技术领域technical field
本发明属于材料表面科学和生物医用高分子材料技术领域，具体涉及一种含有氨基的磷酰胆碱聚合物与戊二醛仿生涂层增密的方法。The invention belongs to the technical fields of material surface science and biomedical polymer materials, and in particular relates to a method for densifying phosphorylcholine polymers containing amino groups and glutaraldehyde bionic coatings.
背景技术Background technique
壳聚糖具有可降解性、抗菌性、无毒、无刺激、pH响应性等优点(CarbohydratePolymers2010,79:724-730)，已经被广泛应用于生物医学等领域。越来越多的研究表明：壳聚糖及其衍生物材料可以用于血液净化。壳聚糖分子上的氨基有助于对血液中多种毒素的吸附，可以用于血液灌流材料(高等学校化学学报2002,23:75-77；Journal ofMicroencapsulation 1993,10:475-486)。壳聚糖膜具有高的透析率，选择性和强度，可以用作血液透析材料(Journal of Applied Polymer Science 1992,46:255-261；263-269)。虽然壳聚糖及其衍生物作为血液净化材料具有许多优点，但是也存在着蛋白质吸附，血小板黏附，最终导致凝血，形成血栓等问题，所以提高壳聚糖及其衍生物材料的血液相容性迫在眉睫(Applied Surface Science 2005,241:485-492；Biomaterials 2002,23:2561-2568；Biomaterials 2003,24:3213-3220)。Chitosan has the advantages of degradability, antibacterial, non-toxic, non-irritating, and pH responsiveness (Carbohydrate Polymers 2010, 79:724-730), and has been widely used in biomedicine and other fields. More and more studies have shown that chitosan and its derivatives can be used for blood purification. The amino group on the chitosan molecule helps to adsorb various toxins in the blood, and can be used as a hemoperfusion material (Chemical Journal of Chinese Universities, 2002, 23:75-77; Journal of Microencapsulation 1993, 10:475-486). Chitosan membrane has high dialysis rate, selectivity and strength, and can be used as hemodialysis material (Journal of Applied Polymer Science 1992, 46:255-261; 263-269). Although chitosan and its derivatives have many advantages as blood purification materials, there are also problems such as protein adsorption and platelet adhesion, which eventually lead to blood coagulation and thrombus formation, so the blood compatibility of chitosan and its derivative materials should be improved. Imminent (Applied Surface Science 2005, 241:485-492; Biomaterials 2002, 23:2561-2568; Biomaterials 2003, 24:3213-3220).
磷酰胆碱(phosphorylcholine，PC)是组成细胞膜基本单元卵磷脂的亲水端基，是细胞外层膜中的外层官能团，同时带有正、负异种电荷，具有较强的结合水的能力和亲水性能，这种结构和组成的表面与生理环境相互作用不仅不会吸附和沉积蛋白质，也不会引发血小板激活、导致凝血等不良反应，具有良好生物相容性。近几年来的研究表明，采用磷酰胆碱基团及其聚合物在材料表面构建具有仿细胞外层膜结构，可以显著改善材料的血液相容性。Phosphorylcholine (PC) is the hydrophilic end group of lecithin, the basic unit of cell membrane, and is the outer functional group in the outer membrane of the cell. It also has positive and negative heterogeneous charges and has a strong ability to bind water. And hydrophilic properties, the surface of this structure and composition interacts with the physiological environment not only will not adsorb and deposit proteins, but also will not cause adverse reactions such as platelet activation and blood coagulation, and has good biocompatibility. Studies in recent years have shown that using phosphorylcholine groups and their polymers to construct a cell-like outer membrane structure on the surface of the material can significantly improve the blood compatibility of the material.
近年来，采用接枝磷酰胆碱小分子的途径(Carbohydrate Polymers 2007，70:82-88；Biomacromolecules 2007,8:3169-3176；Biomacromolecules 2006,7:3151-3156；Journal of Applied Polymer Science 2003,88:489-493；Polymer International2003,52:81-85；Journal of biomaterials science,Polymer edition 2002,13:501-510；Colloids and Surfaces B:Biointerfaces2009,71:268-274)改性壳聚糖，使得壳聚糖的血液相容性显著提高。但是，这些方式往往在材料表面的磷酰胆碱基团的密度不高，限制了其在生物医用材料改性领域的应用和血液相容性的进一步提高。In recent years, the approach of grafting phosphorylcholine small molecules (Carbohydrate Polymers 2007,70:82-88; Biomacromolecules 2007,8:3169-3176; Biomacromolecules 2006,7:3151-3156; Journal of Applied Polymer Science 2003, 88:489-493; Polymer International2003,52:81-85; Journal of biomaterials science, Polymer edition 2002,13:501-510; Colloids and Surfaces B: Biointerfaces2009,71:268-274) modified chitosan, making The blood compatibility of chitosan is significantly improved. However, these methods often have a low density of phosphorylcholine groups on the surface of the material, which limits its application in the field of biomedical material modification and further improvement of blood compatibility.
