CN108969805B - 一种可催化一氧化氮释放的抗凝血水凝胶涂层及其制备方法 - Google Patents
一种可催化一氧化氮释放的抗凝血水凝胶涂层及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种可催化一氧化氮释放的抗凝血水凝胶涂层及其制备方法。该方法包括以下步骤：((1)预处理基底材料；(2)室温下，将预处理后的基底材料依次置于聚阳电解质、多酚与化合物A的混合溶液、聚阴电解质中反应1～20min后，清洗3～5次；(3)室温下，以步骤(2)所得产物为底物，重复步骤(2)所述操作5～20次，经氮气干燥后，得到可催化一氧化氮释放的抗凝血水凝胶涂层。本发明方法制备得到的水凝胶涂层方法简单、结构稳定、生物相容性好，且具有长期稳定的催化NO释放保护内皮细胞和抗凝血作用，该涂层可用于心血管内支架材料和心脏瓣膜的表面改性。
Description
技术领域
本发明属于医用材料技术领域，具体涉及一种可催化一氧化氮释放的抗凝血水凝胶涂层及其制备方法，该涂层可用于心血管内支架材料和心脏瓣膜的表面改性。
背景技术
随着人们生活压力增大、作息与饮食的不规律，我国心血管疾病已经远远超过肿瘤及其他疾病对生命健康的威胁，成为疾病导致死亡的第一杀手。心血管材料主要包括血管支架、人工血管、心血管生物瓣膜、血液透析器等。一直以来，心血管材料面临着抗凝血的重大问题，具有优异的表面抗凝血性能是血液接触材料的关键。一氧化氮具有调节血管舒张、抑制血管内平滑肌细胞增生以及血小板聚集、激活也起到重要的作用。目前报道的可以释放一氧化氮的材料主要通过表面的可降解高分子涂层中装载一氧化氮供体亚硝基硫醇(RSNO)，随着涂层的降解催化一氧化氮释放，然而这种材料由于装载的一氧化氮供体量有限，且高分子涂层降解速率过快，导致不能满足服役周期内的抗凝血功能。随着一氧化氮供体的消耗会出现晚期凝血问题，缩短了材料的使用寿命。因此，制备一种具有可控装载一氧化氮供体和持续释放的表面涂层材料是心血管材料亟待解决的问题。
发明内容
针对现有技术中的上述不足，本发明提供一种可催化一氧化氮释放的抗凝血水凝胶涂层及其制备方法，可有效解决现有涂层材料中一氧化氮释放量不可控，不能满足服役周期内的抗凝血功能的问题。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)预处理基底材料；
(2)将预处理后的基底材料置于0.001～2mM，pH值为5～12的聚阳电解质中，于室温下反应1～2min后，清洗3～5次；
(3)将步骤(2)所得产物置于pH值为5～12的混合溶液中，于室温下反应5～20min后，清洗3～5次；所述混合溶液包括0.001～20mM的多酚化合物和0.001～0.1mM化合物A；其中，所述化合物A为诱导催化一氧化氮释放，且二硫键或二硒键的两端含有胺基或巯基的化合物；
(4)将步骤(3)所得产物置于0.001～1mM，pH值为5.5～12的聚阴电解质中，于室温下反应1～20min后，清洗3～5次；
(5)于室温下，以步骤(4)所得产物为底物，重复步骤(2)～(4)所述操作5～20次，经氮气干燥后，得到抗凝血水凝胶涂层。
进一步地，步骤(1)中基底材料为金属基生物材料、陶瓷基生物材料或高分子基生物材料。
其中，金属基生物材料为镁及其合金、钛及其合金、铁及其合金、不锈钢、钴基合金、金或锌合金；
陶瓷基生物材料TiO2薄膜、硅、SiO2、羟基磷灰石、热解碳LTIC、磷酸钙、金刚石或类金刚石；
高分子基生物材料为聚己内酯、聚乳酸、聚甲醛、聚氨酯、聚四氟乙烯、硅橡胶、涤纶、乙交酯-丙交酯共聚物或聚三亚甲基碳酸酯。
进一步地，步骤(2)中聚阳电解质为聚-L-赖氨酸氢溴酸盐、聚-L-精氨酸盐酸盐或壳聚糖。
进一步地，步骤(3)中多酚化合物为表没食子儿茶素没食子酸酯、白藜芦醇、姜黄素、丹参酮、表儿茶素、表没食子儿茶素、表儿茶素没食子酸酯、没食子酸、维生素B2、黄酮化合物或单宁酸。
进一步地，化合物A为半胱氨酸、胱氨酸、硒代胱氨酸、硒代胱胺、胱胺、依布硒淋、谷胱甘肽过氧化酶或双(邻胺甲酰基苯基)二硒化物。
进一步地，步骤(4)中聚阴电解质为聚谷氨酸、聚天冬氨酸、胶原蛋白、透明质酸、葡聚糖、硫酸软骨素、硫酸皮肤素、硫酸角质素、肝素、聚谷氨酸或或脱氧核糖核酸。
进一步地，聚阳电解质浓度为0.1mM，pH值为6.5；所述混合溶液中多酚化合物浓度为10mM，化合物A浓度为0.1mM；所述聚阴电解质浓度为0.01mM，pH值为7.4。
