CN110604132B - 空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法 - Google Patents
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Abstract
本发明的空心核‑卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，属于纳米粒子制备技术领域。将2‑甲基咪唑和六水合硝酸锌的甲醇溶液混合，在室温下搅拌反应12小时，离心、洗涤、再分散，得到ZIF‑8纳米颗粒；再与多巴胺的甲醇溶液混合，加热到60℃反应1～7小时，离心、洗涤、再分散，获得空心聚多巴胺纳米颗粒；加入硝酸银的甲醇溶液，室温反应1小时，离心、洗涤、真空干燥，得到空心核‑卫星状聚多巴胺载银抗菌纳米胶囊。本发明制备条件温和，环境友好，操作简便，制备的聚多巴胺载银空心纳米胶囊具有很好的抗菌效果。
Description
技术领域：
本发明属于纳米粒子制备技术领域，具体涉及制备“空心核-卫星”状聚多巴胺载银抗菌纳米胶囊，并研究其在抗菌领域的应用。
技术背景：
近年来，人类的健康受到公共卫生风险的严重威胁，迫切需要制药和医学研究人员开发新的抗菌剂以应对公共卫生保健中不断增加的卫生要求。银纳米颗粒作为最有效的抗菌剂之一，具有强大的杀菌活性和广泛的抑制性杀菌，以及对人类和其他动物相对较低的毒性。然而它在实际应用中经常受到聚集的影响，将导致它们的抗菌活性减少甚至完全丧失。为了解决这个问题，科研人员已经花费大量努力将银纳米颗粒固定到各种基质上，以增强它们的稳定性和抗菌活性。迄今为止，许多材料被报道作为支持银纳米颗粒的基质。尽管它们可以保持银纳米颗粒的稳定性和抗菌活性，但合成过程复杂并伴随其他试剂的加入，如稳定剂、还原剂等，会导致环境毒性或生物危害。因此，迫切需要研究一种简单直接且环境友好的合成具有良好的生物相容性和优异的抗菌活性的银基纳米复合材料。
发明内容：
本发明要解决的技术问题是克服背景技术存在的问题，提供一种制备工艺简单高效，生物相容性好，抗菌活性高的银基纳米复合材料的制备方法。
本发明的技术方案如下：
一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，有以下步骤：
(1)将摩尔比为2:1的2-甲基咪唑的甲醇溶液和六水合硝酸锌的甲醇溶液混合，在室温下搅拌反应12小时，离心、洗涤、再分散，得到ZIF-8纳米颗粒；所述的ZIF-8纳米颗粒是一种由过渡金属与有机配体相互结合而成的具有立体网络结构的晶体材料；
(2)将多巴胺的甲醇溶液与步骤(1)配制的ZIF-8纳米颗粒混合，加热到60℃反应1～7小时，离心、洗涤、再分散，获得空心聚多巴胺(PDA)纳米颗粒；所述的多巴胺的甲醇溶液的浓度为10～50mmol/L；按摩尔计，多巴胺的用量与步骤(1)中的六水合硝酸锌的用量比例为0.08～0.4:1；
(3)向步骤(2)制备的空心聚多巴胺纳米颗粒中加入硝酸银的甲醇溶液，室温反应1小时，离心、洗涤、真空干燥，得到空心核-卫星状聚多巴胺载银抗菌纳米胶囊；按摩尔计，硝酸银的用量与步骤(1)中六水合硝酸锌用量比例为0.07～0.28:1。
在步骤(1)中，所述的2-甲基咪唑溶液浓度优选40mmol/L；所述的六水合硝酸锌溶液浓度优选20mmol/L；所述的搅拌速度优选500转/分钟；
在步骤(2)中，按摩尔计，多巴胺的用量与步骤(1)中的六水合硝酸锌的用量比例优选为0.26:1。
在步骤(3)中，所述的硝酸银的浓度优选8mg/mL；按摩尔计，硝酸银的用量与步骤(1)中六水合硝酸锌用量比例优选0.07:1。
步骤(1)、(2)、(3)中所述的离心、洗涤条件具体是在6000～9000转/分钟条件下离心5～15分钟、无水乙醇洗涤3～5次；步骤(3)中所述的真空干燥具体是指30℃～40℃真空干燥12～24小时。
有益效果：
1、本发明制备了“空心核-卫星”状聚多巴胺载银纳米胶囊，制备条件温和，环境友好，操作简便。
2、本发明采用的自刻蚀模板ZIF-8是一种金属有机框架，随着聚多巴胺壳逐渐形成，ZIF-8中的Zn2+与聚多巴胺发生配位作用，ZIF-8模板不断被刻蚀，最终消失，形成空心聚多巴胺纳米胶囊。
3、本发明采用聚多巴胺原位还原硝酸银，在空心聚多巴胺核上生长了一层单分散性好、稳定性高的银纳米颗粒。
4、本发明聚多巴胺载银空心纳米胶囊具有很好的抗菌效果，银粒子和Zn2+共同作用发挥抑菌功能。
附图说明：
图1为本发明实施例1制备的空心聚多巴胺纳米结构的透射电子显微镜照片。
图2为本发明实施例2制备的“空心核-卫星”状聚多巴胺载银纳米胶囊的透射电子显微镜照片
