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Development and Characterization of a Collagen-Based Three-Dimensional In Vitro Model to Mimic Biofilm Formation in a
Kameel Zuniga1, Marc Thompson1, Preeti J Muire1
1Combat Wound Care, U.S. Army Institute of Surgical Research, JBSA, Ft. Sam Houston, Texas 78234-7767, United States.
ACS Applied Bio Materials
|May 28, 2025
Summary
This study developed a 3D hydrogel model to mimic wound biofilms, showing increased bacterial growth and resistance. Rifampin reduced bacterial load but did not fully eradicate the biofilm in this advanced in vitro model.
Area of Science:
- Biomaterials Science
- Microbiology
- Wound Healing Research
Background:
- Current in vitro biofilm models do not fully replicate the host environment.
- Understanding bacterial behavior in a complex wound microenvironment is crucial for effective treatment.
Purpose of the Study:
- To develop a 3D hydrogel scaffold that mimics the in vivo wound biofilm microenvironment.
- To investigate biofilm formation and antibiotic efficacy in this novel model.
Main Methods:
- A collagen-based hydrogel was created and infected with Staphylococcus aureus (UAMS-1).
- Biofilm formation was assessed using plating, SEM, and PCR analysis.
- Antibiotic efficacy of vancomycin and rifampin was evaluated.
Main Results:
- The hydrogel supported robust biofilm formation with increased polysaccharide production and adhesion gene expression.
- Vancomycin was ineffective, while rifampin significantly reduced bacterial CFUs but increased small colony variants (SCVs).
- The 3D model demonstrated greater similarity to the host environment than static biofilm models.
Conclusions:
- The developed 3D hydrogel model effectively recapitulates key aspects of wound biofilm formation.
- Antibiotic treatment showed varied efficacy, highlighting the challenges of biofilm eradication in complex environments.
- This model offers a more relevant platform for studying persistent wound infections and testing therapeutic strategies.
