Utilization of Glycosaminoglycans/Proteoglycans as Carriers for Targeted Therapy Delivery

Figure 7

Uptake of pSV-β-galactosidase/Tf-PEG-PEI (nanoparticles) into target cell. Delivery of pSV-β-gal nanoparticles in colon cancer cells (adapted from [43]). (a) In situ  β-galactosidase expression in the target cell. The Apc10.1-HAS2 clone and the CT26 cells were treated for 48 h with the pSV-β-gal alone (panel 1), with the pSV-β-gal with liposome (panel 2), or with the pSV-β-gal/nanoparticles (35 nm average diameter, 8 μg of pSV-β-gal/mL) (panel 3). The average size of the pSV-β-gal/nanoparticles is ~35 nm ± 20 nm. The transfected cells were fixed in 0.2% glutaraldehyde in PBS and washed twice in PBS. The cells were treated with a β-galactosidase staining solution and digitally photographed. (b) Transferrin-dependent uptake and in situ  β-galactosidase expression in the target cell. The Apc 10.1-HAS2 cells were transfected with the pSV-β-gal with liposome (panel 1), treated with Tf-R antibody and followed by transfection with the pSV-β-gal/nanoparticles (8 μg pSV-β-gal/mL) (panel 2), or treated with the pSV-β-gal/nanoparticles (8 μg pSV-β-gal/mL) alone (panel 3). The transfected cells were fixed in 0.2% glutaraldehyde in PBS and washed twice in PBS. β-galactosidase expressions in the cells were digitally photographed. (c) Cell-free extracts of parallel cultures were prepared in 10 mM CHAPS buffer and assayed for β-galactosidase activity using o-nitrophenyl β-D-galactopyranoside as substrate. The results are expressed as micromoles of o-nitrophenol formed per min/mg protein and represent ± S.D. of triplicate assays from the untransfected, liposome-transfected, or nanoparticles-treated cultures for each cell type.

(a) In situ  β-galactosidase expression in the target cell

(b) Tf-dependent uptake and in situ  β-galactosidase expression in the target cell

(c) β-galactosidase enzyme activity in the target cell