Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, 710062, Shaanxi, China.
College of Chemistry and Materials Science, Jiangsu Key Laboratory of Biofunctional Materials, Nanjing Normal University, Wenyuan Road No.1, Nanjing, 210046, China.
Photodiagnosis Photodyn Ther. 2018 Mar;21:316-326. doi: 10.1016/j.pdpdt.2018.01.003. Epub 2018 Jan 4.
Bacterial infection is a common clinical problem. Community-associated Staphylococcus aureus (S. aureus) infections can cause extensive tissue damage and necrosis. Photodynamic antimicrobial chemotherapy (PACT) has recently attracted attention as a feasible bacterial therapy. Octa-cationic zinc phthalocyanines are newly identified photosensitizers derived from phthalocyanines bearing 1, 2-ethanediamine groups and quaternized derivatives with different numbers of positive charges (ZnPc, n = 4 or 8). Here we report the antimicrobial effects of ZnPc-mediated PACT on planktonic and biofilm cultures of S. aureus.
ZnPc uptake was detected by photometry after alkaline lysis. Dark-toxicity and light-mediated antimicrobial effects of the drug was determined by the plate count method. The production of intracellular reactive oxygen species (ROS) was detected by flow cytometry. SYTO 9 and propidium iodide (PI) were used to detect the bacterial cell membrane permeability. DNA damage after ZnPc-PACT was analyzed by flow cytometry and PI staining. The destruction of biofilm was evaluated by scanning electron microscope (SEM).
The study of uptake showed that the relative fluorescence intensity of ZnPc in S. aureus peaked at 15 min. Generation of reactive oxygen species (ROS) by ZnPc was enhanced in PACT treatment groups. SYTO 9 and PI staining indicated that cell membrane was damaged. Flow cytometry and PI staining revealed DNA damage. Biofilms were damaged in PACT treatment groups.
Our results suggest that light-activated ZnPc can efficiently inhibit planktonic and biofilm cultures of S. aureus.
细菌感染是一种常见的临床问题。社区相关性金黄色葡萄球菌（S. aureus）感染可导致广泛的组织损伤和坏死。光动力抗菌化疗（PACT）最近作为一种可行的细菌治疗方法引起了关注。八正锌酞菁是新发现的光敏剂，源自带有 1,2-乙二胺基团的酞菁和具有不同正电荷数的季铵化衍生物（ZnPc，n=4 或 8）。在这里，我们报告了 ZnPc 介导的 PACT 对金黄色葡萄球菌浮游和生物膜培养物的抗菌作用。
通过碱性裂解后测光检测 ZnPc 的摄取。通过平板计数法测定药物的暗毒性和光介导的抗菌作用。通过流式细胞术检测细胞内活性氧（ROS）的产生。SYTO 9 和碘化丙啶（PI）用于检测细菌细胞膜通透性。通过流式细胞术和 PI 染色分析 ZnPc-PACT 后的 DNA 损伤。通过扫描电子显微镜（SEM）评估生物膜的破坏。
摄取研究表明，ZnPc 在金黄色葡萄球菌中的相对荧光强度在 15 分钟时达到峰值。ZnPc 在 PACT 处理组中产生的活性氧（ROS）增强。SYTO 9 和 PI 染色表明细胞膜受损。流式细胞术和 PI 染色显示 DNA 损伤。生物膜在 PACT 处理组中受损。
我们的结果表明，光激活的 ZnPc 可以有效地抑制浮游和生物膜培养的金黄色葡萄球菌。
