TY - JOUR

T1 - Hydrogel-based microfluidics for vascular tissue engineering

AU - Koroleva, Anastasia

AU - Deiwick, Andrea

AU - Nguyen, Alexander

AU - Narayan, Roger

AU - Shpichka, Anastasia

AU - Kufelt, Olga

AU - Kiyan, Roman

AU - Bagratashvili, Victor

AU - Timashev, Peter

AU - Scheper, Thomas

AU - Chichkov, Boris

N1 - Funding information: The authors would like to acknowledge Dr. Daria Guseva (MHH) for the assistance in confocal microscopy imaging. Anastasia Shpichka thanks the DAAD and the Ministry of Education and Science of the Russian Federation (Michail Lomonosov program) for the research scholarship. This work was supported by the DFG Cluster of Excellence REBIRTH and Low Saxony project Biofabrication for Nife. Grant of the Government of the Russian Federation for the Support of Scientific Investigations under the Supervision of Leading Scientists Contract No. 14.B25.31.0019 is also acknowledged.

PY - 2016/5/1

Y1 - 2016/5/1

N2 - In this work, we have explored 3-D co-culture of vasculogenic cells within a synthetically modified fibrin hydrogel. Fibrinogen was covalently linked with PEG-NHS in order to improve its degradability resistance and physico-optical properties. We have studied influences of the degree of protein PEGylation and the concentration of enzyme thrombin used for the gel preparation on cellular responses. Scanning electron microscopy analysis of prepared gels revealed that the degree of PEGylation and the concentration of thrombin strongly influenced microstructural characteristics of the protein hydrogel. Human umbilical vein endothelial cells (HUVECs) and human adipose-derived stem cells (hASCs), used as vasculogenic co-culture, could grow in 5:1 PEGylated fibrin gels prepared using 1:0.2 protein to thrombin ratio. This gel formulation supported hASCs and HUVECs spreading and the formation of cell extensions and cell-to-cell contacts. Expression of specific ECM proteins and vasculogenic process inherent cellular enzymatic activity were investigated by immunofluorescent staining, gelatin zymography, western blot and RT-PCR analysis. After evaluation of the optimal gel composition and PEGylation ratio, the hydrogel was utilized for investigation of vascular tube formation within a perfusable microfluidic system. The morphological development of this co-culture within a perfused hydrogel over 12 days led to the formation of interconnected HUVEC-hASC network. The demonstrated PEGylated fibrin microfluidic approach can be used for incorporating other cell types, thus representing a unique experimental platform for basic vascular tissue engineering and drug screening applications.

AB - In this work, we have explored 3-D co-culture of vasculogenic cells within a synthetically modified fibrin hydrogel. Fibrinogen was covalently linked with PEG-NHS in order to improve its degradability resistance and physico-optical properties. We have studied influences of the degree of protein PEGylation and the concentration of enzyme thrombin used for the gel preparation on cellular responses. Scanning electron microscopy analysis of prepared gels revealed that the degree of PEGylation and the concentration of thrombin strongly influenced microstructural characteristics of the protein hydrogel. Human umbilical vein endothelial cells (HUVECs) and human adipose-derived stem cells (hASCs), used as vasculogenic co-culture, could grow in 5:1 PEGylated fibrin gels prepared using 1:0.2 protein to thrombin ratio. This gel formulation supported hASCs and HUVECs spreading and the formation of cell extensions and cell-to-cell contacts. Expression of specific ECM proteins and vasculogenic process inherent cellular enzymatic activity were investigated by immunofluorescent staining, gelatin zymography, western blot and RT-PCR analysis. After evaluation of the optimal gel composition and PEGylation ratio, the hydrogel was utilized for investigation of vascular tube formation within a perfusable microfluidic system. The morphological development of this co-culture within a perfused hydrogel over 12 days led to the formation of interconnected HUVEC-hASC network. The demonstrated PEGylated fibrin microfluidic approach can be used for incorporating other cell types, thus representing a unique experimental platform for basic vascular tissue engineering and drug screening applications.

KW - microfluidic hydrogels

KW - PEGylated fibrin

KW - tissue engineering

KW - vasculogenesis

UR - http://www.scopus.com/inward/record.url?scp=84991518255&partnerID=8YFLogxK

U2 - 10.1515/bnm-2015-0026

DO - 10.1515/bnm-2015-0026

M3 - Article

AN - SCOPUS:84991518255

VL - 17

SP - 19

EP - 32

JO - BioNanoMaterials

JF - BioNanoMaterials

SN - 2193-0651

IS - 1-2

ER -