Disruption of Methicillin-resistant Staphylococcus aureus Biofilms with Enzymatic Therapeutics

Abstract

The purpose of this study was to establish an in vitro Staphylococcus aureus biofilm model that mimics wound-like conditions and employ this model to evaluate the anti-biofilm activity of four enzymatic compounds. Varying concentrations of human plasma from 0-50% were supplemented into growth media to evaluate biomass accumulation in 2 laboratory and 4 clinical multi-drug resistant S. aureus strains. Supplementation with 10% human plasma resulted in the most robust and reproducible biofilms in all 6 strains. The enzymes papain, bromelain, alpha-amylase, and lysostaphin were then tested on the 6 S. aureus strains in the 10% human plasma biofilm model, and biofilm biomass was quantified using the crystal violet assay. All four enzymes significantly reduced biomass compared to controls in all strains after 2 and 24 hours of treatment. Lysostaphin decreased biomass by up to 76%, whereas the other agents reduced biomass by up to 94-98% in the 6 bacterial strains. Visualization of the biofilms with confocal laser scanning microscopy and scanning electron microscopy confirmed that the dispersal agents detached the biofilm matrix and bacteria from the growth surface, and lysostaphin, but not the other enzymes, induced changes in cell morphology indicative of bacterial cell damage. Overall, our results indicate use of enzymes may be an effective means of eradicating biofilms and a promising strategy to improve treatment of multidrug resistant bacterial infections.

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Document Details

Document Type
Technical Report
Publication Date
Apr 29, 2015
Accession Number
ADA621777

Entities

People

  • Chase Watters
  • Dickson Kirui
  • Nancy J. Millenbaugh
  • Tarea Burton

Organizations

  • Naval Medical Research Center

Tags

DTIC Thesaurus Topics

  • Bacteria
  • Bacterial Infections
  • Cellular Structures
  • Confocal Laser Scanning Microscopy
  • Debridement
  • Electron Microscopy
  • Health Services
  • Indicator Dyes
  • Infection
  • Medical Personnel
  • Microscopes
  • Microscopy
  • Scanning Electron Microscopy
  • Staphylococcus Aureus
  • Therapy
  • Wound Infections
  • Wounds And Injuries

Fields of Study

  • Biology

Readers

  • Microbial Pathology
  • Molecular and Cellular Biochemistry

Technology Areas

  • Biotechnology
  • Directed Energy
  • Microelectronics