Abstract
This work reports on the properties of natural bovine hydroxyapatite (HAp) scaffolds calcined at temperatures of 600 and 1000 °C and on the effects of apatite formation on these calcined scaffolds during immersion experiments in Hank's Balanced Salt Solution (HBSS) from 0 to 28 days. The study systematically identifies different mechanisms that determine the formation of the apatite layer. First, compositional analysis of the scaffolds by inductively couple plasma (ICP) reveals significant differences in macromineral content, which includes calcium, phosphorus, magnesium, and sodium. Subsequent X-ray diffraction (XRD) analysis reveals the presence of hydroxyapatite in both scaffolds, while the 1000 °C sample has additional β-whitlockite due to the higher calcination temperature. Further investigation by Fourier Transform Infrared Spectroscopy (FTIR) and Raman analyzes revealed carbonated HAp in the 600 °C scaffolds and characteristic bands of HAp in the 1000 °C scaffolds. Morphological changes during immersion, documented by Scanning Electron Microscopy (SEM) images, display leaf-like and cauliflower-like structures in the 600 °C sample and rash-like features leading to leaf-type apatite formation in the 1000 °C sample. High-Resolution Transmission Electron Microscopy (HR-TEM) showed elongated crystals in both samples, confirming exclusive HAp presence in the 600 °C sample and the coexistence of HAp and β-whitlockite phases in the 1000 °C sample. After 28 days of immersion, TEM images at 600 °C reveal ordered apatite particles, while the 1000 °C images exhibit random apatite formation, confirmed by Energy Dispersive X-ray Spectroscopy in TEM (EDS-TEM). In addition, both scaffolds calcined at 600 and 1000 °C showed biocompatibility, as cytotoxicity tests confirmed a remarkable 100 % cell viability, underlining their non-toxicity. The scaffolds calcined at 600 and 1000 °C therefore exhibit high bioactivity and viability, supporting their suitability for bone tissue engineering applications.
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