CN113332424A - 光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用 - Google Patents
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Abstract
本发明公开了一种光热剂PACP‑MnO2纳米微球在制备抗菌药物中的应用，属于抗菌材料的应用技术领域。光热剂PACP‑MnO2纳米微球对大肠杆菌或金黄葡萄球菌有很好的抑制效果，在近红外光照射条件下，细菌可全部杀死。本发明制备的PACP‑MnO2纳米微球具有很好的中空结构，无近红外光照条件下展示良好的生物相容性，当有近红外光照条件下，细菌被快速杀死。
Description
技术领域
本发明属于抗菌材料的应用技术领域，具体涉及光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用。
背景技术
纳米材料是至少一个方向的尺寸为1-1000 nm的材料，在实际中应用较多的为1-400 nm范围的材料。纳米由于具有较小的尺寸而致使其具有优异的表面效应，如材料的亲和性好、生物相容性好、以及在生物体内易吸收、易游走等特性。
目前研究较多的抗菌材料主要有以下几类：一是Ag+等金属型纳米抗菌材料，其主要利用Ag+等金属离子对细胞膜的通透性，使胞细菌体内酶蛋白失活，从而杀死细菌；二是是ZnO、TiO2等光催化型纳米抗菌材料，主要利用此类材料的光催化作用，与H2O2或OH-反应生成的具有强氧化性的羟基自由基（·OH）和超氧自由基（·O2 -）来杀死细菌；三是季铵盐或季磷盐修饰改性无机纳米颗粒，如纳米蒙脱土（MMT）或SiO2，此类无机纳米颗粒内部有特殊的结构而带有不饱和负电荷，从而具有强烈的阳离子交换能力，经季铵盐或季磷盐修饰后，对细菌有强的吸附固定作用，从而起到抗菌作用。以上技术对正常细胞也有杀伤，而光热型抗菌剂，是将光热剂定位到细菌表面，在细菌上产生局域性的过高热，从而通过热消融机制加速细菌死亡的途径来杀死细菌。
发明内容
针对现有技术中存在的问题，本发明要解决的技术问题在于提供光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用。
为了解决上述问题，本发明所采用的技术方案如下：
光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用。
所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，所述细菌为大肠杆菌或金黄葡萄球菌。
所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，将光热剂PACP-MnO2纳米微球与大肠杆菌或金黄葡萄球菌混合，在37℃条件下培养24h；然后再用近红外光照射0~10min。
所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，具体步骤如下：
（1）将金黄色葡萄球菌或大肠杆菌的单个菌落分别接种到无菌液体LB培养基中，在37°C的恒温培养摇床中以每分钟180转的速度孵育一夜；
（2）使用无菌磷酸盐缓冲盐水将细菌稀释为1×106 CFU/mL，用PACP-MnO2配制成2mg/m L的混悬液；
（3）将200μL细菌溶液和20μL PACP-MnO2震荡摇匀，对细菌样品施加近红外辐射。
所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，近红外光照射时间为10min，每5min摇动一次。
所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，所述近红外光的波长为808nm，辐射功率为1W。
有益效果：与现有的技术相比，本发明的优点包括：
本发明制备的PACP-MnO2纳米微球具有很好的中空结构，无近红外光照条件下展示良好的生物相容性，当有近红外光照条件下，细菌被快速杀死。
附图说明
图1为实施例1制备的PACP-MnO2的扫描电子显微镜照片图；
图2为808nm近红外光照600s后大肠杆菌的扫描电子显微镜照片图；
图3为808nm近红外光照600s后金黄葡萄球菌发热扫描电子显微镜照片图；
图4为808nm近红外光照600s后细菌细胞的荧光图像。
具体实施方式
为使本发明的上述目的、特征和优点能够更加明显易懂，下面结合具体实施例对本发明的具体实施方式做详细的说明。
实施例1
1、吸附剂PACP的制备
将0.06g聚乙二醇辛基苯基醚(TritonX-100)添加到60mL的去离子水中形成均匀溶液，吸取0.38mL苯胺与0.29mL吡咯添加至溶液中，在室温下预冷1h；将过硫酸铵（（NH4)2S2O8）水溶液添加至预冷混合物中0℃反应12h，离心超声得到沉淀，去离子水洗至无色，取黑色固体放入真空干燥箱中60℃下干燥24h，得到聚苯胺聚吡咯（PACP）空心球。
2、吸附剂PACP-MnO2的制备
称取0.13g的PACP，用45mL去离子水溶解配置成溶液，用盐酸将pH调整至1，随后超声15min。将12.5mL的0.05mol·L-1高锰酸钾（KMnO4）溶液加入至PACP溶液中，磁力搅拌器下反应4h，离心得到黑色沉淀，放入鼓风干燥箱中干燥24h，得到PACP-MnO2。图1为实施例1制备的PACP-MnO2的扫描电子显微镜照片图，由图1可知，该纳米微球具有很好的中空结构，无近红外光照条件下展示良好的生物相容性。
