CN115554480A - 用于骨和牙体表面抗菌的纳米涂敷材料及其制备方法 - Google Patents
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Abstract
本发明公开了一种用于骨和牙体表面抗菌的纳米涂敷材料及其制备方法,涉及纳米抗菌材料技术领域。本发明制备方法包括多酚/氯己定纳米抗菌颗粒的制备、在制得的纳米抗菌颗粒表面接枝羟基磷灰石识别分子;多酚为表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚。本发明制得的纳米材料中的氯己定具有高效、广谱的抗菌能力;氯己定和多酚的复合纳米球可以长期释放氯己定实现长久抗菌和避免氯己定的细胞毒性;羟基磷灰石识别分子可以靶向结合在牙体和骨表面,减少对其它脏器的毒副作用。
Description
技术领域
本发明属于纳米抗菌材料技术领域,特别是涉及一种用于骨和牙体表面抗菌的纳米涂敷材料及其制备方法。
背景技术
口腔中的大量细菌是导致牙体和牙周等部位的口腔疾病的重要致病因素,牙体疾病中有代表性的是龋病,是指以细菌为主的多种因素影响下牙体硬组织发生慢性、进行性破坏的一种疾病,第四次全国口腔健康流行病学调查结果显示5岁儿童乳牙的患龋率高达66%,12岁儿童恒牙的患病率为28.9%,35岁以上的人群龋病的患病率甚至高于88%,并随着年龄增长患病率逐渐增高。牙周疾病中代表性的是牙周炎,其患病率在40%-75%,成年人发病率高达70%,是人类最常见的慢性病,其最主要致病因素是牙菌斑。另外,骨科手术部位感染是院内感染的第二大原因,占院内感染的38%,感染所引发的骨不连是骨科手术的重要并发症,其导致的致死率是无感染患者的2倍以上。故控制细菌是预防感染类疾病发生的最重要手段。
长期以来,抑菌方式主要采用抗生素类药物,但存在以下问题:1)抗菌药物的滥用,加速了细菌的进化产生耐药性(如耐药金黄色葡萄球菌),成为危害公众用药安全的突出问题;2)抗生素多不具有广谱抗菌能力,对部分致病菌抑制效果不佳;3)抗生素无法富集在病灶部位,导致治疗效果差,且分布到其它脏器会产生毒副作用。故寻找更好的抗菌药物及更有效的药物载体成为科学家关注的焦点。但存在以下问题亟待解决:1)如何选择有效的抗菌物质,保证广谱、高效的抗菌能力;2)如何实现抗菌物质的有效释放;3)如何将抗菌物质富集到病灶部位。
发明内容
本发明的目的在于提供一种用于骨和牙体表面抗菌的纳米涂敷材料,该纳米涂敷材料中的氯己定具有高效、广谱的抗菌能力;氯己定和多酚的复合纳米球可以长期释放氯己定实现长久抗菌和避免氯己定的细胞毒性;羟基磷灰石识别分子可以靶向结合在牙体和骨表面,减少对其它脏器的毒副作用。
本发明的目的还在于种用于骨和牙体表面抗菌的纳米涂敷材料制备方法,本发明制备过程在溶液中进行,反应条件温和,无需复杂仪器,制备简单,易于工业化生产。
为解决上述技术问题,本发明是通过以下技术方案实现的:
本发明为一种用于骨和牙体表面抗菌的纳米涂敷材料,其特征在于:包括纳米抗菌颗粒以及接枝在纳米抗菌颗粒外表面的羟基磷灰石识别分子;所述羟基磷灰石识别分子包括阿仑膦酸盐(ALN)或氨基封端聚乙二醇修饰的阿仑膦酸盐(NH2-PEG-ALN)或巯基封端聚乙二醇修饰的阿仑膦酸盐(SH-PEG-ALN);所述纳米抗菌颗粒为多酚/氯己定纳米抗菌颗粒。
一种纳米涂敷材料的制备方法,包括如下步骤:
步骤1、将多酚和氯己定均匀分散在中性或碱性溶液中,反应2-24小时;分离纯化得到多酚/氯己定纳米抗菌颗粒;将得到的多酚/氯己定纳米抗菌颗粒均匀分散在中性或碱性溶液中得到组分A,待用;
步骤2、在步骤1制得多酚/氯己定纳米抗菌颗粒表面接枝羟基磷灰石识别分子,具体包括:
取羟基磷灰石识别分子分散在中性或碱性溶液中得到组分B;
取步骤1中组分A和组分B混合均匀得到混合溶液,反应12-24小时;
分离纯化得到纳米涂敷材料颗粒。
进一步地,所述氯己定浓度0.5-5mg/mL,且所述多酚与氯己定的质量份数比为1:0.2-1:2。
进一步地,在所述步骤2中,混合溶液中羟基磷灰石识别分子与纳米颗粒的质量浓度比范围是1:10-1:100。
进一步地,所述步骤1和步骤2中的中性或碱性溶液的pH值为7-10,且采用去离子水溶液或去离子水/乙醇混合溶液。
进一步地,所述步骤1和步骤2中,分离纯化为:通过高速离心弃上清、去离子水清洗、再离心;或为通过透析去除杂质后冷冻干燥。
进一步地,所述步骤1中的多酚为表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚。
本发明具有以下有益效果:
1、本发明通过多酚中酚羟基(带负电)的强反应能力,可以与氯己定中胍基(带正电)形成电荷作用和共价作用,形成纳米复合抗菌颗粒;电荷作用赋予纳米复合材料释放氯己定的能力,实现纳米复合颗粒表面和周围的长期抗菌。
2、本发明采用中性或碱性环境制备纳米复合颗粒,该条件下可以增加多酚的反应活性,可以形成更多的交联,减缓氯己定的释放,且多酚的良好生物相容性和负电荷可以中和氯己定高正电荷导致的细胞毒性。
