Microdilution Assay for Nanosilver MIC

A microdilution protocol was applied to determine the minimum inhibitory concentration (MIC) of the synthesized nanosilver. Briefly, binary serial dilutions from the nanopowder were prepared in Luria–Bertani (LB) broth (Thermo Fischer Scientific) using sterile 96-well Nunc plates (Thermo Fischer Scientific) and then inoculated with microbial suspensions of 10⁶ (colony-forming units) CFU/mL (in phosphate-buffered saline (PBS), Sigma/Merck). After standard incubation for 24 h, the MIC values were macroscopically evaluated (through naked-eye analysis) and spectrophotometrically determined (as the lowest concentration of AgNPs that inhibited microbial growth) [142 (link),143 (link)].

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Constantinescu S., Niculescu A.G., Hudiță A., Grumezescu V., Rădulescu D., Bîrcă A.C., Irimiciuc S.A., Gherasim O., Holban A.M., Gălățeanu B., Oprea O.C., Ficai A., Vasile B.Ș., Grumezescu A.M., Bolocan A, & Rădulescu R. (2023). Silver/Graphene Oxide Nanostructured Coatings for Modulating the Microbial Susceptibility of Fixation Devices Used in Knee Surgery. International Journal of Molecular Sciences, 25(1), 246.

Publication 2023

Nunc plates

Corresponding Organization : Academy of Romanian Scientists

Other organizations : Carol Davila University of Medicine and Pharmacy, University of Bucharest, National Institute for Laser Plasma and Radiation Physics

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Variable analysis

independent variables

Concentration of synthesized nanosilver (binary serial dilutions)

dependent variables

Minimum inhibitory concentration (MIC) of the synthesized nanosilver, evaluated macroscopically (through naked-eye analysis) and spectrophotometrically

control variables

Luria–Bertani (LB) broth (Thermo Fischer Scientific) used for dilutions
Sterile 96-well Nunc plates (Thermo Fischer Scientific) used for dilutions
Microbial suspensions of 10^6 colony-forming units (CFU)/mL (in phosphate-buffered saline (PBS), Sigma/Merck)
Standard incubation time of 24 hours

controls

No positive or negative controls were explicitly mentioned in the protocol.

Annotations

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