Influence of carboxymethyl chitin on stability and biocompatibility of 3D nanohydroxyapatite/gelatin/carboxymethyl chitin composite for bone tissue engineering

Abstract

A novel three-dimensional (3D) scaffold has been developed from the unique combination of nanohydroxyapatite/gelatin/carboxymethyl chitin (n-HA/gel/CMC) for bone tissue engineering by using the solvent-casting method combined with vapor-phase crosslinking and freeze-drying. The surface morphology and physiochemical properties of the scaffold were investigated by dissolvability test, infrared absorption spectra (IR), X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), mechanical testing, and soaking in simulated body fluid (SBF). An optimized (composition and processing parameters) ratio of n-HA:gel:CMC (1:2:1), exhibited ideal porous structure with regular interconnected pores (75-250 μm) and higher mechanical strength. Result suggested that the divalent (Ca(++)), carboxyl (COO(-)), amino (NH4(+)), and phosphate (PO4(3-)) groups created favorable ionic interactions which facilitated structural stability and integrity of the composite scaffold. The SBF soaking experiment confirmed the apatite nucleation ability, induced by CMC incorporation. Furthermore, hemocompatibility (hemolysis, platelet adhesion, and protein adsorption) and biocompatibility with MG63 osteoblast cells (MTT assay, cell morphology, and confocal studies from within the 3D scaffold) indicated that the structural and dimensional stability of composite scaffold provided an optimal mechanosensory environment for enhancement of cell adhesion, proliferation, and network formation. The n-HA/gel/CMC composite, therefore, may serve as a promising composite scaffold for guided bone regeneration.