Title Information
Title
Application of a Novel Software for Analysis and Visualization of Biofilms on Implant Materials
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.
Name
Name Part
Turcu, Adrian
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Berns, Ellis
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Barrett, Caitlin
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Spake, Carole
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Leong, Jacqueline
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Born, Christopher
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name
Name Part
Garcia, Dioscaris
Role
Role Term (marcrelator) (authorityURI="http://id.loc.gov/vocabulary/relators", valueURI="http://id.loc.gov/vocabulary/relators/aut")
Author
Name: Corporate
Name Part
Brown University. Alpert Medical School. Scholarly Concentration Program. Non-Scholarly Concentrator
Role
Role Term: Text
research program
Subject (fast) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01048565")
Topic
Orthopedics
Subject (fast) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01150331")
Topic
Three-dimensional imaging
Subject (fast) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/00832066")
Topic
Biofilms
Language
Language Term: Text (ISO639-2B)
English
Origin Information
Date Created (keyDate="yes", encoding="w3cdtf")
2020
Note (displayLabel="Scholarly concentration")
Non-Scholarly Concentrator
Access Condition: use and reproduction (href="")
All rights reserved
Access Condition: logo (href="")
Identifier: DOI
10.26300/wy0t-ya73