Interactions of Fibroblast with Cytotoxic and Invasive Strains of Pseudomonas aeruginosa on ECM Mimicking Fibers

It is estimated that approximately 2 million fires which occur in United States each year result in 1.2 million burn victims. Fibroblasts are responsible for responding to this tissue damage by breaking down the damaged extracellular matrix (ECM) and secreting a new ECM which aids in wound repair and supports the migration of immune cells. Pseudomonas aeruginosa is an opportunistic pathogen commonly associated with health-care infections (HCAIs) due to its ability to take advantage of immunocompromised hosts. However, little research has investigated how wound invading P. aeruginosa interacts with wound repairing fibroblasts. To address this lack of understanding, this thesis focuses on quantifying changes in fibroblast morphology, migratory behavior, and force exertion to investigate this host cell's response to representative cytotoxic (PAO1) and invasive (PA14) strains of P. aeruginosa. These assays study host cell-pathogen interactions on highly aligned nanofibers of varied spacing and diameter, which mimic the fibroblast deposited ECM and dictate fibroblast morphology. We discovered that the cytotoxic strain of P. aeruginosa induced significantly shorter fibroblast death times. Furthermore, two modes of death, sharp and gradual, were identified and found to be dependent on both fiber configuration and strain of P. aeruginosa. In addition, fibroblasts exposed to PAO1 migrating on the parallel formation were found to be significantly slower and less persistent than those exposed to PA14, however, fibroblasts exposed to both strains of bacteria were shown to exert similar forces. Lastly, exposure to PA14 led to the greatest change in actin, evident by increased actin punctae and less prominent actin stress fiber formation.

Name thesis:degree_name

Master of Science

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

Mechanical Engineering

Department dc:contributor.department

Mechanical Engineering

Grantor dc:publisher

Virginia Tech

Year dc:date.issued

2021

Author dc:creator

• Berman, Lauren Kathryn

Chair dc:contributor.committeechair

• Behkam, Bahareh

Committee members dc:contributor.committeemember

• Kale, Sohan
• Nain, Amrinder

• mechanobiology
• host cell-pathogen interaction
• fibroblast
• pseudomonas aeruginosa
• extracellular matrix
• nanofibers
• would infection

Statement dc:rights

• In Copyright

Licence dc:rights.uri

• http://rightsstatements.org/vocab/InC/1.0/

Dc Identifier Other

vt_gsexam:32227

OAI identifier oai:identifier

oai:vtechworks.lib.vt.edu:10919/114117