CN116510058B - 一种抗菌型细菌纤维素基创伤敷料的制备方法和应用 - Google Patents
一种抗菌型细菌纤维素基创伤敷料的制备方法和应用 Download PDFInfo
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Abstract
一种抗菌型细菌纤维素基创伤敷料的制备方法,涉及一种创伤敷料的制备方法。制备方法:将预处理的细菌纤维素膜经过TEMPO氧化处理浸泡到MOF(ZIF‑8)经过单宁酸(TA)改性处理后配置成一定浓度的溶液中,再次浸泡在一定浓度的MXene纳米片水分散液中,得到抗菌型细菌纤维素基创伤敷料。本发明所制备的细菌纤维素基抗菌型创伤敷料具有很好的抗菌效果(抑菌率>99%),且ZIF‑8和MXene光热协同抗菌的方法可以有效避免细菌的耐药性,且温和的光热效果还能够促进细胞的分化、增殖和迁移,加快血管再生等;本发明制备的抗菌型敷料具有很好的抗氧化性、组织湿粘附性、生物可降解性、可裁剪性、良好的吸水性能、优异的生物相容性、止血性能和机械强度等性能。
Description
技术领域
本发明涉及一种抗菌型细菌纤维素基创伤敷料的制备方法和应用。
背景技术
糖尿病患者发生慢性糖尿病性溃疡的风险很大,且伤口愈合差,复发率高,易导致截肢。糖尿病慢性皮肤创伤的愈合是一个极其复杂的过程。慢性炎症主要是由于大量活性氧(ROS)的积累、促炎趋化因子和细菌感染引起的,是慢性伤口长期无法治愈的主要原因。慢性伤口往往停留在炎症期,超过细胞的抗氧化能力,从而阻止伤口从炎症期过渡到增殖期。一般来说,在敷料中添加抗生素、消炎药和生长因子是消除慢性炎症的主要方式。虽然已经开发出了用于治疗细菌感染伤口的抗生素敷料,但抗生素引起的副作用和抗生素本身的细胞毒性不可忽视。此外,由于微生物生物膜的存在,抗生素对细菌感染创面的渗透能力较弱,影响了治疗效果。近年来,利用敷料材料调节创面炎症微环境的系统已被开发出来,包括去除ROS、吸附炎症因子。因此,具有创面炎症微环境内在协同调节机制的创伤敷料在治疗慢性糖尿病创面方面具有巨大潜力。
细菌纤维素(BC)是一种天然的纳米纤维水凝胶,主要由木质醋酸菌产生,广泛用于创伤敷料,由于BC具有较高的保水能力、柔韧性、高结晶度、良好的力学性能、生物相容性等,BC具有创面愈合能力,但缺乏抗菌活性和体内可降解性,阻碍了其在该领域的广谱应用。为了解决这一问题,功能化BC是开发新型BC基敷料的重要策略。
金属有机框架(Metal-organicframeworks,MOFs)是近年来备受关注的一类新兴纳米材料。MOFs的大表面积、高孔隙率和分散良好的活性位点使其在多个领域具有突出的性能。近年来,多项研究报道了锌MOF(ZIF-8)具有优异的抗菌性能,开辟了MOF作为抗菌药物的新应用。此外,考虑到其固有的生物降解性和生物相容性,ZIF-8已被应用于伤口敷料以防止细菌感染。ZIF-8在伤口愈合过程中可以缓慢释放Zn2+,Zn2+不仅可以通过破坏微生物膜有效杀死细菌,还可以通过促进细胞迁移、血管生成和胶原沉积来促进伤口愈合。众所周知,Zn2+作为血液中第二丰富的金属离子,在血液凝固中起着关键作用,它可以激活因子XII和VII,启动凝血级联,加速凝血酶和纤维蛋白的生成。此外,ZIF-8具有高孔隙率和大表面积,可以加速对血液中水的吸收和红细胞/血小板的凝结。
光热治疗(PTT)因其机制新颖,避免了传统抗生素容易产生细菌耐药的缺点而备受关注。PTT通过近红外激光照射光热试剂,将光能迅速转化为热能,从而对细菌造成不可逆的损伤。与传统治疗方法相比,PTT具有穿透力强、疗程短、副作用少等优点。MXene作为一种新型二维纳米材料,具有良好的光热稳定性,是一种很有前途的光热试剂。由于MXene的主要元素是C、N和过渡金属(如Ti、Nb、Ta),具有良好的生物相容性,可以被降解并从体内排出,同时,MXene具有良好的抗菌性能,可同时杀灭革兰氏阳性和革兰氏阴性细菌,从而使得其在生物医疗领域具有很好的应用前景。
发明内容
针对现有技术中存在的问题,本发明的目的在于提供一种抗菌型细菌纤维素基创伤敷料的制备方法和应用,该敷料具有良好的力学性能、粘附性能和抗氧化性能,ZIF-8和MXene(光热抗菌)协同抗菌,而且制备工艺简单,制备出的敷料性能优良。
