Author Archives: Chad Moles

Chad Moles

About Chad Moles

My name is Chad Moles and I am a rising senior at Tulane University's School of Public Health and Tropical Medicine. I have worked endlessly to understand the science and art of preventing disease, prolonging life and promoting health. Ultimately, you will find I have extensive lab experience and a solid understanding of research strategies.

The End

This past friday was the final day of my Summer Undergraduate Research Fellowship with the Laboratory for Regenerative Wound Healing at Cincinnati Children’s Hospital Medical Center. I gave a poster presentation along with over 150 other students at 9:00am, which detailed my summers work and what I was able to accomplish. I am proud to say that I was awarded honorable mention, but the true award to me was being able to share my project with so many other people. In only a couple hours, I had over a dozen students, parents, research faculty, and doctors visit my poster. Now, I would like to share with you what I have done and find so interesting.

Capstone Symposium 2014

INTRODUCTION:

Looking at the big picture, we know that each year in the developed world about 100 million patients acquire scars, and the global value is undoubtedly higher. Scarring can be an emotional burden and lead to significant financial costs, which can be the result of aesthetic, physical, or psychological consequences. Currently, there is no effective scar therapy. With that in mind, our lab uses the fetal response to injury as a roadmap to achieve regenerative wound healing. What does that mean? If you were to make an incision or the fetus suffered an injury, it would heal without a scar. However, we obviously can’t do that, so what is the difference between the fetus and adults? Our lab has shown a significant role for IL-10 in the fetal regenerative response. Additionally, we have shown that viral over-expression of IL-10 in adult wounds results in scarless wound healing. However, there are some translational issues with this. Hence, our lab developed a sustained delivery hydrogel (HH10) that overcomes the translatable issues of gene therapy. The last thing that I want to mention is that currently, there is no quantifiable method to assess scars on a histologic scale. Wound healing relies heavily on histology, which is basically staining cells and looking at them under a microscope.

HYPOTHESIS:

If we develop a novel method to histologically assess dermal scarring, we can validate the properties of HH10 in cutaneous wounds AIM 1: Develop a new computational method of scar analysis compared to the regenerative benchmark of skin AIM 2: Validation of the extended release hydrogel using an innovative scar assessment tool

METHODS: AIM 1

My first priority for the summer was to develop a new method for analyzing scars histologically. To do this, I ordered mice and (following all ethical guidelines and standard protocol) made 4mm wounds on the backs of the mice. After 28 days, I collected the wounds, and looked at the differences between skin and scars (n=20/group). With these images, I was able to establish 5 parameters or statistical differences between skin and scar. These were height of the epidermis, orientation of keratinocytes (the main cell line in the epidermis), scar area, the number of dermal appendages (functional aspect of the skin, includes hair follicles, sweat glands), and the vascular density (presence of vessels).

METHODS: AIM 2

After I discovered these parameters, I wanted to test the labs hydrogel using this scar assessment scale. I followed the same basic protocol, but this time, after I made the wounds, I inserted the various treatment groups. Gel Control has all of the components in HH10, except IL-10. After 28 days, I collected the wounds and analyzed them using these parameters.

Treatment Groups

RESULTS:

I was able to quantifiably define a scar based on 5 parameters, as the skin and scar are statistically different. When you look histologically, you will find that HH10 restores scar parameters to levels observed in normal unwounded skin. Also, HH10 is statistically similar to gene transfer by viral over-expression. HH10 showed improved collagen arrangement, number of dermal appendages, and epidermal structure compared to Gel Control and scar.

CONCLUSION:

This scar assessment tool is a novel method, which has the potential to be used as the future benchmark in determining the efficacy of preclinical anti-scarring therapeutics to achieve regenerative repair. HH10 is a safe, non-viral mediated delivery system that restores epidermal and dermal scar parameters to the levels observed in normal unwounded skin. Lastly, the potential therapeutic benefits of HH10 extend beyond cosmetic benefit, but may apply to a broad range of diseases characterized by excessive fibroplasia, including post-surgical wounds, intra-abdominal adhesions, pathologic scars and organ fibrosis.

