Modeling and optimization of gelatin-chitosan micro-carriers preparation for soft tissue engineering: Using Response Surface Methodology

Electrospray ionization is a wide spread technique for producing polymeric microcarriers (MCs) by applying electrostatic force and ionic cross-linker, simultaneously. In this study, fabrication process of gelatin-chitosan MCs and its optimization using the Response Surface Methodology (RSM) is reported. Gelatin/chitosan (G/C) blend ratio, applied voltage and feeding flow rate, their individual and interaction effects on the diameter and mechanical strength of the MCs were investigated. The obtained models for diameter and mechanical strength of MCs have a quadratic relationship with G/C blend ratio, applied voltage and feeding flow rate. Using the desirability curve, optimized G/C blend ratios that are introduced, include the desirable quantities for MCs diameter and mechanical strength. MCs of the same desirable diameter (350 μm) and different G/C blend ratio (1, 2, and 3) were fabricated and their elasticity was investigated via Atomic Force Microscopy (AFM). The biocompatibility of the MCs was evaluated using MTT assay. The results showed that human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) could attach and proliferate on fabricated MCs during 7 days of culturing especially on those prepared with G/C blend ratios of 1 and 2. Such gelatin-chitosan MCs may be considered as a promising candidate for injectable tissue engineering scaffolds, supporting attachment and proliferation of hUCMSCs. - Highlights: • Gelatin-chitosan Micro-carriers fabricated by electrospray ionization method. • The effects of blend ratio, the syringe feeding rate, and voltage on micro-carrier optimization were investigated via RSM. • Both diameter and mechanical strength of Micro-carriers have a quadratic relationship with selected parameters. • The optimum conditions with fixed diameter of 350μm and maximized strength in different blend ratios were achieved. • The elasticity and biocompatibility of desirable fabricated micro-carriers characterized.