为此，将含有磷酰胆碱基团的甲基丙烯酸-甲基丙烯酰氧乙基磷酰胆碱二元共聚物(PMA)聚阴离子，与壳聚糖(聚阳离子)进行层层静电自组装，获得了具有仿细胞外层膜结构的涂层表面(Colloids and Surfaces B:Biointerfaces 2011,85:48-55)。蛋白质吸附和血小板黏附的实验结果表明：改性后表面的血液相容性有了显著提高。鉴于这种改性方法的种种优势，必将为提升生物医用材料的血液相容性提供技术支撑。然而，以物理吸附方式结合在移植器件表面的仿细胞外层膜结构聚合物涂层，在体内复杂环境中难免发生溶解、脱落。为此，Lewis和徐建平等(Biomaterials 2001,22:99-111；Biomaterials 2004,25:3099-3108；European Polymer Journal 2004,40:291-298)分别对含有三甲氧基硅基团和磷酰胆碱基团的聚合物涂层进行了研究。结果表明，涂层中聚合物分子链上三甲氧基硅基团遇水会发生水解、交联，也可与基材表面的活性基团形成共价键，从而使磷酰胆碱类聚合物涂层的性能得到显著提高。由此可见，聚合物之间的交联及其与基材表面官能团的反应，是提高磷酰胆碱类聚合物涂层性能的关键因素。For this reason, the methacrylic acid-methacryloyloxyethyl phosphorylcholine binary copolymer (PMA) polyanion containing phosphorylcholine group is subjected to layer-by-layer electrostatic autolysis with chitosan (polycation). Assembled, a coated surface with a cell-like outer membrane structure was obtained (Colloids and Surfaces B: Biointerfaces 2011, 85:48-55). The experimental results of protein adsorption and platelet adhesion show that the hemocompatibility of the modified surface has been significantly improved. In view of the various advantages of this modification method, it will certainly provide technical support for improving the blood compatibility of biomedical materials. However, the polymer coating imitating the membrane structure of the outer layer of the cell bound to the surface of the implanted device by physical adsorption will inevitably dissolve and fall off in the complex environment in the body. For this reason, Lewis and Xu Jianping (Biomaterials 2001,22:99-111; Biomaterials 2004,25:3099-3108; European Polymer Journal 2004,40:291-298) respectively studied the Choline-based polymer coatings were studied. The results show that the trimethoxysilyl group on the polymer molecular chain in the coating will undergo hydrolysis and crosslinking when it meets water, and can also form a covalent bond with the active groups on the surface of the substrate, so that the phosphorylcholine polymer The performance of the coating is significantly improved. It can be seen that the crosslinking between polymers and the reaction with the functional groups on the surface of the substrate are the key factors to improve the performance of the phosphorylcholine polymer coating.
将可交联基团引入到磷酰胆碱聚合物可形成相对稳定的仿细胞外层膜结构涂层，但这种构建稳定涂层的磷酰胆碱聚合物合成复杂、应用比较困难、稳定性不理想。另外，受聚合物中磷酰胆碱含量的限制及涂层干燥、储存过程中表面磷酰胆碱基团取向内迁移，致使涂层表面磷酰胆碱基团密度较低，不易有效地模拟细胞外层膜结构，血液相容性还有待提高。因此，构建结构稳定、磷酰胆碱基团密度高的仿细胞外层膜结构涂层具有重要的研究意义及应用前景。The introduction of crosslinkable groups into phosphorylcholine polymers can form a relatively stable coating imitating the outer membrane structure of cells, but the synthesis of phosphorylcholine polymers for building stable coatings is complicated, difficult to apply, and stable Sex is not ideal. In addition, due to the limitation of phosphorylcholine content in the polymer and the internal migration of phosphorylcholine groups on the surface during coating drying and storage, the density of phosphorylcholine groups on the surface of the coating is low, making it difficult to effectively simulate Cell outer membrane structure, blood compatibility needs to be improved. Therefore, it is of great research significance and application prospect to construct a structure-stable coating with a high density of phosphorylcholine group imitating the outer layer of the cell membrane.
发明内容Contents of the invention
本发明所要解决的技术问题在于针对上述现有技术的不足，提供一种含有氨基的磷酰胆碱聚合物与戊二醛仿生涂层增密的方法。本发明的稳定、磷酰胆碱基团密度高的仿细胞外层膜结构涂层的制备方法简单、条件温和，为获得性能优异的仿细胞外层膜结构的涂层表面提供了一种新的途径。The technical problem to be solved by the present invention is to provide a method for densifying biomimetic coatings of phosphorylcholine polymers containing amino groups and glutaraldehyde in view of the deficiencies in the prior art above. The preparation method of the stable and high-density phosphorylcholine imitation cell outer layer membrane structure coating of the present invention is simple and the conditions are mild, and provides a new method for obtaining the coating surface of the imitation cell outer layer membrane structure with excellent performance. way.
