The essential oils Carvone, Eugenol, Geraniol, and Nerolidol are included in the viricidal spray AV2 in equal volumes diluted 50% in olive oil. The spray is topically administered to the cervix while the subject is in the lithotomic position and fitted with a speculum. Two pumps are administered with each pump delivering 100 micro-L of the solution.

HepG2 cells were cultured in minimum essential medium (MEM) Eagle (with 2 mM L-glutamine and Earle's buffered saline solution (BSS) adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM nonessential amino acids, and 1 mM sodium pyruvate), containing 10% fetal bovine serum. Cells were seeded in a 24-well plate and allowed to attach and recover for 1 day prior to drug treatment. Nerolidol was added at concentrations of 1.2 and 2.4 microM for proliferation assays and 10, 50, and 100 microM for cell viability assays. Cells were cultured for up o 3 days without media change or drug replenishment. At various time points, cells were washed, fixed and stained with sulforhodamine B to determine cell proliferation. Cytotoxicity of nerolidol was also evaluated through MTT assay. Nerolidol was toxic to HepG2 cells after 24 hours with a very strong inhibition of cell proliferation present even at the lowest concentrations tested (10 microM).