CN113197211A - 光动力微生物消杀剂 - Google Patents
光动力微生物消杀剂 Download PDFInfo
- Publication number
- CN113197211A CN113197211A CN202110480578.1A CN202110480578A CN113197211A CN 113197211 A CN113197211 A CN 113197211A CN 202110480578 A CN202110480578 A CN 202110480578A CN 113197211 A CN113197211 A CN 113197211A
- Authority
- CN
- China
- Prior art keywords
- photodynamic
- microbe
- biocide
- amine compound
- photosensitizer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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Abstract
本发明公开了一种光动力微生物消杀剂,包含光敏剂和胺类化合物,两者在光动力光源的照射下反应得到消杀剂。本发明的消杀剂在光动力光源的照射下通过胺类化合物的加入调控并增强光敏剂的活性氧生成效率,从而增强光动力消杀性能;该消杀剂具有可调控、成本低、工艺简单且对医用材料表面生物膜具有高效、广谱高效杀菌等优点,解决了传统的光敏剂在使用过程中光稳定性差、活性氧生成效率低等不足之处。
Description
技术领域
本发明涉及一种消杀剂,尤其涉及一种光动力微生物消杀剂。
背景技术
传统的消毒/杀菌技术,包括化学消毒剂和紫外线照射杀菌,存在能耗较高、细菌易产生耐药性或引发臭氧污染等缺点。同时,抗生素滥用及由此产生的细菌抗生素耐药已成为人类面临的重大公共卫生问题,然而传统抗生素的研发过程却大大滞后于细菌耐药性的产生,如果抗生素耐药性趋势得不到有效控制,到2050年,世界经济的累计损失可能达到100万亿美元。因此,探索新型、廉价、有效的有害微生物消杀物质及方法显得尤为重要。近年来,人们已经设计和开发了大量的非抗生素类抗菌试剂(如抗菌肽、阳离子聚合物和金属纳米粒子等)来应对这一公共卫生难题。然而,这些抗菌试剂同样面临一些挑战,如抗菌肽的化学合成成本昂贵、药物半衰期短且存在特异性毒性;阳离子聚合物易引起免疫原性和炎症反应;金属纳米粒子由于其比表面积大而发生聚集,其稳定性的下降严重影响其抗菌性能,极大地限制了其临床应用范围。
光动力疗法主要利用光照激活光敏感物质(即光敏剂),促使其产生活性氧分子(ROS)以杀伤细菌。活性氧如单线态氧、超氧阴离子和羟基自由基都具有广谱抗菌活性,可以破坏大部分的微生物分子靶点(如蛋白质、脂质和核酸),因此微生物对光敏剂不太可能产生耐药性。
传统的光敏剂材料虽然具有良好的生物相容性和较低的暗毒性,但大多数光敏剂光稳定性差,在光照下较快的分解很大程度上影响了活性氧的生成效率,限制了在光动力抗菌疗法中的使用,导致难以及时且彻底地清除细菌或由此引发的生物膜感染(ChemicalEngineering Journal 355(2019)130–136)。
发明内容
发明目的:本发明提供了一种能广谱高效灭杀微生物、成本低、工艺简单的光动力微生物消杀剂。
技术方案:本发明所述的光动力微生物消杀剂,包含光敏剂和胺类化合物,两者在光动力光源的照射下反应得到消杀剂。
本发明的光敏剂为可见光活性物质,优选的,所述可见光活性物质为核黄素和/或其衍生物、二氧化钛、g-C3N4、孟加拉红或类分子筛咪唑骨架材料(如ZIF-8)。
其中,光敏剂的浓度为0.001mM~1000mM;进一步地,光敏剂的浓度为0.001mM~100mM;更进一步地,光敏剂的浓度为0.001mM~10mM;更优选的,光敏剂的浓度为0.002mM~1mM。
本发明的胺类化合物为水溶性胺类化合物,其中,所述水溶性胺类化合物为脂肪族、芳香族仲胺或叔胺类小分子,或含有仲胺或叔胺结构的聚合物。
其中,水溶性胺类化合物为如下结构式(1)-(3)中的任意一种:
其中,x,y,z=0~3000;
更进一步地,所述水溶性胺类化合物为如下8种结构式中的任意一种:
本发明胺类化合物的浓度为0.01mM~1000mM;进一步地,其浓度为1mM~100mM。