为解决上述技术问题，本发明采用的技术方案是：In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
一种含有氨基的磷酰胆碱聚合物与戊二醛仿生涂层增密的方法，其特征在于，包括以下步骤：A method for densification of amino-containing phosphorylcholine polymers and glutaraldehyde biomimetic coatings, is characterized in that it comprises the following steps:
步骤一、将含有两性离子磷酰胆碱基团的乙烯基单体和含有氨基的乙烯基单体在引发剂作用下进行溶液自由基聚合，合成含有氨基的磷酰胆碱聚合物；Step 1, carrying out solution free radical polymerization under the action of an initiator of vinyl monomers containing zwitterionic phosphorylcholine groups and vinyl monomers containing amino groups to synthesize phosphorylcholine polymers containing amino groups;
步骤二、将含有氨基的磷酰胆碱聚合物与戊二醛溶于极性溶剂中混合均匀，涂覆在需要改性的材料表面，晾干；其中含有氨基的磷酰胆碱聚合物中的氨基与戊二醛中醛基的摩尔比约为(100∶5)～(100∶20)；Step 2, dissolve the phosphorylcholine polymer containing amino groups and glutaraldehyde in a polar solvent and mix evenly, apply it on the surface of the material to be modified, and dry it; the phosphorylcholine polymer containing amino groups The molar ratio of amino groups in glutaraldehyde to aldehyde groups in glutaraldehyde is about (100:5) to (100:20);
步骤三、将步骤二中晾干后的待改性材料置于含有两性离子磷酰胆碱基团的乙烯基单体的溶液中，在20℃～40℃条件下反应5h～12h；Step 3, placing the material to be modified after drying in step 2 in a solution of vinyl monomers containing zwitterionic phosphorylcholine groups, and reacting at 20°C to 40°C for 5h to 12h;
步骤四、将步骤三中处理的膜置于蒸馏水中，在80℃～95℃条件下处理2h～12h，再用溶剂洗涤，即可得到表面磷酰胆碱密度高的仿细胞外层膜结构涂层。Step 4: Place the membrane treated in step 3 in distilled water, treat it at 80°C to 95°C for 2h to 12h, and then wash it with a solvent to obtain a cell-like outer membrane structure with a high surface phosphorylcholine density coating.
步骤一中，所述含有两性离子磷酰胆碱基团的乙烯基单体为2-甲基丙烯酰氧乙基磷酰胆碱单体，所述含有氨基的乙烯基单体为2-氨乙基甲基丙烯酸酯盐酸盐单体，所述引发剂为过硫酸钾。In step 1, the vinyl monomer containing zwitterionic phosphorylcholine group is 2-methacryloyloxyethylphosphorylcholine monomer, and the vinyl monomer containing amino group is 2-amino Ethyl methacrylate hydrochloride monomer, the initiator is potassium persulfate.
步骤一中，所述含有两性离子磷酰胆碱基团的乙烯基单体与含有氨基的乙烯基单体摩尔比为(90:10)～(60:40)。In step 1, the molar ratio of the vinyl monomer containing the zwitterionic phosphorylcholine group to the vinyl monomer containing the amino group is (90:10)-(60:40).
步骤一中，所述含有两性离子磷酰胆碱基团的乙烯基单体与含有氨基的乙烯基单体及引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。In step 1, the vinyl monomer containing the zwitterionic phosphorylcholine group, the vinyl monomer containing the amino group and the initiator are polymerized at 70°C for 24 hours under the protection of nitrogen, dialyzed after the reaction, and then in Freeze drying at -50°C to obtain amino group-containing phosphorylcholine polymers.
步骤二中，所述极性溶剂为甲醇或乙醇。In step 2, the polar solvent is methanol or ethanol.
步骤二中，含有氨基的磷酰胆碱聚合物与戊二醛混合溶液中，含有氨基的磷酰胆碱聚合物的浓度为0.5～5mg/mL，涂覆体积为10微升/cm2/面。In step 2, in the mixed solution of phosphorylcholine polymers containing amino groups and glutaraldehyde, the concentration of phosphorylcholine polymers containing amino groups is 0.5-5 mg/mL, and the coating volume is 10 microliters/cm2 / noodle.
步骤三中，所述含有两性离子磷酰胆碱基团的乙烯基单体的溶液，浓度为5～20mg/mL，溶剂为甲醇。In step 3, the solution of the vinyl monomer containing the zwitterionic phosphorylcholine group has a concentration of 5-20 mg/mL, and the solvent is methanol.
