CN115137750A - 一种可热熔流延型纳米复合凝胶抗菌剂及其制备方法 - Google Patents

一种可热熔流延型纳米复合凝胶抗菌剂及其制备方法 Download PDF

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CN115137750A
CN115137750A CN202210634786.7A CN202210634786A CN115137750A CN 115137750 A CN115137750 A CN 115137750A CN 202210634786 A CN202210634786 A CN 202210634786A CN 115137750 A CN115137750 A CN 115137750A
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antibacterial agent
hot
antibacterial
dispersion system
radiation
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李月生
饶璐
刘东亮
杨金玉
王亚洋
王智俊
付娌丽
杨小洁
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Hubei University of Science and Technology
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Hubei University of Science and Technology
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Abstract

本发明提供了一种可热熔流延型纳米复合凝胶抗菌剂及其制备方法，属于抗菌材料技术领域。将70％～80％的热熔胶、2％～10％的纳米粒子、1％～4.0％的辐照敏化剂、0.2％～4.0％的pH调节剂混合，超声搅拌制成均匀分散体系后，放入油浴锅约为6～8h，完全溶解至溶胶状态；然后加入2.0％～4.0％的乳化剂制成分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻‑解冻3～5次，保存备用；置于伽马射线或电子射线产生的电离射线下进行辐射交联反应，选择的电子束能量为1～5MeV，辐射剂量为5～60kGy，剂量率为5～20kGy/pass；用去离子水浸泡去除未交联的单体，最后所得的产物即为水凝胶抗菌剂。无毒价廉、低温熔融性、耐热性强、不易分解、杀菌力高等特点。

Description

一种可热熔流延型纳米复合凝胶抗菌剂及其制备方法
技术领域
本发明属于抗菌材料技术领域，涉及一种可热熔流延型纳米复合凝胶抗菌剂及其制备方法，特别涉及一种以热熔胶和纳米材料为基材、进行辐射制备具有抗菌性能的凝胶基抗菌剂的方法。
背景技术
根据世界卫生组织(WHO)的数据，细菌引起的疾病正严重威胁着人类的健康。全世界每年有1300万患者的健康受到来自于细菌感染的威胁。抗生素的出现让抗菌进入了一个快速发展期，然而，抗生素的过度使用和长期使用导致了多重耐药菌(MDR)的出现，并严重威胁着人类的生命安全。在某些情况下，大多数耐药细菌感染病例需要通过组织清创，进行长时间的抗生素治疗，导致患者负担过高的医疗保健费用。随着时间的推移，各种抗生素的持续选择性压力，最终导致生物体增强辅助耐药机制，从而引发多药耐药、新型青霉素结合蛋白、酶依赖性药物改变、膜通透性改变、药物靶点突变和外排泵表达增加等副作用。因此，研发新型、高效、无毒副作用的抗菌剂，在抗菌抗感染方面有着不可替代的作用。随着科学技术的发展，对抗菌机制研究的深入，一些学者们制备出了更加高效的抗菌新材料。
