对于血小板的黏附率越高，材料的促凝血效果越好。
从表2可以看出，本发明实施例1与实施例2得到的包覆多酚/聚合物促凝涂层的材料的血小板黏附率明显高于对比例1～3、对比例9与对比10，说明多酚/聚合物促凝涂层改性的材料具有更好的血小板黏附性能，通过多酚/聚合物促凝涂层改性可以通过提高血小板黏附率来提高材料的促凝血性能。
具体说明如下：
对比例1是将实施例1中的葡聚糖添加量由5mg/mL提高为10mg/mL，实施例1的血小板黏附率为44.1％，对比例1的血小板黏附率为39.5％。结果表明，提高葡聚糖添加量得到的多酚/聚合物复合涂层的材料血小板黏附率偏低，对应其BCI较高，促凝血效果不好。因为复合涂层中葡聚糖的含量的提高会增强对于多酚基团与凝血蛋白强作用力的屏蔽作用，使表面与蛋白产生有效相互作用偏弱，黏附的血小板偏少。
对比例2是将实施例1中的葡聚糖添加量由5mg/mL降低为1mg/mL，实施例1的血小板黏附率为44.1％，对比例2的血小板黏附率为33.9％。结果表明，降低葡聚糖添加 量得到的多酚/聚合物复合涂层的材料血小板黏附率较低，对应其BCI较高，促凝血效果不好。因为复合涂层中葡聚糖的含量的降低会减弱对于多酚基团与凝血蛋白强作用力的屏蔽作用，使蛋白不易活化，无法形成有利于血小板黏附的蛋白构象，从而难以黏附血小板。
对比例3是将实施例1中的不带电多羟基聚合物(葡聚糖)去掉，实施例1的血小板黏附率为44.1％，对比例3的血小板黏附率为24.3％。结果表明，仅使用多酚进行包覆，不添加不带电聚合物(葡聚糖)组分时，得到涂层的材料血小板黏附率较低，对应其BCI较高，促凝血效果不好。因为直接形成多酚涂层，纯多酚涂层中的酚羟基和凝血蛋白相互作用力过强，抑制其活化，无法形成有利于血小板黏附的蛋白构象。
对比例9与对比例10对应实施例1与实施例2中不做任何改性的基材，明胶海绵的血小板黏附率为23.1％，改性后血小板黏附率为44.1％；明胶海绵颗粒血小板黏附率为20.4％，改性后血小板黏附率为37.1％。结果表明：本发明通过在市售止血材料上包覆多酚/聚合物促凝涂层可以增加血小板黏附率，从而提高促凝血性能。
测试例3大鼠股动脉止血效果测试
对于明胶海绵基材料：选取体重在160-190g的SD大鼠通过腹腔注射水合氯醛(10％去离子水，0.5mL/100g体重)麻醉。用外科剪刀切开左后肢上皮组织，暴露大鼠股动静脉。用手术刀迅速切断股动脉(连同难以与动脉分离的静脉和周围神经)。切断股动脉后使其自由出血10秒，同时使用预先称重的纱布(m1)收集前10s失血量，并称重得到m1’。自由出血10秒后，迅速将预先称重的材料(m2，明胶海绵基材料预先裁成1.5×2cm的大小)覆盖在出血部位，并立即用标准砝码(100g)覆盖。止血过程中及时用预先称重的滤纸(m3)擦掉溢出的血液，先止血1.5分钟，观察其是否出血，如果仍在出血，每隔1分钟观察，直至停止出血。记录出血时间，收集和称重得到m2’与m3’。实验中前失血异常(>400mg或<200mg)的大鼠样本被排除在失血量和止血时间的最终分析之外，以减少大鼠个体差异和操作失误的影响。
通过以下公式计算前失血量与失血量：
前失血量M1＝m1’-m1.................................................................................式3