图3聚多巴胺载银纳米胶囊对金黄葡萄球菌的抑菌效果
图4聚多巴胺载银纳米胶囊对大肠杆菌的抑菌效果
具体实施方法：
以下是本发明的实施例用到的基础条件，但本发明能实施的范围并不限于这些条件，也不限于这些实施例：
环境温度30℃，1个大气压；
六水和硝酸锌，分子量297.49g/mol；
二甲基咪唑，分子量82.10g/mol；
无水甲醇，密度0.791g/mL，分子量32.04g/mol；
无水乙醇，密度0.789g/mL，分子量46.07g/mol
多巴胺(HDA)，分子量189.64g/mol；
硝酸银，分子量169.87g/mol。
实施例1：
将5ml的2-甲基咪唑(40mM)和5ml的六水合硝酸锌(20mM)溶液混合，在室温下500rpm反应12h。通过8000rpm离心10min收集产物，用乙醇洗涤数次，并最终分散在3.0mL甲醇中。得到ZIF-8纳米颗粒。
将上述3.0mL ZIF-8甲醇溶液加入到5.0mL甲醇中，与0.8mL多巴胺甲醇溶液(32.5mM)混合。将混合物搅拌并在60℃下回流5h，其颜色逐渐从乳白色变为黑色。通过8000rpm离心5min收集所得物，用乙醇洗涤三次，并最终分散在6.0mL甲醇中。得到的空心聚多巴胺纳米结构如图1所示。
取2mL上述分散液并加入1mL甲醇稀释，然后与50μL硝酸银甲醇溶液(8mg/mL)混合，在室温下搅拌1h。将产物离心并用乙醇冲洗数次。随后，将聚多巴胺载银空心纳米胶囊冷冻干燥，得到聚多巴胺载银空心纳米胶囊，如图2所示。
实施例2：
采用摇瓶法评价实施案例1中制备的聚多巴胺载银空心纳米胶囊对金葡萄球菌(革兰氏阳性菌)和大肠杆菌(革兰氏阴性菌)的抑菌效果，测试结果如图3、图4所示。
将金葡萄球菌和大肠杆菌在LB(Luria-Berrani)培养液中37℃培养12h。取一定量10mg/mL聚多巴胺载银空心纳米胶囊分散在5mL培养液中，分别加入50μL浓度为107CFU/mL的金色葡萄球菌和大肠杆菌悬浮液。在37℃中孵育，通过监测600nm处的光密度，来测量细菌的生长速度。图3是金色葡萄球菌的抑菌效果，图4是大肠杆菌的抑菌效果；从图中明显看出，当聚多巴胺载银空心纳米胶囊的终浓度为16μg/mL时，可完全抑制两种细菌的生长。
Claims (5)
1.一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，有以下步骤：
(1)将摩尔比为2:1的2-甲基咪唑的甲醇溶液和六水合硝酸锌的甲醇溶液混合，在室温下搅拌反应12小时，离心、洗涤、再分散，得到ZIF-8纳米颗粒；所述的ZIF-8纳米颗粒是一种由过渡金属与有机配体相互结合而成的具有立体网络结构的晶体材料；
(2)将多巴胺的甲醇溶液与步骤(1)配制的ZIF-8纳米颗粒混合，加热到60℃反应1～7小时，离心、洗涤、再分散，获得空心聚多巴胺纳米颗粒；所述的多巴胺的甲醇溶液的浓度为10～50mmol/L；按摩尔计，多巴胺的用量与步骤(1)中的六水合硝酸锌的用量比例为0.08～0.4:1；
(3)向步骤(2)制备的空心聚多巴胺纳米颗粒中加入硝酸银的甲醇溶液，室温反应1小时，离心、洗涤、真空干燥，得到空心核-卫星状聚多巴胺载银抗菌纳米胶囊；按摩尔计，硝酸银的用量与步骤(1)中六水合硝酸锌用量比例为0.07～0.28:1。
2.根据权利要求1所述的一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，其特征在于，在步骤(1)中，所述的2-甲基咪唑溶液浓度为40mmol/L；所述的六水合硝酸锌溶液浓度为20mmol/L；所述的搅拌，速度为500转/分钟。
3.根据权利要求1所述的一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，其特征在于，在步骤(2)中，按摩尔计，多巴胺的用量与步骤(1)中的六水合硝酸锌的用量比例为0.26:1。
4.根据权利要求1所述的一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，其特征在于，在步骤(3)中，所述的硝酸银的浓度为8mg/mL；按摩尔计，硝酸银的用量与步骤(1)中六水合硝酸锌用量比例为0.07:1。
5.根据权利要求1～4任一所述的一种空心核-卫星状聚多巴胺载银抗菌纳米胶囊的制备方法，其特征在于，步骤(1)、(2)、(3)中所述的离心、洗涤条件是在6000～9000转/分钟条件下离心5～15分钟、无水乙醇洗涤3～5次；步骤(3)中所述的真空干燥是指30～40℃真空干燥12～24小时。
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