抗菌剂是对病原菌有杀灭作用的功能性材料，通过抑制或杀灭细菌的代谢活性，从而将病原菌的致病作用最小化，但又不妨碍机体正常生长。抗菌剂在医学实践中用于预防或抵抗浅表组织的细菌感染，并对器械和被感染的材料进行消毒。抗菌机理主要包括：通过静电作用破坏细菌的细胞壁，从而将细菌的结构破坏，导致细菌的死亡；或者是扰乱DNA和RNA的合成；亦或扰乱核糖体的正常功能，进而阻碍蛋白质的合成。
抗菌剂种类繁多，目前常见的抗菌剂可分为无机抗菌剂，有机抗菌剂，天然抗菌剂等三类。无机抗菌剂中的金属离子型抗菌剂，主要是将对病原菌具有抑制特性的金属原子氧化成离子(如:将金属银(Ag)氧化成银离子(Ag+))，随后将其负载在多孔的无机材料上。虽然金属离子型抗菌剂的抗菌性能具有持久性，但是由于能发挥抗菌作用的基本都是重金属，所以在使用时会造成严重的环境污染。除此之外，金属离子型抗菌材料也因其高昂的价格限制其广泛应用。纳米技术提供了一种开发新型无机抗菌剂的方法，纳米SiO2等氧化物颗粒所特有的高比表面积等特性，使其具有减少细菌污染、广谱抗菌高活性、无毒副作用。但纳米型抗菌材料的生产工艺冗杂，制备过程能耗高，对生产设备的要求也极高。以上严苛的生产要求，也极大地限制了此类抗菌剂的推广。
有机抗菌剂已经运用于各种不同行业中，其应用范围远优于其他类型的抗菌剂。有机抗菌剂可以吸附在细菌或霉菌的细胞膜表面，通过扩散作用与细菌或霉菌内部的一些基团(如巯基)等作用，从而破坏细胞结构的完整性，同时抑制细胞内部某些酶的活性，达到抗菌效果。与其它抗菌剂相比较，有机抗菌剂具有广谱性且抗菌性能优异。但是有些有机抗菌剂的耐热性差，不适宜用在塑料等对耐热性要求较高的行业。目前来说，有机抗菌剂的安全性能尚在研究之中，还普遍存在耐热性较差，易水解、有效期短等不足之处。
天然抗菌剂，主要来自天然物质的提取物，大致可分为三大类，动物类、植物类和矿物类，天然抗菌剂的抗菌作用有限，而且耐热性较差，杀菌力较低。因此，使用的范围严重受限。
中国专利CN113907090A公开了一种SiO2水溶胶基抗菌剂的制备方法，是将微米单质银分散液和壳聚糖溶液搅拌混合，沉淀，洗涤过滤后获得壳聚糖/微米单质银复合抗菌剂，加入表面活性剂和烷基硅氧烷，继续搅拌反应，从而获得SiO2水溶胶基抗菌剂。虽然制备过程简单，但是壳聚糖耐热性差，容易分解，使用寿命短，银粒子未能控制为纳米级别，抗菌能力较弱。而且单质银氧化呈灰色或褐色，影响制品颜色。并且银原料贵重稀少，致使制造成本增加。
中国专利CN114085428A介绍了一种塑料改性用抗菌剂及其制备方法，所述抗菌剂利用类流体技术，由聚硅氧烷季铵盐为内冠、长链烷基醚钾盐为外冠，通过表面自组装技术和离子交换技术对负载银、锌离子的磷酸氢锆层状介孔材料进行包覆，并通过季铵盐的改性制得。这种方法合成步骤繁多复杂，制造困难，会大大增加材料成本，不利于工业化生产，限制其推广与广泛应用。
中国专利CN113875776A介绍了一种可溶性银锌复合物、可溶性银锌抗菌剂、片剂及制备方法，涉及水、物体表面微生物处理领域。具体为螯合剂，银盐和可溶性锌盐制成。克服了银离子见光变色，影响产品的性能和外观，降低抗菌性能的缺陷。但是可溶性锌盐，银盐均会对水体及水游生物造成污染，而银，锌等可能增加健康安全风险，且螯合剂容易造成二次污染，不符合绿色环保理念。
目前，水凝胶抗菌剂成分、形态和制备方法都过于单一，特别是制备方法还是局限在传统的冷冻和化学交联等方法。传统的水凝胶普遍采用化学交联法，制备的水凝胶不仅易在水体中释放残余的有毒交联剂或氧化还原试剂，会出现不均匀交联，并且化学交联难以控制交联度；其次，水凝胶不易获得均匀分布的凝胶骨胶体系，而且纳米粒子和药物与凝胶分子间的作用力太弱甚至没有，导致其易从凝胶孔洞中流失,从而影响了水凝胶的功能性。为克服化学法带来的不足，辐射合成可热熔流延型热熔胶的纳米复合凝胶抗菌剂的方法应运而生。辐射法相对于化学法实现了有效的补充和完善，具体表现为：①反应过程不需要添加任何对人体有毒的引发剂和催化剂，纯度高，效率高，产品均一性强；②反应条件温和，在室温下即可进行，操作简便，反应周期短；③通过控制组分配比和辐照条件可以准确调控水凝胶的相关性能，能及时针对抗菌剂的应用要求做出调整；④凝胶制备、塑形和灭菌等过程可同步完成。因此，辐射合成技术已成为智能水凝胶制备极富前景的研究途径。
