Thea viridis derived carbon dots for drug-resistant bacterial infections by photodynamic therapy

{ "defaultlang": "zh", "titlegroup": { "articletitle": [ { "lang": "zh", "data": [ { "name": "text", "data": "绿茶衍生碳点用于光动力治疗耐药菌感染" } ] }, { "lang": "en", "data": [ { "name": "italic", "data": [ { "name": "text", "data": "Thea viridis" } ] }, { "name": "text", "data": " derived carbon dots for drug-resistant bacterial infections by photodynamic therapy" } ] } ] }, "contribgroup": { "author": [ { "name": [ { "lang": "zh", "surname": "张", "givenname": "云秀", "namestyle": "eastern", "prefix": "" }, { "lang": "en", "surname": "ZHANG", "givenname": "Yunxiu", "namestyle": "eastern", "prefix": "" } ], "stringName": [], "aff": [ { "rid": "aff1", "text": "1" }, { "rid": "aff3", "text": "3" } ], "role": [ "first-author" ], "bio": [ { "lang": "zh", "text": [ "张云秀（1990－），女，山东聊城人，在读博士生，主要从事光诊疗材料设计及抗肿瘤应用。E-mail：zhangyunxiu114@mails.ucas.ac.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196870&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196872&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196871&type=", "width": "19.13724518", "height": "24.58345032", "fontsize": "" } ], "data": [ [ { "name": "text", "data": "张云秀" }, { "name": "text", "data": "（1990－），女，山东聊城人，在读博士生，主要从事光诊疗材料设计及抗肿瘤应用。E-mail：" }, { "name": "text", "data": "zhangyunxiu114@mails.ucas.ac.cn" } ] ] } ], "email": "zhangyunxiu114@mails.ucas.ac.cn", "deceased": false }, { "name": [ { "lang": "zh", "surname": "贾", "givenname": "庆岩", "namestyle": "eastern", "prefix": "" }, { "lang": "en", "surname": "JIA", "givenname": "Qingyan", "namestyle": "eastern", "prefix": "" } ], "stringName": [], "aff": [ { "rid": "aff1", "text": "1" }, { "rid": "aff2", "text": "2" } ], "role": [ "corresp" ], "corresp": [ { "rid": "cor2", "lang": "en", "text": " iamqyjia@nwpu.edu.cn", "data": [ { "name": "text", "data": " iamqyjia@nwpu.edu.cn" } ] } ], "bio": [ { "lang": "zh", "text": [ "贾庆岩（1990－），男，山东聊城人，博士，副教授，2018年毕业于中国科学院理化技术研究所，主要从事光电诊疗材料与器件在重大疾病中的应用研究。 E-mail: iamqyjia@nwpu.edu.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196873&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196875&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196874&type=", "width": "19.66516304", "height": "24.87277222", "fontsize": "" } ], "data": [ [ { "name": "text", "data": "贾庆岩" }, { "name": "text", "data": "（1990－），男，山东聊城人，博士，副教授，2018年毕业于中国科学院理化技术研究所，主要从事光电诊疗材料与器件在重大疾病中的应用研究。 E-mail: " }, { "name": "text", "data": "iamqyjia@nwpu.edu.cn" } ] ] } ], "email": "iamqyjia@nwpu.edu.cn", "deceased": false }, { "name": [ { "lang": "zh", "surname": "葛", "givenname": "介超", "namestyle": "eastern", "prefix": "" }, { "lang": "en", "surname": "GE", "givenname": "Jiechao", "namestyle": "eastern", "prefix": "" } ], "stringName": [], "aff": [ { "rid": "aff1", "text": "1" }, { "rid": "aff3", "text": "3" } ], "role": [ "corresp" ], "corresp": [ { "rid": "cor1", "lang": "en", "text": "E-mail： jchge2010@mail.ipc.ac.cn", "data": [ { "name": "text", "data": "E-mail： jchge2010@mail.ipc.ac.cn" } ] } ], "bio": [ { "lang": "zh", "text": [ "葛介超（1970－ ），男，山东临沂人，博士，研究员，2008年于山东师范大学获得博士学位，主要从事新型光响应纳米材料的设计及其在抗肿瘤、病毒、细菌、炎症或创伤修复等领域应用的研究。E-mail： jchge2010@mail.ipc.ac.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196876&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196878&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196877&type=", "width": "20.77372360", "height": "25.89210510", "fontsize": "" } ], "data": [ [ { "name": "text", "data": "葛介超" }, { "name": "text", "data": "（1970－ ），男，山东临沂人，博士，研究员，2008年于山东师范大学获得博士学位，主要从事新型光响应纳米材料的设计及其在抗肿瘤、病毒、细菌、炎症或创伤修复等领域应用的研究。E-mail： " }, { "name": "text", "data": "jchge2010@mail.ipc.ac.cn" } ] ] } ], "email": "jchge2010@mail.ipc.ac.cn", "deceased": false } ], "aff": [ { "id": "aff1", "intro": [ { "lang": "zh", "label": "1", "text": "中国科学院理化技术研究所，光化学转换与功能材料重点实验室， 北京　100190", "data": [ { "name": "text", "data": "中国科学院理化技术研究所，光化学转换与功能材料重点实验室， 北京　100190" } ] }, { "lang": "en", "label": "1", "text": "Key Laboratory of Photochemical Conversion and Optoelectronic Materials and CityU-CAS Joint Laboratory of Functional Materials and Devices， Technical Institute of Physics and Chemistry， Chinese Academy of Sciences， Beijing， 100190， China", "data": [ { "name": "text", "data": "Key Laboratory of Photochemical Conversion and