TY - JOUR
T1 - EndOxy
T2 - Mid-term stability and shear stress resistance of endothelial cells on PDMS gas exchange membranes
AU - Hellmann, Ariane
AU - Klein, Sarah
AU - Hesselmann, Felix
AU - Djeljadini, Suzana
AU - Schmitz-Rode, Thomas
AU - Jockenhoevel, Stefan
AU - Cornelissen, Christian G.
AU - Thiebes, Anja Lena
N1 - Funding Information:
This study was funded by the Aachen Interdisciplinary Center for Clinical Research (IZKF) at RWTH Aachen University Hospital (project T12) and by the Excellence Initiative of the German Federal and State Governments in the framework of the i3tm Seed Fund and STEP2 projects. We thank the Clinic for Gynaecology and Obstetrics, RWTH Aachen University Hospital, headed by Univ.‐Prof. Dr med Elmar Stickeler for providing umbilical cords. The authors thank Clara Sabine Grezella and Sophia Halbe (both BioTex) for technical assistance.
Funding Information:
This study was funded by the Aachen Interdisciplinary Center for Clinical Research (IZKF) at RWTH Aachen University Hospital (project T12) and by the Excellence Initiative of the German Federal and State Governments in the framework of the i3tm Seed Fund and STEP2 projects. We thank the Clinic for Gynaecology and Obstetrics, RWTH Aachen University Hospital, headed by Univ.-Prof. Dr med Elmar Stickeler for providing umbilical cords. The authors thank Clara Sabine Grezella and Sophia Halbe (both BioTex) for technical assistance.
Publisher Copyright:
© 2020 The Authors. Artificial Organs published by International Center for Artificial Organ and Transplantation (ICAOT) and Wiley Periodicals LLC
PY - 2020/10
Y1 - 2020/10
N2 - Endothelialized oxygenator devices (EndOxy) with a physiological, nonthrombogenic, and anti-inflammatory surface offer the potential to overcome current shortcomings of conventional extracorporeal membrane oxygenation such as complications like thromboembolism and bleeding that deteriorate adequate long-term hemocompatibility. The approach of endothelialization of gas exchange membranes, and thus the formation of a nonthrombogenic and anti-inflammatory surface, is promising. In this study, we investigated the mid-term shear stress resistance as well as gas transfer rates and cell densities of endothelial cells seeded on RGD-conjugated polydimethylsiloxane (RGD-PDMS) gas exchange membranes under dynamic conditions. Human umbilical vein endothelial cells were seeded on RGD-PDMS and exposed to defined shear stresses in a microfluidic bioreactor. Endothelial cell morphology was assessed by bright field microscopy and immunocytochemistry. Furthermore, gas transfer measurement of blank, RGD-conjugated, and endothelialized PDMS oxygenator membranes was performed. RGD-PDMS gas exchange membranes proved suitable for the dynamic culture of endothelial cells for up to 21 days at a wall shear stress of 2.9 dyn/cm(2). Furthermore, the cells resisted increased wall shear stresses up to 8.6 dyn/cm(2) after a previous dynamic preculture of each one hour at 2.9 dyn/cm(2) and 5.7 dyn/cm(2). Also, after a longer dynamic preculture of three days at 2.9 dyn/cm(2) and one hour at 5.7 dyn/cm(2), increased wall shear stresses of 8.6 dyn/cm(2) were tolerated by the cells and cell integrity could be remained. Gas transfer (GT) tests revealed that neither RGD conjugation nor endothelialization of RGD-PDMS significantly decrease the gas transfer rates of the membranes during short-term trials. Gas transfer rates are stable for at least 72 hours of dynamic cultivation of endothelial cells. Immunocytochemistry showed that the cell layer stained positive for typical endothelial cell markers CD31 and von Willebrand factor (VWF) after all trials. Cell density of EC on RGD-PDMS increased between 3 and 21 days of dynamic culture. In this study, we show the suitability of RGD-PDMS membranes for flow resistant endothelialization of gas-permeable membranes, demonstrating the feasibility of this approach for a biohybrid lung.
AB - Endothelialized oxygenator devices (EndOxy) with a physiological, nonthrombogenic, and anti-inflammatory surface offer the potential to overcome current shortcomings of conventional extracorporeal membrane oxygenation such as complications like thromboembolism and bleeding that deteriorate adequate long-term hemocompatibility. The approach of endothelialization of gas exchange membranes, and thus the formation of a nonthrombogenic and anti-inflammatory surface, is promising. In this study, we investigated the mid-term shear stress resistance as well as gas transfer rates and cell densities of endothelial cells seeded on RGD-conjugated polydimethylsiloxane (RGD-PDMS) gas exchange membranes under dynamic conditions. Human umbilical vein endothelial cells were seeded on RGD-PDMS and exposed to defined shear stresses in a microfluidic bioreactor. Endothelial cell morphology was assessed by bright field microscopy and immunocytochemistry. Furthermore, gas transfer measurement of blank, RGD-conjugated, and endothelialized PDMS oxygenator membranes was performed. RGD-PDMS gas exchange membranes proved suitable for the dynamic culture of endothelial cells for up to 21 days at a wall shear stress of 2.9 dyn/cm(2). Furthermore, the cells resisted increased wall shear stresses up to 8.6 dyn/cm(2) after a previous dynamic preculture of each one hour at 2.9 dyn/cm(2) and 5.7 dyn/cm(2). Also, after a longer dynamic preculture of three days at 2.9 dyn/cm(2) and one hour at 5.7 dyn/cm(2), increased wall shear stresses of 8.6 dyn/cm(2) were tolerated by the cells and cell integrity could be remained. Gas transfer (GT) tests revealed that neither RGD conjugation nor endothelialization of RGD-PDMS significantly decrease the gas transfer rates of the membranes during short-term trials. Gas transfer rates are stable for at least 72 hours of dynamic cultivation of endothelial cells. Immunocytochemistry showed that the cell layer stained positive for typical endothelial cell markers CD31 and von Willebrand factor (VWF) after all trials. Cell density of EC on RGD-PDMS increased between 3 and 21 days of dynamic culture. In this study, we show the suitability of RGD-PDMS membranes for flow resistant endothelialization of gas-permeable membranes, demonstrating the feasibility of this approach for a biohybrid lung.
KW - biohybrid lung
KW - extracorporeal membrane oxygenation
KW - human umbilical vein endothelial cells
KW - polydimethylsiloxane
KW - tissue engineering
KW - VON-WILLEBRAND-FACTOR
KW - OXYGENATION USE
KW - LUNG
KW - PLATELETS
KW - ADHESION
KW - ECMO
KW - RGD
U2 - 10.1111/aor.13712
DO - 10.1111/aor.13712
M3 - Article
C2 - 32320079
SN - 0160-564X
VL - 44
SP - E419-E433
JO - Artificial Organs
JF - Artificial Organs
IS - 10
ER -