Bioprinting and Biofilm Mimicry: A Student Science Adventure


Biofilms, complex extracellular structures created by bacterial colonies, pose a serious threat to human health. They are responsible for survival and antibiotic resistance of many bacteria, and lead to serious infectious diseases such as cystic fibrosis and endocarditis.
Preventing or destroying biofilms is an important area of research, and could help cure these diseases. A major challenge in studying biofilms is our ability to create them reproducibly and precisely in the laboratory so that experimental results are reliable. The r3bEL Bioprinter can mimic biofilms through culturing and printing bacteria in an alginate medium onto a petri dish or another substrate. These highly reproducible biofilm mimicries can then be tested for effects of key variables  and of antibacterial substances.

About me
I am Shruteek Mairal, a sophomore at Irvington High School in Fremont. Science and technology have always fascinated me, and in the spring of 2016, I came to know of SE3D’s 3D bioprinting technology. Intrigued, I approached Dr. Mayasari Lim and Prof. Prashanth Asuri, co-founders of SE3D, for an internship opportunity. Over the summer that year, I experimented with novel applications for the r3bEL 3D bioprinter. In the fall, my dad introduced me to his friend’s daughter, Nikita Salunke. She was interested in microbiology and had been considering topics for a science fair project. We both soon realized that our interests were complementary, and we decided to combine our two fields of interest by printing bacterial biofilms with the r3bEL bioprinter. Dr. Lim agreed to let us borrow r3bEL and supervise the project, and the rest was history.

The Project

Both Nikita and I were motivated to do something well-defined that could be submitted to a science fair. After much discussion, we decided on the Synopsys Alameda County Science and Engineering Fair (ACSEF) as the forum for our project. However, a major challenge lay ahead - our schools were located in two different counties, mine in Alameda county and Nikita’s in Santa Clara county. We wanted to enter as a team, but most teams came from the same schools. After some back-and-forth with science fair officials in both counties, we secured permission to submit as a team into the Synopsys Science Fair.
In this project, we proposed a novel approach - the use of 3D bioprinting technology - to mimic the formation of biofilms so that the effects of various antimicrobial solutions can be studied. Specifically, we investigated the effects of varying concentrations of alginate, a natural sugar, and Penicillin Streptomycin, a known antibiotic, on the bacteria of a 3D-printed biofilm. We conducted experiments by taking Escherichia Coli from a pre-existing culture, then placing the bacteria into test tubes containing media broth with varying concentrations of alginate, and incubating the test tubes overnight.  The next day, we would use the r3bEL Bioprinter to print assays of the bacteria, along with varying concentrations of penicillin streptomycin in an indicator solution of X-GAL and Isopropyl β-D-1-thiogalactopyranoside (IPTG). The 3D bioprints would exhibit varying levels of blue dye from the X-GAL/IPTG solution depending on how much of the bacteria had survived. A time curve study was also conducted in the lab to ensure the viability of the 16-hour incubation period, in which we monitored the amounts of released indicator dye in the bacteria over time based on different concentrations of alginate, to ensure that the bacteria reached its peak survival phase at 16 hours. The petri dishes from both the time curve study and the normal experiments were then analyzed and compared to find trends of survival across concentrations of alginate and antibiotics.

The Outcome
Overall, the bacterial survival indicated that alginate increased the antimicrobial resistance of bacteria when added to biofilms, and penicillin streptomycin resulted in reduced viability of cultures, which supported our hypotheses. The project earned us a first place prize in the category Biology Microbiology Molecular Biology (BMMB). However, more than the award and the scientific discovery, this project was also a valuable experience for my partner and me. By showing us what it was like to work in a laboratory setting, the experiments piqued our interest in biology and biotechnology. 3D bioprinting proved to be a sophisticated technology, involving  hands-on interaction with physical and biological systems, making science accessible, interesting, and often even fun. Throughout the project, we also learned many valuable lessons, including the importance of discipline, consistency, and safe laboratory practices when working in a laboratory, conducting scientific experiments, or analyzing data. This experience will likely influence the choices we make for the rest of our educational careers, including the selection of majors and the use of technology in further pursuits, especially the r3bEL 3D bioprinter.

Concluding Thoughts and Advice for Prospective Students
I feel like I have found my passion. This application of biotechnology to the microbiology field was amazing to me, and over the course of the project, I have grown in this area. I am fortunate to have an invite from Dr. Lim to return to continue my work with SE3D. I have a number of ideas that I wish to explore as next steps, but as I have learned over the last 5 months, those steps will come one at a time, with discipline and rigor.

My advice for high school students who are interested in bioscience fields is to explore and experiment. I was able to get the opportunity to do this project because I pursued my interests in different fields, including 3D bioprinting technology. If I hadn't followed my interest in biology and modern technology, I would have never acquired the valuable experiences that came with doing this project. Don't be afraid to explore new fields, even if it's just to gain experience.

SE3D bioprinting platform helps students explore multiple fields with one device. 3D printing has been used primarily to make solid objects for many years now, but SE3D has extended that technology in a way that allows applications to scientific fields. This cross-disciplinary thinking enabled our project, and will help others like us become interested in technology and biology.


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