Sep 18, 2015 | By Kira

A group of nine TU Delft students participating in the upcoming iGem Synthetic Biology Competition have built the first 3D printer capable of creating customizable and reproducible bacterial biofilm, and they’ve made it almost entirely out of the children’s toy K’nex. This development could allow scientists to more effectively study the disease-causing bacteria and devise better methods for removing it. spoke with Hector Sanguesa Ferrer, one of the students who developed the Biolinker 3D printer, in order to get a better understanding of their goals and how they were accomplished.

Though you may never have heard of them, biofilms are all around—and probably even inside—us. Produced by different species of bacteria, algae and fungi, biofilms are microorganisms that live in cell clusters on surfaces. They are formed when the bacteria adhere to a surface in a watery environment and begin to excrete a slimy, glue-like substance that can stick to anything from metals to plastics to soil and even biological tissues such as our organs and teeth. Dental plaque, for example, is one of the most common forms of biofilm.

What makes biofilm such an important area of study is that once formed, they become highly resistant to antibiotics and chemical control. In terms of human health alone, bacterial biofilm are responsible for a host of extremely prevalent and fatal diseases, including acute ear infection, bacterial endocarditis, cystic fibrosis and hospital-acquired infections. Within the both the medical and environmental sciences, gaining a better understanding of biofilm and finding an efficient method for their removal is a vital concern. 

3D printed lines of bacteria in varying layer thicknesses

According to Ferrer, one of the biggest difficulties in studying biofilm is that they can grow in many different states—from relatively harmless formations on rocks, to the above-mentioned diseases—and therefore have a host of variables. If these variables are not carefully and precisely recreated in a laboratory environment, then scientists aren’t getting the full picture.

In order to solve this, the Biolink team, comprised of multinational biotechnology, bionanoscience and applied physics students, set about finding a way to create reproducible, automated, cheap and customizable bacterial biofilm for testing purposes. Through months of research and testing, they made an important finding: “What we discovered is that the main component of biofilms is one strain of proteins, known as amyloids,” Ferrer explaned to us. “Using biotechnology techniques, we managed to make our homemade bacteria using these proteins.” The problem of making them automated and reproducible, however, still remained.

The team thus turned to 3D printing technology, which they knew had already been widely used in synthetic biology for printing organs and bone implants. At first, they tried adapting a normal inkjet printer for their purposes, but quickly realized this wouldn’t work. So, although none ofthe members were engineers or had any experience with 3D printing, they decided to design a printer themselves. “First of all we managed to get a pump…then we needed a structure that could be moved for printing in the 3rd dimension,” said Ferrer. “One of our team members suggested that we could use the K’nex in his basement. He brought everything to our office, and in one week we had our first prototype.”

Time-lapse video of the K'nex 3D printer being built

In case you didn’t grow up in the 90’s, K’nex is a popular construction toy system consisting of interlocking plastic rods, connectors, gears and wheels, which can be pieced together to create just about any model or machine you could imagine. Although they are designed for the 5-12 year old age group, makers of all ages and skill sets have used it for a variety of advanced projects, such as this Mec’Nex 3D printer or this filament spool holder. For the biofilm 3D printer, which they have called the Biolinker, K’nex allowed the students to create a completely customized machine quickly and at almost no cost.

Even then, things didn’t go entirely smoothly. During construction, the Biolinks realized they had run out of K’nex pieces, and had to turn to the public for support. “We had an interview with a radio station to explain the project…we also asked for people’s collaboration, so if anyone had a couple pieces of K’nex lying around like we did,” said Ferrer. “Unexpectedly in two days, a lot of kids and different people were calling our office and coming to our workplace.” They ended up with way more K’nex then they knew what to do with, but were incredibly grateful for the show of support. “We were really happy people were getting involved with our project…it shows people really care about that kind of scientific research.” As they’ve approached the deadline, the team has also used Facebook to make urgent appeals, such as when two of their K’nex motors broke down.

While the first prototype used only 300 pieces, it was simply a static structure that could support the petri dish. The final version, capable of 3D movement and powered by several motors, is made of 689 individual K’nex components. Per their desired goal, it can successfully print up to 4 layers of bacterial biofilm in a reproducible and automated way. “The bioink is made out of our engineered bacterial cells, which we successfully immobilized into an alginate hydrogel,” they explain. “With this bioink we can print lines with a resolution (thickness) of down to 1 mm. We were able to successfully print and image of up to 4 layers of bioink with only little mixing between the layers.” From start to finish, their project took roughly six months, including research and development.

The first continuous hydrogel line made by the K'NEX 3D printer

The Biolink team will be bringing the 3D printer and their findings to the 2015 iGem Giant Jamboree, the premiere synthetic biology competition for both university and high school level students. Taking place from September 24-28 in Boston, the Jamboree gathers over 4600 participants on 280 teams from countries around the world to solve real-world challenges and advance the synthetic biology field. The nine students involved hail from the Netherlands to Swizterland, Romania and Spain, and study at Delft University of Technology and Rotterdam University of Applied Sciences. The participants are Max van’t Hof, Tudor Vlas, Stefan Marsden, Samantha Basalo Vasquez, Michelle Post, Liana Uilecan, Marit van der Does, Anne Rodenburg, and of course Hector Sanguesa Ferrer.

When we last spoke with Hector, he and the team were frantically writing content, modifying the wiki page, and preparing the materials to bring to the U.S., yet he remained confident in the project: “It was really challenging… we have been working hard to get good results in the synthetic biology parts, and also with designing the 3D printer, but fortunately, we’re a very motivated team.” We’re wishing team Biolink all the best at the competition, and hope that their research and invention can contribute to biofilm research within the medical and environmental sciences.



Posted in 3D Printers



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