Optoelectronic Materials and CityU-CAS Joint Laboratory of Functional Materials and Devices， Technical Institute of Physics and Chemistry， Chinese Academy of Sciences， Beijing， 100190， China" } ] } ] }, { "id": "aff2", "intro": [ { "lang": "zh", "label": "2", "text": "西北工业大学，柔性电子前沿科学中心， 陕西 西安　710129", "data": [ { "name": "text", "data": "西北工业大学，柔性电子前沿科学中心， 陕西 西安　710129" } ] }, { "lang": "en", "label": "2", "text": "Frontiers Science Center for Flexible Electronics （FSCFE）， Xi’an Institute of Flexible Electronics （IFE） and Xi’an Institute of Biomedical Materials & Engineering （IBME）， Northwestern Polytechnical University， 127 West Youyi Road， Xi’an 710072， China", "data": [ { "name": "text", "data": "Frontiers Science Center for Flexible Electronics （FSCFE）， Xi’an Institute of Flexible Electronics （IFE） and Xi’an Institute of Biomedical Materials & Engineering （IBME）， Northwestern Polytechnical University， 127 West Youyi Road， Xi’an 710072， China" } ] } ] }, { "id": "aff3", "intro": [ { "lang": "zh", "label": "3", "text": "中国科学院大学，未来技术学院， 北京　100049", "data": [ { "name": "text", "data": "中国科学院大学，未来技术学院， 北京　100049" } ] }, { "lang": "en", "label": "3", "text": "School of future technology， University of Chinese Academy of Sciences， Beijing， 100049， China", "data": [ { "name": "text", "data": "School of future technology， University of Chinese Academy of Sciences， Beijing， 100049， China" } ] } ] } ] }, "abstracts": [ { "lang": "zh", "data": [ { "name": "p", "data": [ { "name": "text", "data": "随着细菌耐药性的不断播散，尤其是“超级细菌”的出现，临床可用抗生素药物越来越少，迫切需要发展新的高效、低毒和无耐药性的抗菌材料和技术。本研究采用生物质绿茶作为碳源，通过溶剂热法，成功制备了具有光动力治疗（Photodynamic therapy， PDT）性能的绿茶衍生碳点（T-CDs）。在660 nm激光照射下，所制备的T-CDs 能够有效产生活性氧。细胞和活体实验表明，T-CDs具有优异的生物相容性，且产生的活性氧能杀死耐甲氧西林金黄色葡萄球菌，从而通过降低细菌引起的伤口炎症，加速伤口愈合。本研究所制备的T-CDs能够通过PDT杀灭致病菌，促进感染伤口的愈合，为开发抗生素替代药物提供了新的思路，同时对探索耐药菌感染伤口临床治疗新方案具有重要参考价值。" } ] } ] }, { "lang": "en", "data": [ { "name": "p", "data": [ { "name": "text", "data": "With the spread of bacterial drug resistance， especially the emergence of \"superbugs\"， it’s an urgent need to develop new antibacterial materials and technologies with high efficiency， low toxicity and no drug resistance. In this study， " }, { "name": "italic", "data": [ { "name": "text", "data": "thea viridis " } ] }, { "name": "text", "data": "derived carbon dots （T-CDs） were successfully prepared by solvothermal method. Under 660 nm laser irradiation， the prepared T-CDs could effectively produce reactive oxygen species （ROS）. " }, { "name": "italic", "data": [ { "name": "text", "data": "In vitro" } ] }, { "name": "text", "data": " and" }, { "name": "italic", "data": [ { "name": "text", "data": " in vivo " } ] }, { "name": "text", "data": "experiments showed that T-CDs have excellent biocompatibility， and can produce ROS under laser irradiation to kill methicillin-resistant " }, { "name": "italic", "data": [ { "name": "text", "data": "Staphylococcus aureus" } ] }, { "name": "text", "data": "， thereby reducing wound inflammation caused by bacteria and accelerating wound healing. The prepared T-CDs can kill pathogenic bacteria through PDT and promote the healing of infected wounds， provide a new idea for the development of antibiotic replacement drugs， and have important value for exploring new clinical treatment schemes of drug-resistant bacteria infected wounds." } ] } ] } ], "keyword": [ { "lang": "zh", "data": [ [ { "name": "text", "data": "碳点" } ], [ { "name": "text", "data": "光动力治疗" } ], [ { "name": "text", "data": "耐药菌感染" } ], [ { "name": "text", "data": "生物质" } ] ] }, { "lang": "en", "data": [ [ { "name": "text", "data": "carbon dots" } ], [ { "name": "text", "data": "photodynamic therapy" } ], [ { "name": "text", "data": "drug-resistant bacteria" } ], [ { "name": "text", "data": "biomass" } ] ] } ], "highlights": [], "body": [ { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "1　引　　言" } ], "level": "1", "id": "s1" } }, { "name": "p", "data": [ { "name": "text", "data": "细菌耐药性（antimicrobial resistance，AMR）是指细菌对于抗菌药物作用的耐受性，一旦产生，药物的治疗作用会明显下降" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "1", "type": "bibr", "rid": "R1", "data": [ { "name": "text", "data": "1" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。AMR的产生是细菌不断进化和人类过度使用抗生素共同导致的后果。