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Hydrothermal Magnesium Alloy Extracts Modulate MicroRNA Expression in RAW264.7 Cells: Implications for Bone
Viviana Costa1, Lavinia Raimondi1, Daniele Bellavia1
1Surgical Sciences and Technologies-SS Omics Science Platform for Personalized Orthopedics, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy.
Journal of Functional Biomaterials
|August 27, 2025
Summary
Hydrothermal treatment of Magnesium AZ31+SPF alloy (Mg AZ31+SPF+HT) reduces pre-osteoclast activation, promoting balanced bone remodeling for orthopedic applications. This Mg alloy modification influences cellular processes and miRNA expression, aiding bone healing.
Area of Science:
- Biomaterials Science
- Orthopedic Engineering
- Cell Biology
Background:
- Magnesium (Mg) alloys, especially Mg AZ31, are promising biodegradable biomaterials for orthopedic uses.
- Hydrothermal (HT) treatment of Mg AZ31+SPF enhances bioactivity and supports osteogenic differentiation of human mesenchymal stem cells (hMSCs).
- Previous studies showed Mg AZ31+SPF+HT modulates signaling pathways crucial for osteogenesis, hypoxia, exosome biogenesis, and lipid metabolism.
Purpose of the Study:
- To evaluate the biocompatibility and effects of Mg AZ31+SPF+HT and Mg AZ31+SPF extracts on murine pre-osteoclasts (RAW 264.7 cells).
- To investigate the impact of these Mg alloy extracts on osteoclastogenesis induction and miRNA expression profiles.
- To assess the potential of Mg AZ31+SPF+HT in modulating cellular responses relevant to bone remodeling.
Main Methods:
- Assessed cytocompatibility, metabolic activity, DNA integrity, and cell morphology of RAW 264.7 cells.
- Quantified osteoclast differentiation via TRAP staining and measured osteoclastogenic markers using qRT-PCR and ELISA.
- Analyzed immunomodulatory properties through multiplex BioPlex assays and performed global miRNA expression profiling on 82 key microRNAs.
Main Results:
- Mg AZ31+SPF+HT extract demonstrated high biocompatibility with no adverse effects on cell viability.
- A significant reduction in TRAP-positive and multinucleated cells was observed, with downregulated osteoclast-specific gene expression and decreased MMP9 protein.
- Cytokine profiling indicated Mg AZ31+SPF+HT extract promoted earlier release of key bone healing cytokines, and miRNA profiling revealed distinct regulatory signatures.
Conclusions:
- Hydrothermal treatment of Mg AZ31+SPF (Mg AZ31+SPF+HT) effectively attenuates pre-osteoclast activation.
- This attenuation occurs through modulation of cellular morphology, gene/protein expression, and post-transcriptional regulation via miRNAs.
- Results highlight the dual importance of promoting bone formation and modulating resorption for optimal implant integration and long-term bone health in orthopedic surgery.
