Riwaldt, Stefan and Monici, Monica and Petersen, Asbjorn Graver and Jensen, Uffe Birk and Evert, Katja and Pantalone, Desire and Utpatel, Kirsten and Evert, Matthias and Wehland, Markus and Krueger, Marcus and Kopp, Sascha and Frandsen, Sofie and Corydon, Thomas and Sahana, Jayashree and Bauer, Johann and Luetzenberg, Ronald and Infanger, Manfred and Grimm, Daniela (2017) Preparation of A Spaceflight: Apoptosis Search in Sutured Wound Healing Models. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 18 (12): 2604. ISSN 1422-0067,
Full text not available from this repository. (Request a copy)Abstract
To prepare the ESA (European Space Agency) spaceflight project "Wound healing and Sutures in Unloading Conditions", we studied mechanisms of apoptosis in wound healing models based on ex vivo skin tissue cultures, kept for 10 days alive in serum-free DMEM/F12 medium supplemented with bovine serum albumin, hydrocortisone, insulin, ascorbic acid and antibiotics at 32 degrees C. The overall goal is to test: (i) the viability of tissue specimens; (ii) the gene expression of activators and inhibitors of apoptosis and extracellular matrix components in wound and suture models; and (iii) to design analytical protocols for future tissue specimens after post-spaceflight download. Hematoxylin-Eosin and Elastica-van-Gieson staining showed a normal skin histology with no signs of necrosis in controls and showed a normal wound suture. TdT-mediated dUTP-biotin nick end labeling for detecting DNA fragmentation revealed no significant apoptosis. No activation of caspase-3 protein was detectable. FASL, FADD, CASP3, CASP8, CASP10, BAX, BCL2, CYC1, APAF1, LAMA3 and SPP1 mRNAs were not altered in epidermis and dermis samples with and without a wound compared to 0 day samples (specimens investigated directly post-surgery). BIRC5, CASP9, and FN1 mRNAs were downregulated in epidermis/dermis samples with and/or without a wound compared to 0 day samples. BIRC2, BIRC3 were upregulated in 10 day wound samples compared to 0 day samples in epidermis/dermis. RELA/FAS mRNAs were elevated in 10 day wound and no wound samples compared to 0 day samples in dermis. In conclusion, we demonstrate that it is possible to maintain live skin tissue cultures for 10 days. The viability analysis showed no significant signs of cell death in wound and suture models. The gene expression analysis demonstrated the interplay of activators and inhibitors of apoptosis and extracellular matrix components, thereby describing important features in ex vivo sutured wound healing models. Collectively, the performed methods defining analytical protocols proved to be applicable for post-flight analyzes of tissue specimens after sample return.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | NF-KAPPA-B; THYROID-CANCER CELLS; EXTRACELLULAR-MATRIX; SIMULATED WEIGHTLESSNESS; ENDOTHELIAL-CELLS; DEATH RECEPTORS; ALTERED GRAVITY; ORGAN-CULTURE; PROTEINS; GROWTH; skin; wound healing; suture; apoptosis; caspases; extracellular matrix proteins |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:18 |
| Last Modified: | 28 Feb 2019 11:12 |
| URI: | https://pred.uni-regensburg.de/id/eprint/1726 |
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