世界卫生组织发布的报告显示，到2050年，AMR每年会导致1000万人丧生，相当于每3秒钟就有1人因 AMR失去生命，其危害程度将超过癌症" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "2", "type": "bibr", "rid": "R2", "data": [ { "name": "text", "data": "2" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。近年来，全球药物研发支出不断增加，但抗生素的研究与开发停滞不前。与其他药物相比，虽然抗生素市场定价极低，但开发、试验并将药物推向市场的成本却非常高。候选抗生素从临床前阶段发展到上市应用大约需10-15年，耗资约10亿美元，且大多数新药在应用后的2-3年内就会出现耐药性" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "3", "type": "bibr", "rid": "R3", "data": [ { "name": "text", "data": "3" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。因此，亟需寻求AMR问题的替代治疗方法" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "4", "type": "bibr", "rid": "R4", "data": [ { "name": "text", "data": "4" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "光动力治疗（Photodyanmic therapy，PDT）是一种利用适当波长光源激发光敏剂产生活性氧（Reactive oxygen species，ROS）杀灭致病微生物的临床治疗方法，表现出非侵入性和高时空选择性等优势" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "5", "type": "bibr", "rid": "R5", "data": [ { "name": "text", "data": "5" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。更重要的是，由于ROS主要通过氧化破坏细菌的脂质体、蛋白质和核酸等生物大分子，无需特定靶点，PDT不会引发细菌耐药，可用于重复治疗" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "6", "type": "bibr", "rid": "R6", "data": [ { "name": "text", "data": "6" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。因此，PDT可作为一种潜在的抗生素替代方案，在抗菌治疗领域表现出广阔的应用前景。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "碳点（Carbon dots，CDs）作为碳基纳米材料的成员之一，具有原料来源广、成本低、发光可调、环境友好和生物安全性高等特性，在肿瘤和细菌感染PDT应用上受到广泛关注" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "7", "type": "bibr", "rid": "R7", "data": [ { "name": "text", "data": "7" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "10", "type": "bibr", "rid": "R10", "data": [ { "name": "text", "data": "10" } ] } } ], "rid": [ "R7", "R8", "R9", "R10" ], "text": "7-10", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。研究表明，CDs可依赖于静电作用和ROS产生等多种复杂的机制，通过破坏细菌细胞壁诱导细胞质泄漏，扰乱细菌内DNA/RNA的二级结构，抑或制基因表达等，从而实现对细菌的高效灭杀" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "11", "type": "bibr", "rid": "R11", "data": [ { "name": "text", "data": "11" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。此外，已有证据证明，CDs的物理化学性质受到碳源的影响" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "12", "type": "bibr", "rid": "R12", "data": [ { "name": "text", "data": "12" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。生物质碳源具有环保、成本低、来源广泛的优点" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "13", "type": "bibr", "rid": "R13", "data": [ { "name": "text", "data": "13" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "，还可以一定程度上替代昂贵的化学试剂。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "卟啉类物质（如：叶绿素a、焦脱镁叶绿酸-a、二氢卟吩e6、菌绿素等）是目前临床常见的光敏剂，而叶绿素类化合物是一类富含卟啉类结构的天然产物。因此，选择合适的富含叶绿素类化合物的生物质作为碳源，可制备出含卟啉残基的碳点，从而有望赋予碳点优异的PDT性能。绿茶作为一种生物质，含有多种不同叶绿素化合物，且原产自中国，来源广泛。基于此，本工作选择以绿茶为碳源，通过溶剂热法成功制备了具有PDT抗菌功能的CDs（T-CDs）。在660 nm激光照射下，制备的T-CDs可有效产生ROS，在细胞和活体水平均能够实现对耐药菌的有效灭杀，为耐药菌感染的PDT提供了一种新选择。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2　实　　验" } ], "level": "1", "id": "s2" } }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.1　实验材料" } ], "level": "2", "id": "s2a" } }, { "name": "p", "data": [ { "name": "text", "data": "1,3-二苯基异苯并呋喃、KBr（AR，" }, { "name": "text", "data": ">" }, { "name": "text", "data": " 99.0%）、无水乙醇（AR，" }, { "name": "text", "data": ">" }, { "name": "text", "data": " 99.7%）、二甲基亚砜（AR，" }, { "name": "text", "data": ">" }, { "name": "text", "data": " 99.0%）和丙酮（" }, { "name": "text", "data": ">" }, { "name": "text", "data": " 99.0%）采购自Adamas-beta（中国）。磷酸缓冲液（PBS）购自广州赛国生物科技有限公司。Luria-Bertani (LB) ager购自Sigma-Aldrich（美国）。AlamarBlue检测试剂盒和活/死染色试剂盒购自美国赛默飞世尔科技公司。DMEM培养基、胎牛血清和青霉素-链霉素溶液订购自上海源培生物科技股份有限公司。耐甲氧西林金黄色葡萄球菌（" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "，ATCC BAA-40）和小鼠胚胎成纤维细胞（NIH3T3，ATCC CRL-1658）源自美国菌种保藏中心（ATCC，USA）。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.2　T-CDs的制备" } ], "level": "2", "id": "s2b" } }, { "name": "p", "data": [ { "name": "text", "data": "利用破碎机将产自陕西汉中的成品绿茶破碎为粉末。将2 g粉末分散于40 mL丙酮中，并转移至含有聚四氟乙烯内衬的高压反应釜。160 ℃下加热12 h后，冷却至室温，得到 T-CDs溶液。将上述溶液在10000 rpm/min下离心5 min后，取上清液，进行旋蒸和真空干燥（60 ℃，12 h）处理，得到最终产物T-CDs粉末。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.3　T-CDs的形貌表征" } ], "level": "2", "id": "s2c" } }, { "name": "p", "data": [ { "name": "text", "data": "透射电镜（FEI Talos F200X）对T-CDs的形貌进行表征。