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首页> 外文会议>Plasma for bio-decontamination, medicine and food security >Inactivation of Candida Strains in Planktonic and Biofilm Forms Using a Direct Current, Atmospheric-Pressure Cold Plasma Micro-Jet
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Inactivation of Candida Strains in Planktonic and Biofilm Forms Using a Direct Current, Atmospheric-Pressure Cold Plasma Micro-Jet

机译：使用直流，常压冷等离子体微射流灭活浮游生物和生物膜形式的念珠菌

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摘要

A direct-current, atmospheric-pressure, He/O_2 (2%) cold plasma microjet is applied to Candida species (C. glabrata, C. albicans and C. krusei). Effective inactivation is achieved both in air and in water within 5 min of plasma treatment. Same plasma treatment also successfully inactivated Candida biofilms on Petri dish. The inactivation was verified by cell viability test (XTT assay). Severe deformation of Candida biofilms after the plasma treatment was observed through scanning electron microscope (SEM). Optical emission spectroscopy shows strong atomic oxygen emission at 777 nm. Hydroxyl radical (·OH), superoxide anion radical (·O_2-) and singlet molecular oxygen (~1O_2) are detected by electron spin resonance (ESR) spectroscopy. The sessile minimal inhibitory concentrations (SMICs) of fluconazole, amphotericin B, and caspofungin against the Candida spp. biofilms were decreased to 2-6 fold dilutions in plasma microjet treated group in comparison with the controls. This novel approach may become a new tool for the treatment of clinical dermatosis Candidiasis, commonly referred to as yeast infection, is caused by Candida species (spp) [1, 2]. It typically encompasses mucocutaneous disorders such as oral candidiasis, vaginal and vulvovaginal candidiasis, as well as systemic infections and potentially life-threatening diseases (often referred to as candidemia or fungemia). Although Candida species are the microorganism exhibiting planktonic unicellular form, filamentous growth or complex multicellular structure is observed mainly in the infected tissues [3]. Candida biofilms are structured microbial communities and are much more resistant than the free-living planktonic cells. The fact that they can attach to surfaces or encase a matrix of exopolymeric materials [4, 5], has raised serious problems for various implanted medical devices such as vascular and urinary catheters, joint prostheses, cardiac valves, artificial vascular bypass devices, or more commonly used contact lens and dentures [6, 7], The complex structure of the biofilms makes them resistant to both host defense and commonly used antifungal drugs [5, 8, 9]. The cells of Candida species can constantly split from the structured microbial communities, spread and further cause antifungal treatment failure, devices failure or persistent infections [10]. Therapeutic methods for Candida bionlm infections are very limited. On the other hand, although Candida albicans infection is most commonly seen, the incidence of Candida non-albicans is increasing in recent years due to (among other reasons) the frequent prophylactic use of antifungal chemicals. Several non-albican species, such as Candida glabrata (C. glabrata) and Candida krusei (C. krusei), may be resistant to azole antifungal therapy. In recent years, atmospheric pressure non-thermal plasmas (NTP) have drawn much attention for their potential applications in medicine and biomedicine. They have been reported to be effective in bacterial inactivation [11], blood coagulation [12], tooth whitening [13], tumor treatment [14] and