The Electrospinner

What if every time you went for a drive, you had to build the road in front of you?

Not only would that suck, but it would take forever to get anywhere. But, this is exactly how wound healing works. Cells have to lay down the "road" in order to grow. Cells can not automatically start growing in a wound. In fact, they require a structure (or matrix) to grow and move on. A release of matrix proteins creates this matrix structure, but this only happens one fragment at a time since the proteins are released as the cells move. This is then continued until the wound is closed. As is the case with the driving analogy, this process is slow. Very slow.

But what if this process could somehow be accelerated? What if the matrix was already in place before the cells started moving? Since I'm writing about this, you've probably realized that there must be a way to do just that. And yes, there is! With the help of an electrospinner, a scaffold can be created. From there, the scaffold can be inserted inside a wound, which will give the cells an open highway to start moving on. So now you're probably wondering, what's an electrospinner? To put it simply, it's a fancy machine that creates scaffolds:*

Electrospinner setup. A) syringe pump. B) Electrospinning cage for maintenance of humidity and electrical current. C) syringe needle for distribution of gelatin fibers. D) target. E) power supply.

But of course, the electrospinner can not make a scaffold from out of thin air! As the photo caption above hints, gelatin is used to create these scaffolds. Currently, two different gelatin's are being used: bovine and porcine. Bovine, which comes from cows and porcine which comes from pigs. The cool thing about gelatin is that it contains absolutely zero cells, which means that it will be easily accepted into our body regardless of the source. 

Why does this matter? 

Because, anything non-biological is considered really foreign to the body. So the best option is to use something biological. And sure, cells are biological, but if they don't belong to us, they're pretty foreign. So here's gelatin to save the day! At the moment, we have not determined which gelatin (bovine or porcine) is better for creating the scaffolds, so both are being used. 

Now, the actual creation (or spinning) of the scaffold takes about an hour. This means, an hour of waiting and watching. It would be nice to leave the machine to do it's thing, but every now and then a falling fiber has to be moved or cleaned off the syringe needle. It may be a bit hard tell, but in the photo below the fibers are coming out of needle and landing on the target (a circular piece of aluminum foil):*

  Once this is all set and done, we have ourselves a scaffold!* 

I can now say I've made a highway. A highway for cells.

-Tudor

*A big thanks to Martha and Tatum for the photos!

Scratch Assay pt. 2

And we're back! This time, with an actual experiment. 

Since the cells from last time didn't grow to full density, new cells had to be grown. This took about a week. Now, the experimental set-up remains the same as previously stated: scratch assays will be used to observe the effects of arsenic and estrogen in a wound. Theoretically, the arsenic contaminated wounds should take longer to close than the control, while the estrogen treated wounds should close around the same time as the control. 

I have to pause here and give credit to Bronson and Oscar. I would love to say that I performed this experiment, but let's be real here, that did not happen. Together, they did the work while I sat back and observed (but don't worry, I took notes). Now just as before, we had to check the growth density of our cells. Fortunately, nothing was stopping us this time! From here on, the actual step by step process of what happened is not worth noting, but here's a summary: The cells were moved from their original container to a "plate" consisting of 12 different wells. Since the plate can be divided into 3 rows, each row represented a different variable (control, arsenic contamination, and estrogen treatment of arsenic contaminated wounds). Once all of this was taken care of, Bronson scratched the cells with a pipette tip. From there, the wells were placed in the incubator to grow. And then we waited.

Every 4 hours, over a 20 hour time period, photos where taken of the wells. The results:

So as expected, the arsenic contamination slowed the wound closure. But, as shown above, estrogen treatment almost reversed the contamination's obstruction to the healing process. Now that's pretty cool.

-Tudor

Edit: I almost forgot...all of this started at 6 in the morning. Bronson...