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Meyd873 – The Whispering Code

2.5. Physicochemical Characterization

• Size & ζ‑potential: Dynamic light scattering (DLS, Malvern Zetasizer Nano ZS) at 25 °C.
• Morphology: Transmission electron microscopy (TEM, JEOL 2100) after negative staining with phosphotungstic acid.
• pH‑responsive release: NPs (1 mg mL⁻¹) were incubated at 37 °C in phosphate‑buffered saline (PBS) at pH 7.4 and acetate buffer at pH 5.5. DOX release was measured at predetermined intervals.
• Redox‑responsive degradation: NPs were exposed to 10 mM GSH (pH 7.4) and monitored for size changes by DLS.

2.6. In‑Vitro Studies

• Cell lines: MDA‑MB‑231 (TNBC) and NIH‑3T3 fibroblasts (ATCC).
• Cytotoxicity: MTT assay after 72 h exposure to free DOX, DOX‑MEYD873 NPs, and blank NPs (0–20 µg mL⁻¹ DOX equivalent).
• Cellular uptake: Confocal laser scanning microscopy (CLSM) after 4 h incubation with DOX‑labelled NPs (5 µg mL⁻¹).
• Apoptosis: Annexin V/PI flow cytometry.

Title:
MEYD873 – A Novel Stimuli‑Responsive Polymeric Platform for Targeted Drug Delivery

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