背景:语音假体上的生物膜形成破坏了语音假体的功能并限制了其寿命。仍然没有有效的临床策略来抑制或去除这些生物膜。磺胺嘧啶银(SSD),作为外源性抗菌剂,已广泛用于预防和治疗感染,然而,其对语音假体生物膜的影响是未知的。这项研究的目的是探讨SSD对语音假体上存在的成熟混合细菌生物膜的影响。
方法:定量和定性方法,包括平板计数法,实时荧光定量PCR,结晶紫染色,2,3-双(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-甲酰苯胺(XTT)还原测定,扫描电子显微镜,和激光共聚焦显微镜,用于确定SSD对细菌菌落数量的影响,生物膜形成能力,代谢活动,和成熟混合细菌(金黄色葡萄球菌,粪链球菌和白色念珠菌)语音假体生物膜模型。结果在来自患者的成熟混合细菌语音假体生物膜上进行了体外验证,并探讨了可能的作用机制。
结果:磺胺嘧啶银减少成熟混合细菌语音假体生物膜上的细菌菌落数量,显著抑制成熟语音假体生物膜的生物膜形成能力和代谢活性,抑制了语音假体生物膜复杂空间结构的形成,并抑制生物膜细胞外基质中多糖和蛋白质的合成。抑制程度和去除效果随SSD浓度的增加而增加。
结论:磺胺嘧啶银能有效抑制和去除成熟的混合细菌语音假体生物膜,降低生物膜形成能力和代谢活性;SSD可能通过抑制语音假体生物膜胞外聚合物中多糖和蛋白质的合成发挥上述作用。
BACKGROUND: Biofilm formation on voice prostheses disrupts the function and limits the lifespan of voice prostheses. There is still no effective clinical strategy for inhibiting or removing these biofilms. Silver sulfadiazine (SSD), as an exogenous antibacterial agent, has been widely used in the prevention and treatment of infection, however, its effect on voice prosthesis biofilms is unknown. The purpose of this study was to explore the effect of SSD on the mature mixed bacterial biofilms present on voice prostheses.
METHODS: Quantitative and qualitative methods, including the plate counting method, real-time fluorescence quantitative PCR, crystal violet staining, the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) (XTT) reduction assay, scanning electron microscopy, and laser confocal microscopy, were used to determine the effect of SSD on the number of bacterial colonies, biofilm formation ability, metabolic activity, and ultrastructure of biofilms in a mature mixed bacterial (Staphylococcus aureus, Streptococcus faecalis and Candida albicans) voice prosthesis biofilm model. The results were verified in vitro on mature mixed bacterial voice prosthesis biofilms from patients, and the possible mechanism of action was explored.
RESULTS: Silver sulfadiazine decreased the number of bacterial colonies on mature mixed bacterial voice prosthesis biofilm, significantly inhibited the biofilm formation ability and metabolic activity of mature voice prosthesis biofilms, inhibited the formation of the complex spatial structure of voice prosthesis biofilms, and inhibited the synthesis of polysaccharides and proteins in the biofilm extracellular matrix. The degree of inhibition and removal effect increased with SSD concentration.
CONCLUSIONS: Silver sulfadiazine can effectively inhibit and remove mature mixed bacterial voice prosthesis biofilms and decrease biofilm formation ability and metabolic activity; SSD may exert these effects by inhibiting the synthesis of polysaccharides and proteins among the extracellular polymeric substances of voice prosthesis biofilms.