步骤四中，所述溶剂洗涤是指依次用甲醇、蒸馏水洗涤。In step 4, the solvent washing refers to washing with methanol and distilled water in sequence.
步骤二中，所述需要改性的材料为壳聚糖膜。In step 2, the material to be modified is a chitosan film.
本发明与现有技术相比，具有以下优点：Compared with the prior art, the present invention has the following advantages:
本发明将含有两性离子磷酰胆碱基团的乙烯基单体、含有氨基乙烯基单体通过简单的溶液自由基聚合，合成含有氨基的磷酰胆碱聚合物，并将其与戊二醛溶于极性溶剂混合均匀，涂覆在壳聚糖膜表面、晾干，然后接枝两性离子磷酰胆碱基团的乙烯基单体。含氨基的磷酰胆碱聚合物是通过含有磷酰胆碱的乙烯基单体与含有氨基的乙烯基单体采用自由基聚合法合成的二元随机共聚物，该二元共聚物与戊二醛混合均匀，涂覆在壳聚糖膜表面、晾干，然后接枝两性离子磷酰胆碱基团的乙烯基单体。通过迈克尔加成将两性离子磷酰胆碱基团的乙烯基单体接枝到涂层表面，再通过温度控制磷酰胆碱聚合物中氨基及壳聚糖膜表面氨基与戊二醛反应，使涂层内部交联的同时锚定基材，使得涂层厚度降低显著提高涂层表面磷酰胆碱的密度，即可获得表面磷酰胆碱密度高的仿细胞外层膜结构涂层。In the present invention, vinyl monomers containing zwitterionic phosphorylcholine groups and vinyl monomers containing amino groups are polymerized by simple solution free radicals to synthesize phosphorylcholine polymers containing amino groups, and combine them with glutaraldehyde Dissolved in polar solvents and mixed evenly, coated on the surface of chitosan film, dried, and then grafted with vinyl monomers of zwitterionic phosphorylcholine groups. Amino-containing phosphorylcholine polymers are binary random copolymers synthesized by free radical polymerization of phosphorylcholine-containing vinyl monomers and amino-containing vinyl monomers. The aldehyde is mixed evenly, coated on the surface of the chitosan film, dried, and then grafted with vinyl monomers of zwitterionic phosphorylcholine groups. The vinyl monomer of the zwitterionic phosphorylcholine group is grafted to the surface of the coating by Michael addition, and then the amino group in the phosphorylcholine polymer and the amino group on the surface of the chitosan film react with glutaraldehyde through temperature control, The substrate is anchored while the coating is internally crosslinked, so that the thickness of the coating is reduced and the density of phosphorylcholine on the surface of the coating is significantly increased, so that a coating with a structure imitating the outer layer of the cell membrane with a high density of phosphorylcholine on the surface can be obtained.
本发明的稳定、磷酰胆碱基团密度高的仿细胞外层膜结构涂层的制备方法简单、条件温和，为获得性能优异的仿细胞外层膜结构的涂层表面提供了一种新的途径。本发明的稳定、磷酰胆碱基团密度高的仿细胞外层膜结构涂层将在血液净化，体内植入材料，组织工程，药物缓释及生物传感器等领域具有广阔的应用前景。The preparation method of the stable and high-density phosphorylcholine imitation cell outer layer membrane structure coating of the present invention is simple and the conditions are mild, and provides a new method for obtaining the coating surface of the imitation cell outer layer membrane structure with excellent performance. way. The stable and high-density phosphorylcholine group imitation cell outer membrane structure coating of the present invention will have broad application prospects in the fields of blood purification, material implantation in vivo, tissue engineering, drug sustained release, biosensor and the like.
附图说明Description of drawings
图1为壳聚糖膜(CS)和改性壳聚糖膜(CS-PMH20GA-MPC)的动态接触角对比图。Figure 1 is a comparison diagram of the dynamic contact angle of chitosan film (CS) and modified chitosan film (CS-PMH20GA-MPC).
图2为壳聚糖膜(CS)和改性壳聚糖膜(CS-PMH20GA-MPC)的表面元素精细能谱图。Fig. 2 is the fine energy spectrum of surface elements of chitosan film (CS) and modified chitosan film (CS-PMH20GA-MPC).
图3为壳聚糖膜(CS)和改性壳聚糖膜(CS-PMH20GA-MPC)的AFM图。Fig. 3 is the AFM image of chitosan film (CS) and modified chitosan film (CS-PMH20GA-MPC).
图4为壳聚糖膜(CS)和改性壳聚糖膜(CS-PMH20GA-MPC)的荧光蛋白吸附量。Fig. 4 is the fluorescent protein adsorption amount of chitosan membrane (CS) and modified chitosan membrane (CS-PMH20GA-MPC).
