Photodynamic inactivation of Candida albicans by BAM-SiPc.

• Other Authors: So, Cheung Wai.
• Format: Others
• Language: English; Chinese
• Published: 2008
• Subjects: Candida albicans; Photochemotherapy; Photosensitizing Agents
• Online Access: http://library.cuhk.edu.hk/record=b5896832; http://repository.lib.cuhk.edu.hk/en/item/cuhk-326528

So, Cheung Wai. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. === Includes bibliographical references (leaves 106-117). === Abstracts in English and Chinese. === Acknowledgements --- p.i === Abstract --- p.iii === 摘要 --- p.v === List of Abbreviations --- p.vii === List of Figures --- p.viii === List of Tables --- p.x === Chapter Chapter 1 --- Introduction === Chapter 1.1 --- Candida albicans and candidiasis === Chapter 1.1.1 --- Historical background --- p.1 === Chapter 1.1.2 --- C. albicans infections --- p.2 === Chapter 1.1.3 --- Current challenges in the treatment of C. albicans --- p.3 === Chapter 1.2 --- Photodynamic therapy --- p.11 === Chapter 1.2.1 --- Historical aspects and development --- p.11 === Chapter 1.2.2 --- Basic principle of photodynamic therapy --- p.13 === Chapter 1.2.3 --- Light applicator --- p.16 === Chapter 1.2.4 --- Generations of photosensitizer --- p.17 === Chapter 1.2.5 --- Characteristics of phthalocyanines --- p.20 === Chapter 1.2.6 --- Photodynamic antimicrobial chemotherapy (PACT) --- p.22 === Chapter 1.3 --- Aim of present study --- p.24 === Chapter Chapter 2 --- Materials and Methods === Chapter 2.1 --- Synthesis of BAM-SiPc --- p.27 === Chapter 2.2 --- Preparation of BAM-SiPc solution --- p.27 === Chapter 2.3 --- Yeast strains and culture conditions --- p.28 === Chapter 2.4 --- Light source --- p.29 === Chapter 2.5 --- Assays of PDT with planktonic C. albicans === Chapter 2.5.1 --- Photodynamic treatment on planktonic cells --- p.30 === Chapter 2.5.2 --- Clonogenic assay --- p.32 === Chapter 2.5.3 --- Cellular uptake of BAM-SiPc --- p.32 === Chapter 2.5.4 --- Distribution of BAM-SiPc in planktonic cells === Chapter 2.5.4.1 --- Fluorescence microscopic analyses --- p.33 === Chapter 2.5.4.2 --- Confocal laser scanning microscopic (CLSM) analyses --- p.34 === Chapter 2.5.5 --- Determination of ROS level in planktonic cells --- p.34 === Chapter 2.5.6 --- Distribution of ROS in planktonic cells --- p.35 === Chapter 2.5.7 --- Effect of ROS inhibitors --- p.35 === Chapter 2.5.8 --- Membrane integrity assay --- p.36 === Chapter 2.6 --- Assays of PDT with C. albicans biofilm === Chapter 2.6.1 --- Biofilm formation --- p.37 === Chapter 2.6.2 --- Photodynamic treatment on C. albicans biofilm --- p.38 === Chapter 2.6.3 --- Viability assays === Chapter 2.6.3.1 --- XTT reduction assay --- p.38 === Chapter 2.6.3.2 --- Molecular probes staining --- p.40 === Chapter 2.6.4 --- Determination of ROS level in biofilm --- p.41 === Chapter 2.6.5 --- Distribution of BAM-SiPc in biofilm --- p.41 === Chapter 2.6.6 --- Photodynamic treatment on C. albicans from resuspended biofilm --- p.42 === Chapter 2.7 --- Statistical analysis --- p.42 === Chapter Chapter 3 --- Results === Chapter 3.1 --- BAM-SiPc mediated PDT on planktonic C. albicans === Chapter 3.1.1 --- Antifungal effect of BAM-SiPc on C. albicans === Chapter 3.1.1.1 --- PDT activities on different strains of C. albicans --- p.43 === Chapter 3.1.1.2 --- Effect of different densities of cells --- p.47 === Chapter 3.1.1.3 --- Effect of a washing step before illumination --- p.47 === Chapter 3.1.2 --- Optimization of PDT conditions with BAM-SiPc on C. albicans === Chapter 3.1.2.1 --- Time course study --- p.50 === Chapter 3.1.2.2 --- Light dose study --- p.50 === Chapter 3.1.3 --- Uptake of BAM-SiPc --- p.53 === Chapter 3.1.4 --- Distribution of BAM-SiPc in the planktonic cells === Chapter 3.1.4.1 --- Analysis with fluorescence microscopy --- p.56 === Chapter 3.1.4.2 --- Analysis with CLSM --- p.56 === Chapter 3.1.5 --- ROS production upon PDT treatment === Chapter 3.1.5.1 --- ROS level in the planktonic cells --- p.59 === Chapter 3.1.5.2 --- Distribution of ROS production --- p.61 === Chapter 3.1.5.3 --- Effect of different ROS inhibitors on BAM-SiPc's potency --- p.64 === Chapter 3.1.6 --- Membrane integrity --- p.66 === Chapter 3.2 --- BAM-SiPc mediated PDT on C. albicans biofilm === Chapter 3.2.1 --- Establishment of the biofilm model with 192887g --- p.69 === Chapter 3.2.2 --- Photodynamic treatment on 192887g biofilm === Chapter 3.2.2.1 --- Viability assay - XTT assay --- p.72 === Chapter 3.2.2.2 --- Viability assay ´ؤ LIVE/DEAD BacLight Bacterial Viability kit --- p.72 === Chapter 3.2.3 --- ROS level in the biofilm after PDT treatment --- p.75 === Chapter 3.2.4 --- Distribution of BAM-SiPc in the biofilm --- p.75 === Chapter 3.2.5 --- Photodynamic treatment on C. albicans from resuspended biofilm --- p.79 === Chapter Chapter 4 --- Discussion --- p.81 === Chapter 4.1 --- Antifungal effect of BAM-SiPc on the planktonic C. albicans --- p.81 === Chapter 4.2 --- Effects of different conditions on the photodynamic treatment with BAM- SiPc --- p.83 === Chapter 4.3 --- Mechanistic study of the antifungal effect of BAM-SiPc --- p.86 === Chapter 4.3.1 --- Interaction between BAM-SiPc and C. albicans --- p.86 === Chapter 4.3.2 --- ROS as mediator of cell damage --- p.89 === Chapter 4.3.3 --- Analysis of membrane integrity upon photodynamic treatment --- p.91 === Chapter 4.4 --- Establishment of the biofilm model of C. albicans --- p.92 === Chapter 4.5 --- In vitro effect of BAM-SiPc mediated PDT on C. albicans biofilm --- p.95 === Chapter Chapter 5 --- Conclusion and Future Perspectives === Chapter 5.1 --- Conclusion --- p.101 === Chapter 5.2 --- Future perspectives --- p.102 === References --- p.106