CN113244436A - 一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和应用 - Google Patents

一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和应用 Download PDF

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CN113244436A
CN113244436A CN202110309134.1A CN202110309134A CN113244436A CN 113244436 A CN113244436 A CN 113244436A CN 202110309134 A CN202110309134 A CN 202110309134A CN 113244436 A CN113244436 A CN 113244436A
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李向阳
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Abstract

本发明公开了一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和应用,涉及抗菌涂层技术领域。本发明的抗菌涂层是由氯己定和儿茶酚胺自组装形成纳米粒子,纳米粒子在多种基底表面粘附,形成一层致密的聚合涂层。本发明通过制备的抗菌涂层与现有技术中公开的含氯己定的抗菌材料相比具有相同的抗菌功能,针对现有技术中抗菌材料具有毒性,生物相容性差的技术问题,选取儿茶酚胺类物质作为基底,显著改善了其细胞毒性,尤其是褐藻多酚与氯己定聚合形成的涂层;同时本抗菌涂层的稳定性强,在模拟体液浸泡多日天的情况下仍然具有一定的抗菌效果;工艺简单,反应条件易控,成本低。

Description

一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和 应用
技术领域
本发明属于抗菌涂层技术领域,特别是涉及一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和应用。
背景技术
具有生物安全性且可以稳定抗菌的材料在生物医用器械领域一直是最亟需的材料之一。实现材料表面的稳定抗菌可以有效减少医用器械表面感染的发生、减少抗生素的使用量,大幅度延长器械的服役时间。目前主要的抗菌表面改性方式均存在一定的弊端,如:在抗菌的同时具有较强的细胞毒性、抗菌谱窄以及抗菌时效较短、涂层不稳定等。
氯己定,化学名为双氯苯双胍己烷,系阳离子表面活性剂,具有相当强的广谱抑菌、杀菌作用,是一种较好的杀菌消毒药,对革兰阳性和阴性菌均具有抗菌效果。氯己定的杀菌作用主要因为其强正电性,可以吸附在细菌胞浆膜的渗透屏障,使细胞内容物漏出,低浓度时呈抑菌作用,高浓度时呈杀菌作用。然而达到杀菌浓度的氯己定具有一定的细胞毒性,且氯己定在材料及体内的吸附时间较短,一般为24小时左右,现有技术并不能有效改善其细胞相容性和持续稳定性。
儿茶酚胺是一类具有单个或多个酚羟基的有机物,其特点是可以通过氧化聚合、金属离子配位以及含胺基有机物聚合形成非材料依赖性的涂层,目前已经是材料科学领域广泛应用的非材料依赖涂层的构建单体之一。儿茶酚胺类多呈负电,且均含有大量酚羟基,可以与具有正点性的氯己定以电荷作用以及共价作用,实现稳定的结合。同时酚类可以和多种基底进行非材料依赖性的吸附,能够实现氯己定、儿茶酚聚合物的有效非材料依赖性的固定,进而完成对多种材料的表面改性。
本发明通过氯己定和儿茶酚胺的自组装性质以及儿茶酚胺的自聚合构建了一种抗菌涂层,申请人惊奇地发现,构建的抗菌涂层除了具有优异的抗菌效果外,并不具有明显的细胞毒性,这是由于负电性的儿茶酚胺和正电性的氯己定中和,降低了整体的表面电位,从而降低了整体的细胞毒性。同时,形成的纳米粒子具有一定的氯己定释放能力,从而起到原位抗菌作用。
发明内容
本发明的目的在于提供一种氯己定和儿茶酚胺共聚合的抗菌涂层及其制备方法和应用,其可用于对生物医用材料表面改性,解决了现有抗菌表面改性不能持续稳定抗菌且生物相容性差的问题,为材料表面抗菌材料提供新的选择。
为解决上述技术问题,本发明是通过以下技术方案实现的:
本发明为一种氯己定和儿茶酚胺共聚合的抗菌涂层,所述抗菌涂层是由氯己定和儿茶酚胺自组装形成纳米粒子,所述纳米粒子在多种基底表面粘附,形成一层致密的聚合涂层。
进一步地,所述抗菌涂层的制备方法,包括以下步骤:
