Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.

As oxygen carriers, perfluorocarbon emulsions might be useful to decrease hypoxia of pancreatic islets before transplantation. However, their hydrophobicity prevents their homogenisation in culture medium. To increase the surface of contact between islets and Perfluorooctyl bromide (PFOB), and cons...

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Main Authors: Maillard, E, Juszczak, M, Langlois, A, Kleiss, C, Sencier, M, Bietiger, W, Sanchez-Dominguez, M, Krafft, M, Johnson, P, Pinget, M, Sigrist, S
Format: Journal article
Language:English
Published: 2012
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author Maillard, E
Juszczak, M
Langlois, A
Kleiss, C
Sencier, M
Bietiger, W
Sanchez-Dominguez, M
Krafft, M
Johnson, P
Pinget, M
Sigrist, S
author_facet Maillard, E
Juszczak, M
Langlois, A
Kleiss, C
Sencier, M
Bietiger, W
Sanchez-Dominguez, M
Krafft, M
Johnson, P
Pinget, M
Sigrist, S
author_sort Maillard, E
collection OXFORD
description As oxygen carriers, perfluorocarbon emulsions might be useful to decrease hypoxia of pancreatic islets before transplantation. However, their hydrophobicity prevents their homogenisation in culture medium. To increase the surface of contact between islets and Perfluorooctyl bromide (PFOB), and consequently oxygen delivery, we tested effect of a PFOB emulsion in culture medium on β-cell lines and rat pancreatic islets. RINm5F β-cell line or pancreatic rat islets were incubated for 3 days in the presence of PFOB emulsion in media (3.5% w/v). Preoxygenation of the medium was performed before culture. Cell viability was assessed by apoptotic markers (Bax and Bcl-2) and by staining (fluoresceine diacetate and propidium iodide). β-Cell functionality was determined by insulin release during a glucose stimulation test and. Hypoxia markers, HIF-1α and VEGF, were studied at days 1 and 3 using RT-PCR, Western blotting, and ELISA. PFOB emulsions preserved viability and functionality of RINm5F cells with a decrease of HIF-1α and VEGF expression. Islets viability was preserved during 3 days of culture. Secretion of VEGF was higher in untreated control (0.09 ± 0.041 μg VEGF/mg total protein) than in PFOB emulsion incubated islets (0.02 ± 0.19 μg VEGF/mg total protein, n = 4, p < 0.05) at day 1. At day 3, VEGF secretion was increased as compared to day 1 in control (0.23 ± 0.04 μg VEGF/mg total protein) but it was imbalance by the presence of PFOB emulsion (0.09 ± 0.03 μg VEGF/mg total protein, n = 5, p < 0.05). While insulin secretion was maintained in response to a glucose stimulation test until day 3 when islets were incubated in the presence of PFOB emulsion preoxygenated (0.81 ± 0.16 at day 1 vs. 0.75 ± 0.24 at day 3), the ability to secrete insulin in the presence of high glucose concentration was lost in islets controls (0.51 ± 0.18 at day 1 vs. 0.21 ± 0.13 at day 3). Atmospheric oxygen delivery by PFOB emulsion might be sufficient to decrease islets hypoxia. However, to improve islets functionality, overoxygenation is needed. Finally, maintenance of islet viability and functionality for several days after isolation could improve the outcome of islets transplantation.
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spelling oxford-uuid:ea6267e2-327e-4142-b4d1-e6c9547cd0cc2022-03-27T11:01:54ZPerfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:ea6267e2-327e-4142-b4d1-e6c9547cd0ccEnglishSymplectic Elements at Oxford2012Maillard, EJuszczak, MLanglois, AKleiss, CSencier, MBietiger, WSanchez-Dominguez, MKrafft, MJohnson, PPinget, MSigrist, SAs oxygen carriers, perfluorocarbon emulsions might be useful to decrease hypoxia of pancreatic islets before transplantation. However, their hydrophobicity prevents their homogenisation in culture medium. To increase the surface of contact between islets and Perfluorooctyl bromide (PFOB), and consequently oxygen delivery, we tested effect of a PFOB emulsion in culture medium on β-cell lines and rat pancreatic islets. RINm5F β-cell line or pancreatic rat islets were incubated for 3 days in the presence of PFOB emulsion in media (3.5% w/v). Preoxygenation of the medium was performed before culture. Cell viability was assessed by apoptotic markers (Bax and Bcl-2) and by staining (fluoresceine diacetate and propidium iodide). β-Cell functionality was determined by insulin release during a glucose stimulation test and. Hypoxia markers, HIF-1α and VEGF, were studied at days 1 and 3 using RT-PCR, Western blotting, and ELISA. PFOB emulsions preserved viability and functionality of RINm5F cells with a decrease of HIF-1α and VEGF expression. Islets viability was preserved during 3 days of culture. Secretion of VEGF was higher in untreated control (0.09 ± 0.041 μg VEGF/mg total protein) than in PFOB emulsion incubated islets (0.02 ± 0.19 μg VEGF/mg total protein, n = 4, p < 0.05) at day 1. At day 3, VEGF secretion was increased as compared to day 1 in control (0.23 ± 0.04 μg VEGF/mg total protein) but it was imbalance by the presence of PFOB emulsion (0.09 ± 0.03 μg VEGF/mg total protein, n = 5, p < 0.05). While insulin secretion was maintained in response to a glucose stimulation test until day 3 when islets were incubated in the presence of PFOB emulsion preoxygenated (0.81 ± 0.16 at day 1 vs. 0.75 ± 0.24 at day 3), the ability to secrete insulin in the presence of high glucose concentration was lost in islets controls (0.51 ± 0.18 at day 1 vs. 0.21 ± 0.13 at day 3). Atmospheric oxygen delivery by PFOB emulsion might be sufficient to decrease islets hypoxia. However, to improve islets functionality, overoxygenation is needed. Finally, maintenance of islet viability and functionality for several days after isolation could improve the outcome of islets transplantation.
spellingShingle Maillard, E
Juszczak, M
Langlois, A
Kleiss, C
Sencier, M
Bietiger, W
Sanchez-Dominguez, M
Krafft, M
Johnson, P
Pinget, M
Sigrist, S
Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title_full Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title_fullStr Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title_full_unstemmed Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title_short Perfluorocarbon emulsions prevent hypoxia of pancreatic β-cells.
title_sort perfluorocarbon emulsions prevent hypoxia of pancreatic β cells
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