Hydrothermal Surface Treatment of Mg AZ31 SPF Alloy: Immune Cell Biocompatibility and Antibacterial Potential for Orthopaedic Applications

Biodegradable magnesium (Mg) alloys are promising materials for temporary orthopaedic implants, combining favourable mechanical properties and superplastic behaviour with in vivo resorption. This enables (i) prolonged implant duration, (ii) fabrication of complexshaped prostheses via superplastic forming (SPF), (iii) elimination of removal surgery, and (iv) reduced risk of long-term complications. However, rapid corrosion under physiological conditions remains a major limitation, highlighting the need for surface treatments that slow degradation while preserving implant integrity. This study investigates the effects of hydrothermal surface treatment on MgAZ31-SPF alloys, focusing on immunomodulatory responses, antibacterial potential, and degradation behaviour. Hydrothermally treated MgAZ31-SPF (MgAZ31-SPF-HT) extracts released lower Mg2+ concentrations (29.2 mg/dL) compared to untreated MgAZ31-SPF (47.5 mg/dL) while maintaining slightly alkaline pH (7–8.7), indicating improved control of early degradation. In vitro assays with human peripheral blood mononuclear cells (hPBMCs) and normal human dermal cells (NHDCs) showed that MgAZ31-SPF-HT extracts maintained higher cell viability over 24–72 h. Gene expression analysis revealed significant downregulation of pro-inflammatory markers CTSE and TNF-α, while protein quantification via ELISA and BioPlex confirmed reduced secretion of TNF-α, TGF-β1, TGF-β2, IL-6, and IL-8, suggesting mitigation of early immune activation. Antibacterial assays demonstrated limited Staphylococcus aureus colonisation on both MgAZ31-SPF and MgAZ31-SPF-HT scaffolds, with CFU counts (~105–106) well below the threshold for mature biofilm formation (~108), and SEM analysis confirmed sparse bacterial distribution without dense EPS-rich layers. Overall, hydrothermal treatment improves Mg alloy biocompatibility by controlling Mg2+ release, modulating early immune responses, and limiting bacterial adhesion, highlighting its potential to enhance clinical performance of Mg-based implants.