上述方法制备得到的可催化一氧化氮释放的抗凝血水凝胶涂层。
本发明的有益效果为：
1、本发明方法制备得到的水凝胶涂层通过聚阳电解质和聚阴电解质与多酚化合物间的静电作用、氢键、π-π作用和少量的共价键协同作用，具有致密的结构和优异的稳定性，为药物分子长期稳定的扩散和持续催化一氧化氮释放提供了可能。
2、通过层层自组装原理和技术将聚阳电解质、夹心层、聚阴电解质制备成“三明治”夹心结构的原位水凝胶，通过组装调控组元的浓度、pH、组装层数来调节涂层厚度、夹心层的含量，可以获得不同装载量的材料，其中，夹心层为多酚类化合物与化合物A结合组成，而化合物A为二硫键或二硒键的两端含胺基或巯基的化合物，能够有效地催化一氧化氮供体释放一氧化氮，由此，本发明法制备得到的水凝胶涂层能在缓慢降解的过程中催化释放一氧化氮，避免水凝胶涂层在降解晚期出现凝血的问题。
3、多酚具有清除体内氧自由基的功能，因此发明方法制备得到的水凝胶涂层具有良好的抗氧化性，能够起到延缓机体衰老的作用；另外，多酚化合物具有良好的抗炎、止痛作用，可以调节血管内细胞的功能，因此，该水凝胶涂层可广泛地应用于抗凝血材料表面，尤其是用于心血管内支架材料和心脏瓣膜的表面改性。
附图说明
图1是本发明制备得到的抗凝血水凝胶涂层的原子力显微镜的形貌图；
图2是本发明制备得到的抗凝血水凝胶涂层的扫描电镜形貌图。
具体实施方式
下面对本发明的具体实施方式进行描述，以便于本技术领域的技术人员理解本发明，但应该清楚，本发明不限于具体实施方式的范围，对本技术领域的普通技术人员来讲，只要各种变化在所附的权利要求限定和确定的本发明的精神和范围内，这些变化是显而易见的，一切利用本发明构思的发明创造均在保护之列。
实施例1
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对不锈钢基底材料进行超声清洗、干燥；
(2)将清洗后的不锈钢置于0.1mM，pH值为6的壳聚糖溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为11，含有1mM单宁酸和0.01mM依布硒淋的混合溶液中，于室温下反应5min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.1mM，pH值为11的透明质酸溶液中，于室温下反应1min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例2
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对钛合金基底材料进行清洗、干燥；
(2)将清洗后的钛合金置于0.01mM，pH值为7.4的聚-L-精氨酸盐酸盐溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.2，含有0.1mM表没食子儿茶素没食子酸酯和0.01mM胱胺的混合溶液中，于室温下反应5min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.1mM，pH值为6.8的聚谷氨酸溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例3
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对聚四氟乙烯材料进行清洗、干燥；
(2)将清洗后的聚四氟乙烯置于2mM，pH值为7.4的聚-L-赖氨酸氢溴酸盐溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为6.5，含有0.5mM表没食子儿茶素和0.001mM硒代胱氨酸的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.05mM，pH值为6.9的葡聚糖溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例4
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对需要改性的纯铁材料进行抛光、清洗、干燥；