3、本发明通过多酚中酚羟基共价连接羟基磷灰石识别分子中的氨基或巯基,使得最终构建的纳米材料具有与牙体和骨上羟基磷灰石的高亲合性,实现纳米复合颗粒快速靶向结合到牙体和骨表面,降低治疗所需时间和纳米材料分布于其它脏器可能导致的毒副作用。
4、本发明制备过程在溶液中进行,反应条件温和,无需复杂仪器,制备简单,易于工业化生产;同时本发明制备的纳米材料分散于溶液并涂敷在骨或牙体表面,使用方法简单,对牙体和骨没有损伤,且可以有效预防这些部位及周边细菌所致并发症,大大降低患者的痛苦和治疗成本。
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为实施例6中牙片表面纳米材料的黏附结构示意图;
图2为实施例6中牙片表面细菌黏附结构示意图。
具体实施方式
实施例1
一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,包括:
步骤1、表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒的制备;
1)、将表没食子儿茶素没食子酸酯(2.5mg/mL)和氯己定(0.5mg/mL)均匀分散在pH=7的去离子水溶液,反应2小时;
2)、通过高速离心弃上清、去离子水清洗、再离心,得到纳米表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒;
3)、表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒均匀分散在pH=7的去离子水溶液中,待用;
步骤2、在步骤1制得纳米抗菌颗粒表面接枝羟基磷灰石识别分子;
1)将表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒去离子水溶液(pH=7)与阿仑膦酸去离子水溶液(pH=7)均匀混合,阿仑膦酸与表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒的质量浓度比是1:10,反应12小时;
2)、通过高速离心弃上清、去离子水清洗、再离心,分离纯化纳米颗粒,既得纳米涂敷材料颗粒;
实施例2
一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,包括:
步骤1、表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒的制备;
1)、将表没食子儿茶素没食子酸酯(2.5mg/mL)和氯己定(5mg/mL)均匀分散在去离子水/乙醇混合溶液(pH=10),反应24小时;
2)、通过透析去除杂质后冷冻干燥,得到表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒;
3)、表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒均匀分散在pH=10去离子水中,待用;
步骤2、在步骤1制得纳米抗菌颗粒表面接枝氨基封端聚乙二醇修饰的阿仑膦酸盐(NH2-PEG-ALN)。
1)、将表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒(pH=10)与NH2-PEG-ALN去离子水溶液(pH=10)均匀混合,NH2-PEG-ALN与表没食子儿茶素没食子酸酯/氯己定纳米抗菌颗粒的质量浓度比是1:100,反应24小时;
2)、通过透析去除杂质后冷冻干燥,分离纯化纳米颗粒,既得纳米涂敷材料颗粒;
实施例3
一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,包括:
步骤1、没食子酸/氯己定纳米抗菌颗粒的制备;
1)、将没食子酸(2.5mg/mL)和氯己定(2.5mg/mL)均匀分散在去离子水溶液(pH=8),反应12小时;
2)、通过高速离心弃上清、去离子水清洗、再离心,分离纯化得到没食子酸/氯己定纳米抗菌颗粒;
3)、没食子酸/氯己定纳米抗菌颗粒均匀分散去离子水溶液(pH=7),待用;
步骤2、在步骤1制得纳米抗菌颗粒表面接枝巯基封端聚乙二醇修饰的阿仑膦酸盐(SH-PEG-ALN)。
1)、将没食子酸/氯己定纳米抗菌颗粒均匀分散去离子水溶液(pH=7)与SH-PEG-ALN去离子水溶液(pH=7)均匀混合,SH-PEG-ALN与没食子酸/氯己定纳米抗菌颗粒的质量浓度比是1:50,反应20小时;
2)、通过透析去除杂质后冷冻干燥,分离纯化纳米颗粒,既得纳米涂敷材料颗粒;
实施例4
一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,包括:
步骤1、单宁酸/氯己定纳米抗菌颗粒的制备;
1)、将单宁酸(2mg/mL)和氯己定(2mg/mL)均匀分散在去离子水溶液(pH9),反应15小时;
2)、通过高速离心弃上清、去离子水清洗、再离心,分离纯化得到单宁酸/氯己定纳米抗菌颗粒;