具体实施方式Detailed ways
本发明一种表面磷酰胆碱密度高的仿细胞外层膜结构涂层的制备方法为：将摩尔比约为90:10～60:40含有两性离子磷酰胆碱基团的乙烯基单体与含有氨基的乙烯基单体进行聚合，合成含有氨基的磷酰胆碱聚合物，并将其与戊二醛溶于甲醇溶剂中混合均匀，涂覆在壳聚糖膜表面，然后置于含有两性离子磷酰胆碱基团的乙烯基单体溶液中，在20℃～40℃条件下反应5h～12h。之后，经80℃～95℃条件下蒸馏水加热处理2h～12h，依次用大量的甲醇、蒸馏水洗涤，即可得到表面磷酰胆碱密度高的仿细胞外层膜结构涂层。The preparation method of the imitation cell outer layer membrane structure coating with high phosphorylcholine density on the surface of the present invention is as follows: the molar ratio is about 90:10～60:40 vinyl monolayer containing zwitterionic phosphorylcholine group Polymerized with amino-containing vinyl monomers to synthesize amino-containing phosphorylcholine polymers, and mixed with glutaraldehyde in methanol solvent, coated on the surface of chitosan membrane, and then placed in In the vinyl monomer solution containing the zwitterionic phosphorylcholine group, react for 5h-12h under the condition of 20°C-40°C. Afterwards, heat treatment with distilled water at 80°C to 95°C for 2h to 12h, and wash with a large amount of methanol and distilled water in sequence to obtain a membrane-like coating with a high density of phosphorylcholine on the surface.
下面结合附图和实施例，对本发明的技术方案做进一步的详细说明。The technical solutions of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.
实施例1Example 1
1)称取16mmol 2-甲基丙烯酰氧乙基磷酰胆碱和4mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物PMH20(20代表聚合物合成过程中氨基的乙烯单体投料摩尔比为20％)。1) Weigh 16mmol 2-methacryloyloxyethylphosphorylcholine and 4mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as initiator, under nitrogen protection, 70°C Polymerization was performed for 24 hours, dialyzed after the reaction, and then freeze-dried at -50°C to obtain amino group-containing phosphorylcholine polymer PMH20 (20 represents 20% molar ratio of amino group vinyl monomer in the polymer synthesis process).
用400MHz核磁共振仪以D2O为溶剂测试聚合物的氢核磁。在5～7ppm处未见出峰，表明所得共聚物中没有残余单体，并成功合成了该聚合物，以3.28ppm处为-N+(CH3)3特征峰，0.9～2.2ppm处为主链上亚甲基和侧链甲基的峰计算聚合物组成，可知该聚合物组成与投料比基本一致。The hydrogen nuclear magnetic resonance of the polymer was tested with a 400MHz nuclear magnetic resonance instrument and D2 O as a solvent. There is no peak at 5-7ppm, indicating that there is no residual monomer in the resulting copolymer, and the polymer has been successfully synthesized, with the characteristic peak of -N + (CH3 ) at 3.28ppm and the characteristic peak of -N+ (CH3 ) at 0.9-2.2ppm. The polymer composition is calculated from the peaks of methylene and side chain methyl groups on the main chain, and it can be known that the polymer composition is basically consistent with the feed ratio.
2)将含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶10溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为1.0mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料(CS)表面晾干，然后经蒸馏水中90度处理6h，之后依次用大量的甲醇、蒸馏水洗涤，即可得对比样品(CS-PMH20/GA)，然后将其浸泡于25mL，10mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在25℃反应10h，之后经蒸馏水中90度处理6h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层(CS-PMH20/GA-MPC)。2) Dissolving the amino group-containing phosphorylcholine polymer and glutaraldehyde in a methanol solvent in a molar ratio of 100:10 to the amino group and mixing them uniformly, so that the concentration of the amino group-containing phosphorylcholine polymer is 1.0 mg/mL, the coating volume is 10 microliters/cm2 /face, dry on the surface of chitosan material (CS), and then treat it in distilled water at 90 degrees for 6 hours, and then wash it with a large amount of methanol and distilled water in turn to obtain the comparison Sample (CS-PMH20/GA), then soak it in 25mL, 10mg/mL methanol solution of 2-methacryloyloxyethylphosphorylcholine, react at 25°C for 10h, then distilled water at 90°C After treatment for 6 hours, followed by washing with a large amount of methanol and distilled water, a high-density coating imitating the outer membrane structure of cells (CS-PMH20/GA-MPC) can be obtained.
