CN113827771A - 医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法 - Google Patents
医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法 Download PDFInfo
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Abstract
本发明公开了一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,涉及医用抗菌材料制备技术领域。包括将医用金属浸没于银/多酚酸性混合溶液,以获得表面负载有银/多酚纳米复合颗粒的医用金属中间产物;将医用金属中间产物进行在50‑250℃高温条件下处理即可;银/多酚酸性混合溶液包括多酚溶液和溶质,多酚溶液选自多巴胺、表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚、去甲基肾上腺素中的一种;溶质选自硝酸银或氟化银。本发明通过在医用金属表面螯合多酚与金属银离子形成的复合物,并通过高温处理提高了纳米颗粒在医用金属表面的结合强度,解决了银颗粒在表面的结合稳定性不足的问题。
Description
技术领域
本发明属于医用抗菌材料制备技术领域,特别是涉及一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法。
背景技术
不锈钢和钛基合金等医用金属基材广泛应用于骨科和牙科植入材料,但细菌易定植于金属表面并形成生物膜,导致感染、炎症等并发症,严重影响治疗效果。传统的金属材料表面抗菌主要通过表面装载抗生素或有机抗菌物质实现,但抗菌谱窄、易导致细菌耐受、结合不稳定等问题,严重影响其抗菌效果,甚至大剂量使用会导致严重的细胞、组织和器官毒性。
由于具有广谱、高效的抗菌能力,银成为极具价值的抗菌物质,但其强细胞毒性制约其临床应用。大量研究采用等离子体浸没离子注入,脉冲抽真空电弧沉积,物理气相沉积等技术,将银装载在金属表面,但高耗能、可控性差制约其应用。多酚具有优异的抗炎、抗氧化、自由基清除能力,也具有螯合和还原金属的能力,被广泛用于制药、纳米颗粒制备、材料表面修饰等,如最近,一种简单的液相原位还原纳米银的方法被广泛报道,即在多种材料表面沉积多巴胺涂层,利用其在碱性环境中的螯合和还原金属能力,原位还原硝酸银水溶液中的银离子,在多巴胺涂层表面形成纳米银颗粒并表现出优异的抗菌性能。但由于银完全暴露在表面,导致严重的细胞毒性且银颗粒在表面的结合稳定性不足。故亟需开发一种强结合、低毒性、高效抗菌的载银表面制备方法,以解决以下问题:1)如何将银稳定结合在材料表面,2)如何避免银的细胞毒性,3)如何保证银的抗菌能力,4)如何实现制备方法简单、可控。
发明内容
本发明的目的在于提供一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,通过在医用金属表面螯合获得多酚与金属银离子形成的复合物,解决了银颗粒在表面的结合稳定性不足的问题。
为解决上述技术问题,本发明是通过以下技术方案实现的:
本发明为一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,包括将医用金属浸没于银/多酚酸性混合溶液,以获得表面负载有银/多酚纳米复合颗粒的医用金属中间产物;将医用金属中间产物进行在50-250℃高温条件下处理即可。
上述50-250℃高温,该温度范围低于多酚的热分解温度,且支持多酚的继续反应形成稳定的键合状态而保持结构稳定和降低银的释放速率,保证长效抗菌和降低细胞毒性
作为本发明的一种优选技术方案,所述医用金属包括钴铬合金、不锈钢、钛及钛合金,这些金属广泛用于骨科和牙科植入材料且极易因手术携带细菌、周围组织细菌侵入而发生细菌所致感染、炎症等并发症。多酚可与金属表面的金属原子或离子螯合而结合在金属表面。
作为本发明的一种优选技术方案,所述银/多酚酸性混合溶液包括多酚溶液和溶质,所述多酚溶液选自多巴胺、表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚、去甲基肾上腺素中的一种;这些多酚具有大量酚羟基可以银离子螯合,也可与医用金属表面的金属离子或原子螯合。
所述溶质选自硝酸银、氟化银,硝酸银、氟化银在水溶液中可以电离成银离子和酸根,其中银离子可以与多酚相互作用并形成银离子和多酚均匀复合的纳米颗粒;
其中,所述银/多酚酸性混合溶液中和多酚的质量比范围为1:200-1:5,该物质配比范围可以保证银含量较低而避免银的细胞毒性,且可以将银/多酚颗粒结合在医用金属表面,所述银/多酚酸性混合溶液中银的浓度范围为0.01-0.5mg/mL,在0.01-0.5mg/mL浓度下,保证了体系中较低的银浓度,进而调控颗粒维持在纳米尺度,且避免银的浪费和未反应的银所导致的环境污染。
其中,所述银/多酚酸性混合溶液的PH为3-6,在酸性条件下,多酚保留了螯合金属的能力,且银离子不会快速聚合成氧化银颗粒,最终形成银和多酚均匀复合的颗粒,避免多酚完全包裹银颗粒形成核壳结构而影响银的抗菌能力,且避免表面银离子过度暴露在表面而导致细胞毒性。
本发明适用于骨科和牙科植入金属材料表面修饰,以防止细菌所致感染、炎症等并发症的发生。
本发明具有以下有益效果:
本发明通过在医用金属表面螯合多酚与金属银离子形成的复合物,通过高温处理提高了纳米颗粒在医用金属表面的结合强度,解决了银颗粒在表面的结合稳定性不足的问题。
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为钛表面制备银/多巴胺涂层前后检测结果附图;
图2为材料抗菌试验结果附图;
图3为材料表面细胞相容性试验结果附图;
图4为材料表面细菌-细胞竞争黏附的活死细菌/细胞荧光染色附图。