将T-CDs分散于无水乙醇溶液中，用1 mL的注射器取少量样品溶液滴在超薄碳基铜网上，置于烘箱（60 ℃）中干燥5 min，随后进行透射电镜测试." } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.4　T-CDs的结构表征" } ], "level": "2", "id": "s2d" } }, { "name": "p", "data": [ { "name": "text", "data": "采用傅里叶变换红外光谱（Fourier transform infrared spectroscopy，FTIR）（Bruker Tensor II）和X射线光电子能谱（X-ray photoelectron spectroscopy, XPS）（Kratos, Britain）表征T-CDs的化学组成。取1 mg T-CDs样品和150 mg KBr进行研磨，通过加压机加压制备透明薄片，用于测定FTIR。直接将T-CDs粉末压片用于测定 XPS，结合能根据284. 6 eV 的 C 1s 峰校准。运用CasaXPS软件对XPS光谱进行分峰拟合，以对不同元素的化学价态进行解析。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.5　T-CDs的光吸收特性表征" } ], "level": "2", "id": "s2e" } }, { "name": "p", "data": [ { "name": "text", "data": "采用紫外可见光谱（Hitachi, Japan）测试T-CDs的吸收光谱。配置0.1 mg/mL T-CDs的DMSO溶液，取2 mL置入石英比色皿中，以DMSO为测试背景，测试400-800 nm的吸收光谱。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.6　T-CDs的ROS产生性能表征" } ], "level": "2", "id": "s2f" } }, { "name": "p", "data": [ { "name": "text", "data": "以1,3-二苯基异苯并呋喃（1,3-Diphenylisobenzofuran，DPBF）作为检测ROS捕获剂，用于检测T-CDs光照后产生ROS的能力。首先，将T-CDs溶液在660 nm处的吸光度调到0.1。以T-CDs溶液为测试背景，加入20 μL DPBF溶液（1 mg/mL）后，在660 nm激光（15 mW/cm" }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "）照射下，收集350-500 nm的DPBF的吸收光谱。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.7　T-CDs的体外抗菌性能表征" } ], "level": "2", "id": "s2g" } }, { "name": "p", "data": [ { "name": "text", "data": "抗菌实验：选取耐甲氧西林金黄色葡萄球菌（Methicillin resistant " }, { "name": "italic", "data": [ { "name": "text", "data": "Staphylococcus aureus" } ] }, { "name": "text", "data": "，" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "）测试T-CDs抗菌性能。37 ℃下，将" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "接种到LB培养基，培养至对数期（OD" }, { "name": "sub", "data": [ { "name": "text", "data": "600" } ] }, { "name": "text", "data": " = 1.0）。取1 mL " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌液离心，5000 rpm/min，5 min，随后用PBS缓冲液重悬。重复清洗三次后，将" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌液进行稀释，并将其浓度控制在10" }, { "name": "sup", "data": [ { "name": "text", "data": "6" } ] }, { "name": "text", "data": "~10" }, { "name": "sup", "data": [ { "name": "text", "data": "7" } ] }, { "name": "text", "data": " CFU/mL。然后，将100 µL " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌液加入装有不同浓度的100 µL T-CDs的96孔板中，在37 ℃恒湿培养箱中培养4 h后，用100 mW/cm" }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "的660 nm激光照射10 min，继续在培养箱中培养0.5 h。取100 µL菌液于新的96孔板中进行10倍稀释，将稀释好的菌液取5 µL滴于LB琼脂板进行点板实验，以计算杀菌率。另外，取 100 µL菌液涂布于 LB 琼脂板上观察细菌存活数量。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "活/死染色：按照上述实验过程，利用100 µg/mL T-CDs对" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "进行PDT处理。随后，离心，获取" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "，并用50 µL含有绿色荧光核酸染色剂（SYTO" }, { "name": "sup", "data": [ { "name": "text", "data": "TM " } ] }, { "name": "text", "data": "9）和碘化丙啶（Propidium iodide，PI）染料的PBS重悬，进一步孵育10 min。然后，将上述菌液在5000 rpm/min下离心5 min， 10 µL PBS重悬后，加入到共聚焦培养皿中，利用共聚焦激光扫描显微镜对细菌进行荧光成像观察。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "扫描电子显微镜（Scanning electron microscope，SEM）形貌观察：将经100 µg/mL T-CDs处理的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌液在5000 rpm/min下离心5 min，除去上清液，用4 %多聚甲醛重悬，4 ℃固定过夜。随后，将固定的细菌离心浓缩后滴至硅片上自然风干，并使用不同浓度梯度的乙醇（20 %、40 %、50 %、60 %、80 %、90 %、95 %、100 %）进行脱水处理。将脱水完毕的样品自然风干，使用离子溅射仪对样品硅片进行喷金处理，再用SEM对细菌形貌进行表征。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.8　T-CDs的细胞毒性表征" } ], "level": "2", "id": "s2h" } }, { "name": "p", "data": [ { "name": "text", "data": "将NIH3T3接种于96孔板（每孔约1 × 10" }, { "name": "sup", "data": [ { "name": "text", "data": "4" } ] }, { "name": "text", "data": "个细胞）中，加入100 µL含10% 胎牛血清和1%青霉素-链霉素溶液的DMEM培养基。在37 ℃、5% CO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "培养箱中培养24 h后，加入不同浓度T-CDs并继续培养24 h。随后，除去含T-CDs培养液，加入100 μL溶解有Alamar Blue 的 DMEM 培养液，继续孵育6 h。使用多功能微孔板检测仪测定590 nm处的荧光强度（" }, { "name": "italic", "data": [ { "name": "text", "data": "λ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "ex" } ] }, { "name": "text", "data": "=530 nm），并计算 NIH3T3 细胞存活率。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "将NIH3T3接种于共聚焦皿（每孔约1 × 10" }, { "name": "sup", "data": [ { "name": "text", "data": "6" } ] }, { "name": "text", "data": "个细胞）中，加入1 mL含10% 胎牛血清和1%青霉素-链霉素溶液的DMEM培养基。在37 ℃、5% CO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "培养箱中培养24 h后，加入不同浓度T-CDs并继续培养24 h。随后，除去含T-CDs培养液，加入1 mL溶解有钙黄绿素乙酰甲酯（Calcein-AM）/PI的 DMEM 培养液，继续孵育0.5 h。使用多功能微孔板检测仪测定590 nm处的荧光强度（" }, { "name": "italic", "data": [ { "name": "text", "data": "λ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "ex" } ] }, { "name": "text", "data": "=530 nm），并采用智能全自动活细胞荧光显微成像系统观察细胞状态。