本发明光动力光源的光照时间为0.1~480min;进一步地,光动力光源的光照时间为0.1~120min;更进一步地,光照时间为0.5~60min;再进一步地,光照时间为1~30min。
本发明中,光动力光源的光照强度为1~10000mW/cm2;进一步地,光照强度为2~1000mW/cm2;更进一步地,光照强度为5~200mW/cm2。
本发明的消杀剂用于消杀细菌和真菌,其中,所述细菌为革兰氏阳性菌和革兰氏阴性菌。
上述革兰氏阳性菌包括金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌或表皮葡萄球菌;所述革兰氏阴性菌包括大肠杆菌、粪肠球菌、耐氨苄大肠杆菌、肺炎克雷伯氏菌、铜绿假单胞菌、鲍曼不动杆菌、耐多药鲍曼不动杆菌或白色念珠菌;所述真菌包括白粉菌、赤霉菌、黑星菌、核盘菌、粉痂菌或根霉菌。
本发明的消杀剂用于织物表面抗菌、家具板材表面抗菌、皮肤表面抗菌、金属基以及高分子基植/介入医疗器械表面抗菌及环境中有害微生物的消杀。
发明机理:本发明利用特定波长的光辐照无毒的光敏剂(PS),基态的PS吸收一个光子后被激发变为短暂的单重激发态1PS*,继而通过系统间的交换变为长期三重态3PS*,通过发射荧光或释放能量重回基态。同时,激发态的3PS*可转移电子至周边物质继而生成自由基离子,如超氧自由基、羟基自由基或者转移能量至氧分子继而产生单线态氧。生成的自由基和单线态氧可通过氧化损伤破坏细菌细胞膜和DNA,而将胺类化合物和光敏剂配合使用,可有效提高总活性氧水平,进而提高对细菌、真菌等有害微生物的消杀效果。
有益效果:与现有技术相比,本发明的消杀剂在光动力光源的照射下通过胺类化合物的加入调控并增强光敏剂的活性氧生成效率,从而增强光动力消杀性能,该消杀剂具有可调控、成本低、工艺简单且对医用材料表面生物膜具有高效、广谱高效杀菌等优点,解决了传统的光敏剂在使用过程中光稳定性差、活性氧生成效率低等不足之处;同时,本发明消杀剂制备使用的原料来源广泛、价格低廉、生物相容性好;此外,本发明的消杀剂杀灭有害微生物效率高,可实现对织物表面、家具板材表面、皮肤表面、金属基以及高分子基植/介入医疗器械表面及环境中其它有害微生物的消杀,具有广阔的应用前景。
附图说明
图1为本发明的消杀剂对金黄色葡萄球菌杀菌实验的定量结果;
图2为本发明的消杀剂对耐甲氧西林金黄色葡萄球菌杀菌实验的定量结果;
图3为本发明的消杀剂体外总活性氧生成量测试结果;
图4A和图4B分别是本发明的消杀剂对两种不同细菌的消杀结果。
具体实施方式
下面结合附图对本发明的技术方案作进一步说明。
实施例1 4-羟乙基哌嗪乙磺酸(HEPES)/核黄素对医用聚氨酯表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在医用聚氨酯表面,经48小时培养形成生物被膜。将含有6.25mM HEPES的核黄素溶液(核黄素浓度:0.001mM)喷洒到医用聚氨酯表面。在光强为150mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为99.9%。
实施例2 4-羟乙基哌嗪乙磺酸(HEPES)/核黄素衍生物对钛基表面大肠肝菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的大肠杆菌培养在钛基表面,经48小时培养形成生物被膜。将含有100mM HEPES的核黄素衍生物溶液(核黄素衍生物浓度:0.005mM)喷洒到钛表面。在光强为50mW/cm2的可见光下照射10min,经检测,杀菌率为96.2%。
实施例3 4-羟乙基哌嗪乙磺酸(HEPES)/二氧化钛对医用聚氨酯表面耐甲氧西林金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在医用聚氨酯表面,经48小时培养形成生物被膜。将含有1000mM HEPES的二氧化钛溶液(二氧化钛浓度:0.05mM)喷洒到医用聚氨酯表面。在光强为20mW/cm2的可见光下照射30min,经检测,杀菌率为99.9%。
实施例4 2-[[三(羟甲基)甲基]氨基]乙磺酸/g-C3N4对硅橡胶表面大肠杆菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的大肠杆菌培养在硅橡胶表面,经48小时培养形成生物被膜。将含有25mM 2-[[三(羟甲基)甲基]氨基]乙磺酸的g-C3N4溶液(g-C3N4浓度:0.005mM)喷洒到硅橡胶表面。在光强为10000mW/cm2的模拟太阳光下照射0.1min,经检测,杀菌率为97.6%。
