Influencing Factors Investigation and In vitro Biological Performance Characterization of Nano β-TCP Synthesized via Microwave-ultrasonic-hydrothermal Multifield Coupling

Abstract

This study employed a microwave-ultrasonic-hydrothermal multifield coupling method to synthesize nano β-Tricalcium phosphate (β-TCP) powder, systematically evaluating the impact of various parameters, including reaction temperature, time, sintering temperature, reactant types and concentrations, and graphene oxide (GO) concentration, on the physicochemical properties of the nano β-TCP powder. The synthesized powder was characterized using X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), inductively coupled plasma optical emission spectroscopy (ICP-OES), and thermogravimetric-differential scanning calorimetry (TG-DSC). The experimental results indicate that the optimal synthesis conditions are achieved with a 0.6 mol/L Ca(NO₃)₂·4H₂O solution and a 0.4 mol/L (NH₄)₂HPO₄ solution at a reaction temperature of 35 °C for 40 minutes, followed by sintering at 720 °C for 2 hours with 1×10⁻¹g/L GO. The prepared β-TCP powder exhibits high crystallinity, a pure phase, good dispersibility, no significant aggregation, and uniform particle size of 59.75±12.84 nm. In vitro cytotoxicity tests show excellent biocompatibility and no cytotoxic effects on bone marrow stromal cells (BMSCs) even at concentrations up to 0.8 mg/mL. Furthermore, results from live-dead staining and nuclear membrane staining of cells co-cultured with the material demonstrate that the β-TCP can promote the proliferation and differentiation of BMSCs to a certain extent, highlighting its potential as a safe and effective material for bone tissue engineering.

Keywords

nanoparticles / β-TCP / microwave-ultrasonic-hydrothermal / biomaterials / in vitro

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