The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids
We evaluated the redox status, precisely glutathione levels, which have a major impact in cellular detoxification and antioxidant defence in IBD-derived and healthy intestinal organoids. Therefore, we wanted to explore the differences in terms of their redox balance and mitochondrial fitness. To thi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-09-01
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| Series: | Organoids |
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| Online Access: | https://www.mdpi.com/2674-1172/2/3/12 |
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| author | Georg Csukovich Janina Huainig Selina Troester Barbara Pratscher Iwan Anton Burgener |
| author_facet | Georg Csukovich Janina Huainig Selina Troester Barbara Pratscher Iwan Anton Burgener |
| author_sort | Georg Csukovich |
| collection | DOAJ |
| description | We evaluated the redox status, precisely glutathione levels, which have a major impact in cellular detoxification and antioxidant defence in IBD-derived and healthy intestinal organoids. Therefore, we wanted to explore the differences in terms of their redox balance and mitochondrial fitness. To this end, we introduced a Grx1-roGFP2 construct into the organoids by lentiviral transduction before performing a stress assay by treating the organoids with hydrogen peroxide and examined the GSH/GSSG ratio using confocal imaging. Using ratio imaging, we could detect statistically significant differences between healthy and IBD-derived samples. To gain more insight, we also performed a GSH/GSSG assay, which directly measured glutathione levels. This analysis revealed that both organoid lines had higher levels of oxidized glutathione due to the stress treatment demonstrated by a lower GSH/GSSG ratio compared to the untreated control. Nevertheless, the results showed no significant difference between healthy and IBD-derived organoids. We further challenged organoids with hydrogen peroxide after incubation with MitoTracker<sup>®</sup> to see if mitochondrial fitness might be different in IBD-derived organoids. However, these results were also very comparable. In summary, our preliminary findings indicate that both organoid lines demonstrate a well-functioning system in terms of analysis but show no clear difference between healthy and IBD-derived samples. |
| first_indexed | 2024-03-10T22:19:01Z |
| format | Article |
| id | doaj.art-105032b36f4d40369aa9b756e02c4459 |
| institution | Directory Open Access Journal |
| issn | 2674-1172 |
| language | English |
| last_indexed | 2024-03-10T22:19:01Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Organoids |
| spelling | doaj.art-105032b36f4d40369aa9b756e02c44592023-11-19T12:21:02ZengMDPI AGOrganoids2674-11722023-09-012315616410.3390/organoids2030012The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal OrganoidsGeorg Csukovich0Janina Huainig1Selina Troester2Barbara Pratscher3Iwan Anton Burgener4Small Animal Internal Medicine, Vetmeduni, 1210 Vienna, AustriaSmall Animal Internal Medicine, Vetmeduni, 1210 Vienna, AustriaInstitute for Medical Biochemistry, Vetmeduni, 1210 Vienna, AustriaSmall Animal Internal Medicine, Vetmeduni, 1210 Vienna, AustriaSmall Animal Internal Medicine, Vetmeduni, 1210 Vienna, AustriaWe evaluated the redox status, precisely glutathione levels, which have a major impact in cellular detoxification and antioxidant defence in IBD-derived and healthy intestinal organoids. Therefore, we wanted to explore the differences in terms of their redox balance and mitochondrial fitness. To this end, we introduced a Grx1-roGFP2 construct into the organoids by lentiviral transduction before performing a stress assay by treating the organoids with hydrogen peroxide and examined the GSH/GSSG ratio using confocal imaging. Using ratio imaging, we could detect statistically significant differences between healthy and IBD-derived samples. To gain more insight, we also performed a GSH/GSSG assay, which directly measured glutathione levels. This analysis revealed that both organoid lines had higher levels of oxidized glutathione due to the stress treatment demonstrated by a lower GSH/GSSG ratio compared to the untreated control. Nevertheless, the results showed no significant difference between healthy and IBD-derived organoids. We further challenged organoids with hydrogen peroxide after incubation with MitoTracker<sup>®</sup> to see if mitochondrial fitness might be different in IBD-derived organoids. However, these results were also very comparable. In summary, our preliminary findings indicate that both organoid lines demonstrate a well-functioning system in terms of analysis but show no clear difference between healthy and IBD-derived samples.https://www.mdpi.com/2674-1172/2/3/12intestinal organoidredox biologyIBDglutathioneoxidative stressROS |
| spellingShingle | Georg Csukovich Janina Huainig Selina Troester Barbara Pratscher Iwan Anton Burgener The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids Organoids intestinal organoid redox biology IBD glutathione oxidative stress ROS |
| title | The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids |
| title_full | The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids |
| title_fullStr | The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids |
| title_full_unstemmed | The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids |
| title_short | The Intricacies of Inflammatory Bowel Disease: A Preliminary Study of Redox Biology in Intestinal Organoids |
| title_sort | intricacies of inflammatory bowel disease a preliminary study of redox biology in intestinal organoids |
| topic | intestinal organoid redox biology IBD glutathione oxidative stress ROS |
| url | https://www.mdpi.com/2674-1172/2/3/12 |
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