TY - JOUR

T1 - Surface modification of decellularized bovine carotid arteries with human vascular cells significantly reduces their thrombogenicity

AU - Keshi, Eriselda

AU - Tang, Peter

AU - Weinhart, Marie

AU - Everwien, Hannah

AU - Moosburner, Simon

AU - Seiffert, Nicolai

AU - Lommel, Michael

AU - Kertzscher, Ulrich

AU - Globke, Brigitta

AU - Reutzel-Selke, Anja

AU - Strücker, Benjamin

AU - Pratschke, Johann

AU - Sauer, Igor Maximillian

AU - Haep, Nils

AU - Hillebrandt, Karl Herbert

N1 - Funding Information: This work was supported by the BMWi (Bundesministerium für Wirtschaft und Energie) project KF2852504CS4. Hannah Everwien was a BIH-Medical Doctoral Research scholarship holder funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health. Dr. Karl Hillebrandt and Dr. Simon Moosburner are participants in the BIH-Charité Junior Clinician Scientist Program funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health. Priv.-Doz. Dr. Struecker was participant in the BIH-Charité Clinician Scientist Program funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health. The author acknowledges the support of the Cluster of Excellence »Matters of Activity. Image Space Material« funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2025–390,648,296.

PY - 2021/11/24

Y1 - 2021/11/24

N2 - Background: Since autologous veins are unavailable when needed in more than 20% of cases in vascular surgery, the production of personalized biological vascular grafts for implantation has become crucial. Surface modification of decellularized xenogeneic grafts with vascular cells to achieve physiological luminal coverage and eventually thromboresistance is an important prerequisite for implantation. However, ex vivo thrombogenicity testing remains a neglected area in the field of tissue engineering of vascular grafts due to a multifold of reasons. Methods: After seeding decellularized bovine carotid arteries with human endothelial progenitor cells and umbilical cord-derived mesenchymal stem cells, luminal endothelial cell coverage (LECC) was correlated with glucose and lactate levels on the cell supernatant. Then a closed loop whole blood perfusion system was designed. Recellularized grafts with a LECC > 50% and decellularized vascular grafts were perfused with human whole blood for 2 h. Hemolysis and complete blood count evaluation was performed on an hourly basis, followed by histological and immunohistochemical analysis. Results: While whole blood perfusion of decellularized grafts significantly reduced platelet counts, platelet depletion from blood resulting from binding to re-endothelialized grafts was insignificant (p = 0.7284). Moreover, macroscopic evaluation revealed thrombus formation only in the lumen of unseeded grafts and histological characterization revealed lack of CD41 positive platelets in recellularized grafts, thus confirming their thromboresistance. Conclusion: In the present study we were able to demonstrate the effect of surface modification of vascular grafts in their thromboresistance in an ex vivo whole blood perfusion system. To our knowledge, this is the first study to expose engineered vascular grafts to human whole blood, recirculating at high flow rates, immediately after seeding.

AB - Background: Since autologous veins are unavailable when needed in more than 20% of cases in vascular surgery, the production of personalized biological vascular grafts for implantation has become crucial. Surface modification of decellularized xenogeneic grafts with vascular cells to achieve physiological luminal coverage and eventually thromboresistance is an important prerequisite for implantation. However, ex vivo thrombogenicity testing remains a neglected area in the field of tissue engineering of vascular grafts due to a multifold of reasons. Methods: After seeding decellularized bovine carotid arteries with human endothelial progenitor cells and umbilical cord-derived mesenchymal stem cells, luminal endothelial cell coverage (LECC) was correlated with glucose and lactate levels on the cell supernatant. Then a closed loop whole blood perfusion system was designed. Recellularized grafts with a LECC > 50% and decellularized vascular grafts were perfused with human whole blood for 2 h. Hemolysis and complete blood count evaluation was performed on an hourly basis, followed by histological and immunohistochemical analysis. Results: While whole blood perfusion of decellularized grafts significantly reduced platelet counts, platelet depletion from blood resulting from binding to re-endothelialized grafts was insignificant (p = 0.7284). Moreover, macroscopic evaluation revealed thrombus formation only in the lumen of unseeded grafts and histological characterization revealed lack of CD41 positive platelets in recellularized grafts, thus confirming their thromboresistance. Conclusion: In the present study we were able to demonstrate the effect of surface modification of vascular grafts in their thromboresistance in an ex vivo whole blood perfusion system. To our knowledge, this is the first study to expose engineered vascular grafts to human whole blood, recirculating at high flow rates, immediately after seeding.

KW - Bypass

KW - Decellularization

KW - Recellularization

KW - Vascular graft

UR - http://www.scopus.com/inward/record.url?scp=85119821321&partnerID=8YFLogxK

U2 - 10.1186/s13036-021-00277-2

DO - 10.1186/s13036-021-00277-2

M3 - Article

AN - SCOPUS:85119821321

VL - 15

JO - Journal of biological engineering

JF - Journal of biological engineering

SN - 1754-1611

IS - 1

M1 - 26

ER -