Zhu Liguo, Li Changqing, Wang Deqing
Department of Blood Transfusion, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University, Qingdao, China.
Institute of Blood Transfusion, Peking Union Medical College and Chinese Academy of Medical Sciences, Chengdu, China.
Front Microbiol. 2024 Jul 2;15:1404468. doi: 10.3389/fmicb.2024.1404468. eCollection 2024.
Treating bacteremia caused by antibiotic-resistant bacteria is a global concern. Antibacterial photodynamic inactivation is a promising strategy to combat it. However, it's challenging to achieve the inactivation of antibiotic-resistant bacteria in whole blood because of its opacity and complexity. We investigated a riboflavin photodynamic method to effectively inactivate antibiotic-resistant bacteria in whole blood. Four strains of antibiotic-resistant bacteria were isolated, identified, and cultured in this research: methicillin-resistant (MRSA), pan-drug-resistant (PDRAB), ESBLs-producing (EPEC) and pan-drug-resistant (PDRKP). To simulate bacteremia, antibiotic-resistant bacteria was added into whole blood. Whole blood was treated using riboflavin photodynamic method with ultraviolet irradiation (308 nm and 365 nm). The ultraviolet irradiation dose was divided into 18 J/cm, 36 J/cm, and 54 J/cm. Microbial count of antibiotic-resistant bacteria in whole blood was used for evaluating inactivation effectiveness. The roles of red blood cells, lymphocytes, coagulation factors, and platelets in whole blood were assessed. In results, inactivation effectiveness increased as the ultraviolet dose increased from 18 J/cm to 54 J/cm. At the dose of 18 J/cm, inactivation effectiveness of four antibiotic-resistant bacteria were more than 80%, while only 67% of MRSA. The antibacterial effect was enhanced by the combination of riboflavin photodynamic treatment and antibiotic. The red blood cell function was susceptible to ultraviolet dose. At the dose of 18 J/cm, hemolysis rate was less than 0.8% and there was no change in levels of ATP and 2,3-DPG. At the same dose, the proliferation, cell killing, and cytokine secretion activities of lymphocytes decreased 20-70%; Factor V and Factor VIII activities decreased 50%; Fibrinogen and platelet function loss significantly but reparable. Consequently, we speculated that riboflavin photodynamic method with a ultraviolet dose of 18 J/cm was effective in inactivating four antibiotic-resistant bacteria in whole blood while whole blood function was preserved. We also provided a novel extracorporeal circulation phototherapy mode for treating bacteremia caused by antibiotic-resistant bacteria.
治疗由耐抗生素细菌引起的菌血症是一个全球关注的问题。抗菌光动力灭活是对抗菌血症的一种有前景的策略。然而，由于全血的不透明性和复杂性，要实现对全血中耐抗生素细菌的灭活具有挑战性。我们研究了一种核黄素光动力方法，以有效灭活全血中的耐抗生素细菌。本研究分离、鉴定并培养了四株耐抗生素细菌：耐甲氧西林金黄色葡萄球菌（MRSA）、泛耐药鲍曼不动杆菌（PDRAB）、产超广谱β-内酰胺酶大肠埃希菌（EPEC）和泛耐药肺炎克雷伯菌（PDRKP）。为了模拟菌血症，将耐抗生素细菌添加到全血中。使用核黄素光动力方法并结合紫外线照射（308nm和365nm）处理全血。紫外线照射剂量分为18J/cm²、36J/cm²和54J/cm²。通过全血中耐抗生素细菌的微生物计数来评估灭活效果。评估了全血中红细胞、淋巴细胞、凝血因子和血小板的作用。结果显示，随着紫外线剂量从18J/cm²增加到54J/cm²，灭活效果增强。在18J/cm²的剂量下，四种耐抗生素细菌的灭活效果均超过80%，而耐甲氧西林金黄色葡萄球菌仅为67%。核黄素光动力治疗与抗生素联合使用可增强抗菌效果。红细胞功能对紫外线剂量敏感。在18J/cm²的剂量下，溶血率小于0.8%，三磷酸腺苷（ATP）和2,3-二磷酸甘油酸（2,3-DPG）水平无变化。在相同剂量下，淋巴细胞的增殖、细胞杀伤和细胞因子分泌活性降低20%-70%；凝血因子Ⅴ和Ⅷ活性降低50%；纤维蛋白原和血小板功能显著丧失但可恢复。因此，我们推测紫外线剂量为18J/cm²的核黄素光动力方法在灭活全血中四种耐抗生素细菌的同时能够保留全血功能。我们还提供了一种用于治疗由耐抗生素细菌引起的菌血症的新型体外循环光疗模式。