本发明所要解决的问题是公开一种可热熔流延型纳米复合凝胶抗菌剂及其辐射制备方法，可通过简单加热的方式改变其物理状态，通过引入抗菌纳米粒子，拓展了热熔胶的功能性，以克服现有技术存在的上述缺陷。
经检索，国内尚未有与本发明相同的专利申请。
发明内容
本发明的目的是针对现有的技术存在的上述问题，提供一种可热熔流延型纳米复合凝胶抗菌剂，本发明所要解决的技术问题是使可热熔流延型凝胶抗菌剂材料满足抗菌疾病治疗。
本发明的目的可通过下列技术方案来实现：一种可热熔流延型纳米复合凝胶抗菌剂，其特征在于，所述抗菌剂是通过热熔胶和抗菌纳米材料基材进行辐射交联反应获得的抗菌材料。
通过电离辐射技术，对热熔胶和抗菌纳米材料基材进行辐射交联反应，通过控制温度从而调节其流动性，满足与任何异形界面的充分接触，热熔胶引入抗菌粒子，拓展热熔胶的功能性。针对抗菌剂的临床要求来调整水凝胶抗菌剂的韧性、溶胀度、流动性、微观结构、机械强度以及生物相容性等性能，从而制备满足抗菌疾病治疗的可热熔流延型凝胶抗菌剂材料。
一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，包括以下步骤：
①将制备水凝胶抗菌剂的材料为：以质量百分比计，热熔胶(70％～80％)、纳米粒子(2％～10％)、辐照敏化剂(1％～4.0％)、pH调节剂(0.2％～4.0％)混合，超声搅拌制成均匀分散体系后，放入油浴锅约为6～8h，完全溶解至溶胶状态；
②将步骤①的样品加入乳化剂(2.0％～4.0％)制成分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻-解冻3～5次，保存备用；
③将步骤②所得样品并置于伽马射线或电子射线产生的电离射线下进行辐射交联反应，选择的电子束能量为1～5MeV，辐射剂量为5～60kGy，剂量率为5～20kGy/pass；
④样品用去离子水浸泡去除未交联的单体，最后所得的产物即为水凝胶抗菌剂。通过简单加热方式可将抗菌剂调整为流动状态，满足与任何异形界面的充分接触实现充分抗菌。
进一步的，所述的热熔胶为聚氨酯、聚烯烃、聚酰胺等中的一种或几种。一种或几种热熔胶在辐照的过程中，可以发生自交联或彼此相互交联，保证了水凝胶具有一定的网络结构，也具有一定的流变性、粘弹性、韧性等特性，保证水凝胶抗菌剂有效应用于抗菌治疗的基本物化性能。
进一步的，所述的纳米粒子为羟基磷灰石、氧化石墨烯、石墨烯、二氧化钛等中的一种或几种。一种或几种超分子在辐照的过程中，可以与热熔胶相互交联，互为贯穿，保证了水凝胶抗菌性，良好的稳定性等特性，有效提高水凝胶抗菌剂的流动性和抗菌性能。
进一步的，所述的辐照敏化剂为胶原蛋白、透明质酸、聚乙烯吡咯烷酮、聚丙烯酸钠等中的一种或几种。辐照敏化剂的引入，缩短了辐照交联所需的时间，提高了生产效率，确保了水凝胶抗菌剂具有适当的流体力学性能和稳定的网络结构，也保证了水凝胶抗菌剂的生物相容性和整体性能的均一性。
进一步的，所述的pH调节剂为磷酸氢二钠、磷酸一氢钠、碳酸氢钠、己二酸、乙酸、Tris-HCl、亚氨基二乙酸、三乙醇胺中的一种或几种。pH调节剂的引入，可以调节高分子混合体系的分散性，确保高分子自身不缠结，有利于后期辐射交联的顺利进行。
进一步的，所述的乳化剂为吐温20、吐温40和吐温60中的一种或几种。吐温系列乳化剂易溶于水、甲醇、乙醇等溶剂，具有良好的分散性和稳定性。
本发明所述的抗菌剂是通过改变细菌的离子通道，而使其离子溶出细菌体外，从而导致细菌因生理活性丧失而死亡。另外，凝胶本身高保水性使得其具有优良的抗黏附抑菌性能。它可以有效防止蛋白质等生物大分子在材料表面的非特异性黏附，并有效减少细菌初始附着，抑制细菌生物膜的形成，从而达到抗菌的效果。
