Detection of Microbial Catecholamines in a Model Wound Environment
Chronic wounds are defined as an injury that does not heal in a predictable pattern or time frame. The healing process is composed of specific stages and chronic wounds have been interrupted in one or more stages of this process. This project focuses on the bacterial component of an infected wound, specifically on the catecholamines influencing the bacteria in the wound site. The assay developed in this experiment is designed to optimally grow bacteria in a simulated eukaryotic wound environment, and evaluate biofilm expression and catecholamine production of the bacteria. Bacteria used in this study are Staphylococcus aureus and Pseudomonas aeruginosa. These bacteria are commonly found expressing a biofilm phenotype which protects them from the host immune system. Catecholamines, such as dopamine and epinephrine, are quantified by HPLC-ED, while biofilms will be analyzed by confocal microscopy. Cell growth and viability are determined by absorbance reading of live/dead cells by a spectrophotometer. From this data, we will be able to characterize the host/pathogen relationship in regards to catecholamines in-vitro.