步骤1:配置一定浓度的儿茶酚胺水溶液和一定浓度的氯己定溶液;
步骤2:向儿茶酚胺水溶液中缓慢滴入步骤1中配置的氯己定溶液,使儿茶酚胺和氯己定聚合充分;
步骤3:静置24小时,在基底上会沉积一层氯己定和儿茶酚胺的聚合涂层。
进一步地,所述儿茶酚胺为多巴胺、去甲基肾上腺素、单宁酸、没食子酸、褐藻多酚、表没食子儿茶素没食子酸酯。
进一步地,所述儿茶酚胺优选为褐藻多酚。
进一步地,儿茶酚胺的浓度对于纳米粒子的聚合至关重要,对于涂层的构建有着至关重要的影响,不同浓度儿茶酚胺具有不同的电性以及酚羟基含量,因此会影响纳米粒子和涂层的聚合。所述儿茶酚胺的浓度范围为(0.05-10mg/mL)。
进一步地,通过梯度浓度实验筛选,所述儿茶酚胺浓度优选为(0.05-2mg/mL)。
进一步地,氯己定的浓度对于纳米粒子的聚合至关重要,进一步对于涂层的构建有着至关重要的影响,不同浓度氯己定的抗菌能力、电荷量以及官能团量均不同,因此会影响纳米粒子和涂层的功能以及聚合。所述氯己定的浓度范围是(0.05-10mg/mL)。
进一步地,通过梯度浓度实验筛选,所述氯己定的浓度优选为(0.05-1.6mg/mL)。
进一步地,所述步骤1中的褐藻多酚水溶液的浓度为0.25mg/mL;所述氯己定溶液的浓度为0.8mg/mL。
进一步地,所述步骤2中聚合过程的pH为8.5。
进一步地,所述的抗菌涂层应用于制备抗菌敷贴、抗菌骨材料和抗菌口腔材料。
进一步地,所述抗菌涂层可在骨填料或者骨植入物表面沉积,改善其生物相容性。而且本申请的抗菌涂层具有良好的稳定性,在骨填料或者骨植入物发生变形时,本申请的抗菌涂层依旧保持完整,不会使基底暴露。
进一步地,步骤1中的反应液经过冷冻干燥即可得到该纳米粒子,可进一步混入多种水凝胶制备具有抗菌效果的水凝胶制品。
进一步地,所述的抗菌涂层的制备方法,还可将其调整为:将试样至于反应液中,通过干燥手段,使溶液挥发获得表面吸附抗菌纳米粒子的抗菌材料。
进一步地,所述水凝胶制品可用于抗菌敷贴或者隐形眼镜内衬物。
本发明具有以下有益效果:
1、本发明制备的抗菌涂层与现有技术中公开的含氯己定的抗菌材料相比具有相同的抗菌功能,此外,针对现有技术中抗菌材料具有毒性,生物相容性差的技术问题,选取儿茶酚胺类物质作为基底,显著改善了其细胞毒性,尤其是褐藻多酚与氯己定聚合形成的涂层,对细胞增殖具有一定的促进作用。同时本抗菌涂层的稳定性强,在模拟体液浸泡多日天的情况下仍然具有一定的抗菌效果。
2、该抗菌涂层具有非材料依赖性,即在不同的材料表面均可以黏附聚合,不同材料包括钛、不锈钢、硅、玻璃等。
3、本发明抗菌涂层的制备方法,工艺简单,反应条件易控,成本低。
当然,实施本发明的任一产品并不一定需要同时达到以上所述的所有优点。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例描述所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本抗菌涂层对钛表面改性前后钛表面接触角变化结果统计图;
图2为本抗菌涂层对钛表面改性前后钛表面扫描电镜结果展示图;
图3为氯己定和儿茶酚胺聚合前后X射线光电子能谱图;
图4为氯己定和儿茶酚胺改性样品周围金黄色葡萄球菌抑菌圈的展示图;
图5为多种酚类和氯己定复合后周围抑菌圈的展示图;
图6为不同浓度的氯己定和儿茶酚胺改性样品和金黄色葡糖球菌共培养后的菌液吸光度值的结果统计图;
图7为不同氯己定和儿茶酚胺改性样品的细胞毒性测定结果统计图;
图8为不同氯己定和儿茶酚胺改性样品表面细胞培养一天后细胞骨架及细胞核(鬼笔环肽/DAPI)染色的的结果展示图;
图9为不同氯己定和儿茶酚胺改性样品表面细胞细菌共培养一天、三天后罗丹明123染色的结果展示图。
附图中,control为钛,0、0.05、0.1、0.2、0.4、0.8、1.6等数字为氯己定和儿茶酚胺的浓度(mg/mL)。
根据附图,可以得出以下结论:
由图1可知,钛表面经过改性,钛表面接触角发生了显著的变化,从而在一定程度上证明了改性的成功。
由图2,结果显示,该氯己定和儿茶酚胺聚合物可以以纳米粒子形式聚合,大量纳米粒子的聚集,可以构建出氯己定和儿茶酚胺涂层。
由图3,结果显示,引入氯己定之后,基底的钛元素特征峰被掩盖,说明氯己定和儿茶酚胺可以在钛表面聚合形成完整的涂层。
由图4,说明氯己定和儿茶酚胺涂层具有抗菌效果,随着氯己定投料浓度的增加,其抗菌效果不断增强。抑菌圈的出现也说明涂层表面的氯己定具有较强的扩散能力。
由图5,结果显示,多种酚类均可以和氯己定在钛表面聚合形成涂层,并且均具有有效的扩散性抗菌能力。
