Description
- Abstract:
- Introduction: Understanding how infection develops following surgery is of paramount importance for orthopaedic surgeons. Infection is particularly problematic in arthroplasty and fracture implants, where colonization by opportunistic pathogens can occur. These organisms form biofilms, dense colonies of bacteria largely resistant to antibiotics and the immune system. Research to formulate materials and irrigation methods that reduce or remove biofilm formation on implants is being carried out, but no reproducible platform to visualize and quantify biofilm formation is available. This study describes a technique to digitally analyze a visual, and quantitative biofilm model, which allows for the evaluation of irrigation techniques and biofilm development. Methods: A computational tool was developed in the Python coding language that consistently identifies and measures the presence of bacterial biofilm on implant materials. This tool takes a series of confocal microscopy images corresponding to vertical slices of the biofilm and calculates the amount of protein, extracellular DNA (eDNA), and exopolysaccharide which have been stained with Sypro Ruby, TOTO-1, and Concanavalin A dyes. The relative ratio and number of pixels covered by each component is rapidly output and a three-dimensional plot is created depicting biofilm generation in a spacio-temporal manner. Results: Our studies have shown this program to be successful by consistently and accurately analyzing complex biofilms, and generating three-dimensional plots for explant materials and in-vitro testing. This tool, in conjunction with an in vitro biofilm platform and hospital-acquired explant materials, has been able to quantify the components of biofilms in approximately 30-120 seconds depending on number of stacks. In experiments aimed at showing the efficacy of irrigation agents on pre-established biofilms, this technology visualized a significant decrease in biofilm coverage by a three-dimensional and easy-to-read output. Discussion: With this technique, various interventions designed to limit biofilm formation on implant materials can be tested and compared to determine the most effective form of treatment. The data acquired by this high-throughput, quantitative, and visual tool will prove critical to the study of biofilms and removal methodologies.
- Notes:
- Scholarly concentration: Non-Scholarly Concentrator
Access Conditions
- Use and Reproduction
- All rights reserved
Citation
Turcu, Adrian, Berns, Ellis, Barrett, Caitlin, et al.,
"Application of a Novel Software for Analysis and Visualization of Biofilms on Implant Materials"
(2020).
Warren Alpert Medical School Academic Symposium.
Brown Digital Repository. Brown University Library.
https://doi.org/10.26300/wy0t-ya73
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Application of a Novel Software for Analysis and Visualization of Biofilms on Implant Materials
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Application of a Novel Software for Analysis and Visualization of Biofilms on Implant Materials
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Warren Alpert Medical School Academic Symposium
The Warren Alpert Medical School Academic Symposium is an annual event at Warren Alpert Medical School of Brown University that provides Year II medical students a venue to present their summer research in a poster format. Participation in the Symposium …...