Week 1 - Natalie

Due to a research collaboration requiring travel to Australia from June 9 – June 26, I began my first immersion week from June 1 – June 8. I will be performing my immersion in the lab of Dr. Mathias Bostrom at the Hospital for Special Surgery (HSS). One of my main goals, besides gaining a greater appreciation for the clinical side of orthopedics, is to learn the technique of quantitative reverse transcription polymerase chain reaction (qRT-PCR). Wednesday was mostly dedicated to securing an ID for HSS and meeting Dr. Bostrom's surgical assistant and starting the process of getting a scrub card. Thursday morning I attended grand rounds for the adult reconstruction and joint replacement service. This meeting is split into two segments. The first half hour is dedicated to case presentations, and the last hour is a research or clinical presentation on a specific topic. This week, the case presentations focused on the use of fluoroscopy in the operating room and several cases that could have benefited from this technology were presented. The clinical presentation was by two physicians, Dr. Alexiades and Dr. Sculco. Each surgeon presented their preferred method of minimally invasive total hip replacement surgery, with Dr. Alexiades championing the posterior approach, and Dr. Sculco focusing on the anterior approach. Even though each surgeon uses different approaches, both agreed that either technique could be successful and that a surgeon should use whatever approach they are most comfortable with. This meeting was especially interesting to me, as it was very clinically focused. I participated in a lab meeting for Dr. Bostrom's group on Friday morning. At this meeting, several current projects in the lab were discussed. From Monday-Wednesday I was able to get involved with lab work by assisting an orthopedic resident who is taking a year off for research in the first steps of the qRT-PCR protocol. This was great, since it is a main goal of mine to learn this technique this summer. The study that Dr. Kitay is performing qRT-PCR for was a fracture model in young and old mice. The bone tissue had already been harvested from the mice. The first steps of the protocol involved pulverizing the bone sample and putting the sample in Trizol, which is used to isolate the RNA. This process took one day, for twelve samples. The next day, we extracted the RNA from the trizol solution by adding chloroform and performing successive buffer washes and spin downs to extract the RNA. We then examined the RNA concentration, 280/260 peak, and 280/230 peak using a NanoDrop machine to get an idea of how successful we were at extracting RNA from the samples. We had excellent concentrations (much higher than expected) and adequate 280/260 and 280/230 peaks for all of the samples. Although this is a helpful measure and likely means that we performed the protocol successfully, the samples still need to be analyzed with an Agilent machine to get the RNA integrity number (RIN), which will confirm that our RNA extraction was successful. It was great to be able to get involved in the lab so quickly, and I look forward to performing the next steps of the protocol and seeing surgeries and attending clinic when I return from Australia!