失血量M＝m2’-m2+m3’-m3......................................................................式4
表3大鼠股动脉止血的失血量和止血时间
样品 GS1(实施 GS6(对 GS对比
  例1) 比例3) 例9)
失血量(mg) 33.9 146.2 353.1
止血时间(min) 2 13.3 7.5
大鼠股动脉止血测试是通过测定材料在大鼠股动脉损伤模型中治疗损伤位置时的止血时间与失血量来模拟评估材料在真实应用场景的止血效果，通常材料的促凝血效果越好，在治疗过程中止血所需的时间越短，损伤位置的失血量越少。
从表3与表4中可以看出，本发明实施例1得到的包覆多酚/聚合物促凝涂层的材料在大鼠股动脉创伤止血中失血量与止血时间低于对比例3、对比例9，说明多酚/聚合物促凝涂层改性的材料在大鼠股动脉损伤模型的治疗中具有更好的治疗效果。具体说明如下：
对比例3是将实施例1中的亲水不带电聚合物(葡聚糖)去掉，直接形成多酚涂层，实施例1的失血量为(33.9mg)小于对比例3的失血量为(146.2mg)，实施例1的止血时间(2min)短于对比例3止血时间(13.3min)。
结果表明：仅包覆纯多酚涂层，不添加不带电聚合物组分时，得到的海绵在止血过程中的失血量更大，止血所需的时间更长，说明其在损伤中无法起到较好的止血效果。
对比例10对应实施例1中不做任何改性的基材，市售明胶海绵的失血量(353.1mg)大于实施例1的失血量(33.9mg)，市售明胶海绵止血时间(7.5min)长于实施例1的止血时间(2min)。
结果表明：医用基材按照本发明发明进行涂层包覆后，具备优异的足够促凝能力，在大鼠股动脉损伤模型中失血量小，止血时间短，在损伤时可以起到较好的止血效果。
以下是蛋白差异化黏附功能相关案例。
实施例9
将盐酸多巴胺50mg，葡聚糖500mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将玻璃片浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，氮气吹干得到GL1。
本对比例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:10。
实施例10
将盐酸多巴胺50mg，葡聚糖250mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将市售明胶海绵浸泡在该溶液中，25℃浸泡5h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺与葡聚糖，冷冻干燥得到GS1。
本实施例中，多酚羟基类物质为多巴胺，亲水不带电聚合物为葡聚糖，质量比为1:5。
对比例14
将盐酸多巴胺50mg溶于50mL Tris缓冲溶液(10mM)中，调节pH为8.5，将玻璃片浸泡在该溶液中，25℃浸泡1h后取出，使用大量去离子水洗涤三次，除去未形成涂层的多巴胺，氮气吹干得到GL2。
本对比例中，多酚羟基类物质为多巴胺，不添加聚合物组分。
对比例15