3)将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶10溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为1.0mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于25mL，10mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在25℃反应10h，之后经蒸馏水中90度处理6h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到稳定仿细胞外层膜结构的涂层表面(CS-PMH20GA-MPC)。3) The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in methanol solvent at a molar ratio of amino group and aldehyde group of 100:10 and mixed uniformly, and then the amino group-containing phosphorylcholine polymer The concentration is 1.0mg/mL, and the coating volume is 10 microliters/cm2 /face. Dry it on the surface of the chitosan material, and then soak it in 25mL, 10mg/mL of 2-methacryloyloxyethylphosphorylcholine In the methanol solution of alkali, react at 25°C for 10h, then treat it in distilled water at 90°C for 6h, and then wash it with a large amount of methanol and distilled water in turn to obtain a stable coating surface imitating the outer membrane structure of cells (CS-PMH20GA- MPC).
如图1所示，本实施例经涂层处理的壳聚糖材料与经对照涂层处理的壳聚糖材料相比，经涂层处理的壳聚糖的表面前进角较低，这是因为含有氨基的磷酰胆碱聚合物与戊二醛涂层表面剩余的氨基通过希夫碱反应接枝2-甲基丙烯酰氧乙基磷酰胆碱单体进一步提高涂层表面磷酰胆碱基团的密度，使得涂层表面亲水性较高，前进角较低。As shown in Figure 1, compared with the chitosan material processed by the coating in the present embodiment, the surface advancing angle of the chitosan processed by the coating is lower, and this is because Phosphorylcholine polymers containing amino groups and the remaining amino groups on the surface of glutaraldehyde coatings were grafted with 2-methacryloyloxyethylphosphorylcholine monomers through the Schiff base reaction to further improve the phosphorylcholine content of the coating surface. The density of the group makes the surface of the coating more hydrophilic and the advancing angle is lower.
表1.壳聚糖膜(CS)和改性壳聚糖膜(CS-PMH20GA-MPC)的表面元素含量。Table 1. Surface element content of chitosan film (CS) and modified chitosan film (CS-PMH20GA-MPC).
表1Table 1
如图2和表1所示，本实施例经涂层处理的壳聚糖材料与经对照涂层处理的壳聚糖材料相比，经改性处理的壳聚糖膜表面磷酰胆碱基团上N和P特征吸收峰较大，表面磷酰胆碱基团密度较高。As shown in Figure 2 and Table 1, compared with the chitosan material processed by the coating in the present embodiment, the phosphorylcholine group on the surface of the modified chitosan film The characteristic absorption peaks of N and P on the group are larger, and the density of phosphorylcholine groups on the surface is higher.
如图3所示，本实施例经涂层处理的壳聚糖材料与经对照涂层处理的壳聚糖材料相比，经涂层处理的壳聚糖的表面形貌显著不同。As shown in FIG. 3 , compared with the chitosan material treated by the coating in this embodiment and the chitosan material treated by the control coating, the surface morphology of the chitosan treated by the coating is significantly different.
如图4所示，本实施例经涂层处理的壳聚糖材料与经对照涂层处理的壳聚糖材料相比，经处理的壳聚糖荧光蛋白吸附量有明显的降低，血液相容性显著提高。As shown in Figure 4, compared with the chitosan material processed by the control coating in the present embodiment, the adsorption capacity of the processed chitosan fluorescent protein is significantly reduced, and blood compatibility Significantly improved.
实施例2Example 2
称取17mmol 2-甲基丙烯酰氧乙基磷酰胆碱和3mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 17mmol 2-methacryloyloxyethylphosphorylcholine and 3mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶5溶于乙醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为0.5mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于20mL，5mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在20℃反应12h，之后经蒸馏水中80度处理12h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in an ethanol solvent in a molar ratio of 100:5 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 0.5mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, then soak it in 20mL, 5mg/mL of 2-methacryloyloxyethylphosphorylcholine In methanol solution, react at 20°C for 12h, then treat in distilled water at 80°C for 12h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例3Example 3
称取12mmol 2-甲基丙烯酰氧乙基磷酰胆碱和8mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 12mmol 2-methacryloyloxyethyl phosphorylcholine and 8mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶15溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为1.5mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于25mL，15mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在30℃反应8h，之后经蒸馏水中85度处理10h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in methanol solvent in a molar ratio of 100:15 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 1.5mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, then soak it in 25mL, 15mg/mL of 2-methacryloyloxyethylphosphorylcholine In methanol solution, react at 30°C for 8h, then treat in distilled water at 85°C for 10h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例4Example 4
