Details
| Original language | English |
|---|---|
| Pages (from-to) | 19-32 |
| Number of pages | 14 |
| Journal | BioNanoMaterials |
| Volume | 17 |
| Issue number | 1-2 |
| Early online date | 18 Apr 2016 |
| Publication status | Published - 1 May 2016 |
Abstract
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.
Keywords
- microfluidic hydrogels, PEGylated fibrin, tissue engineering, vasculogenesis
ASJC Scopus subject areas
- Chemical Engineering(all)
- Bioengineering
- Engineering(all)
- Biomedical Engineering
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In: BioNanoMaterials, Vol. 17, No. 1-2, 01.05.2016, p. 19-32.
Research output: Contribution to journal › Article › Research › peer review
}
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 -