具体实施方式
实施例1
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将钴铬合金浸没于0.01mg/mL的硝酸银和2mg/mL的多巴胺混合溶液(pH值=3)以在表面负载具有广谱抗菌的银/多巴胺纳米复合颗粒;
步骤2、然后将样品在50°烘箱中加热,既得;
实施例2
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将316L不锈钢浸没于0.5mg/mL的氟化银和2.5mg/mL的表没食子儿茶素没食子酸酯混合溶液(pH值=6)以在表面负载具有广谱抗菌的银/表没食子儿茶素没食子酸酯纳米复合颗粒;
步骤2、然后将样品在250°烘箱中加热,既得;
实施例3
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将316L不锈钢浸没于0.5mg/mL的氟化银和2.5mg/mL的没食子酸混合溶液(pH值=6)以在表面负载具有广谱抗菌的银/没食子酸纳米复合颗粒;
步骤2、然后将样品在250°烘箱中加热,既得;
实施例4
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将纯钛浸没于0.25mg/mL的硝酸银和2.5mg/mL的单宁酸混合溶液(pH值=5)以在表面负载具有广谱抗菌的银/单宁酸纳米复合颗粒;
步骤2、然后将样品在150°烘箱中加热,既得;
实施例5
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将Ni-Ti形状记忆合金浸没于0.3mg/mL的氟化银和3mg/mL的褐藻多酚混合溶液(pH值=5)以在表面负载具有广谱抗菌的银/褐藻多酚纳米复合颗粒;
步骤2、然后将样品在200°烘箱中加热,既得;
实施例5
一种医用金属表面负载银/多酚纳米复合抗菌颗粒的制备方法,包括以下步骤:
步骤1、将多孔纯钛浸没于0.02mg/mL的硝酸银和2mg/mL的去甲基肾上腺素混合溶液(pH值=4)以在表面负载具有广谱抗菌的银/去甲基肾上腺素纳米复合颗粒;
步骤2、然后将样品在150°烘箱中加热,既得。
在图1中,A是钛表面制备银/多巴胺涂层(Ti-Ag/DA)前后的表面形貌,结果显示制备涂层表面黏附大量纳米颗粒,插图为背散射结果,照片显示纳米颗粒没有明显衬度变化,证明多巴胺和银均匀复合;B是钛表面制备银/多巴胺涂层(Ti-Ag/DA)前后的X射线光电子能谱结果,左图为全谱扫描显示涂层表面有大量的银元素和氮元素,证明多巴胺和银都结合在钛表面,右图为Ag3d高分辨扫描结果,显示表面结合大量的银,这些结果证明纳米复合颗粒成功制备在钛表面。
在图2中,材料抗菌结果,钛表面和涂层表面黏附金黄色葡萄球菌(A)和伴放线菌放线杆菌(B)的活/死细菌荧光染色照片,钛表面和涂层表面黏附的金黄色葡萄球菌(C)和伴放线菌放线杆菌(D)超声分散在生理盐水后涂板结果,这些结果显示制备涂层具有广谱、高效的抗菌效果;E是将涂层样品浸泡于细菌培养基中,然后每3天加入不同稀释倍数的菌液,观察培养溶液体系的浑浊情况,结果显示当第5次加入高浓度细菌后,培养体系开始浑浊,证明涂层具有良好的抗菌稳定性,可以在细菌多次攻击下,保持良好的抗菌效果。
在图3中,材料表面细胞相容性结果,钛表面和涂层表面黏附小鼠前成骨细胞系MC-3T3-E1(A)和小鼠成纤维细胞3T3(B)的荧光染色照片,结果显示两种表面的细胞均随着培养时间延长而明显增殖,涂层并没有表现出明显的细胞毒性。
在图4中,材料表面细菌-细胞竞争黏附的活死细菌/细胞荧光染色照片,钛表面和涂层表面在金黄色葡萄球菌/小鼠前成骨细胞系MC-3T3-E1(A)和金黄色葡萄球菌/小鼠成纤维细胞3T3(B)培养体系中培养,结果显示钛表面均随着培养时间延长而被细菌覆盖,涂层表面均随着培养时间延长而被细胞覆盖,证明材料在有菌环境中保持了良好的细胞相容性。
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 (7)
1.一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于:包括将医用金属浸没于银/多酚酸性混合溶液,以获得表面负载有银/多酚纳米复合颗粒的医用金属中间产物;将医用金属中间产物进行高温处理即可。
2.根据权利要求1所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述医用金属包括钴铬合金、不锈钢、钛及钛合金。
3.根据权利要求1所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述银/多酚酸性混合溶液包括多酚溶液和溶质,所述多酚溶液选自多巴胺、表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚、去甲基肾上腺素中的一种;所述溶质选自硝酸银或氟化银。
4.根据权利要求3所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述银/多酚酸性混合溶液中和多酚的质量比范围为1:200-1:5。
5.根据权利要求1所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述银/多酚酸性混合溶液的PH为3-6。
6.根据权利要求1所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述银/多酚酸性混合溶液中银的浓度范围为0.01-0.5mg/mL。
7.根据权利要求1所述的一种医用金属表面负载银多酚纳米复合抗菌颗粒的制备方法,其特征在于,所述高温处理的处理温度为50-250℃。
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