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.9　T-CDs的活体抗菌性能表征" } ], "level": "2", "id": "s2i" } }, { "name": "p", "data": [ { "name": "text", "data": "选用6-8周、体重为25 ± 5 g的雄性昆明小鼠，异氟醚气体麻醉后，对其背部进行脱毛处理。随后，将直径为8 mm的实心铜棒置于 90 °C水浴中加热10分钟，取出后迅速放置于小鼠背部并施加一定压力保持10 s，以形成皮肤烫伤伤口。在构建小鼠烫伤伤口后，在创面中滴加" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "溶液（2.5 x 10" }, { "name": "sup", "data": [ { "name": "text", "data": "6" } ] }, { "name": "text", "data": " CFU/mL, 10 μL）。细菌接种6 h后，在创面涂抹上T-CDs溶液，并在4 h后利用660 nm激光（100 mW/cm" }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "）光照20 min。以仅T-CDs处理伤口为对照组。每2天称重一次并更换一次敷料，治疗和观察周期共14天。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "在创面愈合过程中，观察实验组和对照组的创面，每隔两天拍照记录，对伤口面积进行量化统计处理。第5天时，评估" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "在创面中的生存情况。收集皮肤伤口组织样本，匀浆后用PBS稀释至1 mL。然后，利用移液枪以10倍的浓度梯度进行梯度稀释，并涂板和计数。此外，采集创面组织，用4%多聚甲醛固定，进行H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E染色和炎症因子染色（白介素-1β（Interleukin-1β，IL-1β）、肿瘤坏死因子（Tumor necrosis factor，TNF-α））。在术后第15天收集器官组织样本进行苏木精/伊红染色实验（Hematoxylin-eosin staining，H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E）染色。" } ] } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3　结果与讨论" } ], "level": "1", "id": "s3" } }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.1　T-CDs的结构和性能" } ], "level": "2", "id": "s3a" } }, { "name": "p", "data": [ { "name": "text", "data": "首先，通过透射电子显微镜（Transmission electron microscopy，TEM）观察了T-CDs的形貌。如" }, { "name": "xref", "data": { "text": "图1", "type": "fig", "rid": "F1", "data": [ { "name": "text", "data": "图1" } ] } }, { "name": "text", "data": "a所示，T-CDs为分散良好的类球形纳米点，平均尺寸约为2.6 nm。高分辨率TEM（High resolution transmission electron microscopy，HRTEM）显示出T-CDs晶格间距为0.21 nm，证明其石墨化结构的存在" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "15", "type": "bibr", "rid": "R15", "data": [ { "name": "text", "data": "15" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。此外，利用FTIR光谱分析了T-CDs的化学组成。如" }, { "name": "xref", "data": { "text": "图1", "type": "fig", "rid": "F1", "data": [ { "name": "text", "data": "图1" } ] } }, { "name": "text", "data": "c所示，3000 cm" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "附近存在明显的特征峰，这归因于O-H拉伸振动，表明T-CDs表面含有-OH官能团；1692 cm" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "处的峰值分别对应于共轭碳核中的C=C的拉伸振动；C-N和C-O的拉伸振动分别位于约1455 cm" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "和1031 cm" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "处" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "16", "type": "bibr", "rid": "R16", "data": [ { "name": "text", "data": "16" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。通过XPS进一步分析了T-CDs的化学组成。如" }, { "name": "xref", "data": { "text": "图1", "type": "fig", "rid": "F1", "data": [ { "name": "text", "data": "图1" } ] } }, { "name": "text", "data": "d所示，XPS全谱在281.3 eV和529.5 eV处显示两个典型峰，表明T-CDs主要由C（78.95 %）和O（21.05 %）组成" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "17", "type": "bibr", "rid": "R17", "data": [ { "name": "text", "data": "17" } ] } }, { "name": "text", "data": ", " }, { "name": "xref", "data": { "text": "18", "type": "bibr", "rid": "R18", "data": [ { "name": "text", "data": "18" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。如" }, { "name": "xref", "data": { "text": "图1", "type": "fig", "rid": "F1", "data": [ { "name": "text", "data": "图1" } ] } }, { "name": "text", "data": "e和f所示，高分辨率C1s光谱显示了三种不同类型的C：C=C、C=O和C-O。此外，O1s光谱出现了C=O和C-O两个峰。XPS结果与上述FTIR结果相吻合。推测T-CDs由C=C内核组成；表面包含羟基、醚键、醛基等官能团，可能来自生物质中的糖类和木质素。" } ] }, { "name": "fig", "data": { "id": "F1", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图1" } ], "title": [ { "name": "text", "data": "（a）T-CDs的TEM图（插图：T-CDs的粒径分布图）。（b）T-CDs的HRTEM图。（c）T-CDs的FTIR光谱。（d）T-CDs的XPS能谱。（e）T-CDs的高分辨率C1s能谱。（f）T-CDs的高分辨率O1s能谱。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 1" } ], "title": [ { "name": "text", "data": "（a） TEM image of T-CDs. （b） HRTEM diagram of T-CDs. （c） FTIR spectra of T-CDs. （d） XPS spectra of T-CDs. （e） High resolution C1s spectrum of T-CDs. （f） High resolution O1s spectrum of T-CDs." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196879&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196881&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196880&type=", "width": "162.79998779", "height": "97.82969666", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "随后，检测了T-CDs的紫外-可见吸收光谱（" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "a）。