Green fluorescent protein (GFP) labeled human corneal epithelial cells (HCECs) were purchased from Life Technologies Ltd (Paisley, UK). WST-1 cell proliferation assay reagent was purchased from Roche Diagnostics GmbH (Mannheim, Germany). Keratinocyte serum free medium (KSFM) with supplements-bovine pituitary extract and recombinant EGF (BPE-rEGF), were procured from Life Technologies Ltd.
To assess whether fabricated collagen-based hydrogel scaffolds with Bevacizumab release system inhibit cell proliferation, 1 × 104 cells were seeded on the top of 6 mm hydrogel disc with incorporated GNPs@Bevacizumab in a 96- well plate and cultured for 48h in BPE-rEGF supplemented KSFM at 37 °C in 5% CO2. Blank scaffolds, hydrogels with free drug were also cultured along with HCECs culture without any hydrogel scaffold, which served as control for the experiment. In 48h, the media was aspirated, and WST-1 reagent was added to the medium following the manufacturer's instructions. The absorbance was taken at 450 nm in Victor3 V 1420 Multilabel Plate Counter (PerkinElmer, Waltham, MA, USA). Results represent mean values from triplicate measurements.
To assess whether fabricated collagen-based hydrogel scaffolds with Bevacizumab release system inhibit cell proliferation, 1 × 104 cells were seeded on the top of 6 mm hydrogel disc with incorporated GNPs@Bevacizumab in a 96- well plate and cultured for 48h in BPE-rEGF supplemented KSFM at 37 °C in 5% CO2. Blank scaffolds, hydrogels with free drug were also cultured along with HCECs culture without any hydrogel scaffold, which served as control for the experiment. In 48h, the media was aspirated, and WST-1 reagent was added to the medium following the manufacturer's instructions. The absorbance was taken at 450 nm in Victor3 V 1420 Multilabel Plate Counter (PerkinElmer, Waltham, MA, USA). Results represent mean values from triplicate measurements.