对上述合成的光热剂PACP-MnO2纳米微球与大肠杆菌或金黄葡萄球菌混合，设定温度为37℃，培养24小时；然后再用近红外光(808nm)对培养的溶液照射0-10分钟，通过DAPI和PI荧光分析活/死细菌，绿色表示活细胞，红色表示死细胞。
具体步骤如下：
（1）将耐药的金黄色葡萄球菌和大肠杆菌的单个菌落分别接种到50mL的无菌液体LB 培养基中，将耐药菌悬液在37°C的恒温培养摇床中以每分钟180转的速度孵育一夜；
（2）使用无菌磷酸盐缓冲盐水(PBS)将细菌稀释为1×106 CFU/mL，用PACP-MnO2配制成2 mg/m L的混悬液；
（3）将200μL上述细菌溶液和20μL PACP-MnO2震荡摇匀作为实验组，使用PBS作为空白对照组，对细菌样品施加或不施加近红外辐射，照射时间为10分钟，每5分钟摇动一次；辐射功率为1W，波长808nm，每组平行三次。
（4）反应后溶液加入500μL无菌水高速离心洗涤菌液后留下沉淀，加水稀释至500μL。然后用200μL DAPI(12.5 μg/m L)和PI (1.25 μg/mL)染色15 min，室温避光放置30min后，取10μL溶液用于激光扫描荧光共聚焦显微镜成像。
图2为808nm近红外光照600s后，大肠杆菌的SEM图片，两端破裂说明大肠杆菌在光热条件下死亡。图3为808nm近红外光照600s后，金黄葡萄球菌的SEM图片，两端破裂说明金黄葡萄球菌在光热条件下死亡。图4为808nm近红外光照600s后，大肠杆菌染色后的荧光显微镜照片，红色说明大肠杆菌全部死亡。
Claims (6)
1.光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用。
2.根据权利要求1所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，其特征在于，所述细菌为大肠杆菌或金黄葡萄球菌。
3.根据权利要求1所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，其特征在于，将光热剂PACP-MnO2纳米微球与大肠杆菌或金黄葡萄球菌混合，在37℃条件下培养24h；然后再用近红外光照射0~10min。
4.根据权利要求1所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，其特征在于，具体步骤如下：
（1）将金黄色葡萄球菌或大肠杆菌的单个菌落分别接种到无菌液体LB培养基中，在37°C的恒温培养摇床中以每分钟180转的速度孵育一夜；
（2）使用无菌磷酸盐缓冲盐水将细菌稀释为1×106 CFU/mL，将PACP-MnO2配制成2mg/mL的混悬液；
（3）将200μL细菌溶液和20μL PACP-MnO2震荡摇匀，对细菌样品施加近红外辐射。
5.根据权利要求1~4任一所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，其特征在于，近红外光照射时间为10min，每5min摇动一次。
6.根据权利要求1~4任一所述的光热剂PACP-MnO2纳米微球在制备抗菌药物中的应用，其特征在于，所述近红外光的波长为808nm，辐射功率为1W。
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| CN112023904A (zh) * | 2020-09-18 | 2020-12-04 | 南京林业大学 | 一种快速合成的吸附剂PACP-MnO2纳米微球及其制备方法和应用 |
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2021
- 2021-04-06 CN CN202110368581.4A patent/CN113332424A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112023904A (zh) * | 2020-09-18 | 2020-12-04 | 南京林业大学 | 一种快速合成的吸附剂PACP-MnO2纳米微球及其制备方法和应用 |
Non-Patent Citations (2)
| Title |
|---|
| FENGMING LIN 等: "Conjugated Polymer-Based Photothermal Therapy for Killing Microorganisms", 《ACS APPL. POLYM. MATER.》 * |
| NAVEEN CHANDRA JOSHI 等: "Biological Synthesis, Characterisations and Antimicrobial activities of manganese dioxide (MnO2) nanoparticles", 《RESEARCH J. PHARM. AND TECH.》 * |
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