实施例5三乙醇胺(TEOA)/ZIF-8对棉织物表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在棉织物表面,经48小时培养形成生物被膜。将含有0.01mM TEOA的ZIF-8溶液(ZIF-8溶液浓度:0.008mM)喷洒到棉织物表面。在光强为10mW/cm2的太阳光下照射480min,经检测,杀菌率为80%。
实施例6三乙醇胺(TEOA)/孟加拉红对实木木材表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在实木木材表面,经48小时培养形成生物被膜。将含有25mM TEOA的孟加拉红溶液(孟加拉红浓度:0.002mM)喷洒到实木木材表面。在光强为100mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为95%。
实施例7聚乙烯亚胺(PEI)/核黄素对医用聚氨酯表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在医用聚氨酯膜表面,经48小时培养形成生物被膜。将含有50mM PEI的核黄素溶液(核黄素浓度:0.005mM)喷洒到医用聚氨酯表面。在光强为100mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为99%。
实施例8 2-吗啉乙磺酸(MES)/MOF对医用聚氨酯表面白色念珠菌的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在医用聚氨酯膜表面,经48小时培养形成生物被膜。将含有100mM MES的钌溶液(ZIF-8浓度:1000mM)喷洒到医用聚氨酯表面。在光强为100mW/cm2的可见光下照射20min,经检测,杀菌率为99.8%。
实施例9 2-吗啉乙磺酸(MES)/核黄素对铜铬合金表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在铜铬合金表面,经48小时培养形成生物被膜。将含有1000mM MES的上转换纳米粒溶液(上转换纳米粒浓度:0.5mM)喷洒到铜铬合金表面。在光强为250mW/cm2的模拟太阳光下照射2min,经检测,杀菌率为99.9%。
实施例10N,N-双(2-羟乙基)-2-氨基乙磺酸(BES)/g-C3N4对医用聚氨酯表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在医用聚氨酯膜表面,经48小时培养形成生物被膜。将含有6.25mM BES的二硫化钼溶液(二硫化钼浓度:0.05mM)喷洒到医用聚氨酯表面。在光强为20mW/cm2的可见光下照射30min,经检测,杀菌率为97%。
实施例11N,N-二甲基苯胺/二氧化钛对钛合金表面白粉菌的光动力杀菌效果检测:
将初始密度为108CFU/mL的白粉菌培养在钛合金表面,经48小时培养形成生物被膜。将含有6.25mM N,N-二甲基苯胺的二氧化钛溶液(二氧化钛浓度:0.05mM)喷洒到钛合金表面。在光强为12mW/cm2的太阳光下照射360min,经检测,杀菌率为96.3%。实施例12 4-羟乙基哌嗪乙磺酸(HEPES)/核黄素对医用聚氨酯表面根霉菌的光动力杀菌效果检测:
将初始密度为108CFU/mL的根霉菌培养在医用聚氨酯表面,经48小时培养形成生物被膜。将含有25mM 4-羟乙基哌嗪乙磺酸(HEPES)(核黄素:0.005mM)喷洒到医用聚氨酯表面。在光强为100mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为96.3%。
实施例13N-甲基苯胺/核黄素对KN95口罩表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在KN95口罩表面,经48小时培养形成生物被膜。将含有6.25mM N-甲基苯胺的核黄素溶液(核黄素浓度:0.005mM)喷洒到KN95口罩表面。在光强为50mW/cm2的可见光下照射10min,经检测,杀菌率为89%。
实施例14 4-羟乙基哌嗪乙磺酸(HEPES)/核黄素对KN95口罩表面金黄色葡萄球菌生物膜的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌培养在KN95口罩表面,经48小时培养形成生物被膜。将含有25mM HEPES的核黄素溶液(核黄素浓度:0.005mM)喷洒到KN95口罩表面。在光强为120mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为99.6%。
