Layered Double Hydroxide as a Potent Non-viral Vector for Nucleic Acid Delivery Using Gene-Activated Scaffolds for Tissue Regeneration Applications
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of MgAl-NO3 LDH Nanoparticles
2.2. Complexation of LDH with Nucleic Acid (pDNA, siRNA, miRNA)
2.2.1. Plasmid DNA (pDNA) Propagation
2.2.2. Physicochemical Nanoparticle Characterization
2.2.3. Nucleic Acid Encapsulation
2.3. Mesenchymal Stromal Cell Uptake and Cytocompatibility of LDH–Nucleic Acid Complexes in 2D Monolayer
2.3.1. Cellular Uptake
2.3.2. LDH–Nucleic Acid Cytocompatibility
2.3.3. Assessment of LDH–Nucleic Acid Nanoparticle Efficacy in 2D
2.4. LDH–Nanoparticle Activated Collagen-Nanohydroxyapatite Scaffold Systems
2.5. Confocal Imaging
2.6. Scanning Electron Microscopy
2.7. Statistical Analysis
3. Results
3.1. Efficiency of LDH–pDNA Nanoparticle Complexation
3.2. Efficiency of LDH–siRNA Complexation and Encapsulation
3.3. Efficiency of LDH–miRNA Complexation and Encapsulation
3.4. Uptake and Cytocompatibility of LDH–pDNA Nanoparticles in Monolayer Transfection
3.5. Uptake and Cytocompatibility of LDH–SiRNA Nanoparticles in Monolayer Transfection
3.6. Uptake and Cytocompatibility of LDH–MiRNA Mimic and LDH–MiRNA Inhibitor Nanoparticles in Monolayer Transfection
3.7. SiRNA Delivery from Gene-Activated Collagen-Nanohydroxyapatite Scaffolds
3.8. MicroRNA Delivery from Gene-Activated Collagen-Nanohydroxyapatite Scaffolds
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
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Costard, L.S.; Kelly, D.C.; Power, R.N.; Hobbs, C.; Jaskaniec, S.; Nicolosi, V.; Cavanagh, B.L.; Curtin, C.M.; O’Brien, F.J. Layered Double Hydroxide as a Potent Non-viral Vector for Nucleic Acid Delivery Using Gene-Activated Scaffolds for Tissue Regeneration Applications. Pharmaceutics 2020, 12, 1219. https://doi.org/10.3390/pharmaceutics12121219
Costard LS, Kelly DC, Power RN, Hobbs C, Jaskaniec S, Nicolosi V, Cavanagh BL, Curtin CM, O’Brien FJ. Layered Double Hydroxide as a Potent Non-viral Vector for Nucleic Acid Delivery Using Gene-Activated Scaffolds for Tissue Regeneration Applications. Pharmaceutics. 2020; 12(12):1219. https://doi.org/10.3390/pharmaceutics12121219
Chicago/Turabian StyleCostard, Lara S., Domhnall C. Kelly, Rachael N. Power, Christopher Hobbs, Sonia Jaskaniec, Valeria Nicolosi, Brenton L. Cavanagh, Caroline M. Curtin, and Fergal J. O’Brien. 2020. "Layered Double Hydroxide as a Potent Non-viral Vector for Nucleic Acid Delivery Using Gene-Activated Scaffolds for Tissue Regeneration Applications" Pharmaceutics 12, no. 12: 1219. https://doi.org/10.3390/pharmaceutics12121219