H 6009iv

Take 2 mL of Blank-NPs, SRM-NPs and PTX-NPs suspensions, and use dynamic light scattering (DLS) method to measure the average particle size, particle size distribution and zeta potential on zeta sizer nano ZS (Malvern instruments, UK).
Keep the Blank-NPs, SRM-NPs and PTX-NPs suspensions that have been tested for particle size, and measure the particle size every 6 d to test the stability of NPs.
100 μl Blank-NPs, SRM-NPs and PTX-NPs suspensions were uniformly dispersed in 1 mL distilled water and dropped on the copper mesh. After drying, the morphology of $NP$ s was observed by transmission electron microscopy (TEM). (h-6009iv, Hitachi).
All measurements are made in triplicate. The average particle size is represented by the volume average diameter, and the reported value is represented by the average ± standard deviation (n = 3).

Size and zeta potential measurements of micelles including BMs, DTX-M, and RGD-DTX-M were performed at room temperature through a dynamic light scattering (DLS) method using a Nicomp Particle Sizing system (380ZLS, Santa Barbara, CA, USA). After the samples were diluted by distilled water and negatively stained with phosphotungstic acid, the morphology of micelles was observed by transmission electron microscope (TEM, H-6009IV, Hitachi, Japan). Drug loading (DL) and encapsulation efficiency (EE) of RGD-DTX-M were determined by high-performance liquid chromatography (HPLC) on a Kromasil 100–5C18 column (5 μm, 250×4.6 mm) at 30°C. The mobile phase was prepared with a mixture of acetonitrile and ultrapure water (55:45, v/v). The DTX was detected at the wavelength of 230 nm (flow rate: 1.0 mL/min). The selectivity, linearity, precision, and recovery of this method were fully validated. EE and DL were determined using Eqns. (1) and (2), respectively.
EE (%) = Weight of the drug encapsulated in micelles (mg)/Weight of the total drug added (mg) × 100%(1)
DL (%) = Weight of the drug encapsulated in micelles (mg)/Weight of the total micelles (mg) × 100%(2)

DOX@MTP/HA-EGCG was prepared according to a previous report with some modifications [19] , [27] (link). Briefly, 200 μL of FeCl₃ solution (0.27 mg/mL) was added to 1 mL of distilled water and 400 μg of DOX solution, and 300 μL of EGCG solution (2 mg/mL) was added dropwise to the mixture under stirring. After the formation of DOX@MTP, HA-EGCG solution (10 mg/mL) was added and the mixture was incubated for 1 h and centrifuged to remove free DOX and insoluble materials to obtain DOX@MTP/HA-EGCG nanoreactors. An equivalent volume of water was used instead of DOX to prepare MTP and MTP/HA-EGCG. DOX/HA-EGCG was formed by self-assembly based on DOX and HA-EGCG. DOX solution was added to HA-EGCG solution under magnetic stirring for 1 h to obtain DOX/HA-EGCG.
The sizes of the DOX@MTP, DOX/HA-EGCG, and DOX@MTP/HA-EGCG nanoreactors were measured by dynamic light scattering (DLS) (Nano-ZS 90; Malvern, UK). Transmission electron microscopy (TEM; H-6009IV; Hitachi, Japan) was used to assess the morphology of the nanoreactors. Phosphotungstic acid (2% (w/w)) was used to stain the nanoreactors for 5 min before observation. Atomic force microscopy (AFM) (Bruker, MultiMode) was applied to evaluate the surface morphology of the nanoreactor.

The transmission electron microscope (TEM; H-6009IV; Hitachi Ltd., Tokyo, Japan) was employed to observe the morphology of DMC. The nanoparticles were placed on a copper grid covered with nitrocellulose after diluting with distilled water and then stained negatively with phosphotungstic acid. The particle size and zeta potential of DMC nanoparticles were measured by dynamic light scattering (Nano-ZS 90; Malvern Instruments, Malvern, UK) under 25°C temperature conditions.

DOTAP modified mPEG-PCL micelles (DMP) were prepared according our previous reports.^{30 (link)} Briefly, 45 mg of MPEG-PCL polymer and 5 mg DOTAP were co-dissolved in methylene dichloride (KeLong Chemicals, Chengdu, China) and solvent was removed under rotary evaporation for 45 min. The lipid film was re-hydrated with distilled water under 55 °C to self-assemble the micelles with a final concentration of 10 mg mL⁻¹. The prepared micelles were stored at 4 °C for future use. The size and surface charge of prepared DMP micelles were determined by Malvern ZS90 (Malvern, Worcestershire, UK). Measurements were performed at 25 °C after equilibration. All results were the mean of three test runs. The morphology of DMP micelles were observed under a transmission electron microscope (TEM) (H-6009IV, Hitachi, Japan).