CN108816689A - 一种具有长效抗菌性能的超亲水涂层及其制备方法 - Google Patents
一种具有长效抗菌性能的超亲水涂层及其制备方法 Download PDFInfo
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- CN108816689A CN108816689A CN201810732803.4A CN201810732803A CN108816689A CN 108816689 A CN108816689 A CN 108816689A CN 201810732803 A CN201810732803 A CN 201810732803A CN 108816689 A CN108816689 A CN 108816689A
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Abstract
本发明提供了一种具有长效抗菌性能的超亲水涂层及其制备方法，制备方法包括以下步骤：对基底材料表面进行预处理；将处理后的基底材料置于偏酸性缓冲液中，然后加入多酚化合物、多胺基化合物、抗菌剂和氧化剂进行反应；将反应所得物浸没于去离子水中，进行超声清洗，然后在氮气条件下干燥，制得。该方法操作简单、反应条件温和，制备的涂层材料中含多酚化合物和大量的羧基、酚羟基、醌基及胺基等功能基团和具有抗菌性能的金属离子或药物，此金属离子和抗菌药物通过与酚羟基官能团相互作用或体系π‑π堆积作用实现抗菌剂长期存在。该涂层可用于医用材料领域的伤口敷料材料的制备。
Description
技术领域
本发明属于医用材料技术领域，具体涉及一种具有长效抗菌性能的超亲水涂层及其制备方法，可用于制备具有抗菌功能的伤口敷料和植入器械表面保护材料。
背景技术
临床上的植入器械普遍存在细菌感染的问题，虽然无菌处理植入器械在一定程度上减少了细菌感染的程度，但是，据文献报道，仍然有4％～6％的患者在植入初期因病原体感染而导致植入失效。据统计分析80％的患者是因为细菌感染而失效的。因为在相应手术治疗过程中，由于患者组织或器官长时间暴露在开放性环境中，接触病原菌机会增加，进而引发感染；同时，材料的植入会损伤人体正常皮肤和粘膜，抑制微区免疫系统功能发挥甚至使其失效。再次，材料植入后短时间内，会在其表面聚集大量蛋白，从而引发一系列后续反应，为细菌的繁殖提供丰富的营养物质，给植入体细菌感染创造有利条件。植入器械细菌感染一旦发生，是非常难以治愈的。因为细菌为适应机体复杂的微环境会在器械表面形成生物被膜，生物被膜可以有效的保护细菌避免被免疫系统清除，并在一定的时候可以将细菌释放出来，形成悬浮菌，造成细菌感染复发。细菌形成稳固的生物被膜后，只能通过二次手术移除植体的方法进行根治。这将给患者带来巨大的身体伤害和经济损失。故在植入体表面构建抗菌剂，抑制生物被膜的形成，对植入器械至关重要。此外，在治疗伤口过程中所用的敷料也应具有一定的抗菌性。植入器械以及伤口敷料所用的抗菌剂的种类主要有无机金属离子和抗生素药物。对无机金属离子的抗菌剂，目前使用最为广泛和使用时间最长的是银粒子，并在骨科和牙缺损治疗得到了广泛的应用。目前，银粒子的抗菌机制普遍认为是银粒子入细菌体内的银离子与其DNA碱基对相结合，置换嘌呤和嘧啶中相邻氮的氢键，扰乱DNA的复制和转录，破坏细胞内呼吸系统正常活动，从而使细菌死亡。抗生素是通过病原体刺激高等动植物或微生物(真菌或细菌)所产生的具有抵抗病原体的一种次级代谢产物，对很多细菌都有一直和杀灭作用，可以长期有效的抵抗细菌感染。目前抗生素的使用方法主要有两种，一是术后给病人注射大量抗生素，预防细菌感染；另一种则是在生物材料表面负载抗生素，赋予其抗菌性能。
对于植入器械而言，材料表面亲/疏水性是影响蛋白质吸附的首要因素。同时，由于蛋白质是带有两性电荷的聚电解质，若材料表面也带有两亲性离子结构或亲水基团，通过富集水化层或空间排斥也可以削弱材料与蛋白的相互作用，抑制非特异性蛋白的吸附。而且，目前研究最为广泛的材料表面的亲/疏水性对蛋白吸附和细胞相容性的影响中，比较一致的观点认为亲水性材料比疏水性材料具有更好的生物相容性。因为疏水表面蛋白粘附得较为牢固，不易脱落，且容易导致蛋白质的构象变化；而亲水表面对蛋白的粘附强度较低，有利于蛋白原有自由构象的调整与维持。传统的制备亲水性表面方法，一般是将亲水性功能基团(如羧酸基、磺酸基、胺基、季胺基)或者是具有亲水性的物质接枝或涂覆在材料表面，而无论是以共价固定还是物理共混涂覆到材料表面，都必须依赖分子的尺寸、构象、活性以及其在材料表面发挥亲水性作用的稳定性及持久性。生物分子一旦在服役过程中失活或大量丢失，便不能有效发挥其生物学功能。因此能在涂层表面直接生成大量稳定的亲水基团且不受接枝过程影响显得尤为重要。对于伤口敷料而言，其抗菌性能并不长久，无法实现对伤口部位的长效抗菌功能。
发明内容
针对现有技术中存在的上述问题，本发明提供一种具有长效抗菌性能的超亲水涂层及其制备方法，本发明涂层通过原位负载方式，将抗菌剂负载到整个涂层中，可以在材料表面长期源源不断的释放出适量的抗菌剂，能有效阻抗和杀灭细菌，并实现长效抗菌的效果。
