Oral Delivery of mRNA by Liposomes Functionalized with Cell-Penetrating Peptides
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Methods
2.2.1. Liposome Preparation by Thin-Film Hydration Method
2.2.2. Size and Size Distribution
2.2.3. Zeta Potential
2.2.4. Cryo-Transmission Electron Microscopy (Cryo-TEM)
2.2.5. Encapsulation Efficiency
2.2.6. Long-Term Storage Stability by Freeze-Drying
2.2.7. In Vitro Studies
Cell Culture
- Thawing Cells
- Cultivating Cells
- Cell Counting
Cytotoxicity Studies
Cellular Uptake of Liposomes
3. Results
3.1. Liposome Preparation and Characterization
3.2. Structure, Lamellarity, Encapsulation Efficiency, and Stability in Simulated Gastric and Intestinal Fluids
3.3. Long-Term Storge Stability by Freeze-Drying
3.4. Cytotoxicity of Liposomal Formulations
3.5. Uptake in Caco-2 Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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[mol%] | Control | DOTAP | CPP | GCTE | CPP-GCTE |
---|---|---|---|---|---|
Lecithin | 90 | 85 | 89 | 85 | 84 |
Cholesterol | 10 | 10 | 10 | 10 | 10 |
DOTAP | 0 | 5 | 0 | 0 | 0 |
CPP | 0 | 0 | 1 | 0 | 1 |
GCTE | 0 | 0 | 0 | 5 | 5 |
Lipid mass [mg/mL] | 72.28 | 72.20 | 74.44 | 75.76 | 77.92 |
Parameter | Setting |
---|---|
Number of measurements | 3 |
Run duration | 10 s |
Number of runs | 10 |
Equilibration time | 60 s |
Refractive index solvent | 1.330 |
Refractive index polystyrene cuvette | 1.590 |
Viscosity | 0.8872 mPa∙s |
Temperature | 25 °C |
Dielectric constant | 78.5 F/m |
Backscattering mode | 173° |
Voltage selection | Automatic |
Equation | Smoluchowski equation |
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Dürr, V.; Wohlfart, S.; Eisenzapf, T.; Mier, W.; Fricker, G.; Uhl, P. Oral Delivery of mRNA by Liposomes Functionalized with Cell-Penetrating Peptides. Appl. Nano 2023, 4, 293-308. https://doi.org/10.3390/applnano4040017
Dürr V, Wohlfart S, Eisenzapf T, Mier W, Fricker G, Uhl P. Oral Delivery of mRNA by Liposomes Functionalized with Cell-Penetrating Peptides. Applied Nano. 2023; 4(4):293-308. https://doi.org/10.3390/applnano4040017
Chicago/Turabian StyleDürr, Valerie, Sabrina Wohlfart, Tom Eisenzapf, Walter Mier, Gert Fricker, and Philipp Uhl. 2023. "Oral Delivery of mRNA by Liposomes Functionalized with Cell-Penetrating Peptides" Applied Nano 4, no. 4: 293-308. https://doi.org/10.3390/applnano4040017