本发明的有益效果是：
1、抗菌剂具有无毒价廉，低温熔融性，耐热性强、不易分解，杀菌力高等特点，满足与任何异形界面或异形空间的有效接触与填充，从而实现快速、稳定、充分的抗菌效果。
2、凝胶成型后具有较好的机械强度，能有效避免因强度不够导致内部结构崩塌，从而导致药物中毒现象。
3、辐射交联所制备的热熔凝胶型抗菌剂，具有良好的生物相容性、韧性、均一性、抗菌性、无毒性、无抗原性和安全性。
4、抗菌剂的辐射制备、塑形及灭菌等可同步完成，具有快速、简单、高效、无损、无毒等特点，大大简化了生产工艺，节约成本，综合经济效益显著，尤其适用于日用品、化妆品、医疗器械、生物医药等诸多技术领域。
附图说明
图1中A图是凝胶抗菌剂的热熔流延状；图1中B图为凝胶抗菌剂的抑菌圈。
图2为不同含量TPU/HAP/GO凝胶抗菌剂在同一辐照剂量下的拉伸应力应变曲线。
图3为不同含量TPU/HAP/GO凝胶抗菌剂在同一辐照剂量下的拉伸模量。
图4为同一含量TPU/HAP/GO凝胶抗菌剂在不同辐照剂量下的拉伸应力应变曲线。
图5为同一含量TPU/HAP/GO凝胶抗菌剂在不同辐照剂量下的拉伸模量。
图6为不同时间大肠杆菌在凝胶抗菌剂上培养的菌落数。
图7为不同时间金黄色葡萄球菌在凝胶抗菌剂培养的菌落数。
图8为凝胶抗菌剂培养小鼠成骨细胞MTT图。
具体实施方式
以下是本发明的具体实施例并结合附图，对本发明的技术方案作进一步的描述，但本发明并不限于这些实施例。
为了弥补上述抗菌剂所存在的不足，我们研发了一种可热熔流延型热熔胶纳米复合凝胶抗菌剂。该抗菌剂避免了无机抗菌剂易造成污染、价格昂贵，有机抗菌剂耐热性差、易分解，天然抗菌剂杀菌力低等三类抗菌剂的不足，同时具有熔融性(如图1)，满足与任何异形界面的充分接触，任何异形空间的有效填充，从而实现充分抗菌。且有一定的强度(如图3、5)，实现了凝胶载药抗菌功能和缓慢降解与吸收，避免因强度不够导致内部结构崩塌而发生药物中毒现象。
实施例1：将0.2g氧化石墨烯投入到常温的20mL去离子水中，超声分散均匀，加入7g聚氨酯、0.1g胶原蛋白、0.02g磷酸氢二钠，油浴锅80℃搅拌6h，搅拌速率为150r/min，完全溶解后，加入0.4g吐温20乳化，所得分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻-解冻3～5次先物理交联，再用电子束加速器辐照，选用辐照剂量为5kGy，剂量率为5kGy/pass的条件进行辐照，样品用去离子水浸泡去除未交联的单体，即可制备水凝胶抗菌剂，所得的样品放置常温保存备用。利用万能拉力机测定其拉伸模量为1.682MPa，利用自动菌落计数仪测定其大肠杆菌抗菌率为55％，金黄色葡萄球菌抗菌率为60％，可以有效应用于抗菌疾病的治疗。
实施例2：将0.3g羟基磷灰石投入到常温的20mL去离子水中，超声分散均匀，加入8g聚烯烃、0.15g聚乙烯吡咯烷酮、0.03g乙酸，油浴锅80℃搅拌6h，搅拌速率为150r/min，完全溶解后，加入0.6g吐温40乳化，所得分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻-解冻3～5次先物理交联，再用电子束加速器辐照，选用辐照剂量为20kGy，剂量率为10kGy/pass的条件进行辐照，样品用去离子水浸泡去除未交联的单体，即可制备水凝胶抗菌剂，所得的样品放置常温保存备用。利用万能拉力机测定其拉伸模量为3.624MPa，利用自动菌落计数仪测定其大肠杆菌抗菌率为75％，金黄色葡萄球菌抗菌率为80％，可以有效应用于抗菌疾病的治疗。
