Influence of PEG Chain Length of Functionalized Magnetic Nanoparticles on the Cytocompatibility and Immune Competence of Primary Murine Macrophages and Dendritic Cells
Abstract
:1. Introduction
2. Results
2.1. Characterization of Unfunctionalized and Functionalized MNPSNPs
2.2. Cytotoxicity of Functionalized MNPSNPs
2.3. Effect of Functionalized MNPSNPs on Cytokine Release
2.4. Effect of Functionalized MNPSNPs on the Formation of Reactive Oxygen Species
2.5. Impact of Functionalized MNPSNPs on the Cell Migration of DCs
2.6. Impact of Functionalized MNPSNPs on the Immune Competence of Macrophages
2.7. Intracellular Uptake of Functionalized MNPSNPs
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Synthesis of Magnetic Nanoporous Silica Nanoparticles (MNPSNPs) and Functionalization with mPEG
4.2.1. Synthesis of MNPSNPs
4.2.2. Synthesis of mPEG Silanes with Different Molecular Weights
4.2.3. Functionalization of MNPSNPs with mPEG and FITC
4.3. Characterizations
4.4. Animals
4.5. Generation of Primary Immune Cells
4.6. Cell Viability Assay
4.7. Determination of Cytokine Release
4.8. Measurement of Reactive Oxygen Species
4.9. Migration Assay of Dendritic Cells
4.10. Evaluation of the Immune Competence of Primary Macrophages
4.11. In Vitro Cellular Uptake Assay
4.12. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
Appendix B
MTS DC 24 h 2 kDa mPEG | 10% aqua bidestillata | 15 µg/mL 2 kDa mPEG | 50 µg/mL 2 kDa mPEG | 150 µg/mL 2 kDa mPEG | 5% DMSO | Background: 150 µg/mL 2 kDa mPEG | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.446 | 0.507 | 0.522 | 0.543 | 0.179 | 0.136 | / |
2 | 0.466 | 0.538 | 0.540 | 0.574 | 0.176 | 0.147 | / | |
3 | 0.901 | 1.006 | 1.055 | 1.065 | 0.185 | 0.139 | 0.095 | |
4 | 0.829 | 0.889 | 0.919 | 0.929 | 0.171 | 0.133 | 0.092 | |
5 | 0.517 | 0.562 | 0.587 | 0.575 | 0.175 | 0.122 | 0.069 | |
6 | 0.567 | 0.628 | 0.666 | 0.697 | 0.158 | 0.130 | 0.067 |
MTS DC 48 h 2 kDa mPEG | 10% aqua bidestillata | 15 µg/mL 2 kDa mPEG | 50 µg/mL 2 kDa mPEG | 150 µg/mL 2 kDa mPEG | 5% DMSO | Background: 150 µg/mL 2 kDa mPEG | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.390 | 0.525 | 0.542 | 0.616 | 0.239 | 0.135 | / |
2 | 0.373 | 0.474 | 0.550 | 0.595 | 0.211 | 0.123 | / | |
3 | 0.772 | 0.829 | 0.909 | 0.966 | 0.212 | 0.139 | 0.094 | |
4 | 0.682 | 0.763 | 0.810 | 0.884 | 0.175 | 0.109 | 0.092 | |
5 | 0.471 | 0.560 | 0.636 | 0.653 | 0.180 | 0.115 | 0.102 | |
6 | 0.603 | 0.679 | 0.790 | 0.847 | 0.179 | 0.120 | 0.099 |
MTS DC 24 h 5 kDa mPEG | 10% aqua bidestillata | 15 µg/mL 5 kDa mPEG | 50 µg/mL 5 kDa mPEG | 150 µg/mL 5 kDa mPEG | 5% DMSO | Background: 150 µg/mL 5 kDa mPEG | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.446 | 0.492 | 0.509 | 0.507 | 0.179 | 0.125 | / |
2 | 0.466 | 0.535 | 0.569 | 0.573 | 0.176 | 0.129 | / | |
3 | 0.901 | 0.995 | 1.030 | 1.082 | 0.185 | 0.126 | 0.095 | |
4 | 0.829 | 0.876 | 0.905 | 0.923 | 0.171 | 0.114 | 0.092 | |
5 | 0.517 | 0.613 | 0.599 | 0.597 | 0.175 | 0.114 | 0.069 | |
6 | 0.567 | 0.659 | 0.664 | 0.664 | 0.158 | 0.124 | 0.067 |