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：
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对基底材料表面进行预处理；
(2)将处理后的基底材料置于pH值为3-7的缓冲液中，接着加入多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂，使得多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂的终浓度分别为0.01-7mg/mL、0.01-6mg/mL、0.01-10mg/mL和0.01-10mg/mL，然后在4-50℃反应1-30h；其中，水溶性抗菌剂为硝酸银、氯化银、氯化铜、高锰酸钾、氯化锌、青霉素、头孢霉素、四环素、红霉素、链霉素、氯霉素、头孢氨苄、诺氟沙星、庆大霉素、季铵盐和季铵盐衍生物中的一种或几种混合；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3-4次，每次4-5min，然后在氮气条件下干燥，制得。
进一步地，基底材料为金属基生物材料、陶瓷基生物材料、高分子基生物材料或复合生物材料。
进一步地，缓冲液为乙酸-乙酸盐缓冲液、2-(N-吗啡啉)乙磺酸缓冲液、甘氨酸-盐酸缓冲液、邻苯二甲酸-盐酸缓冲液、邻苯二甲酸氢钾-氢氧化钠缓冲液、磷酸氢二钠-柠檬酸缓冲液、柠檬酸-氢氧化钠-盐酸缓冲液或柠檬酸-柠檬酸钠缓冲液。
进一步地，多酚化合物为单宁酸、没食子酸、丹酚酸B、表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯、表儿茶素、表没食子儿茶素、邻苯二酚、邻苯三酚、黄酮类、花色素苷、鞣花酸和原花色素A2中的一种或几种混合。
进一步地，多胺基化合物为L-精氨酸、乙二胺、戊二胺、2,2,4-三甲基六亚甲基二胺、1,8-二氨基辛烷、甲基环己烷二胺、1,3-二氨基甲基环己烷、2,4,6-三氨甲基环己烷、1,4-双二氨己基环己烷和2-(3,4-二羟基苯基)乙胺中的一种或几种混合。
进一步地，水溶性抗菌剂为头孢霉素和有机硅季铵盐按重量比为1：2混合的混合物。
进一步地，水溶性氧化剂为过氧化氢、过硫酸铵、三氯化铁、浓硝酸、高碘酸钠、高锰酸钾和重铬酸钾中的一种或几种混合。
进一步地，步骤(2)中多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂的终浓度分别为5mg/mL、7mg/mL、3mg/mL和7mg/mL。
进一步地，步骤(2)中反应温度为25℃，反应时间为2h。
本发明提供的具有长效抗菌性能的超亲水涂层及其制备方法，具有以下有益效果：
本发明通过加入多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂，使其相互作用制备改性涂层，在涂层表面生成多种亲水功能基团，从而使得涂层具有超亲水性，因涂层具有超亲水性，便赋予了涂层还具有易洗、防污、抗污的特性，可以有效阻抗非特异性蛋白的黏附，同时由于抗菌剂和多酚官能团的存在，使得涂层具有优异的抗菌性能，此处涂层表面大量的官能团可以为后续生物分子的接枝修饰改性提供较好的平台。
在具体反应过程中，多酚化合物、多胺基化合物与抗菌剂在基底材料表面改性能力主要是基于其可以在不同的材料表面自选择地以共价、氢键、疏水作用力及超分子作用力等多种模式与材料表面发生物理/化学沉积，形成多酚层，并且，多酚化合物与氧化剂反应过程中，被氧化剂的电子攻击，使得邻二醌结构转化为羧基结构，使得涂层含有大量的羧基，从而形成超亲水表面；同时，无机抗菌剂会与多酚化合物通过氧化还原作用和螯合作用参与到涂层中，在增强涂层内聚力的同时还使得涂层具有优异的抗菌能力；抗生素会通过π-π作用和氢键作用与多酚化合物结合，并在酸性条件下释放出来，使得涂层长期具有优异的抗菌能力。
多酚化合物的分子结构中含有大量的邻位酚羟基，这些邻位酚羟基可与金属形成稳定螯合作用，因此在金属表面可形成稳定的结合，此外还能通过分子间疏水作用和氢键作用与陶瓷生物材料、高分子生物材料形成稳定的结合；剩余的多酚化合物还具有优异的抗氧化性和清除氧自由基的能力，对心血管具有保护作用。
综上所述，本发明制备方法操作简单，反应条件温和，制得的涂层具有优异的生物抗污性能和杀菌性能，能有效的阻抗细菌在材料表面的吸附和其生物被膜的形成，使材料具有良好的抗菌性，能有效用于制备具有抗菌功能的伤口敷料和植入器械表面保护材料，且材料表面的多功能基团为后续生物分子的接枝和修饰量提供较好的平台。
附图说明
图1为传统多酚沉积薄膜(A)和新型多酚薄膜(B)的抑菌结果图。
具体实施方式
实施例1
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对医用不锈钢材料进行抛光、清洗和干燥处理；
