A new method developed for 3D printing of living microorganisms to enhance biomaterials

Researchers at Lawrence Livermore National Laboratory use LED light from a stereolithographic device for microbial bioprinting 3D printers developed by LLNL to “capture” microorganisms in 3D structures by suspending bacteria in photosensitive bioresin. bottom. Projection stereolithography machines can print at high resolutions on the order of 18 microns. It is about as thin as the diameter of a human cell.Credit: Illustration by Thomas Reason / LLNL

Lawrence Livermore National Laboratory (LLNL) scientists can use artificial bacteria to recover rare earth metals, purify wastewater, detect uranium, etc. by developing a new method for 3D printing of live microorganisms in controlled patterns. Expanded sex.

The research team has succeeded in printing an artificial biofilm that resembles a thin layer of a microbial community that is widespread in the real world through a new technology that uses light and bacteria-injected resin to generate 3D patterns of microorganisms. bottom. The research team floated bacteria in a photosensitive bioresin and used LED light from a LLNL-developed stereolithography device for microbial bioprinting (SLAM) 3D printer to “capture” microorganisms into a 3D structure. Projection stereolithography machines can print at high resolutions on the order of 18 microns. It is about as thin as the diameter of a human cell.

Articles published online in the journal Nano letter, Researchers have demonstrated that the technology can be effectively used to design structurally defined microbial communities. They demonstrated that such 3D-printed biofilms could be applied to uranium biosensing and rare earth biomining applications, and showed how shape affects printed matter performance.

“We are pushing the cutting edge of 3D microbial culture technology,” said William “Rick” Hines, a senior researcher and LLNL bioengineer. “We think it’s a space that hasn’t been very well investigated and its importance is not yet well understood. We find that microorganisms are geometrically complex yet highly controlled. We are working on the development of tools and techniques that researchers can use to better investigate how they behave under these conditions. Access to a more controlled and applied approach to the 3D structure of microbial populations. By strengthening, it can directly affect the interaction of microbial populations and improve system performance within the biomanufacturing process. “

Although seemingly simple, Hines explained that microbial behavior is actually very complex and driven by the spatiotemporal properties of the environment, including the geometry of members of the microbial community. According to Hines, how microbes are organized affects a variety of behaviors, including how and when they grow, what they eat, how they work together, how they protect themselves from competitors, and the molecules they produce. May be given.

Previous methods for producing biofilms in the laboratory have little control over the microbial tissue within the film, limiting scientists’ ability to fully understand the complex interactions found in natural bacterial communities. Hines explained. With the ability to bioprint microbes in 3D, LLNL scientists better observe how bacteria work in their natural habitats, and “electron-eating” bacteria (electrotrofus) surplus power during off-peak hours. You can investigate technologies such as microbial electrosynthesis that transforms. Produces biofuels and biochemicals.

Currently, the inefficient interface between electrodes (usually wires or 2D surfaces) and bacteria limits the electrosynthesis of microorganisms, Hines added. By 3D printing microorganisms on devices combined with conductive materials, engineers can achieve highly conductive biomaterials with significantly expanded and enhanced electrodes and microorganism interfaces, resulting in a much more efficient electrosynthesis system. It needs to be realized.

Biofilms are of increasing interest in the industry, where they are used as hydrocarbon repairs, important metal recovery, barnacle removal from ships, and as biosensors for a variety of natural and artificial chemicals.Constructed based on the synthetic biology function of LLNL, which is inhabited by bacteria Kauro Bactor Crescents Researchers at LLNL, who extracted rare earth metals and genetically modified them to detect uranium deposits, investigated the effect of bioprinting shape on microbial function in their latest treatise.

In a series of experiments, researchers compared the recovery rates of rare earth metals in different bioprint patterns and found that cells printed on a 3D grid could absorb metal ions much faster than traditional bulk hydrogels. Shown. The team also printed a live uranium sensor and observed an increase in the fluorescence of the manipulated bacteria when compared to a control print.

“The development of these effective biomaterials with enhanced microbial function and mass transfer properties has important implications for many bioapplications,” said co-author and LLNL microbiologist Yongqin Jiao. “The new bioprinting platform not only improves system performance and scalability with an optimized shape, but also maintains cell viability and enables long-term storage.”

LLNL researchers continue to develop more complex 3D grids and create new bioresins with better printing and biological performance. They value conductive materials such as carbon nanotubes and hydrogels for transporting electrons and supply bioprinted electrotrophic bacteria to increase production efficiency in microbial electrosynthesis applications. The team is also deciding how to optimize the shape of the bioprinted electrodes to maximize mass transfer of nutrients and products through the system.

“We are just beginning to understand how structure governs the behavior of microorganisms, and this technology is a step in that direction,” said Monica Moya, a bioengineer and co-author at LLNL. “Manipulating both microorganisms and their physicochemical environment to achieve more advanced functions has a variety of uses, including biomanufacturing, repair, biosensing / detection, and even the development of artificial biomaterials. These materials are autonomously patterned and can be self-healing or sensitive / responsive. To their environment. “

Reference: “Projection Microstereolithographic Microbial Bioprinting for Engineered Biofilms” by Karen Dubbin, Ziye Dong, Dan M. Park, Javier Alvarado, Jimmy Su, Elisa Wasson, Claire Robertson, Julie Jackson, Arpita Bose, Monica L. Moya, Yongqin Jiao, Williams , January 28, 2021 Nano letter..
DOI: 10.1021 / acs.nanolett.0c04100

A research and development program led by the Institute funded the research.

Co-authors include LLNL scientists and engineers, Karen Dabin, Jie Don, Dan Park, Javier Alvarado, Jimmy Sue, Elisa Wasson, Claire Robertson, Julie Jackson, and Washington University in St. Louis. Includes Alvarado Bose. A new method developed for 3D printing of living microorganisms to enhance biomaterials

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