MTS DC 48 h 5 kDa mPEG | 10% aqua bidestillata | 15 µg/mL 5 kDa mPEG | 50 µg/mL 5 kDa mPEG | 150 µg/mL 5 kDa mPEG | 5% DMSO | Background: 150 µg/mL 5 kDa mPEG | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.390 | 0.506 | 0.540 | 0.591 | 0.239 | 0.125 | / |
2 | 0.373 | 0.509 | 0.563 | 0.613 | 0.211 | 0.124 | / | |
3 | 0.772 | 0.834 | 0.868 | 0.955 | 0.212 | 0.126 | 0.094 | |
4 | 0.682 | 0.736 | 0.792 | 0.864 | 0.175 | 0.104 | 0.092 | |
5 | 0.471 | 0.592 | 0.640 | 0.641 | 0.180 | 0.113 | 0.102 | |
6 | 0.603 | 0.725 | 0.778 | 0.800 | 0.179 | 0.119 | 0.099 |
MTS DC 24 h 10 kDa mPEG-FITC | 10% aqua bidestillata | 15 µg/mL 10 kDa mPEG-FITC | 50 µg/mL 10 kDa mPEG-FITC | 150 µg/mL 10 kDa mPEG-FITC | 5% DMSO | Background: 150 µg/mL 10 kDa mPEG-FITC | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.446 | 0.484 | 0.538 | 0.626 | 0.179 | 0.201 | / |
2 | 0.466 | 0.492 | 0.541 | 0.633 | 0.176 | 0.200 | / | |
3 | 0.901 | 0.968 | 1.007 | 1.084 | 0.185 | 0.211 | 0.095 | |
4 | 0.829 | 0.870 | 0.898 | 0.963 | 0.171 | 0.184 | 0.092 | |
5 | 0.517 | 0.575 | 0.610 | 0.671 | 0.175 | 0.206 | 0.069 | |
6 | 0.567 | 0.614 | 0.681 | 0.769 | 0.158 | 0.199 | 0.067 |
MTS DC 48 h 10 kDa mPEG-FITC | 10% aqua bidestillata | 15 µg/mL 10 kDa mPEG-FITC | 50 µg/mL 10 kDa mPEG-FITC | 150 µg/mL 10 kDa mPEG-FITC | 5% DMSO | Background: 150 µg/mL 10 kDa mPEG-FITC | Background: MTS | |
---|---|---|---|---|---|---|---|---|
Biological replicate | 1 | 0.390 | 0.493 | 0.584 | 0.703 | 0.239 | 0.222 | / |
2 | 0.373 | 0.490 | 0.579 | 0.692 | 0.211 | 0.175 | / | |
3 | 0.772 | 0.795 | 0.860 | 0.962 | 0.212 | 0.211 | 0.094 | |
4 | 0.682 | 0.729 | 0.770 | 0.884 | 0.175 | 0.177 | 0.092 | |
5 | 0.471 | 0.580 | 0.640 | 0.759 | 0.180 | 0.206 | 0.102 | |
6 | 0.603 | 0.752 | 0.808 | 0.975 | 0.179 | 0.191 | 0.099 |
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Functionalization | Blank MNPSNP | 2 kDa mPEG-FITC | 5 kDa mPEG-FITC | 10 kDa mPEG-FITC |
---|---|---|---|---|
BET [m2·g−1] | 910 | 730 | 610 | 640 |
d [nm] | 3.6 | 3.4 | 3.0 | 3.4 |
V [cm3 g−1] | 0.7 | 0.5 | 0.3 | 0.4 |
ZP [mV] | −30 | −38 | −35 | −36 |
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Storjohann, R.; Gericke, B.; Reifenrath, J.; Herrmann, T.; Behrens, P.; Oltmanns, H.; Meißner, J. Influence of PEG Chain Length of Functionalized Magnetic Nanoparticles on the Cytocompatibility and Immune Competence of Primary Murine Macrophages and Dendritic Cells. Int. J. Mol. Sci. 2023, 24, 2565. https://doi.org/10.3390/ijms24032565
Storjohann R, Gericke B, Reifenrath J, Herrmann T, Behrens P, Oltmanns H, Meißner J. Influence of PEG Chain Length of Functionalized Magnetic Nanoparticles on the Cytocompatibility and Immune Competence of Primary Murine Macrophages and Dendritic Cells. International Journal of Molecular Sciences. 2023; 24(3):2565. https://doi.org/10.3390/ijms24032565
Chicago/Turabian StyleStorjohann, Ronja, Birthe Gericke, Janin Reifenrath, Timo Herrmann, Peter Behrens, Hilke Oltmanns, and Jessica Meißner. 2023. "Influence of PEG Chain Length of Functionalized Magnetic Nanoparticles on the Cytocompatibility and Immune Competence of Primary Murine Macrophages and Dendritic Cells" International Journal of Molecular Sciences 24, no. 3: 2565. https://doi.org/10.3390/ijms24032565