Thesis
Improving understanding of acute recurrent tonsillitis through 3D bioprinting of bacterial biofilms
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2023
- Thesis identifier
- T16581
- Person Identifier (Local)
- 202153789
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Introduction: Acute recurrent tonsillitis is a chronic, biofilm-related infection which is a significant burden to patients and the NHS. It is often treated with repeated courses of antibiotics, which contributes to antimicrobial resistance. Studying surface associated biofilms is key to understanding this disease process. In vitro modelling of this type of infection using 3D bioprinted hydrogels is a promising approach to achieve this. The aim of this study was to create the first 3D in vitro model of a multi-strain bacterial biofilm, with assessment by morphotype and viability analysis, and antibiotic susceptibility testing. Methodology: This study required development of a hydrogel formulation which displayed mechanical properties suitable for 3D printing and supported bacterial growth and formation of biofilm. Multi-strain bacterial culture of pseudomonas fluorescens and escherichia coli K-12 was performed to allow production of bacterial bioink and bioprinting of bacteria-laden 3D hydrogel construct fabricated using computer-aided design. This construct was cultured to develop a biofilm. The resulting specimens were assessed by morphotype and viability analysis by fluorescence microscopy, and antibiotic sensitivity testing versus a planktonic culture control. Results: A 3D printed bacteria-laden hydrogel construct was successfully fabricated. Biofilms were observed using optical fluorescence microscopy. A Live/Dead cellular staining protocol demonstrated biofilm viability, with high ‘Live’ signal in visualised biofilm colonies. Antibiotic sensitivity testing was inconclusive. Discussion: This study demonstrates first use of 405 nm light-based stereolithography 3D printing of a bacteria-laden bioink to manufacture a hydrogel construct which supports formation of bacterial biofilms. Further development of this biofilm model will increase fidelity and improve understanding of acute recurrent tonsillitis. This could support development of novel therapeutics which will in turn will reduce excessive antibiotic prescribing which drives antimicrobial resistance. Initiating a study with clinically relevant ex vivo tonsil bacteria will be an important next step in improving treatment of this prodigious, impactful, but understudied disease.
- Advisor / supervisor
- Shu, Will
- Resource Type
- DOI
- Date Created
- 2022
- Embargo Note
Relations
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File | 2023-05-30 | University of Strathclyde |