治疗由抗生素耐药菌引起的菌血症是全球关注的问题。抗细菌光动力灭活是对抗它的有希望的策略。然而,由于全血中抗生素抗性细菌的不透明性和复杂性,实现其灭活具有挑战性。我们研究了核黄素光动力学方法,以有效灭活全血中的抗生素抗性细菌。分离出4株耐药菌,已识别,并在本研究中培养:耐甲氧西林金黄色葡萄球菌(MRSA),泛耐药鲍曼不动杆菌(PDRAB),产ESBLs大肠埃希菌(EPEC)和泛耐药肺炎克雷伯菌(PDRKP)。为了模拟菌血症,全血中加入抗生素耐药菌.采用核黄素光动力法处理全血,紫外线照射(308nm和365nm)。紫外线照射剂量分为18J/cm2、36J/cm2和54J/cm2。全血中抗生素抗性细菌的微生物计数用于评估灭活效果。红细胞的作用,淋巴细胞,凝血因子,并对全血中的血小板进行了评估。在结果中,随着紫外线剂量从18J/cm2增加到54J/cm2,灭活效果增加。在18J/cm2的剂量下,4种耐药菌的灭活率均在80%以上,而只有67%的MRSA。核黄素光动力处理和抗生素的组合增强了抗菌作用。红细胞功能易受紫外线剂量的影响。在18J/cm2的剂量下,溶血率小于0.8%,并且ATP和2,3-DPG的水平没有变化。在相同的剂量下,扩散,细胞杀伤,淋巴细胞的细胞因子分泌活性下降20-70%;因子V和因子VIII活性下降50%;纤维蛋白原和血小板功能明显丧失,但可以修复。因此,我们推测,紫外线剂量为18J/cm2的核黄素光动力学方法可有效灭活全血中的四种抗生素抗性细菌,同时保留全血功能。我们还提供了一种新型的体外循环光疗模式,用于治疗由抗生素抗性细菌引起的菌血症。
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 Staphylococcus aureus (MRSA), pan-drug-resistant Acinetobacter baumannii (PDRAB), ESBLs-producing Escherichia coli (EPEC) and pan-drug-resistant Klebsiella pneumoniae (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/cm2, 36 J/cm2, and 54 J/cm2. 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/cm2 to 54 J/cm2. At the dose of 18 J/cm2, 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/cm2, 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/cm2 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.