Determination of Antibacterial Activity of Film Coatings against Four Clinically Relevant Bacterial Strains

SCILLETTA, NATALIA; PEZZONI, MAGDALENA; DESIMONE, MARTIN; SOLER-ILLIA, GALO; BELLINO, MARTIN; CATALANO, PAOLO

BIO-PROTOCOL

bio-protocol

Año: 2021 vol. 11

2331-8325

Antibacterial coatings have currently gained great importance in biomedical technologyinvestigations. Because of the spatial arrangement of the film coatings, evaluation of antibacterialactivity presents a new challenge regarding traditional bacterial counting methods. In this protocol, fourclinically relevant pathogens, Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa,and Staphylococcus aureus 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 countwas determined by the standard plate-count technique. Secondly, bacteria membrane integrity wasevaluated by utilization of two fluorescent dyes, which allow distinction between live (membrane intact)and dead (membrane disrupted) bacteria. Lastly, extracellular DNA was quantified byspectrophotometry. In this manner, the three aforementioned techniques enabled the study of bacterialviability by qualitative and quantitative analyses.