In-situ gelation of a novel hydrogel nanocomposite based on Sodium Alginate/ Sodium Hyaluronate/ Xanthan Gum reinforced with Hydroxyapatite nanoparticles through dual injection of CaCl 2 solution

Nowadays, the prevalence of unintentional accidents and traumas is leading to a global rise in the incidence of bone injuries and fractures. The fabrication and design of effective bone tissue engineering scaffolds have become a central subject of investigation within the realm of science, offering a unique strategy for the treatment of bone disorders. This article represents the fabrication and characterization of a novel hydrogel nanocomposite scaffold based on sodium alginate for bone tissue engineering application. This system was reinforced with sodium hyaluronate, xanthan gum, and hydroxyapatite nanoparticles to boost its efficacy and maximize its cell proliferation and regeneration capability. The prepared mixture was crosslinked by calcium chloride solution and the final hydrogel was characterized and experimented on MG-63 cells. The two solutions can be injected through a double barrel syringe to the damaged tissue site for the in-situ gelation of hydrogel with minimum injury and invasiveness. X-ray diffraction along with Fourier transform infrared spectroscopy were performed to characterize the interactions of the components and to ensure the successful synthesis procedure. Scanning electron microscopy images confirmed the three-dimensional interconnected porous structure of the scaffolds with pore sizes suitable for cell adhesion and proliferation. Viscosity of samples of each step of synthesis and the crosslinked hydrogel was measured. The data confirmed acceptable rheology for injectable samples and suitable solidity and firmness for the final crosslinked hydrogel. Cellular efficacy assessments were performed by alizarin red staining, MTT assay, and alkaline phosphatase activity. The outcomes showed appropriate tissue microenvironment stimulation and promising cell proliferation and regeneration potential with no significant cytotoxicity. Overall, the designed scaffold showed promising potential as a novel hydrogel scaffold for tissue engineering applications, offering an appropriate microenvironment for cellular growth and regeneration.