由图6,结果显示,不同浓度的氯己定和儿茶酚胺涂层均具有较强的原位抗菌能力。
由图7,经MTT检测,所有涂层均不具有细胞毒性,其中低浓度氯己定改性式样具有一定程度的细胞促进作用。
由图8,结果显示,该氯己定和儿茶酚胺涂层对细胞的增值以及细胞骨架的铺展没有明显的影响,不具有明显的细胞毒性。
由图9,结果显示,在细胞细菌共培养的环境中,氯己定和儿茶酚胺改性后试样可以有效抑制细菌,并且没有细胞毒性。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例,都属于本发明保护的范围。
实施例1(褐藻多酚和氯己定复合抗菌涂层的制备)
1、一个清洗基体的步骤,先将经抛光处理后的基体送入超声波清洗机,依次在丙酮和无水酒精中利用15~30kHz的超声波分别清洗5~10min,然后烘干。
2、将褐藻多酚和氯己定分别溶解于pH=8.5的Tris溶液中,使溶液中褐藻多酚最终浓度为0.5mg/mL,然后加入氯己定溶液,其中氯己定浓度分别为0、0.05、0.1、0.2、0.4、0.8、1.6mg/mL。然后把上述溶液加入到试样表面,静置24小时。
3、清洗试样的步骤如下:将试样置于RO水中,隔水超声,利用15~30kHz的超声波分别清洗5~10min,然后在冷风中烘干。
实施例2(褐藻多酚和氯己定复合纳米粒子的制备)
1、将褐藻多酚和氯己定分别溶解于pH=8.5的Tris溶液中,使溶液中褐藻多酚最终浓度为0.5mg/mL,然后加入氯己定溶液,其中氯己定浓度分别为0、0.05、0.1、0.2、0.4、0.8、1.6mg/mL。然后把上述溶液加入到试样表面,静置24小时。
2、将上述液体经3000g离心力离心10分钟后即可得到悬浮于液体中的纳米粒子。
实施例3(多巴胺和氯己定复合抗菌涂层的制备)
1、一个清洗基体的步骤,先将经抛光处理后的基体送入超声波清洗机,依次在丙酮和无水酒精中利用15~30kHz的超声波分别清洗5~10min,然后烘干。
2、将多巴胺和氯己定分别溶解于pH=8.5的Tris溶液中,使溶液中多巴胺最终浓度为0.5mg/mL,然后加入氯己定溶液,其中氯己定浓度分别为0、0.05、0.1、0.2、0.4、0.8、1.6mg/mL。然后把上述溶液加入到试样表面,静置24小时。
3、清洗试样的步骤如下:将试样置于RO水中,隔水超声,利用15~30kHz的超声波分别清洗5~10min,然后在冷风中烘干。
实施例4(去甲基肾上腺素和氯己定复合抗菌涂层的制备)
1、一个清洗基体的步骤,先将经抛光处理后的基体送入超声波清洗机,依次在丙酮和无水酒精中利用15~30kHz的超声波分别清洗5~10min,然后烘干。
2、将去甲基肾上腺素和氯己定分别溶解于pH=8.5的Tris溶液中,使溶液中去甲基肾上腺素最终浓度为0.5mg/mL,然后加入氯己定溶液,其中氯己定浓度分别为0、0.05、0.1、0.2、0.4、0.8、1.6mg/mL。然后把上述溶液加入到试样表面,静置24小时。
3、清洗试样的步骤如下:将试样置于RO水中,隔水超声,利用15~30kHz的超声波分别清洗5~10min,然后在冷风中烘干。
实施例5(单宁酸和氯己定复合抗菌涂层的制备)
1、一个清洗基体的步骤,先将经抛光处理后的基体送入超声波清洗机,依次在丙酮和无水酒精中利用15~30kHz的超声波分别清洗5~10min,然后烘干。
2、将单宁酸和氯己定分别溶解于pH=8.5的Tris溶液中,使溶液中单宁酸最终浓度为0.5mg/mL,然后加入氯己定溶液,其中氯己定浓度分别为0、0.05、0.1、0.2、0.4、0.8、1.6mg/mL。然后把上述溶液加入到试样表面,静置24小时。
3、清洗试样的步骤如下:将试样置于RO水中,隔水超声,利用15~30kHz的超声波分别清洗5~10min,然后在冷风中烘干。
在本说明书的描述中,参考术语“一个实施例”、“示例”、“具体示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中,对上述术语的示意性表述不一定指的是相同的实施例或示例。而且,描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
以上公开的本发明优选实施例只是用于帮助阐述本发明。优选实施例并没有详尽叙述所有的细节,也不限制该发明仅为所述的具体实施方式。显然,根据本说明书的内容,可作很多的修改和变化。本说明书选取并具体描述这些实施例,是为了更好地解释本发明的原理和实际应用,从而使所属技术领域技术人员能很好地理解和利用本发明。本发明仅受权利要求书及其全部范围和等效物的限制。

