September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
The effect of air, SF6 and C3F8 on human corneal endothelial cells
Author Affiliations & Notes
  • Matthias Fuest
    Ophthalmology, RWTH Aachen University, Aachen, Germany
  • Niklas Plange
    Ophthalmology, RWTH Aachen University, Aachen, Germany
  • Sandra Johnen
    Ophthalmology, RWTH Aachen University, Aachen, Germany
  • David Kuerten
    Ophthalmology, RWTH Aachen University, Aachen, Germany
  • Peter Walter
    Ophthalmology, RWTH Aachen University, Aachen, Germany
  • Footnotes
    Commercial Relationships   Matthias Fuest, None; Niklas Plange, None; Sandra Johnen, None; David Kuerten, None; Peter Walter, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1209. doi:
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    • Get Citation

      Matthias Fuest, Niklas Plange, Sandra Johnen, David Kuerten, Peter Walter; The effect of air, SF6 and C3F8 on human corneal endothelial cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1209.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : While air-bubbles remain in the anterior chamber only for few days to aid attachment during posterior lamellar surgery, these periods can be prolonged with gases in non-expansible concentrations. To test the effects of different gases on immortalized human corneal endothelial cells (HCEC-12), we utilized transwell-inserts with semi-permeable membranes completely sunk in medium as an artificial anterior chamber model.

Methods : 4x103 HCEC-12 were cultured in transwell-inserts for 24 hours, then flipped bottom side up, burdened and sunk with titanium rings in medium (M, 95% DMEM, 5% FCS, 1/100 Penicillin-Streptomycin and 1/125 Amphotericin B) and filled with 2 ml of air (A), 20% SF6 (S), or 12% C3F8 (C). After a cultivation time of 24, 48 and 120 hours, the cells were evaluated by live/dead fluorescence staining (5µg/mL fluorescein diacetate, 10µg/mL propidium iodide) and by a luminescent-based cell viability assay.

Results : Cells continued to proliferate on sunken inserts without gas contact, while gas exposition led to an inhibition of proliferation and an increase in the number of dead cells, independent of the type of gas (120 h cell count/2.25cm2: M= 660.8 ± 57.0 cells; A= 125.5 ± 17.4 cells, S=123.5 ± 17.0 cells, C= 118.8 ± 16.6 cells; p-value: M vs. A/S/C<0.001; 120 h dead cells: M= 2.6 ± 1.0 %, A=8.4 ± 2.7 %, S= 9.5 ± 3.2 %, C= 11.3 ± 3.1 %; p-value: M vs. A/S/C<0.01). M also proved significantly higher luminescent viability values in comparison to the gases, which did not differ significantly among them (120 h luminescence: M= 1752.20 ± 91.44, A= 432.97 ± 30.36, S= 507.79 ± 23.34, C= 523.82 ± 20.27; p-value: M vs. A/S/C<0.01).

Conclusions : Gas exposition led to an inhibition of proliferation and an increase in cell death in HCEC-12. However, no differences between different gas compositions were found.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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