Scilletta Natalia A, Pezzoni Magdalena, Desimone Martín F, Soler-Illia Galo J A A, Bellino Martín G, Catalano Paolo N
Departamento de Micro y Nanotecnología, Instituto de Nanociencia y Nanotecnología-Comisión Nacional de Energía Atómica y Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. General Paz 1499, B1650KNA San Martín, Argentina.
Departamento de Radiobiología, Comisión Nacional de Energía Atómica, Av. General Paz 1499, B1650KNA San Martín, Argentina y Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).
Bio Protoc. 2021 Jan 20;11(2):e3887. doi: 10.21769/BioProtoc.3887.
Antibacterial coatings have currently gained great importance in biomedical technology investigations. Because of the spatial arrangement of the film coatings, evaluation of antibacterial activity presents a new challenge regarding traditional bacterial counting methods. In this protocol, four clinically relevant pathogens, , and were incubated on titania mesostructured thin film coatings for 24 h. Then, cell viability was studied considering three methods: counting of the number of colony forming units (CFU), live/dead staining, and quantification of extracellular DNA in suspension. Firstly, bacterial count was determined by the standard plate-count technique. Secondly, bacteria membrane integrity was evaluated by utilization of two fluorescent dyes, which allow distinction between live (membrane intact) and dead (membrane disrupted) bacteria. Lastly, extracellular DNA was quantified by spectrophotometry. In this manner, the three aforementioned techniques enabled the study of bacterial viability by qualitative and quantitative analyses.
抗菌涂层目前在生物医学技术研究中具有重要意义。由于薄膜涂层的空间排列，抗菌活性的评估对传统细菌计数方法提出了新的挑战。在本实验方案中，将四种临床相关病原体，即[此处原文缺失病原体名称]，在二氧化钛介孔结构薄膜涂层上孵育24小时。然后，考虑三种方法研究细胞活力：菌落形成单位（CFU）计数、活/死染色以及悬浮液中细胞外DNA的定量分析。首先，通过标准平板计数技术确定细菌数量。其次，利用两种荧光染料评估细菌膜完整性，这两种染料可区分活细菌（膜完整）和死细菌（膜破裂）。最后，通过分光光度法定量分析细胞外DNA。通过这种方式，上述三种技术能够通过定性和定量分析研究细菌活力。
