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image of Experimental Approaches to Prophylaxis of MRSA Biofilm Formation on 3D Scaffolds Modified with Rough Calcium Phosphate Coating

Abstract

Introduction

Microbial biofilm formation on surgical implants significantly exacerbates implant-associated infections (IAIs) and often leads to implant failure. This study aimed to investigate the ex vivo effects of elevated temperature and antibiotics, or zinc incorporation, on methicillin-resistant Staphylococcus aureus (MRSA) biofilm growth on titanium (Ti)-based 3D plates fabricated by selective laser melting (SLM) and coated with a rough calcium phosphate (CaP) layer using the micro-arc oxidation technique.

Materials and Methods

MRSA strain 43300 broth cultures were exposed to heating at 45°C for 60 min or 56°C for 30 min, with or without vancomycin (VMN). The control culture (250 microbial cells per 1 ml of isotonic sodium chloride) was maintained at 37°C. Ti-based discs (10 mm in diameter and 1 mm in thickness) with structured surfaces were coated with a CaP layer via micro-arc oxidation. The Ti-based samples were added to MRSA broth cultures for 2 hours, then transferred to fresh serum-free medium and incubated for 48 hours at 37 °C. Bacterial biofilm optical density on the CaP coating was quantified by computer morphometry after fixation and Gram staining.

Results

A single ex vivo administration of VMN at its minimum inhibitory concentration (MIC, 2 μg/ml) was ineffective against MRSA biofilm growth. The therapeutic VMN concentration (20 μg/ml) reduced the biofilm optical density by 17%. Heating of MRSA broth cultures showed no significant bacteriostatic effect, and combining hyperthermia with VMN did not enhance antimicrobial efficacy. In contrast, Zn-doped (~0.31 at.%) CaP coatings inhibited MRSA biofilm development at 37°C. The bacteriostatic effect (5%) exceeded that of VMN at MIC (0%) but was lower than that observed for the therapeutic VMN dose (17%).

Discussion

The negatively charged CaP coating on titanium implants may promote MRSA biofilm formation through electrostatic interactions with bacterial cell surfaces. Hyperthermia at 45°C or 56°C, either alone or in combination with vancomycin, showed limited efficacy, possibly due to bacterial thermotolerance and antibiotic resistance. Incorporation of zinc into CaP coatings demonstrated a modest bacteriostatic effect, suggesting potential as an adjunct strategy alongside conventional antibiotics such as vancomycin.

Conclusion

Zn-doped rough CaP coatings on Ti-based implants show promise for use in bone tissue engineering, where there is a risk of implant-associated infection, owing to the bacteriostatic activity of zinc at low concentrations.

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2026-01-09
2026-02-02

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Experimental Approaches to Prophylaxis of MRSA Biofilm Formation on 3D Scaffolds Modified with Rough Calcium Phosphate Coating
Bentham Science Publishers ; https://doi.org/10.2174/0115748855408793251103042650
/content/journals/cdth/10.2174/0115748855408793251103042650
/content/journals/cdth/10.2174/0115748855408793251103042650
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  • Article Type:     Research Article
Keywords: zinc ions ; heating ; titanium substrate ; ex vivo bacteriostatic effect ; vancomycin ; selective laser melting ; micro-arc coating ; Bacterial culture

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