为了实现上述目的,本发明采取以下技术方案:
一种抗菌型细菌纤维素基创伤敷料的制备方法,包括以下步骤:
步骤一:将氧化细菌纤维素膜加入到没食子酸的Tris溶液中,在pH8.5的条件下反应6-24h后,用去离子水洗涤至中性,得到TBC@GA膜;
步骤二:将ZIF-8粉末加入到去离子水中,搅拌均匀得到溶液A,将单宁酸粉末加入到溶液A中继续搅拌反应,然后用去离子水离心清洗掉未反应的单宁酸粉末,真空干燥后得到ZIF-8@TA粉末;
步骤三:将TBC@GA膜加入到ZIF-8@TA分散液中反应6-24h后,用去离子水清洗得到TTZ类水凝胶膜;
步骤四:将TTZ类水凝胶膜浸入到MXene水溶液中,在30℃条件下反应6-24h后,经过去离子水清洗后,得到抗菌型细菌纤维素基创伤敷料TTZM。
进一步的,步骤一中,所述氧化细菌纤维素膜的制备方法为:将细菌纤维素膜用去离子水反复清洗、浸泡,然后再用碱液清洗,继续用去离子水反复浸泡、洗涤至中性,得到细菌纤维素类水凝胶膜;按1-10g:1L的比例将细菌纤维素类水凝胶膜加入到混合溶液中,并在室温下连续氧化反应2h,pH保持在10.5,最后通过加入乙醇终止反应,用去离子水洗涤至中性,得到氧化细菌纤维素膜,其中每升混合溶液包括0.5gNaBr、80mgTEMPO和NaClO,其中NaClO与细菌纤维素类水凝胶膜的比例为5-20mmol/g,溶剂为去离子水。
进一步的,碱液清洗的时间为2-8h。
进一步的,步骤一中,氧化细菌纤维素膜与没食子酸的质量比为1:1。
进一步的,步骤二中,ZIF-8与单宁酸的质量比为2-10:1。
进一步的,步骤三中,ZIF-8@TA分散液的浓度为1-10g/L。
进一步的,步骤四中,所述MXene水溶液的浓度为1-5mg/ml。
一种所述的抗菌型细菌纤维素基创伤敷料的应用,在光热条件下,将抗菌型细菌纤维素基创伤敷料贴至患处。
进一步的,光热条件为0.2-2w/cm2照射2-10min。
与现有技术相比,本发明的有益效果是:
1、本发明采用的细菌纤维素是由木质醋酸菌产生,绿色无毒害,具有优异的生物相容性和细胞亲和性,并对其结构式中C6位置氧化改性后具有一定的生物可降解性,与皮肤具有很好的粘附性,且具有类似水凝胶的结构特性等。没食子酸作为多酚表现出与不同高分子材料的良好粘附能力,为功能材料的设计提供了一个仿生平台。没食子酸(GA)可以在碱性溶液中氧化和自聚合而获得,可以在基底上产生强大的涂层,赋予基底新的功能。没食子酸具有抗氧化、抗菌、抗炎等性能,与氧化细菌纤维素膜(TBC)结合后提高了其抗菌性、抗氧化性和湿粘附性等。
2、MXene的光热抗菌与ZIF-8释放Zn2+负载于细菌纤维素敷料中的协同作用更好地提高了本发明细菌纤维素敷料的抗菌性能及抗菌持久性;由于Zn2+接触空气中的氧气会产生ROS,进一步导致氧化应激,加速细菌死亡;在MXene和ZIF-8的协同作用下,Zn2+离子可以粘附在微生物膜上,影响细胞内化和与细胞内含量的相互作用。在TTZ类水凝胶膜中加入MXene,进一步提高了其抗菌性能,即MXene破坏细胞壁的切割作用和光热性能,可以帮助MXene进入细菌内部,使蛋白质泄漏并最终在近红外照射下死亡。且MXene,ZIF-8@TA和GA赋予了本发明细菌纤维素敷料一定的抗氧化性、湿粘附性,还能够根据创面的形状及大小进行裁剪,能够促进创面细胞的分化、增殖和迁移,从而加快创面的愈合;本发明复合敷料还具有良好的吸水性能、优异的生物相容性以及止血性能,在慢性难愈合创伤修复领域具有潜在的应用前景。TTZM复合水凝胶组,可杀灭98.8%的金黄色葡萄球菌和95.8%的大肠杆菌。近红外辐照后,该复合水凝胶的温度明显升高,从20℃上升到56℃。值得注意的是,在808nm近红外激光的辅助下,MXene负载水凝胶几乎灭活了所有金黄色葡萄球菌(99.9%)和大肠杆菌(99.8%)。
3、本发明方法制备简单、条件温和,原材料成本低廉,易于产品转化,有利于工业规模化生产,并且制备过程中无有毒、有害物质排放,属于绿色无污染加工过程。
附图说明
图1为实施例1TBC膜的SEM图;
图2为实施例1ZIF-8@TA的TEM图;
图3为实施例1TTZ类水凝胶膜的SEM图;
图4为实施例1TTZM膜的SEM图;
