After four incredible weeks of observing and conducting my advisor’s experiments, I was able to design my own novel experiment this week! The goal of the experiment was to study the impact of Ibuprofen (Advil) and Naproxen (Aleve), two non-steroidal anti-inflammatory drugs (NSAIDs), on monocyte morphology, including change in MDW and release of prostaglandin and cytokines.
Ibuprofen (IB) and Naproxen (Nx) minimize pain response through inhibition of the cyclooxygenase 1 (COX1) and cyclooxygenase 2 (COX2) enzymes, which are involved in synthesis of proinflammatory mediators such as prostaglandin (PG). When designing the experiment, I took inspiration from a paper which studied the impacts of Nx on macrophages and synovial fluid cells (https://www.sciencedirect.com/science/article/pii/S1063458420300455?via%3Dihub), however we extended our research to include MDW as an additional metric. We aimed to determine whether there is any correlation between drug dosage and MDW, similar to correlation observed in literature between drug dosage and prostaglandin levels.
Our experiment involved 8 treatments– in addition to an untreated control and a control with only LPS added, each drug had three additional treatments: a sample with only the drug added, a sample with both the drug and LPS added at the same time, and a sample with LPS added 30 minutes before the drug. We chose a drug concentration of 100 µg/mL based on the procedures followed in two other similar experiments. We used whole blood samples, which must be experimented upon quickly due to the coagulation that begins around 5 hours after drawing. Since other studies that employed cell line samples instead of whole blood could incubate for 24+ hours, we increased our LPS concentration to stimulate similar results in a shorter period of time. I was thrilled when I was able to apply my stoichiometry skills learned in 11th grade Chemistry to calculate concentrations and molarity for dilutions and stock solutions.
Surprisingly, the samples treated with Nx and IB didn’t exhibit any MDW changes. We aren’t discouraged by these results, since they indicate that our chosen drug concentration wasn’t high enough. We plan to repeat the experiment next week with a higher concentration in order to generate more representative data. I look forward to reporting back in my week 6 summary with our results!
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