Non-esteri
fied fatty acids
–induced alterations to BOEC
membrane properties might also explain the divergent
results on BOEC viability we have observed. Because the
enzymatic reaction of the trypsin added to estimate this
parameter might very well rupture the NEFA-impaired cell
membranes, causing lysis of otherwise viable cells and thus
induce falsely high viability percentages and masking the
effects of the NEFAs added. To verify these data, in situ
viability assessment and quanti
fication techniques, such as
a neutral red assay [62], can be performed to distinguish
enzyme-induced damages from the actual NEFA effects.
We also noticed a decreased cell number with increased
NEFA concentrations. As NEFA toxicity is primarily attributed
to elevated reactive oxygen species production and as
reactive oxygen species target high-fat cell membranes,
lipid radicals could be generated in BOECs as seen in other