Idoxuridine

Idoxuridine è un analogo nucleosidico, una forma modificata della desossiuridina, sufficientemente simile da essere incorporata nella replicazione del DNA virale, ma l'atomo di iodio aggiunto al componente uracile blocca l'appaiamento delle basi. Questo composto è usato per il trattamento topico delle infezioni oculari erpetiche dovute al virus dell'herpes simplex (HSV)[2].

• Linee cellulari

  Cultured Crandell-Reese feline kidney (CRFK) cells and FHV-1 strain 727.

• Concentrazioni

  5, 10, and 50 µM

• Tempo di incubazione

  24, 48, and 72 h

• Metodo

  The CRFK cells are cultured in 25-cm2 flasks that contained DMEM with 10% FBS. The concentration of idoxuridine is 5, 10, and 50µM. Cells from 1 control flask and 1 flask containing each drug concentration are examined by use of an inverted microscope to detect morphologic changes and evaluate confluence, and cells are then harvested at 24, 48, and 72 hours. Culture medium is decanted, and the monolayer is rinsed once with PBS solution. Cells are then enzymatically detached with trypsin-EDTA solution. Trypsinization is stopped by the addition of DMEM with 10% FBS, and cells are forcefully pipetted to ensure complete detachment from the flasks. Each flask is then rinsed once with DMEM with 10% FBS, and the eluent is combined with the first cell suspension to ensure collection of as many cells as possible from each flask. Cell-containing medium is centrifuged (300 X g for 6 minutes), and the supernatant is decanted. Cells are then resuspended in a known volume of DMEM and counted on a hemacytometer. Cells cultured in each drug concentration at each time point are counted twice, and the total number of cells in each flask is calculated.