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Impressum
Evidence of toxic side effects using Perfluorohexyloktane after vitreoretinal surgery and after incubation with human retinal pigment epithelium and human corneal endothelium in vitro
Mertens S., Bednarz J., Richard G., Engelmann K.
University Eye Clinic Hamburg, Martinistrasse 52, D-20246 Hamburg
Objective: Perfluorohexyloctane (PHO) has recently been used as
an intraoperative tool in difficult surgical procedures. However, in our
clinic 2 out of 3 patients developed ocular irritation with white precipitates.
The aim of this study was to examine and to compare the influence of PHO
and perfluorodecaline (PFD) on human retinal pigment epithelium (RPE)
and human corneal endothelium (HCEC) in vitro.
Methods: Vitality and proliferative capacity of cell cultures were
measured after incubation with PHO and PFD for up to 5 days. Vitality
of cell cultures was evaluated using the Life/Dead Viability Cytotoxicity
assay. Cell proliferation was determined according to incorporation of
5-bromo-2`-deoxyuridine into cellular DNA. Furthermore 2 pairs of human
donor corneas were incubated with PHO for 5 days and documented.
Results: After 5 day incubation period with PHO both cell types
showed significantly lower extinctions (RPE 38,4%, HCEC 3,5%) for vital
cells compared to controls. PFD caused a decrease of vital cells to 55.8%
in cultures of RPE and 29.1% in HCEC. After 3 to 5 days in cultures of
HCEC circumscribed areas of nearly complete cell necrosis appeared at
the interface where PHO was located on the cell layer. After incubation
with PHO HCEC presented a decreasing amount of proliferation from day
1 to 5 (HCEC 56.5%). In contrast cultures of RPE presented a proliferative
capacity of 91.9% after 5 days contact to PHO. Incubation with PFD caused
a decrease of mean proliferation to 90.6% in cultured RPE and 94.4% in
HCEC after 5 days. Endothelium of donor corneas incubated in the presence
of PHO developed circumscribed areas of cell necrosis at the interface
between substance and medium.
Conclusion: Deteriorated extinctions in Life-Dead assay correlated
with decreased amounts of vital cells in areas with contact to PHO and
PFD. Proliferative capacity after incubation revealed that remaining cells
were not irreversibly damaged. Since PHO has a lower specific weight (1.35
g/cm3) than PFD (1.93 g/cm3) cell damage can not only be explained by
mechanical effect. Beside impending metabolic exchanges, a direct toxic
effect such as interaction between PHO and the cellular lipoprotein membrane
has to be considered.