(2)将处理后的基底材料置于pH值为4的乙酸-乙酸钠缓冲液中，接着加入没食子酸、L-精氨酸、硝酸银和质量分数为65％-68％的浓硝酸，没食子酸、L-精氨酸、硝酸银和浓硝酸的终浓度分别为0.01mg/mL、0.01mg/mL、0.01mg/mL和质量分数为34％，然后在40℃反应30h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例2
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对镍钛合金材料进行抛光、清洗和干燥处理；
(2)将处理后的基底材料置于pH值为3.5的邻苯二甲酸-盐酸缓冲液中，接着加入表没食子儿茶素没食子酸酯(EGCG)、乙二胺、氯化锌和质量分数为30％的过氧化氢溶液，表没食子儿茶素没食子酸酯(EGCG)、乙二胺、氯化锌和质量分数为30％的过氧化氢溶液的终浓度分别为1mg/mL、2mg/mL、1mg/mL和质量分数为15％，然后在30℃反应10h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例3
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对聚四氟乙烯材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为6的2-(N-吗啡啉)乙磺酸缓冲液中，接着加入表儿茶素没食子酸酯(ECG)、1,8-二氨基辛烷、氯化铜和高碘酸钠溶液，表儿茶素没食子酸酯(ECG)、1,8-二氨基辛烷、氯化铜和高碘酸钠溶液的终浓度分别为2mg/mL、4mg/mL、2mg/mL和4mg/mL，然后在15℃反应24h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例4
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对聚氨酯材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为6的柠檬酸-柠檬酸钠缓冲液中，接着加入没食子酸、1,4-双二氨己基环己烷、青霉素和过硫酸铵溶液，没食子酸、1,4-双二氨己基环己烷、青霉素和过硫酸铵溶液的终浓度分别为2mg/mL、5mg/mL、1mg/mL和2mg/mL，然后在20℃反应18h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例5
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对各向同性热解碳LTIC材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为4.5的邻苯二甲酸氢钾-氢氧化钠缓冲液中，接着加入表儿茶素、2-(3,4-二羟基苯基)乙胺、头孢霉素、有机硅季铵盐和高碘酸钠溶液，表儿茶素、2-(3,4-二羟基苯基)乙胺、头孢霉素、有机硅季铵盐和高碘酸钠溶液的终浓度分别为5mg/mL、7mg/mL、1mg/mL、2mg/mL和7mg/mL，然后在25℃反应2h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例6
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对医用胶原材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为4的甘氨酸-盐酸缓冲液中，接着加入表没食子儿茶素(EGC)、戊二胺、红霉素和质量分数为65％-68％的浓硝酸，表没食子儿茶素(EGC)、戊二胺、红霉素和质量分数为65％-68％的浓硝酸的终浓度分别为0.1mg/mL、0.5mg/mL、0.5mg/mL和质量分数为34％，然后在25℃反应6h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例7
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对天然橡胶材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为5.5的磷酸氢二钠-柠檬酸缓冲液中，接着加入邻苯三酚、1,3-二氨基甲基环己烷、庆大霉素和高锰酸钾溶液，邻苯三酚、1,3-二氨基甲基环己烷、庆大霉素和高锰酸钾溶液的终浓度分别为7mg/mL、10mg/mL、5mg/mL和10mg/mL，然后在40℃反应1h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
实施例8
一种具有长效抗菌性能的超亲水涂层，其制备方法包括以下步骤：
(1)对生物活性玻璃陶瓷涂层增强的钛合金材料进行清洗和干燥处理；
