Direct Cardiac Epigenetic Reprogramming through Codelivery of 5′Azacytidine and miR-133a Nanoformulation
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis and Characterization of PEI-miR-133a, PLGA, PLGA-AZA, PLGA-PEI-miR-133a, PEI-miR-133a-AZA PLGA Nanoparticles
2.2. Encapsulation and Stability of PEI-miR-133a Polyplexes
2.3. Invitro Release Studies
2.4. Cellular Uptake
2.5. Cytocompatibility and Cell Viability of PLGA NPs
2.6. Live/Dead Cytotoxicity Analysis
2.7. cTnT Expression and Global Methylation Analysis
3. Materials and Methods
3.1. Materials
3.2. Methods
3.2.1. miRNA Polyplexes Preparation and Gel Retardation Assay
3.2.2. Synthesis and Characterization of PEI-miR-133a, PLGA AZA, and PLGA-PEI-miRNA-AZA
3.2.3. Physico-Chemical Characterization
3.2.4. Percentage Yield and Encapsulation Efficiency
3.2.5. Gel Electrophoresis Assay to Confirm Encapsulation and Stability of miRNA Polyplexes to PLGA Nanoparticles
3.2.6. Invitro miRNA/Drug Release
3.2.7. Cell Culture
3.2.8. PLGA-PEI-miR133a Transfection
3.2.9. PrestoBlue Cell Viability Assay
3.2.10. Live/Dead Cell Cytotoxicity Assay
3.2.11. Cellular Uptake
3.2.12. cTnT Cardiac Late Marker Expression
3.2.13. RNA Isolation Quantitative Real-Time PCR (qRT-PCR)
3.2.14. Global Methylation Analysis
3.2.15. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample | Z-Average Diameter (nm) | Polydispersity Index (PDI) | Zeta Potential (mV) | Yield (%) | Encapsulation Efficiency (%) |
---|---|---|---|---|---|
PLGA void | 186 ± 3.1 | 0.063 ± 0.06 | −15.6 ± 0.04 | 37.13% | - |
PEI miRNA | 124 ± 0.6 | 0.301 ± 0.04 | 27.35 ± 0.03 | - | |
PLGA AZA | 298 ± 2.0 | 0.034 ± 0.03 | −15.8 ± 0.86 | 37.35% | 96% |
PLGA AZA-miRNA | 298 ± 2.4 | 0.031 ± 0.02 | −15.7 ± 0.65 | 38.15% | 97% |
PLGA-miRNA | 298 ± 2.6 | 0.032 ± 0.02 | −15.6 ± 0.34 | 37.35% | 96% |
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Muniyandi, P.; Palaninathan, V.; Hanajiri, T.; Maekawa, T. Direct Cardiac Epigenetic Reprogramming through Codelivery of 5′Azacytidine and miR-133a Nanoformulation. Int. J. Mol. Sci. 2022, 23, 15179. https://doi.org/10.3390/ijms232315179
Muniyandi P, Palaninathan V, Hanajiri T, Maekawa T. Direct Cardiac Epigenetic Reprogramming through Codelivery of 5′Azacytidine and miR-133a Nanoformulation. International Journal of Molecular Sciences. 2022; 23(23):15179. https://doi.org/10.3390/ijms232315179
Chicago/Turabian StyleMuniyandi, Priyadharshni, Vivekanandan Palaninathan, Tatsuro Hanajiri, and Toru Maekawa. 2022. "Direct Cardiac Epigenetic Reprogramming through Codelivery of 5′Azacytidine and miR-133a Nanoformulation" International Journal of Molecular Sciences 23, no. 23: 15179. https://doi.org/10.3390/ijms232315179