未进行改性的玻璃片GL。
对比例16
未进行改性的市售明胶海绵GS。
测试例4蛋白黏附效果测试
检测条件：将各实验例、对比例得到的材料进行蛋白黏附效果对比实验，检测用的蛋白为人源纤维蛋白原和人源白蛋白，检测使用BCA蛋白浓度测定试剂盒。
测试方法：将玻璃片材料裁成1×1cm的大小，放入24孔板中待测，明胶海绵材料称取3mg，放入2mL离心管中待测。以人源纤维蛋白为例，将人源纤维蛋白原配制为10mg/mL的生理盐水溶液，取50μL纤维蛋白原溶液加在材料上，将材料置于37℃的恒温水浴锅中孵育，玻璃片材料孵育30分钟，明胶海绵材料孵育5分钟。孵育完成后使用500μL生理盐水清洗玻璃片，取50μL洗涤液于2mL离心管中，加入提前配置好的1mL BCA工作液(试剂A：试剂B＝50:1(v/v))，置于37℃的恒温水浴锅中孵育30分钟，取100μL加入96孔板，测试562nm处的吸光度Abs。空白组是向50μL蛋白溶液中加入500μL生理盐水与材料组同比例稀释后，取50μL稀释液于2mL离心管中，用BCA试剂盒测试溶液中蛋白的含量。
蛋白黏附率η％＝(1－蛋白含量 样品/蛋白含量 空白)×100％............式1
式中：蛋白含量 样品是实施例与对比例通过BCA试剂盒测定的洗涤液液中的蛋白含量；蛋白含量 空白是空白组通过BCA试剂盒测定的稀释液液中的蛋白含量。
表4不同样品对三种蛋白的黏附效果测试
Figure PCTCN2022116742-appb-000003
蛋白黏附率是表征材料表面蛋白黏附作用的重要指标，黏附百分率越大，表明材料的蛋白黏附性能越好。
由表4的数据可以看出，以3-6％蛋白黏附率作为中等/合格蛋白黏附性能看，本发明的实施例9对纤维蛋白原为高黏附，对白蛋白为中等黏附；实施例10对纤维蛋白原为超高黏附，对球蛋白为中等黏附。
由此可见，采用本发明的制备方法，通过向多酚涂层中加入亲水不带电或负电性聚合物，可以实现对材料表面蛋白黏附能力的调控以及对不同蛋白的差异化黏附。具体分析如下：
对比例14是将实施例9中的不带电聚合物去掉，直接形成多酚涂层，相比于对比例15(未改性的玻璃片)，对两种蛋白的黏附率都提高(从6.2％到8.1％、3.1％到6.6％)，表明是纯多酚涂层对蛋白是广谱性提高黏附，是没有选择性。
实施例9以及对比例14的数据可以看出，当加入葡聚糖、形成多酚/聚合物复合涂层后，实施例9对白蛋白的黏附率基本不变(黏附率从3.1％到3.2％)，对纤维蛋白原的黏附率变高了(黏附率从6.2％升高到8.2％)。由此可见，本发明制备的涂层改性的玻璃片可以实现对蛋白的差异化黏附。差异化黏附。
实施例10以及对比例16(未改性的明胶海绵)的蛋白黏附率数据可以看出，当加入葡聚糖、形成多酚/聚合物复合涂层后，改性明胶海绵对纤维蛋白原的黏附率变高了(黏附率从6.4％升高到17.2％)，对白蛋白的黏附率降低了(黏附率从6.6％降低到3.1％)。由此可见，本发明制备的涂层改性海绵可以实现对蛋白的差异化黏附。
最后说明的是，以上优选实施例仅用以说明本发明的技术方案而非限制，尽管通过上述优选实施例已经对本发明进行了详细的描述，但本领域技术人员应当理解，可以在形式上和细节上对其作出各种各样的改变，而不偏离本发明权利要求书所限定的范围。