It has been a fantastic summer full of learning and growth. I just want to take a moment to acknowledge the Center for Engaged Learning and Teaching, because without the CELT Summer Internship Award, I wouldn’t have had the successful summer I wrote about in this post.

The clock is ticking

I have 2 weeks left until my program at Cincinnati Children’s Hospital Medical Center ends, which is when I will present a poster highlighting my work at the annual Capstone Symposium. There will be 100 students presenting their work at this competition. As far as my experience here is concerned, I have since changed projects because my previous experiment did not pan out. As mentioned in my previous post, I had tried to inject fatty tissue loaded with IL-10 (chemical messenger) into wounds to induce wound healing and prevent scar formation. While I had done a literature search on the subject and followed similar protocols to those online, I had difficulties isolating specific components of the fat. Now, I am working on a project where I basically load IL-10 into a gel that was previously developed by the lab, which can be applied topically to wounds. I bought mice from a supplier, created 3 wounds on the back of the mice (located on either side of the spine and behind the head), and then injected the gel into one wound (the other two were used as a controls). After 7 days, I humanely euthanized the mice following standard protocol and collected the wounds. I am currently in the process of staining the samples, and when I finish this and have analyzed the data I will post my findings on here. I also realized I haven’t posted any photos, so I am going to fix that now.

This is my work desk:

My desk

My desk

This is my lab space:

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This is one of the many surgery rooms in the veterinary facility:

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Lastly, this is a picture of the lab:

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Overall, it has been an exciting, fast-paced summer, and I look forward to sharing my findings with the rest of my peers and the scientific community.

-Chad

 

New and Noteworthy

This summer I am interning with the Summer Undergraduate Research Fellowship (SURF) at Cincinnati Children’s Hospital Medical Center (CCHMC) within the Fetal Cellular and Molecular Therapy division. The lab I work for focuses on regenerative medicine, specifically scar prevention and wound healing. As I am going into my second week here, I thought I would fill everybody in on my project and my plans for the summer.

Due to recent advances in cellular biology, the scientific community has developed a particular interest in fat grafting. The term fat grafting refers to when a doctor harvests fat from a patient’s body and then reinserts it at another location. This reconstructive and cosmetic procedure first became popular in the 1980’s when liposuction was first introduced, as fatty tissue is readily available, easy to obtain, and inexpensive. Since then, there have been encouraging results in experiments suggesting that the transfer of adipose tissue or fat to a wound can mitigate the formation of scars and can aid in the process of healing. Take a second to absorb that and consider the possible implications. Since there are over 200 million surgeries occurring each year around the world that end in the formation of a scar, the discovery of a therapeutic targeting this biological process has the potential to significantly decrease the psychological and physical burden that comes with scarring.

Our lab firmly believes that Interleukin-10 (IL-10), a cytokine or chemical messenger, is the driving force behind wound healing. My goal this summer is to show that fatty tissues ability to lessen scar formation is mediated by IL-10. With this in mind, I have ordered C57BL/6mice (this is the standard laboratory mouse) that I will use for my experiment. My methodology, generally speaking,

  1. Harvest fat from the groin area of the mouse (because there is more fat here than any other area)
  2. Process the fat (I have to remove the oils, blood, etc.)
  3. Create two wounds (4mm each) on the back of the mouse (I will cover the wound with a clear “band aid”)
  4. I will take a syringe and inject the fat (with various supplements) onto the bed of the wound
  5. I will humanely euthanize the mice at various time points (for example, day 7 and 28) and harvest the wounds along with the surrounding area
  6. I will then take histological images and run a variety of tests for analysis of different wound healing/scar parameters (for example, the quantity of new vessels formed in the wound bed)

Overall, I have to say it was a great first week. I went through badging easily, met new people in the lab, worked on this experimental plan with my supervisor, and the SURF program hosted a graduate fair and picnic to meet other SURF students and faculty, learn about recent advances in research, and just hang out. I am thrilled to be moving forward with this fat grafting project, and will post again soon.

-Chad