称取15mmol 2-甲基丙烯酰氧乙基磷酰胆碱和5mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 15mmol 2-methacryloyloxyethylphosphorylcholine and 5mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶20溶于乙醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为2mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于30mL，20mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在35℃反应6h，之后经蒸馏水中95度处理2h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in an ethanol solvent in a molar ratio of 100:20 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 2mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, then soak it in 30mL, 20mg/mL 2-methacryloyloxyethylphosphorylcholine methanol In the solution, react at 35°C for 6h, then treat in distilled water at 95°C for 2h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例5Example 5
称取14mmol 2-甲基丙烯酰氧乙基磷酰胆碱和6mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 14mmol 2-methacryloyloxyethylphosphorylcholine and 6mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶8溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为3mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于22mL，7mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在40℃反应5h，之后经蒸馏水中83度处理11h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in methanol solvent in a molar ratio of 100:8 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 3mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, and then soak it in 22mL, 7mg/mL methanol of 2-methacryloyloxyethylphosphorylcholine In the solution, react at 40°C for 5h, then treat in distilled water at 83°C for 11h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例6Example 6
称取13mmol 2-甲基丙烯酰氧乙基磷酰胆碱和7mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 13mmol 2-methacryloyloxyethylphosphorylcholine and 7mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶12溶于乙醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为4mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于27mL，17mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在22℃反应11h，之后经蒸馏水中87度处理8h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in an ethanol solvent in a molar ratio of 100:12 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 4mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, and then soak it in 27mL, 17mg/mL of 2-methacryloyloxyethylphosphorylcholine methanol In the solution, react at 22°C for 11h, then treat in distilled water at 87°C for 8h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例7Example 7
称取16mmol 2-甲基丙烯酰氧乙基磷酰胆碱和4mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 16mmol 2-methacryloyloxyethylphosphorylcholine and 4mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶17溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为5mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于23mL，14mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在32℃反应7h，之后经蒸馏水中92度处理3h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in methanol solvent in a molar ratio of 100:17 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 5mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, then soak it in 23mL, methanol of 14mg/mL 2-methacryloyloxyethylphosphorylcholine In the solution, react at 32°C for 7h, then treat in distilled water at 92°C for 3h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
实施例8Example 8
称取18mmol 2-甲基丙烯酰氧乙基磷酰胆碱和2mmol 2-氨乙基甲基丙烯酸酯盐酸盐，以0.1mmol过硫酸钾为引发剂，在氮气保护下，70℃聚合反应24h，反应结束后透析，然后在-50℃下冷冻干燥，得到含氨基的磷酰胆碱聚合物。Weigh 18mmol 2-methacryloyloxyethylphosphorylcholine and 2mmol 2-aminoethyl methacrylate hydrochloride, use 0.1mmol potassium persulfate as the initiator, and polymerize at 70°C under nitrogen protection After 24 hours, the reaction was dialyzed, and then freeze-dried at -50°C to obtain an amino group-containing phosphorylcholine polymer.
将本实施例含有氨基的磷酰胆碱聚合物与戊二醛按氨基与醛基摩尔比为100∶18溶于甲醇溶剂中混合均匀后，使含有氨基的磷酰胆碱聚合物的浓度为5mg/mL，涂覆体积10微升/cm2/面在壳聚糖材料表面晾干，然后将其浸泡于23mL，14mg/mL的2-甲基丙烯酰氧乙基磷酰胆碱的甲醇溶液中，在32℃反应7h，之后经蒸馏水中92度处理3h，之后依次用大量的甲醇、蒸馏水洗涤，即可得到高密度的仿细胞外层膜结构涂层表面。The amino group-containing phosphorylcholine polymer and glutaraldehyde in this embodiment are dissolved in a methanol solvent in a molar ratio of 100:18 according to the amino group and the aldehyde group. After mixing uniformly, the concentration of the amino group-containing phosphorylcholine polymer is 5mg/mL, coating volume 10 microliters/cm2 /face on the chitosan material surface to dry, then soak it in 23mL, methanol of 14mg/mL 2-methacryloyloxyethylphosphorylcholine In the solution, react at 32°C for 7h, then treat in distilled water at 92°C for 3h, and then wash with a large amount of methanol and distilled water in turn to obtain a high-density coating surface imitating the outer membrane structure of cells.
