Week 7 was essentially
our (mine and Aarons) last week in the hospital seeing patients/observing
surgeries/etc. which we’ve become so used to the flow of things. Our last week
Dr. Spector would be traveling, and we anticipated us being in lab in our
newfound “free-time”, trying to finish up experiments/make whatever we could
out of our lab work thus far. Coincidentally, our last major procedure was the
same type as we had seen on our very first day of immersion, a mandible
reconstruction with a free fibula flap. When I look back at these two experiences,
it makes me think of just how far we had come/how much we have experienced in
the short amount of time. I walked into the OR with a sense of
confidence/familiarity, having accumulated enough knowledge/experience to give
context to what’s going on in the room at each stage of the procedure. The
surgery played out more or less as was discussed during their video-conference
planning meeting the month prior, down to the minute details drilled over and
over by the respective surgeons (ENT, Oral, and Plastics (Dr. Spector)).
Additionally, for this procedure they were able to hookup the video camera
attachment to the microscope, so we could watch with great detail the barely
visible movement which Dr. Spector and his fellow were performing in order to isolate/anastamose
the respective blood vessels. With the whole room operating smoothly/efficiently,
Dr. Spector’s team finished up the operation hours earlier than our first
reconstruction (roughly 8AM to 10PM at night).
My final week of immersion
was spent in the lab. Initially, I was highly optimistic. After
trouble-shooting my MeHA-Megel photocrosslinking protocol (requiring far
greater concentration of Inrgacure photoinitiatior/UV exposure time), and
undergone training/clearance on the confocal LSM at the college, I was eager to
see how the long term viability of seeded HUVEC/encapsulated Pericytes would
turn out. Admittedly, my approach in preparing whole subsets of my respective
time-points en mass was atypical, and frankly poor experimental design, “placing
all my eggs in one basket” if you will. However, given the limited amount of
time I had left towards the end, I viewed it as my last chance at obtaining
usable data. Alas, it didn’t, and that is all I would like to speak of on that
subject if you will… The high degree of cell death could have been due to a
number of issues given how substantially I had deviated from my lab’s normal
photocrosslinking protocol (e.g. the Irgacure concentration at that point may
have been highly cytotoxic). After learning from one of my lab mates from the
bioprinting project that they’ve recently been coming into similar issues in
photocrosslinking with that newest batch of material I had been using (possibly
due to poor methacrylation) I decided to instead finish teaching several new
long-term lab members the different assays/imaging/etc. which would be required
if Dr. Spector would rather have his students conduct these in vitro studies in
NYC versus me conducting them back in Ithaca after more material has been
produced (methacrylation/purification is a time intensive process). I outlined
the setup/obtained the components necessary for peristaltic pumping systems for
conditioning the tissue-engineered microvessel constructs moving forward, as
the new lab members were unfamiliar with these systems/process involved. I view
it as a necessary component moving forward in preparation of micro-anastamosing
these constructs to their rat model. Additionally, the long term viability of
the cell lining can be tested at physiologically relevant hemodynamic conditions.
I’ll have to wait until after Dr. Spector returns when the three of us (him, my
PI, and myself) can discuss the logistics of how to complete these studies
moving forward. I am greatly appreciative of the experience overall, having
seen so much in the hospital, but am looking forward to getting back into the
full swing of things back in Ithaca.
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