One-Step Pharmaceutical Preparation of PEG-Modified Exosomes Encapsulating Anti-Cancer Drugs by a High-Pressure Homogenization Technique
Abstract
:1. Introduction
2. Results
2.1. Influence of High-Pressure Homogenization Using a Microfluidizer on Physicochemical Properties of Exosomes
2.2. Effects of HPH on Expression of Representative Exosomal Marker Protein and Cellular Uptake of RAW-Exos
2.3. PEG Modification onto RAW-Exos by HPH
2.4. Encapsulation of the Anti-Cancer Drug Doxorubicin into RAW-Exos by HPH
2.5. Anti-Proliferative Effects of DOX-Encapsulated RAW-Exos on Colon-26 Cells
2.6. Suppression of Tumor Growth by Treatment with DOX-PEG-RAW-Exos
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Isolation of Exosomes
4.3. Measurement of Physicochemical Properties and Protein Concentration of RAW-Exos
4.4. High-Pressure Homogenization (HPH) with a Microfluidizer
4.5. Cryo-Transmission Electron Microscopy
4.6. Western Blotting
4.7. Cellular Uptake of RAW-Exos
4.8. Modification of RAW-Exos with PEG-Lipid
4.9. Measurement of Sample Temperature after HPH
4.10. Doxorubicin Encapsulation into RAW-Exos by HPH
4.11. In Vitro Cytotoxicity Assay
4.12. In Vivo Anti-Cancer Experiments
4.13. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample | Particle Size (d.nm) | Polydispersity Index (PDI) | ζ-Potential (mV) |
---|---|---|---|
RAW-Exos | 150.1 ± 4.8 | 0.25 ± 0.03 | −30.0 ± 2.8 |
PEG-RAW-Exos (Incubation) | 125.6 ± 12.2 | 0.20 ± 0.05 | −29.4 ± 5.9 |
PEG-RAW-Exos (HPH) | 113.2 ± 27.6 | 0.29 ± 0.04 | −17.4 ± 1.6 |
Process Pressure (psi) | Before (°C) | After (°C) | Degree of Temperature Change (°C) |
---|---|---|---|
20,000 | 9.6 ± 1.5 | 33.4 ± 0.4 | 24.4 ± 1.8 |
30,000 | 12.0 ± 2.2 | 44.7 ± 0.9 | 32.7 ± 2.8 *** |
Sample | Particle Size (d.nm) | Polydispersity Index (PDI) | ζ-Potential (mV) |
---|---|---|---|
DOX-RAW-Exos (20,000 psi) | 1222.7 ± 187.0 | 0.38 ± 0.15 | −19.8 ± 7.2 |
DOX-RAW-Exos (30,000 psi) | 887.2 ± 445.6 | 0.49 ± 0.16 | −21.3 ± 3.8 |
DOX-PEG-RAW-Exos | 110.8 ± 12.4 | 0.35 ± 0.04 | −27.5 ± 1.3 |
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Fukuta, T.; Ikeda-Imafuku, M.; Kodama, S.; Kuse, J.; Matsui, K.; Iwao, Y. One-Step Pharmaceutical Preparation of PEG-Modified Exosomes Encapsulating Anti-Cancer Drugs by a High-Pressure Homogenization Technique. Pharmaceuticals 2023, 16, 108. https://doi.org/10.3390/ph16010108
Fukuta T, Ikeda-Imafuku M, Kodama S, Kuse J, Matsui K, Iwao Y. One-Step Pharmaceutical Preparation of PEG-Modified Exosomes Encapsulating Anti-Cancer Drugs by a High-Pressure Homogenization Technique. Pharmaceuticals. 2023; 16(1):108. https://doi.org/10.3390/ph16010108
Chicago/Turabian StyleFukuta, Tatsuya, Mayumi Ikeda-Imafuku, Satoshi Kodama, Junko Kuse, Ko Matsui, and Yasunori Iwao. 2023. "One-Step Pharmaceutical Preparation of PEG-Modified Exosomes Encapsulating Anti-Cancer Drugs by a High-Pressure Homogenization Technique" Pharmaceuticals 16, no. 1: 108. https://doi.org/10.3390/ph16010108