图5为实施例1在不同功率下的光热升温情况;
图6为实施例1抗菌型细菌纤维素创伤敷料的抗菌效果图(金黄色葡萄球菌),左为对照组,右为实验组。
图7为实施例1抗菌型细菌纤维素创伤敷料的抗菌效果图(大肠杆菌),左为对照组,右为实验组。
图8为实施例1抗菌型细菌纤维素创伤敷料用于大鼠伤口治疗不同时间点的伤口愈合图片和伤口愈合痕迹的叠加图;
图9抗菌型细菌纤维素敷料用于大鼠伤口治疗不同时间点的伤口愈合面积的统计图。
具体实施方式
下面将结合附图和实施例,对本发明中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅是本发明的一部分实施例,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做出创造性劳动的前提下所获得的所有其他实施例,都属于本发明保护的范围。
一种抗菌型细菌纤维素基创伤敷料的制备方法,涉及一种创伤敷料的制备方法。制备方法包括以下步骤:将预处理的细菌纤维素膜经过TEMPO氧化处理后,与没食子酸反应,然后浸泡到MOF(ZIF-8)中,经过单宁酸(TA)改性处理后配置成一定浓度的溶液中,再次浸泡在一定浓度的MXene纳米片水分散液中,得到抗菌型细菌纤维素基创伤敷料。本发明所制备的细菌纤维素基抗菌型创伤敷料具有很好的抗菌效果(抑菌率>99%),且ZIF-8和MXene光热协同抗菌的方法可以有效避免细菌的耐药性,且温和的光热效果还能够促进细胞的分化、增殖和迁移,加快血管再生等;本发明制备的抗菌型敷料具有很好的抗氧化性、组织湿粘附性、生物可降解性、可裁剪性、良好的吸水性能、优异的生物相容性、止血性能和机械强度等性能。
实施例1
一种抗菌型细菌纤维素基创伤敷料的制备方法,按照以下步骤进行:
步骤一:将细菌纤维素膜用去离子水反复清洗、浸泡,然后再用碱液清洗2-8h,继续用去离子水反复浸泡、洗涤至中性,得到细菌纤维素类水凝胶膜(BC膜),冷藏备用;将BC膜剪成均匀大小,然后将1g细菌纤维素类水凝胶膜加入到1L的混合溶液中(其中每升混合溶液包括0.5gNaBr,80mgTEMPO,NaClO,其中NaClO与细菌纤维素类水凝胶膜的比例为5-20mmol/g,其中溶剂为去离子水),并在室温下连续氧化反应2h,pH保持在10.5。最后通过加100ml乙醇终止反应,用去离子水洗涤至中性,得到氧化细菌纤维素膜(TBC)。然后将TBC加入到没食子酸(GA)的Tris溶液中在pH8.5条件下反应6h后,用去离子水洗涤至中性,得到TBC@GA;
其中,所述BC膜的大小剪裁为3cm×2cm;
所述BC膜的有效纤维素含量为1g;
所述TBC与没食子酸的质量比为1:1;
步骤二:将一定质量的ZIF-8粉末加入到50ml去离子水中,搅拌20min后得到溶液A,将一定质量的单宁酸(TA)粉末加入到溶液A中继续搅拌30min,用去离子水离心清洗掉未反应的TA粉末,真空干燥后得到ZIF-8@TA粉末;
所述ZIF-8与TA的质量比为2:1;
步骤三:将TBC@GA膜加入到一定浓度的ZIF-8@TA分散液中,反应12h后,用去离子水清洗得到TTZ类水凝胶膜;
所述ZIF-8@TA的浓度为2.5g/L;
步骤四:将TTZ类水凝胶膜浸入到一定浓度的MXene水溶液中,在30℃条件下反应12h后,经过去离子水清洗后,最终得到抗菌型细菌纤维素基创伤敷料TTZM;
所述MXene水溶液的浓度为2mg/ml;
图1为实施例1制备的TBC膜的SEM图;图2为实施例1制备的ZIF-8@TA的TEM图,可以看到ZIF-8多面体表面长了一层TA膜;图3为实施例1制备的TTZ类水凝胶膜的SEM图,可以看到ZIF-8@TA可以均匀的长到TBC纤维上;图4为实施例1TTZM膜的SEM图,可以看到MXene纳米片吸附到TBC的纤维之间;图5为实施例1制备的抗菌型细菌纤维素基创伤敷料TTZM在不同功率下的光热升温情况;图6为实施例1制备的抗菌型细菌纤维素基创伤敷料的抗菌效果图(金黄色葡萄球菌),左为对照组,右为实验组。图7为实施例1制备的抗菌型细菌纤维素基创伤敷料的抗菌效果图(大肠杆菌),左为对照组,右为实验组;图8为实施例1制备的抗菌型细菌纤维素基创伤敷料用于大鼠伤口治疗不同时间点的伤口愈合图片和伤口愈合痕迹的叠加图;图9为实施例1制备的抗菌型细菌纤维素基创伤敷料用于大鼠伤口治疗不同时间点的伤口愈合面积的统计图。
