FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies

Preclinical toxicity screening of the new retinal compounds is an absolute requirement in the pathway of further drug development. Since retinal neuron cultivation and in vivo studies are relatively expensive and time consuming, we aimed to create a fast and reproducible ex vivo system for retinal t...

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Autores principales: Smedowski, Adrian, Pietrucha-Dutczak, Marita, Maniar, Ruchi, Ajeleti, Michael, Matuszek, Iwona, Lewin-Kowalik, Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5831603/
https://www.ncbi.nlm.nih.gov/pubmed/29560079
http://dx.doi.org/10.1155/2018/2487473
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author Smedowski, Adrian
Pietrucha-Dutczak, Marita
Maniar, Ruchi
Ajeleti, Michael
Matuszek, Iwona
Lewin-Kowalik, Joanna
author_facet Smedowski, Adrian
Pietrucha-Dutczak, Marita
Maniar, Ruchi
Ajeleti, Michael
Matuszek, Iwona
Lewin-Kowalik, Joanna
author_sort Smedowski, Adrian
collection PubMed
description Preclinical toxicity screening of the new retinal compounds is an absolute requirement in the pathway of further drug development. Since retinal neuron cultivation and in vivo studies are relatively expensive and time consuming, we aimed to create a fast and reproducible ex vivo system for retinal toxicity screening. For this purpose, we used rat retinal explant culture that was retrogradely labeled with the FluoroGold before the isolation. Explants were exposed to a toxic concentration of gentamicin and ciliary neurotrophic factor (CNTF), a known neuroprotective agent. The measured outcomes showed the cell density in retinal ganglion cell layer (GCL) and the activity of lactate dehydrogenase (LDH) in the culture medium. Gentamicin-induced oxidative stress resulted in retinal cell damage and rapid LDH release to the culture medium (p < 0.05). Additional CNTF supplementation minimized the cell damage, and the increase of LDH release was insignificant when compared to LDH levels before gentamicin insult (p > 0.05). As well as this, the LDH activity was directly correlated with the cell count in GCL (R = −0.84, p < 0.00001), making a sensitive marker of retinal neuron damage. The FLOREC protocol could be considered as a fast, reproducible, and sensitive method to detect neurotoxicity in the screening studies of the retinal drugs.
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spelling pubmed-58316032018-03-20 FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies Smedowski, Adrian Pietrucha-Dutczak, Marita Maniar, Ruchi Ajeleti, Michael Matuszek, Iwona Lewin-Kowalik, Joanna Oxid Med Cell Longev Research Article Preclinical toxicity screening of the new retinal compounds is an absolute requirement in the pathway of further drug development. Since retinal neuron cultivation and in vivo studies are relatively expensive and time consuming, we aimed to create a fast and reproducible ex vivo system for retinal toxicity screening. For this purpose, we used rat retinal explant culture that was retrogradely labeled with the FluoroGold before the isolation. Explants were exposed to a toxic concentration of gentamicin and ciliary neurotrophic factor (CNTF), a known neuroprotective agent. The measured outcomes showed the cell density in retinal ganglion cell layer (GCL) and the activity of lactate dehydrogenase (LDH) in the culture medium. Gentamicin-induced oxidative stress resulted in retinal cell damage and rapid LDH release to the culture medium (p < 0.05). Additional CNTF supplementation minimized the cell damage, and the increase of LDH release was insignificant when compared to LDH levels before gentamicin insult (p > 0.05). As well as this, the LDH activity was directly correlated with the cell count in GCL (R = −0.84, p < 0.00001), making a sensitive marker of retinal neuron damage. The FLOREC protocol could be considered as a fast, reproducible, and sensitive method to detect neurotoxicity in the screening studies of the retinal drugs. Hindawi 2018-02-13 /pmc/articles/PMC5831603/ /pubmed/29560079 http://dx.doi.org/10.1155/2018/2487473 Text en Copyright © 2018 Adrian Smedowski et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Smedowski, Adrian
Pietrucha-Dutczak, Marita
Maniar, Ruchi
Ajeleti, Michael
Matuszek, Iwona
Lewin-Kowalik, Joanna
FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title_full FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title_fullStr FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title_full_unstemmed FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title_short FluoroGold-Labeled Organotypic Retinal Explant Culture for Neurotoxicity Screening Studies
title_sort fluorogold-labeled organotypic retinal explant culture for neurotoxicity screening studies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5831603/
https://www.ncbi.nlm.nih.gov/pubmed/29560079
http://dx.doi.org/10.1155/2018/2487473
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