(2)将处理后的基底材料置于pH值为4.5的柠檬酸-氢氧化钠-盐酸缓冲液中，接着加入表没食子儿茶素没食子酸酯(EGCG)、2,2,4-三甲基六亚甲基二胺、头孢氨苄和重铬酸钾溶液，表没食子儿茶素没食子酸酯(EGCG)、2,2,4-三甲基六亚甲基二胺、头孢氨苄和重铬酸钾溶液的终浓度分别为6mg/mL、7mg/mL、4mg/mL和3mg/mL，然后在30℃反应5h；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3次，每次5min，然后在氮气条件下干燥，制得。
对比例1
传统的多酚涂层是多酚在碱性环境中通过自氧化、交联沉积薄膜制得，即将不锈钢基底放在2mg/ml的pH为8.5的多巴胺溶液中静置3h，然后超声清洗3遍，每次5min。
用传统多酚涂层和实施例5制备的新型抗菌涂层做抑菌圈实验，具体过程为：用紫外灭菌样品正、反面各30min待用；取浓度为2*107的金黄色葡萄球菌200ul进行涂板，并将样品放在培养皿涂板的中间位置；将放有样品的细菌培养板倒置放于37℃的烘箱中培养24h；观察，拍照，其结果见图1。由图1可知，本发明涂层的抑菌圈明显大于传统的涂层，说明本发明涂层具有优异的抗菌性能。
本发明制得的涂层通常低于200nm，所得涂层均匀，制备涂层所需原料投入很少，投入量易于调控，该涂层可以在多种材料表面进行修饰而不影响本体材料的性能，与传统的涂层相比，操作简单，成本低，适用于工业化生产。
Claims (10)
1.一种具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，包括以下步骤：
(1)对基底材料表面进行预处理；
(2)将处理后的基底材料置于pH值为3-7的缓冲液中，接着加入多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂，使得多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂的终浓度分别为0.01-7mg/mL、0.01-6mg/mL、0.01-10mg/mL和0.01-10mg/mL，然后在4-50℃反应1-30h；其中，水溶性抗菌剂为硝酸银、氯化银、氯化铜、高锰酸钾、氯化锌、青霉素、头孢霉素、四环素、红霉素、链霉素、氯霉素、头孢氨苄、诺氟沙星、庆大霉素、季铵盐和季铵盐衍生物中的一种或几种混合；
(3)将步骤(2)所得物浸没于去离子水中，超声清洗3-4次，每次4-5min，然后在氮气条件下干燥，制得。
2.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，基底材料为金属基生物材料、陶瓷基生物材料、高分子基生物材料或复合生物材料。
3.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，缓冲液为乙酸-乙酸盐缓冲液、2-(N-吗啡啉)乙磺酸缓冲液、甘氨酸-盐酸缓冲液、邻苯二甲酸-盐酸缓冲液、邻苯二甲酸氢钾-氢氧化钠缓冲液、磷酸氢二钠-柠檬酸缓冲液、柠檬酸-氢氧化钠-盐酸缓冲液或柠檬酸-柠檬酸钠缓冲液。
4.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，多酚化合物为单宁酸、没食子酸、丹酚酸B、表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯、表儿茶素、表没食子儿茶素、邻苯二酚、邻苯三酚、黄酮类、花色素苷、鞣花酸和原花色素A2中的一种或几种混合。
5.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，多胺基化合物为L-精氨酸、乙二胺、戊二胺、2,2,4-三甲基六亚甲基二胺、1,8-二氨基辛烷、甲基环己烷二胺、1,3-二氨基甲基环己烷、2,4,6-三氨甲基环己烷、1,4-双二氨己基环己烷和2-(3,4-二羟基苯基)乙胺中的一种或几种混合。
6.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，水溶性抗菌剂为头孢霉素和有机硅季铵盐按重量比为1：2混合的混合物。
7.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，水溶性氧化剂为过氧化氢、过硫酸铵、三氯化铁、浓硝酸、高碘酸钠、高锰酸钾和重铬酸钾中的一种或几种混合。
8.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，步骤(2)中多酚化合物、多胺基化合物、水溶性抗菌剂和水溶性氧化剂的终浓度分别为5mg/mL、7mg/mL、3mg/mL和7mg/mL。
9.根据权利要求1所述的具有长效抗菌性能的超亲水涂层的制备方法，其特征在于，步骤(2)中反应温度为25℃，反应时间为2h。
10.如权利要求1-9任一项所述的制备方法制得的具有长效抗菌性能的超亲水涂层。
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