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Published on: October 27, 2020
Inhibiting connexin channels protects against cryopreservation-induced cell death in human blood vessels
1Department of Basic Medical Sciences - Physiology Group, Faculty of Medicine & Health Sciences, Ghent University, Ghent 9000, Belgium.
Insights
Blocking connexin channels with Gap27 during cryopreservation significantly reduces vascular cell death in human blood vessels. This method enhances cell viability after freezing and thawing, crucial for successful vascular reconstruction.
Area of Science:
- Vascular Biology
- Cryobiology
- Cell Signaling
Background:
- Cryopreservation of blood vessels is vital for vascular reconstruction but causes significant cell death, potentially leading to graft failure.
- Vascular cells utilize connexin proteins to form gap junction channels and hemichannels, which can mediate cell death signaling.
- Hemichannels, normally closed, can open under stress, promoting cell death, while gap junctions may transmit death signals between cells.
Purpose of the Study:
- To investigate the potential of blocking connexin channels to prevent cell death in cryopreserved human blood vessels.
- To determine if inhibiting gap junctions and hemichannels improves vascular cell viability post-cryopreservation.
Main Methods:
- Human saphenous veins and femoral arteries were cryopreserved and thawed with the inclusion of Gap27, a connexin channel inhibitory peptide.
- Cell death was assessed using terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assays and histological examination.
Main Results:
- Gap27 significantly reduced cell death in both human femoral arteries and saphenous veins during cryopreservation and thawing.
- Smooth muscle cell death decreased by 73% in arteries and 71% in veins.
- Endothelial cell death was reduced by 32% in arteries and 51% in veins.
Conclusions:
- Inhibiting connexin channels during cryopreservation is a promising strategy to enhance vascular cell viability.
- Blocking these channels effectively preserves vascular tissue integrity, potentially reducing graft failure in reconstruction procedures.
Objectives:
Cryopreserved blood vessels are being increasingly employed in vascular reconstruction procedures but freezing/thawing is associated with significant cell death that may lead to graft failure. Vascular cells express connexin proteins that form gap junction channels and hemichannels. Gap junction channels directly connect the cytoplasm of adjacent cells and may facilitate the passage of cell death messengers leading to bystander cell death. Two hemichannels form a gap junction channel but these channels are also present as free non-connected hemichannels. Hemichannels are normally closed but may open under stressful conditions and thereby promote cell death. We here investigated whether blocking gap junctions and hemichannels could prevent cell death after cryopreservation.
Materials And Methods:
Inclusion of Gap27, a connexin channel inhibitory peptide, during cryopreservation and thawing of human saphenous veins and femoral arteries was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assays and histological examination.
Results:
We report that Gap27 significantly reduces cell death in human femoral arteries and saphenous veins when present during cryopreservation/thawing. In particular, smooth muscle cell death was reduced by 73% in arteries and 71% in veins, while endothelial cell death was reduced by 32% in arteries and 51% in veins.
Conclusions:
We conclude that inhibiting connexin channels during cryopreservation strongly promotes vascular cell viability.
