4.3 In vitro evaluation of the repair promoting effect of ANVs and
THB@ANVs
4.3.1 In vitro wound
healing assay
The HaCaT cells were seeded in 24-well plates and incubated to 100%
confluence. Scratches were generated using a 200 μL pipette tip. After
using 500 μL PBS to wash out the non-adherent cells, 500 μL of low serum
medium containing 1% FBS and ANVs or AIE@ANVs was added into each well.
Images were acquired by microscope at 0 h, 12 h, and 24 h. The wound
areas were measured using Image J.
4.3.2 Transwell cell migration assay
The Transwell migration assay were conducted using 24-well Transwell
insert with 8 μm pore (Corning, Milan, Italy), 200 μl cell suspension (1
x 106 cells/ml in basic medium) was added on top of
the filter membrane and incubated for 10 minutes at 37 °C and 5%
CO2 to allow the cells to settle down. Carefully added
600μl medium containing 5% FBS and ANVs or THB@ANVs into the bottom of
the lower chamber without generating bubbles, and incubated for
12~48h. After removing the media and remaining cells on
top of the filter membrane, the cells on the other side of the membrane
were fixed with 4% polyformaldehyde for 20 min, and then stained by
0.2% crystal violet for 5-10 minutes. Images of cells were acquired by
microscope and analyzed by Image J.