VCOM Louisiana Research Day Program

Pharmacology

Stephanie Torres, OMS-III 1 ; Sarah Voth, PhD 1 ; Hassan Ebrahim, PhD 2 ; Kristina McLeod-van Amstel, OMS-II 1 ; Caleb L. Hamilton, PhD 3 ; Khalid A El Sayed, PhD 2 ; K. Adam Morrow, PhD 1 1 Edward Via College of Osteopathic Medicine-Louisiana, Monroe, Louisiana; 2 University of Louisiana Monroe College of Pharmacy, Monroe, Louisiana; 3 Greenville University, Greenville, IL 53 OLIVE OIL DERIVATIVE, OLEOCANTHAL, AS A NOVEL WAY TO FIGHT NOSOCOMIAL PNEUMONIA

Context: Nosocomial Pseudomonas infections place a great burden on healthcare staff and hospitals, complicating inherently difficult cases, elongating length of stay within the hospital, and increasing morbidity for patients. In recent years, healthcare providers have seen a rise of antibiotic-resistance bugs and have expressed the need for alternative approaches to combat infection. Moreover, it has been postulated that Pseudomona s infections can lead to secondary chronic effects of which we do not yet fully comprehend through microvascular and cellular damage to the surrounding tissue environment. A natural olive oil phenolic, (-)-Oleocanthal (OLC) has shown protection against lipopolysaccharide-induced acute lung injury in poultry and downregulated inflammatory markers in vitro . Objective: In this study, we sought to determine whether administration of OLC can provide protection against Pseudomonas aeruginosa induced damage in pulmonary microvascular endothelial cells. Methods: Using doses ranging from 0.1 µM to 50 µM of OLC, we sought to test its safety and effectiveness on both primary infection with Pseudomonas aeruginosa and on exposure to experimentally derived cytotoxic material

in pulmonary microvascular endothelial cells (PMVECs). PMVECs were pre-treated with OLC for 2 hours in a dose range from 0.25 – 1 µM prior to Pseudomonas aeruginosa infection. Images were captured at 6 hours post-infection, at which point supernatant was collected, filter sterilized, and transferred to naïve cell monolayers to investigate the effects of this cytotoxic material derived from infected cells. Images were captured at 24- and 36-hours post transfer. In separate experiments, rather than pre-treatment, we cotreated PMVEC monolayers with a range of OLC (0.1 – 50 µM) at the time of P. aeruginosa infection. Images were captured hourly during the primary infection, supernatant was collected at ~6 hours post-infection, filter sterilized, and transferred to naïve PMVEC monolayers. Results: We visualized significant protection to the cellular monolayers during primary infection and in supernatant transfer using a pretreatment strategy with OLC in a dose-dependent manner. Using the cotreatment approach with OLC, we did not observe protection in the primary infection, but protection against the cytotoxic supernatant was observed. The optimal dose of OLC in our experiments appears to be in the 0.5 – 1.0 µM range, with significant cell death see at doses greater than 5 µM and minimal protection

seen at doses less than 0.5 µM.

Conclusions: In conclusion, we discovered that OLC had a positive effect on cellular health in response to both primary Pseudomonas infections and cytotoxic supernatant exposure. In future studies we plan to use lactate dehydrogenase (LDH) release assays to quantify cell damage and immunoblot analysis to assess the production of beta-amyloid and oligomeric tau species, proteins known to propagate cell damage in response to P. aeruginosa infection.

69 2023 Via Research Recognition Day