Mwco 1000

In vitro tube formation was performed using Matrigel Matrix (354248, Corning, USA). A pre-cooled 96-well plate was coated with 100 μL Matrigel per well and then incubated for 4 at 37°C. The supernatant of treated glioma cells was filtered through a 0.45 µm filter and concentrated using an ultrafiltration tube (Millipore, MWCO 1000). ECs resuspended in concentrated cell supernatants were seeded on Matrigel. After 6 h, images were taken using a digital camera (IX83; Olympus, Toyko, Japan). Quantification was performed using Image J software (NIH Image J system, Bethesda, MD, USA).

LF@EMO-NPs were prepared using a slightly modified dialysis [15 (link)]. In brief, 100 mg LF was dissolved in 40 mL ultrapure water, along with the dissolution of 10 mg EMO in 2 mL DMSO. Then 2 ml of EMO solution was dripped into 40 ml of LF solution. To improve the solubility of EMO, the above solutions were sonicated for 5 min using a probe sonicator (Sonicator XL; Misonix, Melville, NY, USA), kept on a magnetic stirrer for 30 min to ensure uniform distribution and optimum size reduction, and dialyzed in ultrapure water with a dialysis bag (MWCO 1000, Millipore, USA) for 8 h. LF@EMO-NPs were passed through a syringe filter (0.22 μm) to remove impurities and free drug.
HA/LF@EMO-NPs were prepared by electrostatic adsorption [26 (link)]. Briefly, 4 mg of HA was dissolved in 2 mL of ultrapure water, followed by dropwise addition of HA solution to the LF@EMO-NPs solution after filtration. After stirring for 30 min and filtering through a syringe filter (0.22 μm), HA/LF@EMO-NPs were lyophilized until further use. All the above steps were performed under light-proof conditions.