(2)将经步骤(1)处理后的纯铁材料置于0.1mM，pH值为7.4的聚-L-精氨酸盐酸盐溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.2，含有0.5M表儿茶素和0.01mM二硒代胱胺的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.1mM，pH值为6.8的硫酸乙酰肝素溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例5
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对需要改性的石英片进行清洗、干燥；
(2)将经步骤(1)处理后的石英片置于2mM，pH值为6.5的壳聚糖溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.2，含有0.001mM表没食子儿茶素和0.001mM硒代胱胺酸的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.1mM，pH值为6.5的脱氧核糖核酸溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例6
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对需要改性的热解碳LTIC材料进行抛光、清洗、干燥；
(2)将经步骤(1)处理后的热解碳LTIC材料置于0.2mM，pH值为6.8的聚-L-精氨酸盐酸盐溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.2，含有0.5mM表儿茶素没食子酸酯和0.1mM硒代胱胺酸的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.05mM，pH值为6.8的聚谷氨酸溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例7
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对需要改性的涤纶材料进行清洗、干燥；
(2)将经步骤(1)处理后的涤纶材料置于0.02mM，pH值为7.4的聚-L-赖氨酸氢溴酸盐溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.6，含有1mM白藜芦醇和0.005mM硒代胱胺酸的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.9mM，pH值为7.2的硫酸软骨素溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
实施例8
一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，包括以下步骤：
(1)对需要改性的镍钛合金材料进行抛光、清洗、干燥；
(2)将经步骤(1)处理后的镍钛合金置于0.1mM，pH值为6.5的壳聚糖溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(3)将步骤(2)所得产物置于pH值为7.2，含有0.1mM单宁酸和0.1mM二硒代胱胺的混合溶液中，于室温下反应2min，然后取出并用去离子水清洗3次；
(4)将步骤(3)所得产物置0.1mM，pH值为6.8的肝素溶液中，于室温下反应3min，然后取出并用去离子水清洗3次；
(5)于室温下，以步骤(4)所得产物为底物，分别重复步骤(2)～(4)所述操作3次、5次、10次和20次，经氮气干燥后，可得到厚度不同，且可催化一氧化氮释放的抗凝血水凝胶涂层。
检测
分别对实施例1制备得到的抗凝血水凝胶涂层进行原子力显微镜形貌检测，以及扫描电镜形貌图，从图1中可以看出组装图层中出现了局部的交联纤维网络结构，说明多酚分子将聚阳电解质与聚阴电解质交联结合起来了，使得采用该方式组装的涂层结构更稳定；图2中所述抗凝血水凝胶涂层在SD大鼠皮下15天后取出，我们可以看到该图层具有良好的结构，局部出现了体内矿化现象，表明其结构稳定性强。
再分别对实施例1制备得到的抗凝血水凝胶涂层与传统涂层进行耐极端条件下的破坏与长期稳定性的测试，其具体过程如下：
1、极端条件下稳定性测试
将制备好的涂层经过预处理后，称量样品的质量并观察样品SEM形貌。随后将涂层分别在pH值为1的盐酸溶液、pH值为12的氢氧化钠、4M NaCl溶液中浸泡2小时，取出用大量去离子水清洗，氮气干燥后称量样品的质量，并测量SEM形貌。通过质量保留比例及样品形貌的变化来对比改性涂层与传统涂层的区别。
