title: Testing the biocompatibility of a glutathione-containing intra-ocular irrigation solution by using an isolated perfused bovine retina organ culture model - an alternative to animal testing.
abstract: The effects of a glutathione-containing intra-ocular irrigation solution, BSS Plus©, on retinal function and on the survival of ganglion cells in whole-mount retinal explants were studied. Evidence is provided that the perfused ex vivo bovine retina can serve as an alternative to in vivo animal testing. Isolated bovine retinas were prepared and perfused with an oxygen-saturated standard irrigation solution, and an electroretinogram was recorded to assess retinal function. After stable b-waves were detected, the isolated retinas were perfused with BSS Plus for 45 minutes. To investigate the effects of BSS Plus on photoreceptor function, 1mM aspartate was added to the irrigation solution in order to obtain a-waves, and the ERG trace was monitored for 75 minutes. For histological analysis, isolated whole retinal mounts were stored for 24 hours at 4°C, in the dark. The percentages of cell death in the retinal ganglion cell layer and in the outer and inner nuclear layers were estimated by using an ethidium homodimer-1 stain and the TUNEL assay. General swelling of the retina was examined with high-resolution optical coherence tomography. During perfusion with BSS Plus, no significant changes in a-wave and b-wave amplitudes were recorded. Retinas stored for 24 hours in BSS Plus showed a statistically significant smaller percentage (52.6%, standard deviation [SD] = 16.1%) of cell death in the retinal ganglion cell layer compared to the control group (69.6%, SD = 3.9, p = 0.0031). BSS Plus did not seem to affect short-term retinal function, and had a beneficial effect on the survival of retinal ganglion cells. This method for analysing the isolated perfused retina represents a valuable alternative for testing substances for their retinal biocompatibility and toxicity.
authors.affiliation: Centre for Ophthalmology, University Eye Hospital, Schleichstr. 12, 72076 Tuebingen, Germany. Kai.Januschowski@med.uni-tuebingen.de
authors.forename: Kai
authors.initials: K
authors.lastname: Januschowski
authors.forename: Ahmad
authors.initials: A
authors.lastname: Zhour
authors.forename: Albert
authors.initials: A
authors.lastname: Lee
authors.forename: Ramin
authors.initials: R
authors.lastname: Maddani
authors.forename: Sebastien
authors.initials: S
authors.lastname: Mueller
authors.forename: Martin S
authors.initials: MS
authors.lastname: Spitzer
authors.forename: Sven
authors.initials: S
authors.lastname: Schnichels
authors.forename: Maximilian
authors.initials: M
authors.lastname: Schultheiss
authors.forename: Deshka
authors.initials: D
authors.lastname: Doycheva
authors.forename: Karl-Ulrich
authors.initials: KU
authors.lastname: Bartz-Schmidt
authors.forename: Peter
authors.initials: P
authors.lastname: Szurman
mesh_terms: D000826:Animal Testing Alternatives; D000818:Animals; D001639:Bicarbonates / Q000009:adverse effects*; D002417:Cattle; D004338:Drug Combinations; D004596:Electroretinography; D005978:Glutathione / Q000009:adverse effects*; D009883:Ophthalmic Solutions / Q000009:adverse effects*; D009924:Organ Culture Techniques; D012160:Retina / Q000187:drug effects*; D041623:Tomography, Optical Coherence
chemical_list: D001639:Bicarbonates; D004338:Drug Combinations; D009883:Ophthalmic Solutions; C013003:glutathione-bicarbonate-Ringer solution; D005978:Glutathione