T-CDs在415 nm和667 nm处显示出两个强的吸收峰，其归属于叶绿素b的特征吸收峰" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "19", "type": "bibr", "rid": "R19", "data": [ { "name": "text", "data": "19" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "，表明T-CDs中存在叶绿素芳香残基。研究表明，叶绿素分子中含有卟吩结构，可被用作光敏剂。因此，我们以DPBF作为ROS探针，考察了T-CDs在660 nm激光激发下的ROS产生能力。如" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "b所示，随着光照时间延长，DPBF的吸收显著降低，证明DPBF在660 nm激光照射下可以被T-CDs产生的ROS快速氧化降解" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "20", "type": "bibr", "rid": "R20", "data": [ { "name": "text", "data": "20" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "21", "type": "bibr", "rid": "R21", "data": [ { "name": "text", "data": "21" } ] } } ], "rid": [ "R20", "R21" ], "text": "20-21", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。" } ] }, { "name": "fig", "data": { "id": "F2", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图2" } ], "title": [ { "name": "text", "data": "（a）T-CDs和叶绿素的紫外-可见吸收光谱。（b）DPBF的降解曲线。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 2" } ], "title": [ { "name": "text", "data": "（a） Ultraviolet-visible absorption spectra of T-CDs. （b） Degradation curve of DPBF." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41196882&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197004&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197003&type=", "width": "122.88546753", "height": "50.78434753", "fontsize": "" } ] } } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.2　T-CDs的体外抗菌性能" } ], "level": "2", "id": "s3b" } }, { "name": "p", "data": [ { "name": "text", "data": "我们选择了革兰氏阳性耐药菌" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "作为代表性耐药菌菌样，通过标准平板计数法评估T-CDs在660 nm激光光照下灭杀耐药菌的能力，并对处理后的细菌菌落数进行了统计与分析。如" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "a所示，在无光照的情况下，" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "经不同浓度的T-CDs处理后，仍均存活良好，表明T-CDs本身对" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "没有明显毒性。当用100 mW/cm" }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "的660 nm激光照射10 min后，" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "成活率随着T-CDs的浓度逐渐下降，且当T-CDs的浓度为100 µg/mL时，细菌可被100%消除。此外，通过琼脂板光学图像（" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "b）同样可以观察到，T-CDs处理的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "的群落数与对照组无明显差异，而经过光照后，" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "的群落数明显减少，与统计结果一致。" } ] }, { "name": "fig", "data": { "id": "F3", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图3" } ], "title": [ { "name": "text", "data": "（a）T-CDs处理后的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "成活率。（**" }, { "name": "italic", "data": [ { "name": "text", "data": "P" } ] }, { "name": "text", "data": "<" }, { "name": "text", "data": "0.01，***" }, { "name": "italic", "data": [ { "name": "text", "data": "P" } ] }, { "name": "text", "data": "<" }, { "name": "text", "data": "0.001）（b）" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "在琼脂板上存活菌落的光学图像。（c）" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "的活/死染色荧光成像。（d）经T-CDs和660 nm激光处理后的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "细菌的SEM图。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig.3" } ], "title": [ { "name": "text", "data": "（a） Survival rate of " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": " of T-CDs. （b） Optical images of viable colonies of " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": " of T-CDs on AGAR plates. （c） LIVE/DEAD staining fluorescence imaging of " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": " treated by T-CDs. （d） SEM image of " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": " treated by T-CDs." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197005&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197007&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197006&type=", "width": "138.53599548", "height": "102.79900360", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "为了更为直观地证明T-CDs的PDT抗菌效果，我们采用活/死染色法和SEM观察了T-CDs处理后" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "在有无光照情况下的活力和形态。如" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "c所示，在无光照的情况下，T-CDs处理后的细菌主要发射绿色荧光（活菌），表明T-CDs本身对细菌没有影响。而光照10 min后，大量的红色荧光（死菌）信号被观察到。这些结果表明了T-CDs在660 nm激光照射下能够有效诱导细菌死亡。此外，我们还通过SEM证明了T-CDs对细菌的PDT杀伤机制。如" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "d所示，未经光照处理的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "结构形态保持完整、边界清晰。但660 nm激光照射后，" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "的形态发生明显变化，表现为细菌细胞壁的收缩和融合。该结果与已报道的PDT抗菌机制一致。" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "22", "type": "bibr", "rid": "R22", "data": [ { "name": "text", "data": "22" } ] } }, { "name": "text", "data": ", " }, { "name": "xref", "data": { "text": "23", "type": "bibr", "rid": "R23", "data": [ { "name": "text", "data": "23" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": " 具体机制为：光照后T-CDs产生的ROS对不饱和脂类、多聚糖等生物大分子产生毒性，从而造成对细菌细胞壁的破坏，导致其形态明显变化。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.3　T-CDs的细胞生物毒性" } ], "level": "2", "id": "s3c" } }, { "name": "p", "data": [ { "name": "text", "data": "我们利用NIH3T3评价了T-CDs的细胞毒性。如" }, { "name": "xref", "data": { "text": "图4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图4" } ] } }, { "name": "text", "data": "a所示，在T-CDs存在的情况下，细胞能够持续生长，且细胞活力与对照组无显著差异。此外，活/死染色法结果显示，与T-CDs共培养的NIH3T3细胞仍具有正常的梭形形貌，说明细胞状态良好，并保持正常增殖（" }, { "name": "xref", "data": { "text": "图4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图4" } ] } }, { "name": "text", "data": "b）。实验结果清楚地表明，T-CDs对NIH3T3细胞没有毒害作用，不会影响其增殖。因此，制备的T-CDs具有良好的生物相容性，确保了后续活体抗感染应用的生物安全性。" } ] }, { "name": "fig", "data": { "id": "F4", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图4" } ], "title": [ { "name": "text", "data": "（a）经T-CDs处理后的NIH3T3细胞的成活率。（b）经T-CDs处理后的NIH3T3细胞的活/死染色荧光成像。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 4" } ], "title": [ { "name": "text", "data": "（a） Survival rate of NIH3T3 cells treated with T-CDs. （b） LIVE/DEAD staining fluorescence images of NIH3T3 cells treated with T-CDs." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197008&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197010&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197009&type=", "width": "133.25877380", "height": "43.97711563", "fontsize": "" } ] } } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.4　T-CDs的活体抗菌性能" } ], "level": "2", "id": "s3d" } }, { "name": "p", "data": [ { "name": "text", "data": "为了进一步验证T-CDs的PDT抗菌性能，我们建立了" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "感染的小鼠皮肤伤口模型。如" }, { "name": "xref", "data": { "text": "图5", "type": "fig", "rid": "F5", "data": [ { "name": "text", "data": "图5" } ] } }, { "name": "text", "data": "a和b所示，仅T-CDs处理的创面组织中可以观察到大量的" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌落（2.32 × 10" }, { "name": "sup", "data": [ { "name": "text", "data": "7" } ] }, { "name": "text", "data": " CFU g" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "）。当光照10 min后，T-CDs处理伤口组织的活菌数量明显降低，存活菌落数仅为1.61 × 10" }, { "name": "sup", "data": [ { "name": "text", "data": "6" } ] }, { "name": "text", "data": " CFU g" }, { "name": "sup", "data": [ { "name": "text", "data": "-1" } ] }, { "name": "text", "data": "，减少了93%，表明T-CDs在光照作用下能够有效抑制伤口感染。" } ] }, { "name": "fig", "data": { "id": "F5", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图5" } ], "title": [ { "name": "text", "data": "T-CDs激光照射处理后，第5天时伤口处" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "菌落涂板（a），菌落计数（b），伤口组织H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E染色（c）和伤口组织中IL-1β和TNF-α免疫组化（d）。（***" }, { "name": "italic", "data": [ { "name": "text", "data": "P" } ] }, { "name": "text", "data": "<" }, { "name": "text", "data": "0.001）" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 5" } ], "title": [ { "name": "text", "data": "On the day 5th after T-CDs laser irradiation， " }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": " colony coating （a）， colony count （b）， wound tissue H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E staining （c） and immunohistochemistry of IL-1β and TNF-α （d）." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197011&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197013&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197012&type=", "width": "122.74900055", "height": "79.19899750", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "此外，我们还利用H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E染色法观察了第5天时的创面部位的炎症状态。如" }, { "name": "xref", "data": { "text": "图5", "type": "fig", "rid": "F5", "data": [ { "name": "text", "data": "图5" } ] } }, { "name": "text", "data": "c所示，T-CDs处理的伤口组织能够观察到明显的紫色中性粒细胞。与对照组相比，治疗组的中性粒细胞明显减少。