实施例3：将0.4g二氧化钛投入到常温的20mL去离子水中，超声分散均匀，加入9g聚酰胺、0.2g透明质酸、0.04g碳酸氢钠，油浴锅80℃搅拌6h，搅拌速率为150r/min，完全溶解后，加入0.8g吐温60乳化，所得分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻-解冻3～5次先物理交联，再用电子束加速器辐照，选用辐照剂量为60kGy，剂量率为20kGy/pass的条件进行辐照，样品用去离子水浸泡去除未交联的单体，即可制备水凝胶抗菌剂，所得的样品放置常温保存备用。利用万能拉力机测定其拉伸模量为4.044MPa，利用自动菌落计数仪测定其大肠杆菌抗菌率为63％，金黄色葡萄球菌抗菌率为72％，可以有效应用于抗菌疾病的治疗。
本发明声称的技术效果通过附图和附图中体现的参数佐证，具体而言：凝胶抗菌剂的热熔流延状如图1中A图所示；凝胶抗菌剂的抑菌圈如图1中B图所示；不同含量TPU/HAP/GO凝胶抗菌剂在同一辐照剂量下的拉伸应力应变曲线如图2所示；不同含量TPU/HAP/GO凝胶抗菌剂在同一辐照剂量下的拉伸模量如图3所示；同一含量TPU/HAP/GO凝胶抗菌剂在不同辐照剂量下的拉伸应力应变曲线如图4所示；同一含量TPU/HAP/GO凝胶抗菌剂在不同辐照剂量下的拉伸模量如图5所示；不同时间大肠杆菌在凝胶抗菌剂上培养的菌落数如图6所示；不同时间金黄色葡萄球菌在凝胶抗菌剂培养的菌落数如图7所示；凝胶抗菌剂培养小鼠成骨细胞MTT图如图8所示。
本文中所描述的具体实施例仅仅是对本发明精神作举例说明。本发明所属技术领域的技术人员可以对所描述的具体实施例做各种各样的修改或补充或采用类似的方式替代，但并不会偏离本发明的精神或者超越所附权利要求书所定义的范围。

Claims (7)

1.一种可热熔流延型纳米复合凝胶抗菌剂，其特征在于，所述抗菌剂是通过热熔胶和抗菌纳米材料基材进行辐射交联反应获得的抗菌材料，具体组分为：按照质量百分比计，70％～80％的热熔胶、2％～10％的纳米粒子、1％～4.0％的辐照敏化剂、0.2％～4.0％的pH调节剂和2.0％～4.0％的乳化剂。
2.一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，包括以下步骤：
①将制备水凝胶抗菌剂的材料为：将70％～80％的热熔胶、2％～10％的纳米粒子、1％～4.0％的辐照敏化剂、0.2％～4.0％的pH调节剂混合，超声搅拌制成均匀分散体系后，放入油浴锅约为6～8h，完全溶解至溶胶状态；
②将步骤①的样品加入2.0％～4.0％的乳化剂制成分散体系灌注到培养皿，之后静置除泡，置于冰箱循环冷冻-解冻3～5次，保存备用；
③将步骤②所得样品并置于伽马射线或电子射线产生的电离射线下进行辐射交联反应，选择的电子束能量为1～5MeV，辐射剂量为5～60kGy，剂量率为5～20kGy/pass；
④样品用去离子水浸泡去除未交联的单体，最后所得的产物即为水凝胶抗菌剂。
3.根据权利要求2所述一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，所述的热熔胶为聚氨酯、聚烯烃、聚酰胺等中的一种或多种的混合。
4.根据权利要求2所述一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，所述的纳米粒子为羟基磷灰石、氧化石墨烯、石墨烯、二氧化钛等中的一种或多种的混合。
5.根据权利要求2所述一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，所述的辐照敏化剂为胶原蛋白、透明质酸、聚乙烯吡咯烷酮、聚丙烯酸钠等中的一种或或多种的混合。
6.根据权利要求2所述一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，所述的pH调节剂为磷酸氢二钠、磷酸一氢钠、碳酸氢钠、己二酸、乙酸、Tris-HCl、亚氨基二乙酸、三乙醇胺中的一种或或多种的混合。
7.根据权利要求2所述一种可热熔流延型纳米复合凝胶抗菌剂的制备方法，其特征在于，所述的乳化剂为吐温20、吐温40和吐温60中的一种或多种的混合。
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