以上所述，仅是本发明的较佳实施例，并非对本发明做任何限制，凡是根据发明技术实质对以上实施例所作的任何简单修改、变更以及等效结构变化，均仍属于本发明技术方案的保护范围内。The above are only preferred embodiments of the present invention, and do not limit the present invention in any way. All simple modifications, changes and equivalent structural changes made to the above embodiments according to the technical essence of the invention still belong to the technical solution of the present invention. within the scope of protection.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710118250.9ACN106905554B (en) | 2017-03-01 | 2017-03-01 | A method of the phosphoryl choline polymer containing amino and the density of glutaraldehyde bionic coating |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710118250.9ACN106905554B (en) | 2017-03-01 | 2017-03-01 | A method of the phosphoryl choline polymer containing amino and the density of glutaraldehyde bionic coating |
| Publication Number | Publication Date |
|---|---|
| CN106905554A CN106905554A (en) | 2017-06-30 |
| CN106905554Btrue CN106905554B (en) | 2019-08-13 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710118250.9AExpired - Fee RelatedCN106905554B (en) | 2017-03-01 | 2017-03-01 | A method of the phosphoryl choline polymer containing amino and the density of glutaraldehyde bionic coating |
| Country | Link |
|---|---|
| CN (1) | CN106905554B (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210153043A (en)* | 2019-02-15 | 2021-12-16 | 어큐이티 폴리머스, 인크. | Biocompatible polymer coating containing therapeutic agent |
| CN113877006A (en)* | 2021-09-24 | 2022-01-04 | 宁波健世科技股份有限公司 | Modified polymer membrane material and preparation method thereof |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101531740B (en)* | 2009-01-12 | 2012-05-09 | 西北大学 | Method for constructing simulated outer cell membrane structure on surface of cross-linked chitosan |
| CN102888013B (en)* | 2011-07-20 | 2015-03-25 | 西北大学 | Method for constructing simulated outer cell membrane structure coating on material surface by RAFT (reversible addition fragmentation chain transfer) polymerization technology |
| CN105237778B (en)* | 2015-11-16 | 2017-10-13 | 西安科技大学 | It is a kind of to improve the method for chitosan blood compatibility at room temperature |
| CN105670022B (en)* | 2016-02-25 | 2018-05-04 | 西安科技大学 | A kind of preparation method of Phosphorylcholine bionic coating |
| Publication number | Publication date |
|---|---|
| CN106905554A (en) | 2017-06-30 |
| Publication | Publication Date | Title |
|---|---|---|
| CN103881126B (en) | A kind of method for improving material blood compatibility | |
| CN106750450B (en) | Preparation method of cross-linked adhesive biomimetic coating containing epoxy phosphorylcholine polymer and dopamine | |
| CN105670022B (en) | A kind of preparation method of Phosphorylcholine bionic coating | |
| CN108129687B (en) | A kind of surface is the preparation method of the imitating cell outer-layer membrane structure coating of Phosphorylcholine | |
| CN106832382B (en) | Coating method of double-bionic dopamine phosphorylcholine substance | |
| CN105237778B (en) | It is a kind of to improve the method for chitosan blood compatibility at room temperature | |
| CN106380990B (en) | The preparation method of phosphoryl choline polymer containing aldehyde radical and dopamine cross-linked coating | |
| CN105175756B (en) | The preparation and application of keratin/methacrylic acid-beta-hydroxy ethyl ester composite hydrogel with inierpeneirating network structure | |
| CN107722321B (en) | The method of two kinds of phosphoryl choline polymer bionic coating Chitosan films containing epoxy and amino | |
| CN106905554B (en) | A method of the phosphoryl choline polymer containing amino and the density of glutaraldehyde bionic coating | |
| CN105288731B (en) | A kind of method that epoxy radicals prepares bionic coating with sulfydryl crosslinking | |
| CN107674225B (en) | The preparation method of two kinds of phosphoryl choline polymers crosslinking bionic coating containing aldehyde radical and amino | |
| CN105504328B (en) | A kind of room temperature coats the method for improving chitosan film blood compatibility in next step | |
| CN111763284B (en) | A kind of preparation method of phosphorylcholine coating containing amino group and carboxyl group | |
| CN108715643B (en) | A kind of preparation method of epoxy and aminophosphorylcholine polymer bionic adhesion coating | |
| CN108003369B (en) | A kind of preparation method of the coating surface imitating cell outer layer membrane structure | |
| CN108715644B (en) | A kind of preparation method of bionic adhesion coating of aldehyde group and aminophosphorylcholine polymer | |
| CN104258471B (en) | Graphene oxide that a kind of imitating cell outer-layer membrane structure is modified and preparation method thereof | |
| CN108794794B (en) | A method for modifying the surface biocompatibility of a material and a biomimetic coating prepared therefrom | |
| CN108659168B (en) | A kind of double biomimetic polymer and its preparation method and application | |
| CN108484947B (en) | A method for preparing biomimetic coating from low surface tension solution | |
| CN105237790B (en) | A kind of preparation method of double bionic coatings | |
| CN111825799B (en) | Preparation method of phosphorylcholine coating containing catechol, amino and carboxyl | |
| CN111793399B (en) | Preparation method of two phosphorylcholine coatings containing catechol, amino and carboxyl | |
| CN108517046A (en) | The method of two kinds of phosphoryl choline polymer bionic coating Chitosan films containing epoxy and sulfydryl |
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | Effective date of registration:20190710 Address after:710054 Shaanxi province Xi'an Beilin District Yanta Road No. 58 Applicant after:Xi'an University of Science and Technology Applicant after:Shaanxi coalfield geology Group Co Ltd Address before:710054 Shaanxi province Xi'an Beilin District Yanta Road No. 58 Applicant before:Xi'an University of Science and Technology | |
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | Granted publication date:20190813 Termination date:20200301 | |
| CF01 | Termination of patent right due to non-payment of annual fee |