Nanoparticle-reinforced polymer-based scaffolding matrices as artificial bone-implant materials are potential suitors for bone regenerative medicine as they simulate the native bone. In the present work, a series of bioinspired, osteoconductive tricomposite scaffolds made up of nanohydroxyapatite (NHA) embedded xanthan gum−chitosan (XAN−CHI) polyelectrolyte complex (PEC) are explored for their bone-regeneration potential. The Fourier transform infrared spectroscopy studies confirmed complex formation between XAN and CHI and showed strong interactions between the NHA and PEC matrix. The X-ray diffraction studies indicated regulation of the nanocomposite (NC) scaffold crystallinity by the physical cues of the PEC matrix. Further results exhibited that the XAN−CHI/NHA5 scaffold, with a 50/50 (polymer/NHA) ratio, has optimized porous structure, appropriate compressive properties, and sufficient swelling ability with slower degradation rates, which are far better than those of CHI/NHA and other XAN−CHI/NHA NC scaffolds. The simulated body fluid studies showed XAN− CHI/NHA5 generated apatite-like surface structures of a Ca/P ratio ∼1.66. Also, the in vitro cell−material interaction studies with MG-63 cells revealed that relative to the CHI/NHA NC scaffold, the cellular viability, attachment, and proliferation were better on XAN−CHI/NHA scaffold surfaces, with XAN−CHI/ NHA5 specimens exhibiting an effective increment in cell spreading capacity compared to XAN−CHI/NHA4 and XAN−CHI/ NHA6 specimens. The presence of an osteo-friendly environment is also indicated by enhanced alkaline phosphatase expression and protein adsorption ability. The higher expression of extracellular matrix proteins, such as osteocalcin and osteopontin, finally validated the induction of differentiation of MG-63 cells by tricomposite scaffolds. In summary, this study demonstrates that the formation of PEC between XAN and CHI and incorporation of NHA in XAN−CHI PEC developed tricomposite scaffolds with robust potential for use in bone regeneration applications.

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The present frontiers of bone tissue engineering are being pushed by novel biomaterials that exhibit phenomenal biocompatibility and adequate mechanical strength. In this work, we fabricated a ternary system incorporating nano-hydroxyapatite (n-HA)/gum arabic (GA)/κ-carrageenan (κ-CG) with varying concentrations, i.e., 60/30/10 (CHG1), 60/20/20 (CHG2), and 60/10/30 (CHG3). A binary system with n-HA and GA was also prepared with a ratio of 60/40 (HG) and compared with the ternary system. A rapid mineralization of the apatite layer was observed for the ternary systems after incubation in simulated body fluid (SBF) for 15 days as corroborated by scanning electron microscopy (SEM). CHG2 exhibited the maximum apatite layer deposition. Further, the nanocomposites were physicochemically analyzed by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and mechanical testing. Their results revealed a substantial interaction among the components, appropriate crystallinity, and significantly enhanced compressive strength and modulus for the ternary nanocomposites. The greatest mechanical strength was achieved by the scaffold containing equal amounts of GA and κ-CG. The cytotoxicity was evaluated by culturing osteoblast-like MG63 cells, which exhibited the highest cell viability for the CHG2 nanocomposite system. It was further supported by confocal microscopy, which revealed the maximum cell proliferation for the CHG2 scaffold. In addition, enhanced antibacterial activity, protein adsorption, biodegradability, and osteogenic differentiation were observed for the ternary nanocomposites. Osteogenic gene markers, such as osteocalcin (OCN), osteonectin (ON), and osteopontin (OPN), were present in higher quantities in the CHG2 and CHG3 nanocomposites as confirmed by western blotting. These results substantiated the pertinence of n-HA-, GA-, and κ-CG-incorporated ternary systems to bone implant materials.

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