Synthesis, characterization and in-vitro behavior of natural chitosan-hydroxyapatite-diopside nanocomposite scaffold for bone tissue engineering

Composite materials based on a combination of biodegradable polymers and bioactive ceramics, including chitosan and hydroxyapatite are discussed as suitable materials for scaffold fabrication. Diopside is a member of bioactive silicates; it is a good choice for hard tissue engineering because of its biocompatibility with host tissue and high mechanical strength. Chitosan and hydroxyapatite were extracted from shrimp shell and bovine bone, respectively and diopside nanoparticles were prepared by the sol-gel method. The present study reports on a chitosan composite which was reinforced by hydroxyapatite and diopside; the scaffolds were fabricated by the freeze-drying method. The so-produced chitosan-hydroxyapatite-diopside (CS-HA-DP) scaffolds were further cross-linked using tripolyphosphate (TPP) to achieve enhanced mechanical strength. The ratios of the ceramic components in composites were 5-58-37, 10-55-35, and 15-52-33 (diopside-hydroxyapatite-chitosan, w/w %). The physicochemical properties of scaffolds were investigated using Fourier-transform infrared spectrometry (FT-IR), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) techniques. The effect of scaffolds composition on bioactivity and biodegradability were studied well. To investigate mechanical properties of samples, compression test was done. Results showed that the composite scaffold with 5% DP has the highest mechanical strength. The porosity of composites dropped from 92% to 76% by increasing the amount of DP. Cytocompatibility of the scaffolds was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity, and cell attachment studies using human osteoblast cells. Results demonstrated no sign of toxicity and cells were found to be attached to the pore walls within the scaffolds; moreover, results illustrated that the developed composite scaffolds could be a potential candidate for tissue engineering. [GRAPHICS] .