A preliminary study on the interaction between Asn-Gly-Arg (NGR)-modified multifunctional nanoparticles and vascular epithelial cells

Liu, Chunxi; Liu, Tingxian; Yu, Xiaoyue; Gu, Yizhu

To examine whether TPIC could change the distribution of CD13 in a manner similar to that of the anti-CD13 antibody and to examine the behavior of CD13 when cells were treated with TPIC, TPIC without DOX was prepared and used to avoid the mutual interference of red fluorescence of DOX and PE. First, HUVEC was incubated with non-DOX-loaded TPIC for 0-4 h at 37 1C with 5% CO 2 in an incubator. Then, test nanocarriers were aspirated, and cells were washed for three times with cold PBS. After being fixed with 4% paraformaldehyde in 0.1 mol/L PBS for 15 min, the cells were treated with a PE-conjugated anti-CD13 antibody (1:100 dilution) for 1 h at 4 1C, permeabilized with 0.1% Triton X-100 for 5 min, treated with 3% bovine serum albumin for 20 min, and labelled for CAV1 as described above. After extensive washing, the glass bottom dishes were examined under a Zeiss LSM700 laser confocal scanning microscope.