3)、单宁酸/氯己定纳米抗菌颗粒均匀分散去离子水溶液(pH=7),待用;
步骤2、在步骤1制得纳米抗菌颗粒表面接枝巯基封端聚乙二醇修饰的阿仑膦酸盐(SH-PEG-ALN);
1)、将单宁酸/氯己定纳米抗菌颗粒均匀分散去离子水溶液(pH=7)与SH-PEG-ALN去离子水溶液(pH=7)均匀混合,SH-PEG-ALN与单宁酸/氯己定纳米抗菌颗粒的质量浓度比是1:20,反应10小时;
2)、通过透析去除杂质后冷冻干燥,分离纯化纳米颗粒,既得纳米涂敷材料颗粒;
实施例5
一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,包括:
步骤1、褐藻多酚/氯己定纳米抗菌颗粒的制备;
1)、将褐藻多酚(1mg/mL)和氯己定(1mg/mL)均匀分散在去离子水/乙醇混合溶液(pH=8),反应20小时;
2)、通过透析去除杂质后冷冻干燥,分离纯化得到褐藻多酚/氯己定纳米;
3)、褐藻多酚/氯己定纳米均匀分散在pH=8去离子水中,待用;
步骤2、在步骤1制得纳米抗菌颗粒表面接枝氨基封端聚乙二醇修饰的阿仑膦酸盐(NH2-PEG-ALN);
1)、将褐藻多酚/氯己定纳米去离子水溶液(pH=8)与NH2-PEG-ALN去离子水溶液(pH=8)均匀混合,NH2-PEG-ALN与褐藻多酚/氯己定纳米的质量浓度比是1:50,反应12小时;
2)、通过透析去除杂质后冷冻干燥,分离纯化纳米颗粒,既得纳米涂敷材料颗粒;
实施例6
在酸蚀的牙片上涂抹单宁酸@氯己定(TA@CHX)和单宁酸@氯己定-阿仑膦酸(TA@CHX-ALN)并观察牙片表面形貌变化。
如图1所示,SEM结果显示酸蚀牙片表面粗糙且有大量不规则的矿物;TA@CHX涂敷的牙片表面除了粗糙且不规则矿物外,有微量的纳米颗粒(箭头所指);TA@CHX-ALN涂敷的牙片表面除了粗糙且不规则矿物外,大量的纳米颗粒,证明只有TA@CHX-ALN可以快速结合在富含羟基磷灰石的牙片表面。
获得实施例6中的牙片与菌液孵育后,观察牙片表面细菌黏附情况:
如图2所示,SEM结果显示对照牙片(未处理)表面黏附了大量细菌,TA@CHX涂敷的牙片表面有少量细菌黏附,TA@CHX-ALN涂敷的牙片表面未见细菌黏附。
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 (10)
1.用于骨和牙体表面抗菌的纳米涂敷材料,其特征在于:包括纳米抗菌颗粒以及接枝在纳米抗菌颗粒外表面的羟基磷灰石识别分子。
2.根据权利要求1所述的用于骨和牙体表面抗菌的纳米涂敷材料,其特征在于,所述羟基磷灰石识别分子包括阿仑膦酸盐(ALN)或氨基封端聚乙二醇修饰的阿仑膦酸盐(NH2-PEG-ALN)或巯基封端聚乙二醇修饰的阿仑膦酸盐(SH-PEG-ALN)。
3.根据权利要求1或2所述的用于骨和牙体表面抗菌的纳米涂敷材料,其特征在于,所述纳米抗菌颗粒为多酚/氯己定纳米抗菌颗粒。
4.一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,包括如下步骤:
步骤1、将多酚和氯己定均匀分散在中性或碱性溶液中,反应2-24小时;分离纯化得到多酚/氯己定纳米抗菌颗粒;将得到的多酚/氯己定纳米抗菌颗粒均匀分散在中性或碱性溶液中得到组分A,待用;
步骤2、在步骤1制得多酚/氯己定纳米抗菌颗粒表面接枝羟基磷灰石识别分子。
5.根据权利要求4所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,所述氯己定浓度0.5-5mg/mL,且所述多酚与氯己定的质量份数比为1:0.2-1:2。
6.根据权利要求4所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,所述步骤2具体包括:
取羟基磷灰石识别分子分散在中性或碱性溶液中得到组分B;
取步骤1中组分A和组分B混合均匀得到混合溶液,反应12-24小时;
分离纯化得到纳米涂敷材料颗粒。
7.根据权利要求4所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,在所述步骤2中,混合溶液中羟基磷灰石识别分子与纳米颗粒的质量浓度比范围是1:10-1:100。
8.根据权利要求6所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,所述步骤1和步骤2中的中性或碱性溶液的pH值为7-10,且采用去离子水溶液或去离子水/乙醇混合溶液。
9.根据权利要求6所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,所述步骤1和步骤2中,分离纯化为通过高速离心弃上清、去离子水清洗、再离心;或为通过透析去除杂质后冷冻干燥。
10.根据权利要求4所述的一种用于骨和牙体表面抗菌的纳米涂敷材料的制备方法,其特征在于,所述步骤1多酚为表没食子儿茶素、没食子酸酯、没食子酸、单宁酸、褐藻多酚。
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