In vitro evaluation of dual growth factor-incorporated chitosan nanoparticle impregnated collagen–chitosan scaffold for tissue engineering

Artificial tissue constructs require vehicles for controlled release of growth factor to induce cellular signaling in vivo conditions. The objective of this study was to develop a three-dimensional porous tissue engineering scaffold with the capability of carrying nanoparticles for their controlled release. Epidermal growth factor and fibroblast growth factor were encapsulated into chitosan nanoparticles of an average diameter of 50–100 nm. Porous collagen–chitosan scaffolds were prepared by freeze-drying method. The pores of the scaffolds were well interconnected, with a mean diameter of 75–150 µm. The in vitro release kinetics data indicated that nanoparticle impregnated scaffolds released epidermal growth factor and fibroblast growth factor in a sustainable manner. The cytocompatibility, proliferation, and cell attachment characteristics of the biopolymeric scaffolds were evaluated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, flow cytometry, and scanning electron microscope. The results demonstrated that the dual delivery of epidermal growth factor and fibroblast growth factor from chitosan nanoparticles of collagen–chitosan scaffolds significantly enhanced the cellular viability and activity. The in vitro data clearly confirmed that the growth factors incorporated in chitosan nanoparticles and placed in hybrid scaffolds have favorable characteristics for drug delivery and tissue engineering application.