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Biocompatibility of magnesium particles evaluated by in vitro cytotoxicity and genotoxicity assays
Ana L Di Virgilio1, Miguel Reigosa, Monica Fernández Lorenzo de Mele
1Institute of Research in Theoretical and Applied Physical Chemistry (INIFTA), Diag. 113 and 64, La Plata 1900, Argentina.
Abstract:
Mg is a biodegradable biomaterial which may release particles (MP) to the environment. The possible cyto- and genotoxic effects of MP derived from magnesium powder (mesh 325) were analyzed on rat osteosarcoma UMR106 cells in order simulate the effect of Mg debris. Neutral red (NR) incorporation and acridine orange/ethidium bromide (AO/EB) staining techniques were used as endpoints to analyze the cytotoxic effects at 25-1000 μg/mL concentration range. Genotoxicity was estimated according to micronucleus (MN) formation and the Comet assay (CA). Results showed that MP size changes with time due to corrosion. Changes in lysosomal activity were observed after 24 h only at 1000 μg/mL. Accordingly, AO/EB staining showed a significant decrease in the number of living cells at 500 μg/mL. Transmission electronic microscopy showed MP internalization (60 and 200 nm diameter) in cells after 2-h treatment, whereas no MP was detected after 24 h. A significant dose-dependent increase in MN frequencies was observed at 25-100 μg/mL range (nontoxic range). DNA damage induction was assessed by CA only at 500 μg/mL. Results showed dose-dependent cytotoxic and genotoxic effects of MP on UMR106 cells with different threshold values of MP concentration.
Insights
Magnesium particles (MP) show dose-dependent cytotoxic and genotoxic effects on rat osteosarcoma cells. Even at non-toxic concentrations, MPs increase micronucleus formation, indicating potential genotoxicity.
Area of Science:
- Biomaterials Science
- Cell Biology
- Toxicology
Background:
- Magnesium (Mg) is a biodegradable biomaterial with potential environmental release of particles (MP).
- Understanding the cytotoxic and genotoxic effects of Mg-MP is crucial for assessing their biological impact.
Purpose of the Study:
- To investigate the cyto- and genotoxic effects of magnesium powder (mesh 325) derived MP on rat osteosarcoma UMR106 cells.
- To simulate the effects of Mg debris in a cellular environment.
Main Methods:
- Cytotoxicity assessed using Neutral Red (NR) incorporation and Acridine Orange/Ethidium Bromide (AO/EB) staining.
- Genotoxicity evaluated via micronucleus (MN) formation and Comet Assay (CA).
- Transmission electron microscopy (TEM) used to observe MP internalization.
Main Results:
- MP size changed over time due to corrosion.
- Lysosomal activity affected only at 1000 μg/mL after 24h; significant decrease in living cells observed at 500 μg/mL via AO/EB staining.
- MP internalization (60-200 nm) observed within 2h, but not after 24h.
- Dose-dependent increase in MN frequencies at 25-100 μg/mL (non-toxic range).
- DNA damage detected by CA only at 500 μg/mL.
Conclusions:
- Magnesium particles exhibit dose-dependent cytotoxic and genotoxic effects on UMR106 cells.
- Different concentration thresholds exist for cytotoxic versus genotoxic effects.
- Further research is needed to fully elucidate the long-term biological impact of magnesium particles.
