新型季铵盐纳米颗粒对金黄色葡萄球菌和铜绿假单胞菌生物膜抑制作用的研究

A novel nanoparticle in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms

目的:探讨低浓度氯化十六烷基吡啶(CPC)季铵盐纳米颗粒在体外对金黄色葡萄球菌和铜绿假单胞菌生物膜的抑制作用。方法:建立金黄色葡萄球菌ATCC25923和铜绿假单胞菌ATCC15692体外生物膜模型。制备低浓度(0.010%、0.025%和0.050%)CPC-季铵盐纳米颗粒和CPC胶体溶液。用alamarBlue法分别在CPC作用5 min和2 h后检测金黄色葡萄球菌和铜绿假单胞菌游离及其生物膜的活性。应用激光扫描共聚焦显微镜(CLSM)观察CPC-季铵盐纳米颗粒与金黄色葡萄球菌和铜绿假单胞菌生物膜之间的相互作用。结果:低浓度(0.010%、0.025%和0.050%)CPC-季铵盐纳米颗粒和CPC胶体溶液对游离金黄色葡萄球菌和铜绿假单胞菌均有显著的抗菌作用(P<0.05),对金黄色葡萄球菌和铜绿假单胞菌生物膜分别作用5 min和2 h后同样具有显著的抗菌效果(P<0.05)。在CLSM研究中,金黄色葡萄球菌生物膜的面积明显减小,铜绿假单胞菌生物膜中的死菌明显增加。结论:低浓度的CPC-季铵盐纳米颗粒即可对金黄色葡萄球菌和铜绿假单胞菌游离菌及生物膜产生显著的杀伤作用,有望应用于慢性鼻窦炎的治疗。

Objective: To investigate CPC-nanoparticles of low concentrations in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms in vitro. Method: We established specific biofilms of staphylococcus aureus ATCC 25923 and pseudomonas aeruginosa ATCC 15692, and prepared CPC-nanoparticles and CPC micelle solutions of low concentrations(0.010%, 0.025% and 0.050%). AlamarBlue was used to test the viability of both planktonic staphylococcus aureus and pseudomonas aeruginosa and their biofilms after treatment for 5 minutes and 2 hours respectively in the bactericidal efficacy study.The interaction between CPC-nanoparticles and staphylococcus aureus and pseudomonas aeruginosa biofilms was observed by confocal laser scanning microscope(CLSM). Result: 0.010%, 0.025% and 0.050% CPC-nanoparticles and CPC micelle solutions had significant bactericidal effect on planktonic staphylococcus aureus and pseudomonas aeruginosa after five-minute exposure(P<0.05), and staphylococcus aureus and pseudomonas aeruginosa biofilms after both five-minute and two-hour treatments(P<0.05). In CLSM study, the size of staphylococcus aureus biofilms decreased, while dead bacteria of pseudomonas aeruginosa biofilms increased after two-hour treatment. Conclusion: CPC-nanoparticles had significant bactericidal effects on staphylococcus aureus and pseudomonas aeruginosa biofilms, which could be used in treatment of CRS.