2、长期稳定性测试
将制备好的样品浸泡在37℃，pH值为7.4的PBS溶液中进行长期浸泡，每隔7天换一次液体，每隔15天观察样品形貌的变化。
3、药物释放速率检测
涂层中药物释放速率测试：将制备好的涂层置于pH值为7.4的PBS溶液中浸泡，每隔一定时间取一定量的溶液，并补充等体积的PBS溶液，经过一定时间的收集与检测，通过超声将涂层破坏收集其中总的药物含量，计算出涂层中药物的总含量，并计算出各个时间点的释放药物的百分比。
本发明实施例1制备得到的抗凝血水凝胶涂层与传统涂层通过上述检测进行对比，可发现本发明实施例1制备得到的抗凝血水凝胶涂层不管是在稳定性还是药物释放速率方面，其性能均远远优于传统涂层。
Claims (8)
1.一种可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，包括以下步骤：
(1)预处理基底材料；
(2)将预处理后的基底材料置于0.001～2mM，pH值为5～12的聚阳电解质中，于室温下反应1～2min后，清洗3～5次；
(3)将步骤(2)所得产物置于pH值为5～12的混合溶液中，于室温下反应5～20min后，清洗3～5次；所述混合溶液包括0.001～20mM的多酚化合物和0.001～0.1mM化合物A；其中，所述化合物A为诱导催化一氧化氮释放，且二硫键或二硒键的两端含有胺基或巯基的化合物；
(4)将步骤(3)所得产物置于0.001～1mM，pH值为5.5～12的聚阴电解质中，于室温下反应1～20min后，清洗3～5次；
(5)于室温下，以步骤(4)所得产物为底物，重复步骤(2)～(4)所述操作5～20次，经氮气干燥后，得到抗凝血水凝胶涂层。
2.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，步骤(1)中所述基底材料为金属基生物材料、陶瓷基生物材料或高分子基生物材料。
3.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，步骤(2)中所述聚阳电解质为聚-L-赖氨酸氢溴酸盐、聚-L-精氨酸盐酸盐或壳聚糖。
4.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，步骤(3)中所述多酚化合物为表没食子儿茶素没食子酸酯、白藜芦醇、姜黄素、丹参酮、表儿茶素、表没食子儿茶素、表儿茶素没食子酸酯、没食子酸、维生素B2、黄酮化合物或单宁酸。
5.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，所述化合物A为半胱氨酸、胱氨酸、硒代胱氨酸、硒代胱胺、胱胺、依布硒淋、谷胱甘肽过氧化酶或双(邻胺甲酰基苯基)二硒化物。
6.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，步骤(4)中所述聚阴电解质为聚谷氨酸、聚天冬氨酸、胶原蛋白、透明质酸、葡聚糖、硫酸软骨素、硫酸皮肤素、硫酸角质素、肝素、聚谷氨酸或脱氧核糖核酸。
7.根据权利要求1所述的可催化一氧化氮释放的抗凝血水凝胶涂层的制备方法，其特征在于，所述聚阳电解质浓度为0.1mM，pH值为6.5；所述混合溶液中多酚化合物浓度为10mM，化合物A浓度为0.1mM；所述聚阴电解质浓度为0.01mM，pH值为7.4。
8.权利要求1～7任一项所述方法制备得到的可催化一氧化氮释放的抗凝血水凝胶涂层。
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| CN111569160B (zh) * | 2020-05-26 | 2021-05-11 | 四川大学 | 基于重组人源胶原蛋白的多功能支架涂层及其制备方法 |
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| CN115232305B (zh) * | 2022-08-05 | 2023-09-19 | 上海交通大学 | 可释放no气体的聚谷氨酸基水凝胶及其制备方法和应用 |
| CN116763722B (zh) * | 2023-06-19 | 2024-04-26 | 广东省第二人民医院（广东省卫生应急医院） | 水凝胶制备材料、水凝胶材料及其制备方法与应用 |
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