Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings—Targeting NF-κB Signalling Pathway
Abstract
:1. Introduction
2. Results
2.1. Characterization of Physical Properties of Coatings
2.2. Adhesion of Macrophages and Multinucleated Giant Cell Formation
2.3. IL-1β Pro-Inflammatory Cytokine Release
2.4. Immunofluorescence Staining of NF-κB in Macrophages
2.5. Western Blotting
2.6. Association of GAG with Macrophages Studied by Confocal Laser Scanning Microscopy (CLSM)
2.7. Association of FITC-Labelled GAG with Macrophages Studied by Flow Cytometry
3. Discussion
4. Materials and Methods
4.1. Chemicals for Surface Modification
4.2. Substrates and Polyelectrolyte (PEL) Preparation
4.3. Polyelectrolyte Multilayers (PEMs) Formation
4.4. Characterization of Surface Properties and Multilayer Formation
4.4.1. Scanning Electron Microscopy (SEM)
4.4.2. Atomic Force Microscopy (AFM)
4.4.3. Water Contact Angle (WCA)
4.4.4. Measurement of Multilayer Thickness by Ellipsometry
4.5. Studies with THP-1 Derived Macrophages
4.5.1. Cell Culture
4.5.2. Cell Adhesion Studies
4.5.3. Analysis of Multinucleated Giant Cells (MNGCs) Formation
4.5.4. IL-1β Production Measurement
4.5.5. Immunofluorescence (IF) Staining of NF-kB
4.5.6. Cell Lysis for Immunoblotting (IB)
4.5.7. SDS-PAGE and Western Blotting
4.5.8. Association of FITC-GAG with Macrophages Studied by Confocal Laser Scanning Microscopy
4.5.9. Uptake of FITC-GAG by Macrophages Studied with Flow Cytometry
4.6. Statistics
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Alkhoury, H.; Hautmann, A.; Fuhrmann, B.; Syrowatka, F.; Erdmann, F.; Zhou, G.; Stojanović, S.; Najman, S.; Groth, T. Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings—Targeting NF-κB Signalling Pathway. Int. J. Mol. Sci. 2020, 21, 3724. https://doi.org/10.3390/ijms21103724
Alkhoury H, Hautmann A, Fuhrmann B, Syrowatka F, Erdmann F, Zhou G, Stojanović S, Najman S, Groth T. Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings—Targeting NF-κB Signalling Pathway. International Journal of Molecular Sciences. 2020; 21(10):3724. https://doi.org/10.3390/ijms21103724
Chicago/Turabian StyleAlkhoury, Hala, Adrian Hautmann, Bodo Fuhrmann, Frank Syrowatka, Frank Erdmann, Guoying Zhou, Sanja Stojanović, Stevo Najman, and Thomas Groth. 2020. "Studies on the Mechanisms of Anti-Inflammatory Activity of Heparin- and Hyaluronan-Containing Multilayer Coatings—Targeting NF-κB Signalling Pathway" International Journal of Molecular Sciences 21, no. 10: 3724. https://doi.org/10.3390/ijms21103724