wound healing [15]. Early work on the effectiveness of NPT on inactivating bacterial biofilms was performed by a few groups [16, 17]. Compared with traditional therapeutic methods, non-thermal plasmas as a physical method could be more economic and effective [18]. In addition, the gaseous form of plasmas provide the possibility to treat inhomoge-neous surfaces, cavities and fissures down to the micrometer scale, and allow the combination with minimally invasive surgery or with antimicrobial chemotherapy. Several papers have reported using non-thermal plasmas against Candida albicans [19, 20], but few have analyze their effect on the fluconazole-resistant Candida strains. In this work, a direct current atmospheric pressure non-thermal plasma microjet (PMJ), with helium and oxygen gas mixture as working gas, is applied to several strains of Candida species including fluconazole-resistant C. albicans, C. glabrata and C. krusei. Further investigation has been done on the fungicidal capability of PMJ on Candida biofilms, and its effect on antifungal susceptibility of candidal biofilms to common antifungal drug. Cell viability test (XTT assay) and scanning electron microscope (SEM) were used after the plasma treatment to verify the inactivation. Electron Spin Resonance (ESR) spectroscopy and optical emission spectroscopy (OES) are employed to evaluate the reactive species generated.
机译：将直流，大气压，He / O_2（2％）冷等离子体微喷射器应用于念珠菌（C. glabrata，C。albicans和C. krusei）。在等离子处理后的5分钟内，可以在空气和水中实现有效的灭活。相同的血浆处理也成功地灭活了培养皿上的念珠菌生物膜。通过细胞存活力测试（XTT测定）验证了失活。通过扫描电子显微镜（SEM）观察到等离子体处理后念珠菌生物膜的严重变形。发射光谱显示在777 nm处有强原子氧发射。通过电子自旋共振（ESR）光谱检测羟基自由基（·OH），超氧阴离子自由基（·O_2-）和单线态分子氧（〜1O_2）。氟康唑，两性霉素B和卡泊芬净对念珠菌的无梗最低抑菌浓度（SMIC）。与对照组相比，血浆微射流治疗组的生物膜稀释度降低至2-6倍。这种新颖的方法可能成为治疗念珠菌病（spp）引起的临床皮肤病念珠菌病（通常称为酵母菌感染）的一种新工具[1、2]。它通常包括粘膜皮肤疾病，例如口腔念珠菌病，阴道和外阴念珠菌病，以及全身感染和可能威胁生命的疾病（通常称为念珠菌血症或真菌病）。尽管念珠菌是表现出浮游单细胞形式的微生物，但主要在被感染的组织中观察到丝状生长或复杂的多细胞结构[3]。念珠菌生物膜是结构化的微生物群落，比自由活动的浮游细胞更具抵抗力。它们可以附着在表面或包裹着外聚合材料的基质[4、5]，这一事实对各种植入式医疗设备（例如血管导管和导尿管，关节假体，心脏瓣膜，人工血管旁路设备等）提出了严重的问题。常用的隐形眼镜和假牙[6，7]，生物膜的复杂结构使其对宿主防御和常用的抗真菌药物都有抵抗力[5，8，9]。念珠菌物种的细胞可以不断地从结构化微生物群落中分裂出来，扩散并进一步引起抗真菌治疗失败，器械失败或持续感染[10]。念珠菌生化细菌感染的治疗方法非常有限。另一方面，尽管最常见白色念珠菌感染，但由于（除其他原因外）频繁地预防性使用抗真菌化学药品，近年来白色念珠菌的发病率正在增加。几种非白色物种，例如光滑念珠菌（C. glabrata）和克鲁斯念珠菌（C. krusei），可能对唑类抗真菌药耐药。近年来，大气压非热等离子体（NTP）在医学和生物医学中的潜在应用备受关注。据报道，它们在细菌灭活[11]，凝血[12]，牙齿增白[13]，肿瘤治疗[14]和伤口愈合[15]方面有效。 NPT对灭活细菌生物膜的有效性的早期工作由几个小组完成[16，17]。与传统的治疗方法相比，非热等离子体作为一种物理方法可能更经济，更有效[18]。另外，等离子体的气态形式提供了治疗低至微米级的异质表面，空腔和裂缝的可能性，并允许与微创手术或抗菌化学疗法相结合。已有几篇论文报道了使用非热等离子体对白色念珠菌的影响[19，20]，但很少有人分析了它们对耐氟康唑的念珠菌菌株的影响。在这项工作中，将氦气和氧气混合物作为工作气体的直流常压非热等离子体微型喷射器（PMJ）应用于几株念珠菌菌株，包括耐氟康唑的白色念珠菌，光滑念珠菌和C.。克鲁赛关于PMJ对念珠菌生物膜的杀菌能力及其对念珠菌生物膜对常用抗真菌药的抗真菌敏感性的影响，已经进行了进一步的研究。等离子体处理后，使用细胞活力测试（XTT分析）和扫描电子显微镜（SEM）来验证失活。电子自旋共振（ESR）光谱和光发射光谱（OES）用于评估所产生的反应物种。

著录项

  • 来源
  • 会议地点 Demanovska Dolina(SK)
  • 作者

    Wei-Dong Zhu; Peng Sun; Yi Sun; Shuang Yu; Haiyan Wu; Wei Liu; Jue Zhang; Jing Fang;

  • 作者单位

    Department of Applied Science and Technology, Saint Peter's College, 2641 Kennedy Boulevard, Jersey City, NJ 07306, USA;

    Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China;

    Department of Dermatology and Venereology, Peking University 1st Hospital, Beijing, China;

    Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China;

    Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China;

    Department of Dermatology and Venereology, Peking University 1st Hospital, Beijing, China;

    Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China;

    Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物工程学（生物技术）;
  • 关键词

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