Dulbecco's Modified Eagle Medium is neuroprotective when compared to standard vitrectomy irrigation solution

Graefes Arch Clin Exp Ophthalmol. 2013 Jun;251(6):1613-9. doi: 10.1007/s00417-012-2255-6. Epub 2013 Jan 11.

Abstract

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.

MeSH terms

  • Animals
  • Annexin A5 / metabolism
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cattle
  • Cell Count
  • Cell Line
  • Cell Survival / drug effects
  • Culture Media / pharmacology*
  • Electroretinography
  • Enzyme-Linked Immunosorbent Assay
  • Excitatory Amino Acid Agonists / toxicity
  • Flow Cytometry
  • Glutamic Acid / toxicity
  • Neuroprotective Agents / pharmacology*
  • Ophthalmic Solutions / pharmacology*
  • Rats
  • Retina / cytology
  • Retina / drug effects*
  • Retina / enzymology
  • Retinal Ganglion Cells / cytology
  • Retinal Ganglion Cells / drug effects*
  • Retinal Ganglion Cells / enzymology
  • Therapeutic Irrigation
  • Vitrectomy*

Substances

  • Annexin A5
  • Culture Media
  • Excitatory Amino Acid Agonists
  • Neuroprotective Agents
  • Ophthalmic Solutions
  • Glutamic Acid
  • Casp3 protein, rat
  • Caspase 3
  • Caspase 7