Claims (10)

  1. 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤为：
    1)将多酚羟基类物质和亲水不带电或亲水带负电聚合物配制成混合溶液；
    2)将基材在所述的混合溶液中浸泡；
    3)洗涤，干燥，得到功能化医用材料；
    所述的功能化为止血功能和/或蛋白差异化黏附功能；
    实现止血功能条件：步骤1)中的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:2～8和步骤2)中的浸泡时间为0.7～4小时，步骤3)得到具备止血功能的医用材料；
    实现蛋白差异化黏附条件：步骤1)中的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:0.5～15和步骤2)中的浸泡时间为0.5～20小时，步骤3)得到具备蛋白差异化黏附的医用材料。
  2. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，当步骤1)的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:2～5，步骤2)所述的浸泡时间为0.7～1.5小时，步骤3)得到具备止血功能的医用材料。
  3. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，当步骤1)的多酚羟基类物质与亲水不带电或亲水带负电聚合物的质量比为1:5～10，步骤2)的浸泡时间为1～5小时，步骤3)得到的是具备蛋白差异化黏附的医用材料，所述的蛋白为纤维蛋白原、白蛋白、脂蛋白、糖蛋白或球蛋白。
  4. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤1)所述的多酚羟基类物质为多巴胺、单宁酸、没食子酸、儿茶素、表儿茶素、表没食子儿茶素没食子酸酯、茶黄素-3-没食子酸酯、5-羟基多巴胺盐酸盐或黄芩素，步骤1)所述的亲水不带电聚合物为葡聚糖、羟丙基纤维素、羟乙基淀粉、羟乙基纤维素、普罗兰多糖、普兰尼克或聚乙二醇，步骤1)所述的亲水负电聚合物为羧甲基纤维素或海藻酸、海藻酸钠、透明质酸、透明质酸钠或羧甲基淀粉。
  5. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤1)所述的多酚羟基类物质为多巴胺、单宁酸或没食子酸，步骤1)所述的亲水不带电聚合物为葡聚糖、羟丙基纤维素或聚乙二醇，步骤1)所述的亲水负电聚合物为羧甲基纤维素。
  6. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤1)所述的多酚羟基类物质在混合溶液中的浓度为0.25～3mg/mL。
  7. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，止血功能的实现条件为：亲水不带电或亲水负电聚合在混合溶液中的浓度为3～5mg/mL，蛋白差异化黏附的实现条件为：亲水不带电或亲水负电聚合在混合溶液中的浓度为5～10mg/mL。
  8. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤2)所述的基材为高分子医用材料、无机类医用材料、金属类医用材料；所述的高分子医用材料为纱布、聚乙烯醇海绵、聚乙烯醇微米级颗粒、聚乙烯醇微球、壳聚糖海绵、壳聚糖无纺布、明胶海绵、明胶海绵微米级颗粒、海藻酸敷料、海藻酸微球、海藻酸无纺布或胶原蛋白海绵；所述的无机类医用材料是生物玻璃、生物陶瓷；所述的金属类医用材料是镍钛合金器械、钛合金医用器械、形状记忆金属微弹簧圈。
  9. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤2)所述的基材为纱布、聚乙烯醇海绵、聚乙烯醇微米级颗粒、聚乙烯醇微球、明胶海绵、明胶海绵微米级颗粒、胶原蛋白海绵、生物玻璃或钛合金医用器械。
  10. 根据权利要求1所述的一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用，其特征在于，步骤2)所述基材为金属类医用材料时，基材在浸泡混合溶液前可以用多酚羟基类物质进行预处理，在基材表面形成多酚涂层，预处理方法为：将基材在多酚羟基类物质的pH为8～9的碱性水溶液中浸泡4～24小时。
PCT/CN2022/116742 2022-09-02 2022-09-02 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用 WO2024045163A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2022/116742 WO2024045163A1 (zh) 2022-09-02 2022-09-02 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2022/116742 WO2024045163A1 (zh) 2022-09-02 2022-09-02 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用

