Details
| Original language | English |
|---|---|
| Article number | 104746 |
| Journal | Microvascular research |
| Volume | 157 |
| Early online date | 13 Sept 2024 |
| Publication status | E-pub ahead of print - 13 Sept 2024 |
Abstract
The endothelialization of cardiovascular implants is supposed to improve the long-term patency of these implants. In addition, in previous studies, it has been shown, that the conditioning of endothelial cells by dynamic cultivation leads to the expression of an anti-thrombogenic phenotype. For the creation of a tissue-engineered vascular graft (TEVG), these two strategies were combined to achieve optimal hemocompatibility. In a clinical setup, this would require the transfer of the already endothelialized construct from the conditioning bioreactor to the patient. Therefore, the reversibility of the dynamic conditioning of the endothelial cells with arterial-like high shear stress (20 dyn/cm2) was investigated to define the timeframe (tested in a range of up to 24 h) for the perseverance of dynamically induced phenotypical changes. Two types of endothelial cells were compared: endothelial colony-forming cells (ECFCs) and human aortic endothelial cells (HAECs). The results showed that ECFCs respond far more sensitively and rapidly to flow than HAECs. The resulting cell alignment and increased protein expression of KLF-2, Notch-4, Thrombomodulin, Tie2 and eNOS monomer was paralleled by increased eNOS and unaltered KLF-2 mRNA levels even under stopped-flow conditions. VCAM-1 mRNA and protein expression was downregulated under flow and did not recover under stopped flow. From these time kinetic results, we concluded, that the maximum time gap between the TEVG cultivated with autologous ECFCs in future reactor cultivations and the transfer to the potential TEVG recipient should be limited to ∼6 h.
Keywords
- Cell alignment, Dynamic cultivation, Endothelial colony-forming cells, Reversibility of dynamic preconditioning, Specific protein and mRNA expression (KLF-2/Notch-4/Tie2/Thrombomodulin/eNOS/VE-Cadherin/VCAM-1), Tissue-engineered vascular graft
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Medicine(all)
- Cardiology and Cardiovascular Medicine
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Microvascular research, Vol. 157, 104746, 01.2025.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Sustainability of shear stress conditioning in endothelial colony-forming cells compared to human aortic endothelial cells to underline suitability for tissue-engineered vascular grafts
AU - Renzelmann, Jannis
AU - Heene, Sebastian
AU - Jonczyk, Rebecca
AU - Krüger, Jana
AU - Alnajjar, Suhayla
AU - Blume, Cornelia
N1 - Publisher Copyright: © 2024 The Authors
PY - 2024/9/13
Y1 - 2024/9/13
N2 - The endothelialization of cardiovascular implants is supposed to improve the long-term patency of these implants. In addition, in previous studies, it has been shown, that the conditioning of endothelial cells by dynamic cultivation leads to the expression of an anti-thrombogenic phenotype. For the creation of a tissue-engineered vascular graft (TEVG), these two strategies were combined to achieve optimal hemocompatibility. In a clinical setup, this would require the transfer of the already endothelialized construct from the conditioning bioreactor to the patient. Therefore, the reversibility of the dynamic conditioning of the endothelial cells with arterial-like high shear stress (20 dyn/cm2) was investigated to define the timeframe (tested in a range of up to 24 h) for the perseverance of dynamically induced phenotypical changes. Two types of endothelial cells were compared: endothelial colony-forming cells (ECFCs) and human aortic endothelial cells (HAECs). The results showed that ECFCs respond far more sensitively and rapidly to flow than HAECs. The resulting cell alignment and increased protein expression of KLF-2, Notch-4, Thrombomodulin, Tie2 and eNOS monomer was paralleled by increased eNOS and unaltered KLF-2 mRNA levels even under stopped-flow conditions. VCAM-1 mRNA and protein expression was downregulated under flow and did not recover under stopped flow. From these time kinetic results, we concluded, that the maximum time gap between the TEVG cultivated with autologous ECFCs in future reactor cultivations and the transfer to the potential TEVG recipient should be limited to ∼6 h.
AB - The endothelialization of cardiovascular implants is supposed to improve the long-term patency of these implants. In addition, in previous studies, it has been shown, that the conditioning of endothelial cells by dynamic cultivation leads to the expression of an anti-thrombogenic phenotype. For the creation of a tissue-engineered vascular graft (TEVG), these two strategies were combined to achieve optimal hemocompatibility. In a clinical setup, this would require the transfer of the already endothelialized construct from the conditioning bioreactor to the patient. Therefore, the reversibility of the dynamic conditioning of the endothelial cells with arterial-like high shear stress (20 dyn/cm2) was investigated to define the timeframe (tested in a range of up to 24 h) for the perseverance of dynamically induced phenotypical changes. Two types of endothelial cells were compared: endothelial colony-forming cells (ECFCs) and human aortic endothelial cells (HAECs). The results showed that ECFCs respond far more sensitively and rapidly to flow than HAECs. The resulting cell alignment and increased protein expression of KLF-2, Notch-4, Thrombomodulin, Tie2 and eNOS monomer was paralleled by increased eNOS and unaltered KLF-2 mRNA levels even under stopped-flow conditions. VCAM-1 mRNA and protein expression was downregulated under flow and did not recover under stopped flow. From these time kinetic results, we concluded, that the maximum time gap between the TEVG cultivated with autologous ECFCs in future reactor cultivations and the transfer to the potential TEVG recipient should be limited to ∼6 h.
KW - Cell alignment
KW - Dynamic cultivation
KW - Endothelial colony-forming cells
KW - Reversibility of dynamic preconditioning
KW - Specific protein and mRNA expression (KLF-2/Notch-4/Tie2/Thrombomodulin/eNOS/VE-Cadherin/VCAM-1)
KW - Tissue-engineered vascular graft
UR - http://www.scopus.com/inward/record.url?scp=85204371410&partnerID=8YFLogxK
U2 - 10.1016/j.mvr.2024.104746
DO - 10.1016/j.mvr.2024.104746
M3 - Article
C2 - 39278537
AN - SCOPUS:85204371410
VL - 157
JO - Microvascular research
JF - Microvascular research
SN - 0026-2862
M1 - 104746
ER -