PURPOSE: Several iatrogenic risk factors during pars plana vitrectomy (PPV) could cause damage to the retina. One mechanism is excitotoxicity. Therefore, neuroprotective irrigation solutions would be desirable. METHODS: Retinal ganglion cells (RGC-5) and retinal whole mounts were incubated in standard irrigation solution (SIS) and Dulbecco's Modified Eagle Medium (DMEM). Cell viability, cell amount, cell survival and caspase 3/7 activity were measured by MTS-Test, crystal-violet staining, Annexin-V/PI flow cytometry and caspase 3/7 activity assay, respectively. The morphology and the function of retinal whole mounts were analysed by Live/Dead(TM) staining and by the b-wave and a-wave of the electroretinogram (ERG). RESULTS: Under excitotoxic conditions (10 mM and 12 mM glutamate) RGC-5 cells incubated in SIS showed a statistically significant reduction in cell viability, cell amount, cell survival and caspase 3/7 activity compared to DMEM. Furthermore, the incubation of retinal whole mounts in DMEM resulted in a significant decrease of cell death under excitotoxic (250 μM glutamate) and standard conditions compared to SIS. ERG b-wave recordings revealed good functional preservation of retinal whole mounts in DMEM, but loss in SIS. CONCLUSION: DMEM seems to support retinal cells very well and to be strongly protective against excitotoxicity. Therefore, DMEM may be considered as possible neuroprotective irrigation solution for PPV.
PURPOSE: Several iatrogenic risk factors during pars plana vitrectomy (PPV) could cause damage to the retina. One mechanism is excitotoxicity. Therefore, neuroprotective irrigation solutions would be desirable. METHODS: Retinal ganglion cells (RGC-5) and retinal whole mounts were incubated in standard irrigation solution (SIS) and Dulbecco's Modified Eagle Medium (DMEM). Cell viability, cell amount, cell survival and caspase 3/7 activity were measured by MTS-Test, crystal-violet staining, Annexin-V/PI flow cytometry and caspase 3/7 activity assay, respectively. The morphology and the function of retinal whole mounts were analysed by Live/Dead(TM) staining and by the b-wave and a-wave of the electroretinogram (ERG). RESULTS: Under excitotoxic conditions (10 mM and 12 mM glutamate) RGC-5 cells incubated in SIS showed a statistically significant reduction in cell viability, cell amount, cell survival and caspase 3/7 activity compared to DMEM. Furthermore, the incubation of retinal whole mounts in DMEM resulted in a significant decrease of cell death under excitotoxic (250 μM glutamate) and standard conditions compared to SIS. ERG b-wave recordings revealed good functional preservation of retinal whole mounts in DMEM, but loss in SIS. CONCLUSION:DMEM seems to support retinal cells very well and to be strongly protective against excitotoxicity. Therefore, DMEM may be considered as possible neuroprotective irrigation solution for PPV.
Authors: Maximilian Schultheiss; Hannah Ruschenburg; Max Warga; Charlotte Schramm; Kai Januschowski; Sven Schnichels; Tilo Biedermann; Peter Szurman; Martin S Spitzer Journal: Retina Date: 2012-07 Impact factor: 4.256
Authors: Kai Januschowski; Sebastian Mueller; Martin S Spitzer; Matthias Lueke; Karl-Ulrich Bartz-Schmidt; Peter Szurman Journal: Graefes Arch Clin Exp Ophthalmol Date: 2010-09-19 Impact factor: 3.117
Authors: Roselie M H Diederen; Ellen C La Heij; Nicolaas E P Deutz; Aize Kijlstra; Alfons G H Kessels; Hans M H van Eijk; Albert T A Liem; Suzanne Dieudonné; Fred Hendrikse Journal: Exp Eye Res Date: 2006-03-10 Impact factor: 3.467
Authors: Kai Januschowski; Sebastian Müller; Carlo Krupp; Martin S Spitzer; José Hurst; Maximilian Schultheiss; Karl-Ulrich Bartz-Schmidt; Peter Szurman; Sven Schnichels Journal: J Vis Exp Date: 2015-03-22 Impact factor: 1.355