Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application
Abstract
:1. Introduction
2. Results and Discussion
2.1. X-ray Diffraction Analysis of the Nanocomposites
2.2. Determination of Size Distribution and Zeta Potential Measurement of Nanocomposites
2.3. Quantification of Dug Loading and Encapsulation Efficacy
2.4. Surface Properties Analysis
2.5. Protocatechuic Acid and Chlorogenic Acid Drug Release Study of Nanocomposites
2.6. Cell Viability Effect of Nanocomposites on Normal and Cancer Cells
2.7. Plasma Membrane Integrity Analysis
2.8. Morphological Observation of Nanocomposite Cellular Uptake
2.9. In Vitro Localization and Accumulation of Nanocomposite
2.10. Apoptosis Induced by Nanocomoposites in HepG2 Cells via Annexin V-FITC/Propidium Iodide (PI) Staining Analysis
2.11. The Effect of Nanocomposite on Cell Cycle Distribution in HepG2 Cells
2.12. Nanocomposite Stimulates Mitochondrial Membrane Potential in HepG2 Cells
2.13. Nanocomposite Induces Intracellular Reactive Oxygen Species Generation in HepG2 Cells
3. Materials and Methods
3.1. Materials
3.2. Cell Lines
3.3. Synthesis of Graphene Oxide and Conjugation with Polyethylene Glycol
3.4. Protocatechuic Acid and Chlorogenic Acid Loading on Graphene Oxide with PEG and Coated with Folic Acid
3.5. Synthesis of FITC-Labelled GOP-PCA/CA-FA
3.6. Physicochemical Characterization of Nanocomposites
3.7. Encapsulation Efficacy and Drug Loading Content Analysis
3.8. In Vitro Drug Release of Protocatechuic Acid and Chlorogenic Acid from Nanocomposites
3.9. In Vitro Cell Cytotoxicity Assay
3.10. Lactate Dehydrogenase Assay for Plasma Membrane Integrity Analysis
3.11. Morphological Observation of Nanocomposite Cellular Uptake
3.12. In Vitro Localization of Nanocomposite by Fluorescent Microscopy Analysis
3.13. Apoptosis Cell Death Analysis
3.14. Cell Cycle Arrest Analysis Using Propidium Iodide
3.15. Mitochondrial Membrane Potential Analysis
3.16. Measurement of Cellular Reactive Oxygen Species
3.17. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Nanocomposites | ζ-Potential (mV) | ||
---|---|---|---|
Deionized Water | Eagle’s Minimum Essential Medium | Eagle’s Minimum Essential Medium + 10% FBS | |
GOP | −8.99 ± 2.021 | −14.15 ± 1.224 | −6.42 ± 1.302 |
GOP-PCA/CA | −17.10 ± 1.072 | −22.19 ± 1.151 | −15.44 ± 1.932 |
GOP-PCA/CA-FA | −22.72 ± 2.311 | −29.28 ± 1.618 | −19.76 ± 2.041 |
Nanocomposites | Loading Content (%) | Encapsulation Efficiency (%) | ||
---|---|---|---|---|
PCA | CA | PCA | CA | |
GOP-PCA/CA | 23.82 | 19.55 | 75.23 | 72.11 |
GOP-PCA/CA-FA | 24.47 | 23.33 | 79.16 | 75.15 |
Nanocomposites IC50 (μg/mL) | HDFa Dermal Fibroblast Cell | HepG2 Cell |
---|---|---|
Graphene oxide-PEG (GOP) | N.C | N.C |
Protocatechuic acid (PCA) | N.C | 40.78 ± 1.92 |
Chlorogenic acid (CA) | N.C | 43.61 ± 1.74 |
GOP-PCA/CA | N.C | 34.73 ± 1.04 |
GOP-PCA/CA-FA | N.C | 26.79 ± 1.63 |
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Buskaran, K.; Hussein, M.Z.; Moklas, M.A.M.; Masarudin, M.J.; Fakurazi, S. Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application. Int. J. Mol. Sci. 2021, 22, 5786. https://doi.org/10.3390/ijms22115786
Buskaran K, Hussein MZ, Moklas MAM, Masarudin MJ, Fakurazi S. Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application. International Journal of Molecular Sciences. 2021; 22(11):5786. https://doi.org/10.3390/ijms22115786
Chicago/Turabian StyleBuskaran, Kalaivani, Mohd Zobir Hussein, Mohamad Aris Mohd Moklas, Mas Jaffri Masarudin, and Sharida Fakurazi. 2021. "Graphene Oxide Loaded with Protocatechuic Acid and Chlorogenic Acid Dual Drug Nanodelivery System for Human Hepatocellular Carcinoma Therapeutic Application" International Journal of Molecular Sciences 22, no. 11: 5786. https://doi.org/10.3390/ijms22115786