Enhanced In Vitro Antiviral Activity of Ivermectin-Loaded Nanostructured Lipid Carriers against Porcine Epidemic Diarrhea Virus via Improved Intracellular Delivery
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemical, Cells, and Viruses
2.2. Preparation of IVM-NLCs
2.3. Characterization of IVM-NLCs
2.3.1. Hydrodynamic Diameter (HD), Polydispersity Index (PDI), and Zeta Potential (ZP)
2.3.2. Transmission Electron Microscopy (TEM)
2.3.3. X-ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FT-IR)
2.3.4. Entrapment Efficiency (EE) and Drug Loading (DL)
2.4. Cell Culture
2.5. Cell Viability Evaluation
2.6. In Vitro Cellular Uptake
2.7. TCID50 Assay
2.8. Antiviral Assay
2.9. One-Step Growth Curve
2.10. RT-qPCR
2.11. Western Blot Analysis
2.12. IFA
2.13. Determination of Reactive Oxygen Species (ROS) Generation
2.14. Mitochondrial Membrane Potential (MMP) Analysis
2.15. Apoptosis Assay
2.16. Statistical Analysis
3. Results and Discussion
3.1. IVM Inhibited the Infectivity of PEDV
3.2. Effect of IVM in Diverse Stages of PEDV Life Cycle
3.3. Characterization of IVM-Loaded Nanostructured Lipid Carriers
3.4. NLCs Improved Cellular Uptake of IVM
3.5. NLCs Enhanced the Antiviral Activity of IVM against PEDV
3.6. Effect of IVM-NLCs on the Apoptosis Rate in PEDV-Infected Vero Cells
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Xu, X.; Gao, S.; Zuo, Q.; Gong, J.; Song, X.; Liu, Y.; Xiao, J.; Zhai, X.; Sun, H.; Zhang, M.; et al. Enhanced In Vitro Antiviral Activity of Ivermectin-Loaded Nanostructured Lipid Carriers against Porcine Epidemic Diarrhea Virus via Improved Intracellular Delivery. Pharmaceutics 2024, 16, 601. https://doi.org/10.3390/pharmaceutics16050601
Xu X, Gao S, Zuo Q, Gong J, Song X, Liu Y, Xiao J, Zhai X, Sun H, Zhang M, et al. Enhanced In Vitro Antiviral Activity of Ivermectin-Loaded Nanostructured Lipid Carriers against Porcine Epidemic Diarrhea Virus via Improved Intracellular Delivery. Pharmaceutics. 2024; 16(5):601. https://doi.org/10.3390/pharmaceutics16050601
Chicago/Turabian StyleXu, Xiaolin, Shasha Gao, Qindan Zuo, Jiahao Gong, Xinhao Song, Yongshi Liu, Jing Xiao, Xiaofeng Zhai, Haifeng Sun, Mingzhi Zhang, and et al. 2024. "Enhanced In Vitro Antiviral Activity of Ivermectin-Loaded Nanostructured Lipid Carriers against Porcine Epidemic Diarrhea Virus via Improved Intracellular Delivery" Pharmaceutics 16, no. 5: 601. https://doi.org/10.3390/pharmaceutics16050601