本实施例所制备的抗菌型细菌纤维素基创伤敷料TTZM对金黄色葡萄球菌和大肠杆菌的抑菌率达到99%以上,选择的光热时间为10min,强度为1w/cm2;创面愈合率12d后可以达到98%以上。
实施例2
本实施例与实施例1不同的是,步骤四中将MXene纳米片溶液的浓度为1mg/ml。其他步骤和工艺参数与实施例1相同。
本实施例所制备的抗菌型细菌纤维素基创伤敷料TTZM对金黄色葡萄球菌和大肠杆菌的抑菌率达到99%以上。
实施例3
本实施例与实施例1不同的是,步骤四中将MXene纳米片溶液的浓度为3mg/ml。其他步骤和工艺参数与实施例1相同。
本实施例所制备的抗菌型细菌纤维素基创伤敷料TTZM对金黄色葡萄球菌和大肠杆菌的抑菌率达到99%以上,选择的光热时间为10min,强度为0.5w/cm2。
此外,应当理解,虽然本说明书按照实施方式加以描述,但并非每个实施方式仅包含一个独立的技术方案,说明书的这种叙述方式仅仅是为清楚起见,本领域技术人员应当将说明书作为一个整体,各实施例中的技术方案也可以经适当组合,形成本领域技术人员可以理解的其他实施方式。
Claims (9)
1.一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:包括以下步骤:
步骤一:将氧化细菌纤维素膜加入到没食子酸的Tris溶液中,在pH8.5的条件下反应6-24h后,用去离子水洗涤至中性,得到TBC@GA膜;
步骤二:将ZIF-8粉末加入到去离子水中,搅拌均匀得到溶液A,将单宁酸粉末加入到溶液A中继续搅拌反应,然后用去离子水离心清洗掉未反应的单宁酸粉末,真空干燥后得到ZIF-8@TA粉末;
步骤三:将TBC@GA膜加入到ZIF-8@TA分散液中反应6-24h后,用去离子水清洗得到TTZ类水凝胶膜;
步骤四:将TTZ类水凝胶膜浸入到MXene水溶液中,在30℃条件下反应6-24h后,经过去离子水清洗后,得到抗菌型细菌纤维素基创伤敷料TTZM。
2.根据权利要求1所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:步骤一中,所述氧化细菌纤维素膜的制备方法为:将细菌纤维素膜用去离子水反复清洗、浸泡,然后再用碱液清洗,继续用去离子水反复浸泡、洗涤至中性,得到细菌纤维素类水凝胶膜;按1-10g:1L的比例将细菌纤维素类水凝胶膜加入到混合溶液中,并在室温下连续氧化反应2h,pH保持在10.5,最后通过加入乙醇终止反应,用去离子水洗涤至中性,得到氧化细菌纤维素膜,其中每升混合溶液包括0.5gNaBr、80mgTEMPO和NaClO,其中NaClO与细菌纤维素类水凝胶膜的比例为5-20mmol/g,溶剂为去离子水。
3.根据权利要求2所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:碱液清洗的时间为2-8h。
4.根据权利要求1所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:步骤一中,氧化细菌纤维素膜与没食子酸的质量比为1:1。
5.根据权利要求1所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:步骤二中,ZIF-8与单宁酸的质量比为2-10:1。
6.根据权利要求1所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:步骤三中,ZIF-8@TA分散液的浓度为1-10g/L。
7.根据权利要求1所述的一种抗菌型细菌纤维素基创伤敷料的制备方法,其特征在于:步骤四中,所述MXene水溶液的浓度为1-5mg/ml。
8.一种权利要求1-7任一权利要求所述的抗菌型细菌纤维素基创伤敷料的应用,其特征在于:在光热条件下,将抗菌型细菌纤维素基创伤敷料贴至患处。
9.根据权利要求8所述的应用,其特征在于:光热条件为0.2-2w/cm2照射2-10min。
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