实施例15 4-羟乙基哌嗪乙磺酸(HEPES)/核黄素对医用聚氨酯表面根霉菌的光动力杀菌效果检测:
将初始密度为108CFU/mL的根霉菌培养在医用聚氨酯表面,经48小时培养形成生物被膜。将含有1000mM HEPES的核黄素溶液(核黄素:0.005mM)喷洒到医用聚氨酯表面。在光强为100mW/cm2的模拟太阳光下照射3min,经检测,杀菌率为96.3%。实施例16三乙醇胺(TEOA)/核黄素对皮肤表面金黄色葡萄球菌的光动力杀菌效果检测:
将初始密度为108CFU/mL的金黄色葡萄球菌滴加到直径为8mm的大鼠伤口表面以建立细菌感染模型,将含有50mM HEPES的核黄素溶液(核黄素浓度:0.005mM)喷洒到伤口处,在光强为120mW/cm2的模拟太阳光下照射5min,经检测,杀菌率为99.9%。图1为实施例1制备的消杀剂对金黄色葡萄球菌杀菌实验定量结果,实验组1、2、3是加入三种不同的消杀剂(分别由4-羟乙基哌嗪乙磺酸、三乙醇胺、3-二甲氨基丙醇和光敏剂核黄素组成),在模拟太阳光条件下光照5min对金黄色葡萄球菌的杀菌实验CFU定量结果。
图2为实施例1制备的消杀剂对耐甲氧西林金黄色葡萄球菌杀菌实验定量结果,实验组1、2、3是加入三种不同的消杀剂(分别由4-羟乙基哌嗪乙磺酸、三乙醇胺、3-二甲氨基丙醇和光敏剂核黄素组成),在模拟太阳光条件下光照5min对耐甲氧西林金黄色葡萄球菌的杀菌实验CFU定量结果。
图3为实施例1制备消杀剂的体外总活性氧生成量测试(光照核黄素与HEPES/核黄素的总活性氧水平测试)以反映抗菌性能,从图中可以明显看出加入HEPES可以有效提高总活性氧水平。
图4A和图4B分别是实施例1制备的消杀剂对金黄色葡萄球菌处理后细菌的形貌变化,将本发明的消杀剂(由4-羟乙基哌嗪乙磺酸和核黄素组成)均匀喷洒到已形成大量生物膜的聚氨酯表面,光照消杀处理后对表面金黄色葡萄球菌生物膜的形貌的变化(图4A:处理前,图4B:处理后),可以发现:未经处理的细菌生物膜菌体形貌正常,经本发明的消杀剂处理后,细菌菌体破裂,表面褶皱,呈现明显死亡的迹象。
表1是由HEPES和核黄素组成的消杀剂中,不同因素(HEPES浓度、光敏剂核黄素浓度、光照强度、光照时间)对金黄色葡萄球菌消杀率的效果,其中序号1-7是只改变HEPES浓度单一变量的结果,序号8-10是只改变核黄素浓度单一变量的结果,序号11-14是只改变光照时间单一变量的结果,序号15-17是只改变光强单一变量的结果。
表1
Claims (10)
1.一种光动力微生物消杀剂,其特征在于:包含光敏剂和胺类化合物,两者在光动力光源的照射下反应得到消杀剂。
2.根据权利要求1所述的光动力微生物消杀剂,其特征在于:所述光敏剂为可见光活性物质,其中,所述可见光活性物质为核黄素、核黄素衍生物、二氧化钛、g-C3N4、孟加拉红、上转换纳米粒、二硫化钼或类分子筛咪唑骨架材料。
3.根据权利要求2所述的光动力微生物消杀剂,其特征在于:所述光敏剂的浓度为0.001mM~1000mM。
4.根据权利要求1所述的光动力微生物消杀剂,其特征在于:所述胺类化合物为水溶性胺类化合物,其中,所述水溶性胺类化合物为脂肪族、芳香族仲胺或叔胺类小分子,或含有仲胺或叔胺结构的聚合物。
7.根据权利要求5或6所述的光动力微生物消杀剂,其特征在于:所述胺类化合物的浓度为0.01mM~1000mM。
8.根据权利要求1所述的光动力微生物消杀剂,其特征在于:所述消杀剂用于消杀细菌和真菌,其中,所述细菌为革兰氏阳性菌和革兰氏阴性菌。
9.根据权利要求8所述的光动力微生物消杀剂,其特征在于:所述革兰氏阳性菌包括金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌或表皮葡萄球菌;所述革兰氏阴性菌包括大肠杆菌、粪肠球菌、耐氨苄大肠杆菌、肺炎克雷伯氏菌、铜绿假单胞菌、鲍曼不动杆菌、耐多药鲍曼不动杆菌或白色念珠菌;所述真菌包括白粉菌、赤霉菌、黑星菌、核盘菌、粉痂菌或根霉菌。
10.根据权利要求1所述的光动力微生物消杀剂,其特征在于:所述消杀剂用于织物表面抗菌、家具板材表面抗菌、皮肤表面抗菌、金属基以及高分子基植/介入医疗器械表面抗菌及环境中有害微生物的消杀。
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