Effects of Magnesium-Doped Hydroxyapatite Nanoparticles on Bioink Formulation for Bone Tissue Engineering

Bioprinting of nanohydroxyapatite (nHA)-based bioinks has attracted considerable interest in bone tissue engineering. However, the role and relevance of the physicochemical properties of nHA incorporated in a bioink, particularly in terms of its printability and the biological behavior of bioprinted cells, remain largely unexplored. In this study, two bioinspired nHAs with different chemical compositions, crystallinity, and morphologies were synthesized and characterized: a more crystalline, needle-like Mg²⁺-doped nHA (N-HA) and a more amorphous, rounded Mg²⁺- and CO₃^2--doped nHA (R-HA). To investigate the effects of the different compositions and morphologies of these nanoparticles on the bioprinting of human bone marrow stromal cells (hBMSCs), gelatin and gelatin methacryloyl (GelMA) were selected as the bioink backbone. The addition of 1% (w/w) of these bioceramic nanoparticles significantly improved the printability of GelMA in terms of extrudability, buildability, and filament spreading. The biological potential of the bioinks was evaluated by examining the hBMSC viability, metabolic activity, and osteogenic differentiation over 21 days. Both nHAs showed high cell viability, with N-HA showing a significant increase in metabolic activity under nonosteogenic conditions and R-HA showing a notable increase with osteogenic stimulation. These results suggest that the two nHAs interact differently with their environment, highlighting the importance of both the chemistry and morphology in bioink performance. In addition, osteogenic differentiation further highlighted how the physicochemical properties of nHAs influence osteogenic markers at both the RNA and protein levels. Clearly, tailoring the physicochemical properties of hydroxyapatite nanoparticles is critical to developing more biomimetic bioinks with great potential for advancing bone bioprinting applications.