Particle Size, Zeta Potential and Morphological Observations of the C-HA-DOs

According to the determined CMC value, an appropriate amount of C-HA-DOs was dissolved in PBS solutions with pH values of 7.4 and 6.4. The solutions were then subjected to 30 minutes of ultrasonication to ensure uniform dispersion of the micellar particles. These prepared micellar solutions were used for the experimental measurements, while the remaining solutions were stored at 4°C. Two C-HA-DOs sample solutions at a concentration of 0.5 g/L were placed in 2 mL cuvettes. The particle size and surface zeta potential of the C-HA-DOs were analysed at room temperature using a Zetasizer (Malvern, Zetasizer Nano ZS90). The corresponding polydispersity index (PDI) was also recorded. Each sample was measured at least 3 times.

An appropriate amount of C-HA-DOs was weighed and dissolved in deionized water to prepare a solution with a concentration of 5×10⁻⁴ mg/L. Then, 10 μL of the prepared solution was drop-cast onto a copper grid covered with a carbon support membrane. After a few minutes of incubation, the excess liquid was removed using filter paper. Subsequently, 2% phosphotungstic acid staining solution was added, and after 1–2 minutes of staining, the excess negative stain was removed using filter paper. The sample was then dried and observed by transmission electron microscopy (TEM, FEI Tecnai 12).