Team:Aalto-Helsinki/Safety
Safety
We and our universities take safety seriously. “Horror stories” are important: even though low safety level biology laboratories may seem free of dangers, it is not the case. Although the risks differ from the perhaps more evident risks in a chemistry laboratory, for example, it is even more likely that things go wrong when one does not spare the time and effort to consider safety. You can find our filled Safety Form from here. On this page, we will explain in more detail what safety measures we have taken into account, what kind of safety questions we have considered and why we find those areas important.
Safety in Laboratory Work
Safety training for the whole team was provided by Aalto University before starting laboratory work, as we worked in their laboratories. Even though most members of our team had taken several laboratory safety courses before, this training introduced Aalto University practices and waste management to us. We collected safety-related materials to our team Drive folder, so that the members who had no laboratory experience could study the basics, in case that they would need to help out with some basic tasks in the lab.
There were a few procedures which required additional attention in the laboratory work. For example, for quickly freezing samples, liquid nitrogen was used. In our laboratory building, liquid nitrogen is stored in bigger containers in a storage room. Pouring the nitrogen from the large container to a smaller one requires special gloves and boots, so that burns can be avoided if something goes wrong. We handled liquid nitrogen very carefully, and also paid attention to safe usage of ethidium bromide, which was used a few times. Later, however, we used SYBR Safe, instead, as it is documented to be less mutagenic, and Aalto University has started using it more often for that exact reason. The university hears what its employees say and quickly changes practices and substances that pose a safety threat. During the last phases of our project, we managed to ship BioBricks back to iGEM on time, even though we had to resent them after the deadline because of a human error on labeling from Aalto University's shipping department. The newer shipments are declared and labeled as non-hazardous, non-infectious DNA and submitted according to the Registry Submission guidelines.
Our cloning organism was the TOP10 strain of Escherichia coli, whereas for production and antimicrobial activity testing, the T7 (Safety data sheet) express competent E. coli was used. Both of these organisms belong to Risk Group 1. This ensured safe handling in our Safety Level 1 laboratory, as we did not have access to laboratories with higher safety levels. We were originally considering antimicrobial testing of our produced peptide with Staphylococcus Aureus, Propionibacterium acnes and possibly other organisms. However, we did not proceed to those tests, as we could not find a proper safety level laboratory, and we could also verify the activity of our peptide by other means. We successfully demonstrated that the peptide we produced is antimicrobially active against the production strain of E. coli. Additionally, there is already data available of the activity of our peptide against P. acnes, S. Aureus and several other organisms. Furthermore, our construct with the peptide (DCD-1L) linked to the cellulose binding domain (CBM) was not active against E. coli, so it is most likely not active against pathogens, either. Had we had the chance to continue our experiments, we would have tested the activity with liposomes - simulating the lipid bilayer constitution of the pathogens. This was suggested to us by the Estonian team instructors. We are grateful for the brilliant suggestion, though!
Safety in the Concept Design
We developed a concept of an acne treatment hydrogel with our dermcidin peptide (DCD-1L). Considering our concept and product design, the end product needs to be safe to use on the human skin, on an everyday basis. We are lucky in this regard, as the peptide we worked with is already present on the human skin. The only thing that needs to be considered is whether abnormally high concentrations of the peptide risk the development of resistance against it. In that case, the body would lose its natural mechanism of defense against acne. One article [1] hypothesizes, however, that it may be much less likely that resistance develops against this peptide, as the peptide is evolutionarily very recent and resistance would require big changes in the constitution of the bacterial cell wall or membrane.
As dermcidin’s mode of action is currently not known for certain, the defense mechanisms that bacteria may develop are also pure guesses. Some caution should therefore be exercised. However, considering other current methods of treating acne, our peptide seems like the least risky option. Compared to antibiotics, which are used to combat far more dangerous diseases, resistance against our peptide does not pose a huge public health risk. From the patient’s perspective, our peptide is a great solution in the sense that it does not likely induce any side effects, as it only acts on our skin. Although being very effective in diminishing acne, isotretinoin sometimes induces symptoms of depression and is especially difficult to take as a woman: one must not be pregnant, breastfeeding and the likelihood that one gets pregnant must be very low. This is because isotretinoin is teratogenic - it can disturb the development of an embryo or a fetus. In medicine, a perfect solution may not always be available. Thus “good treatment” could be seen as the least risky option.
During the summer, we had two major application areas in mind: acne and wound treatment. In wound treatment, many more safety questions would be relevant. One would need to consider whether dermcidin causes any problems if it enters the bloodstream. We discovered that wound care is a very complex application area, where no perfect product or method exists. We discussed with Kirsi Isoherranen, MD and found out that clinical studies are hardly ever conducted in a reliable manner, because every person and every wound is very different. Our conclusion was that our peptide may not be efficient enough in this application area, as it is really important that no harmful bacteria remain once the wound area is cleaned. DCD-1L might also not be able to compete with silver products that are on the market.
A good side of the DCD-1L peptide is that it is likely that introducing it to the skin, even in much higher concentrations than it naturally is, would not cause significant changes in the skin microbiota. This is of course something that cannot be known for certain, and it would be wise to test it.
References
[1] Schittek, B. (2012). The multiple facets of dermcidin in cell survival and host defense. Journal of innate immunity, 4(4), 349-360.
