Development and characterization of chitosan–alginate scaffolds: Influence of chitosan depolymerization on structural and biological properties

Abstract

Biopolymer-based scaffolds are structures designed to mimic the extracellular matrix and serve as an alternative for cell culture applications. This study aimed to develop and characterize scaffolds composed of chitosan (Ch) and alginate (Alt) and to evaluate their biocompatibility in Vero cell culture. Functional scaffolds were obtained by freeze-dry method mixing depolymerized chitosan (dpCh), Ch and Alt, followed by lyophilization. Physicochemical characterization was conducted using Fourier-Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and swelling behavior analysis. Biocompatibility was assessed through an adhesion assay with Vero cells. FTIR analysis revealed that the depolymerization process altered the absorbance spectrum at 3294 cm⁻¹ but did not affect the characteristic primary amine peaks nor its degree of deacetylation. SEM analysis demonstrated variations in the predominant pore size distribution (100–600 µm) depending on the scaffold formulation. The swelling capacity of most samples was greater at acidic pH (1.4). The dpCh 1% Alt 1% 1:2 and dpCh 0.5% Alt 0.75% dpCh 0.5% scaffolds exhibited the most favorable physicochemical properties and were selected for the Vero cell adhesion assay, which confirmed excellent biocompatibility. These findings suggest that Ch/Alt scaffolds hold significant potential as three-dimensional supports for Vero cell culture applications.