此外，我们还检测了创面的炎症因子TNF-α和IL-1β。结果显示，对照组的TNF-α和IL-1β的表达水平明显高于治疗组，说明细菌的减少能够缓解伤口的炎症响应（" }, { "name": "xref", "data": { "text": "图5", "type": "fig", "rid": "F5", "data": [ { "name": "text", "data": "图5" } ] } }, { "name": "text", "data": "d）。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "我们记录了治疗后感染伤口的愈合过程。如" }, { "name": "xref", "data": { "text": "图6", "type": "fig", "rid": "F6", "data": [ { "name": "text", "data": "图6" } ] } }, { "name": "text", "data": "a所示，仅经T-CDs处理的伤口在第一周内可以观察到黄色脓液，表明" }, { "name": "italic", "data": [ { "name": "text", "data": "MRSA" } ] }, { "name": "text", "data": "感染会引起伤口呈现严重的炎症状态。相比之下，PDT治疗组的伤口环境较干净，无明显组织渗出物，伤口愈合速度快。另外，我们统计了治疗后两周内的伤口愈合率（" }, { "name": "xref", "data": { "text": "图6", "type": "fig", "rid": "F6", "data": [ { "name": "text", "data": "图6" } ] } }, { "name": "text", "data": "b）。治疗后的第3天，治疗组的伤口愈合率为31%，显著高于对照组的12%。第7天时，治疗组创面愈合率上升至43%，明显快于对照组的15%。" } ] }, { "name": "fig", "data": { "id": "F6", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图6" } ], "title": [ { "name": "text", "data": "T-CD处理后，感染伤口在不同时间的光学照片（a）和伤口闭合率（b）。（**" }, { "name": "italic", "data": [ { "name": "text", "data": "P" } ] }, { "name": "text", "data": "<" }, { "name": "text", "data": "0.01，***" }, { "name": "italic", "data": [ { "name": "text", "data": "P" } ] }, { "name": "text", "data": "<" }, { "name": "text", "data": "0.001）" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 6" } ], "title": [ { "name": "text", "data": "Optical photographs of infected wounds at different times after T-CD treatment （a） and wound closure rate （b）." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197014&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197016&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197015&type=", "width": "133.56024170", "height": "46.36933517", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "治疗2周后，光照治疗组伤口几乎完全闭合（98%），而对照组伤口愈合情况仍表现出一定程度的滞后。伤口发生细菌感染会带来一系列不良症状，如过度炎症，大大阻碍了伤口的愈合过程。我们所制备的T-CDs能在光照作用下有效产生ROS，用于杀灭细菌，避免了感染引起的过度炎症，能够促进耐药细菌感染伤口的愈合。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "在治疗过程中，我们监测了小鼠的体重变化。如" }, { "name": "xref", "data": { "text": "图7", "type": "fig", "rid": "F7", "data": [ { "name": "text", "data": "图7" } ] } }, { "name": "text", "data": "a所示，与对照组比较， 治疗组小鼠的体重没有出现明显的变化，表明T-CDs在小鼠体内展现出良好的生物相容性。此外，我们在对治疗后第15天小鼠的主要器官（心、肝、脾、肺、肾）进行了H" }, { "name": "text", "data": "&" }, { "name": "text", "data": "E染色组织学分析（" }, { "name": "xref", "data": { "text": "图7", "type": "fig", "rid": "F7", "data": [ { "name": "text", "data": "图7" } ] } }, { "name": "text", "data": "b），均未观察到明显的炎症和损伤，证明了T-CDs较低的生物毒性。因此，我们所制备的T-CDs表现出具有良好的生物相容性，确保了其抗感染应用的生物安全性。" } ] }, { "name": "fig", "data": { "id": "F7", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图7" } ], "title": [ { "name": "text", "data": "T-CDs处理后，感染小鼠的体重变化（a）和第15天时主要器官的H&E染色图像（b）。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 7" } ], "title": [ { "name": "text", "data": "Body weight changes of infected mice treated by T-CDs （a） and H&E staining images of major organs on the day 15th （b）." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197017&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197019&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=41197018&type=", "width": "96.10729218", "height": "80.76853180", "fontsize": "" } ] } } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "4　结　　论" } ], "level": "1", "id": "s4" } }, { "name": "p", "data": [ { "name": "text", "data": "综上所述，本研究采用绿茶作为碳源，通过溶剂热法，制备了T-CDs。通过对其形貌和结构表征分析可知，T-CDs为类球形，尺寸小于10 nm，表面带有羟基、醚键、醛基等基团。而由于含有叶绿素残基，T-CDs被证明在660 nm激光光照下能够有效产生ROS。体外生物学性能研究表明，T-CDs具有优异的细胞相容性。同时，在660 nm激光光照下，T-CDs能够通过产生ROS实现对革兰氏阳性耐药菌的100%高效杀灭。此外，T-CDs同样可以通过PDT有效灭杀小鼠感染模型伤口部位的耐药菌，同时下调伤口部位炎症因子TNF-α和IL-1β的水平，展现出有效的促伤口愈合效果。因此，该T-CDs提供了一种简单、高效、低毒和非耐药的PDT抗菌策略，对临床耐药菌感染治疗具有潜在的应用价值。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "本文（补充文件及）专家审稿意见和作者回复内容下载地址：" }, { "name": "extlink", "data": { "text": [ { "name": "text", "data": "http://cjl.lightpublishing.cn/thesisDetails#10.37188/CJL.20220" } ], "href": "http://cjl.lightpublishing.cn/thesisDetails#10.37188/CJL.20220" } }, { "name": "text", "data": "***（20220***为文章稿号）." } ] } ] } ], "footnote": [], "reflist": { "title": [ { "name": "text", "data": "参考文献" } ], "data": [ { "id": "R1", "label": "1", "citation": [ { "lang": "en", 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