Claims (6)

1.一种氯己定和儿茶酚胺共聚合的抗菌涂层,其特征在于:所述抗菌涂层是由氯己定和儿茶酚胺自组装形成纳米粒子,所述纳米粒子在多种基底表面粘附,形成一层致密的聚合涂层。
2.根据权利要求1所述的抗菌涂层的制备方法,其特征在于,包括以下步骤:
步骤1:配置一定浓度的儿茶酚胺水溶液和一定浓度的氯己定溶液;
步骤2:向儿茶酚胺水溶液中缓慢滴入步骤1中配置的氯己定溶液,使儿茶酚胺和氯己定聚合充分;
步骤3:静置24小时,在基底上会沉积一层氯己定和儿茶酚胺的聚合涂层。
3.根据权利要求2所述的抗菌涂层的制备方法,其特征在于,所述儿茶酚胺为多巴胺、去甲基肾上腺素、单宁酸、没食子酸、褐藻多酚或者表没食子儿茶素没食子酸酯。
4.根据权利要求3所述的抗菌涂层的制备方法,其特征在于,所述步骤1中的褐藻多酚水溶液的浓度为0.25mg/mL;所述氯己定溶液的浓度为0.8mg/mL。
5.根据权利要求2-4任一项所述的抗菌涂层的制备方法,其特征在于,所述步骤2中聚合过程的pH为8.5。
6.根据权利要求1所述的一种氯己定和儿茶酚胺共聚合的抗菌涂层的应用,其特征在于,其应用于制备抗菌敷贴、抗菌骨材料和抗菌口腔材料。
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