Publications (1)

Publication Number Publication Date
WO2024045163A1 true WO2024045163A1 (zh) 2024-03-07

Family

ID=90100191

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/116742 WO2024045163A1 (zh) 2022-09-02 2022-09-02 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用

Country Status (1)

Country Link
WO (1) WO2024045163A1 (zh)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105218847A (zh) * 2015-09-14 2016-01-06 北京化工大学 一种基于多酚类化合物的材料表界面功能化改性方法
CN106474524A (zh) * 2015-08-31 2017-03-08 山东美泰医药有限公司 一种可降解淀粉止血海绵及其制备方法
CN106794283A (zh) * 2014-08-08 2017-05-31 舒尔默迪克斯公司 包含低聚多酚层的制品涂层及其生物使用方法
US20180147326A1 (en) * 2015-05-26 2018-05-31 The University Of British Columbia Antifouling Polymeric Coating Compositions
WO2021177536A1 (ko) * 2020-03-02 2021-09-10 주식회사 이노테라피 지혈용 기구 및 이의 제조방법
CN114767918A (zh) * 2022-03-21 2022-07-22 东莞市人民医院 一种促凝止血蛋白材料、促凝止血抗菌材料及其制备方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106794283A (zh) * 2014-08-08 2017-05-31 舒尔默迪克斯公司 包含低聚多酚层的制品涂层及其生物使用方法
US20180147326A1 (en) * 2015-05-26 2018-05-31 The University Of British Columbia Antifouling Polymeric Coating Compositions
CN106474524A (zh) * 2015-08-31 2017-03-08 山东美泰医药有限公司 一种可降解淀粉止血海绵及其制备方法
CN105218847A (zh) * 2015-09-14 2016-01-06 北京化工大学 一种基于多酚类化合物的材料表界面功能化改性方法
WO2021177536A1 (ko) * 2020-03-02 2021-09-10 주식회사 이노테라피 지혈용 기구 및 이의 제조방법
CN114767918A (zh) * 2022-03-21 2022-07-22 东莞市人民医院 一种促凝止血蛋白材料、促凝止血抗菌材料及其制备方法

Similar Documents

Publication Publication Date Title
Shi et al. Puff pastry-like chitosan/konjac glucomannan matrix with thrombin-occupied microporous starch particles as a composite for hemostasis
US20210100830A1 (en) Starch-based hemostatic powder and preparation method thereof
Cao et al. Multifunctional dopamine modification of green antibacterial hemostatic sponge
EP2203053B1 (en) Modified starch material of biocompatible hemostasis
Fan et al. Rapid hemostatic chitosan/cellulose composite sponge by alkali/urea method for massive haemorrhage
GB2489541A (en) Wound dressing
Lan et al. Improvement of platelet aggregation and rapid induction of hemostasis in chitosan dressing using silver nanoparticles
CN112972749B (zh) 基于壳聚糖纤维的高效止血材料及其制备方法
JPS587251A (ja) 複合傷用包帯
Song et al. Kaolin-loaded carboxymethyl chitosan/sodium alginate composite sponges for rapid hemostasis
CN113244442B (zh) 多氨基阳离子化合物在制备表面促凝血增强型止血材料上的应用
Zhang et al. An injectable serotonin–chondroitin sulfate hydrogel for bio-inspired hemostatic adhesives with high wound healing capability
Cassano et al. Hemostatic gauze based on chitosan and hydroquinone: Preparation, characterization and blood coagulation evaluation
CN107412843B (zh) 一种具有抗菌性能的淀粉基微孔止血材料及其制备方法和应用
Atashgahi et al. Epinephrine-entrapped chitosan nanoparticles covered by gelatin nanofibers: A bi-layer nano-biomaterial for rapid hemostasis
Panwar et al. In-vitro and In-vivo evaluation of biocompatible and biodegradable calcium-modified carboxymethyl starch as a topical hemostat
JP2752782B2 (ja) 可溶性止血織物
Yan et al. A keratin/chitosan sponge with excellent hemostatic performance for uncontrolled bleeding
CN114767918A (zh) 一种促凝止血蛋白材料、促凝止血抗菌材料及其制备方法
Wang et al. Calcium functioned carboxymethylated cotton fabric for hemostatic wound dressing
Luo et al. Development and evaluation of tilapia skin-derived gelatin, collagen, and acellular dermal matrix for potential use as hemostatic sponges
Wu et al. Strong tissue adhesive polyelectrolyte complex powders based on low molecular weight chitosan for acute hemorrhage control
CN112300418B (zh) 一种可粘附高效止血微球及其制备方法
Fan et al. One-step fabrication of chitosan sponge and its potential for rapid hemostasis in deep trauma
WO2024045163A1 (zh) 一种多酚-聚合物复合涂层的制备方法及其在医用材料功能化中的应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22956985

Country of ref document: EP

Kind code of ref document: A1