Marine Biologist Modifies Bioprinting for the Creation of Bionic Coral

Corals are dying globally. In the face of climate change and global warming, we can expect some severe consequences, which in turn directly affects marine life. In what is panning out to be a mass extinction event, coral reefs have been dangerously threatened by toxic substances and excess carbon dioxide for years, causing the certain death of may of these diverse marine invertebrates. Once the coral is dead, the reefs will also die and erode, destroying important marine life, that would otherwise feed and spawn on it.

Considering that scientists have predicted that nearly all coral reefs will disappear in 20 years, it is crucial that we protect corals and learn from them. For the expanding field of biotechnology, untapped resources like corals hold great potential, as bioactive compounds for cancer research or simply as an inspiration for the production of bioenergy and bioproducts.

In an interview with 3DPrint.com, interdisciplinary marine biologist Daniel Wangpraseurt, from the University of California San Diego (UCSD)’s Department of NanoEngineering, explained how bioprinting technology was a pivotal point in his work to develop bionic 3D printed corals as a new tool for coral-inspired biomaterials that can be used in algal biotechnology, coral reef conservation and in coral-algal symbiosis research. 

“For many years I have been studying how corals optimize light management and discovered that there are lots of interesting evolutionary tricks, such as different growth forms and material properties, so I became interested in copying these strategies and developing artificial materials that could host living microalgae, just like corals do in nature,” revealed Wangpraseurt.

Daniel Wangpraseurt

As one of the most productive ecosystems globally, coral reefs use photosynthesis to convert carbon dioxide into energy that they in turn use for food. Even though light provides the energy that fuels reef productivity, key nutrients such as nitrogen and phosphorus are also required, but are found in very low quantities in warm tropical oceans where coral reefs are generally found, making scientists wonder how these marine animals have managed to create a competitive habitat with such limited resources.

A laser beam is intensely scattered by elastic coral tissue and aragonite skeleton. (Credit: Daniel Wangpraseurt)

Wangpraseurt described that, while different corals have developed a plethora of geometries to achieve such capabilities, they are all characterized by an animal tissue-hosting microalgae, built upon a calcium carbonate skeleton that serves as mechanical support and as a scattering medium to optimize light delivery toward otherwise shaded algal-containing tissues.

“Taking what we learned about corals and biomaterials, we began working on a project to develop a synthetic, symbiotic system using a 3D bioprinting approach. We know corals have both animal cells and algal cells, and, so far, we have mimicked the animal part of the corals, that is, the physical and chemical microhabitat that partially controls the activity of the algal cells.”

At UCSD, Wangpraseurt expects to continue recreating coral-inspired photosynthetic biomaterial structures using a new bioprinting technique and a customized 3D bioprinter capable of mimicking functional and structural traits of the coral-algal symbiosis. Along with fellow researchers from UCSD, the University of Cambridge, the University of Copenhagen and the University of Technology Sydney, and thanks to a grant from the European Union’s Horizon 2020 research and innovation program, and the National Institutes of Health (NIH), the team reported the results of their work on bioinspired materials that was published in the journal Nature Communications earlier this year.

“We want to go further and not just develop similar physical microhabitat but also modulate cellular interactions, by mimicking biochemical pathways of symbiosis. We hope that this allows us to not only optimize photosynthesis and cell growth, but also to gain a deeper understanding of how the symbiosis works in nature. By doing so, we can improve our understanding of stress phenomena such as coral bleaching, which is largely responsible for global coral death.”

Living colonies of Symbiodinium are visible within the 3D bioprinted tissues (Credit: Daniel Wangpraseurt)

So, how did bioprinters become the go-to technology for this project? Wangpraseurt explains that, while working as a researcher at the University of Cambridge’s Department of Chemistry Bio-Inspired Photonics lab, he noticed that scientists were using cellulose as a biomaterial with interesting optical responses. He was wondering how he could use cellulose to develop a material with very defined architectural complexity. 

“In the beginning, the main aim was to develop a coral-inspired biomaterial, that has a similar optical response as natural coral, and then to grow algae on it or within it. Thereby, we started off with simple techniques, using conventional 3D printers; however, it wasn’t very easy to recreate the spatial resolution we needed for corals.”

Inspired by 3D bioprinting research in the medical sciences, Wangpraseurt reached out to scientists at the UCSD NanoEngineering lab that were developing artificial liver models, and who later became collaborators in the project.

A laser beam is intensely scattered by elastic coral tissue and aragonite skeleton (Credit: Daniel Wangpraseurt)

The team went on to develop a 3D printing platform that mimics morphological features of living coral tissue and the underlying skeleton with micron resolution, including their optical and mechanical properties. It uses a two-step continuous light projection-based approach for multilayer 3D bioprinting and the artificial coral tissue constructs are fabricated with a novel bioink solution, in which the symbiotic microalgae are mixed with a photopolymerizable gelatin-methacrylate (GelMA) hydrogel and cellulose-derived nanocrystals (CNC). Similarly, the artificial skeleton is 3D printed with a polyethylene glycol diacrylate-based polymer (PEGDA).

Close up of coral polyps and living photosynthetic biomaterials. Living colonies of Symbiodinium are visible within the 3D bioprinted tissues (Credit: Daniel Wangpraseurt)

Based in San Diego, Wangpraseurt has spent months trying to recreate the intricate structure of the corals with a distinguished symbiotic system that is known to grow as it creates one of the largest ecosystems on the planet. 

“We used a 3D bioprinter that had been developed for medical purposes, which we modulated and further developed a specific bioink for corals. A lot of the work was related to the optimization of the material properties to ensure cell viability. Having the right bioink for our algal strains was crucial as if we were to use mixtures commonly used for human cell cultures, the cells will not grow very well and can die rapidly.”

The implications of the newly developed 3D printed bionic corals capable of growing microalgae are many. Wangpraseurt said he plans to continue working on bionic corals and potentially scale up the process for his startup, called mantaz, as well as for commercial properties; or to develop coral-inspired materials at a larger scale to have a more immediate impact on efforts related to coral reef restoration, and also for biotechnology.

SEM images of the skeleton structure of the coral Stylophora pistillata and the coral-inspired, 3D-printed material (Credit: Daniel Wangpraseurt)

Wangpraseurt is looking to scale the bioprinting system to have a more immediate impact on algae biotechnology, bioenergy, and bioproducts. He claims that he and his colleagues can “customize the environment of the algae and fine-tune the production of a certain bioproduct to potentially tap into the algae bioproduct market and scale the system for bioenergy production.” 

“Another interest of mine is to further develop a 3D bioprinted synthetic coral-algal symbiosis system, which can provide important insight into the mechanisms that lead to coral death, but can also result in the development of future technology for coral reef restoration.”

The researcher talks about coral reefs with a reverent passion that today goes beyond his lab work. When he is not moving the research along at USCD, Wangpraseurt is working with his social enterprise in Panama, as he and his team try to restore coral reef ecosystems to help coastal communities in the tropics, including local fishermen, by harvesting algae biomass that can be sold for different purposes, such as natural fertilizer, which contributes to an organic and sustainable chain of production. Furthermore, the coral-inspired aspects of Wangpraseurt’s research and startup company are really coalescing to enable him and his team to understand how corals work and, in turn, how we can learn from them for the benefit of our planet.

Nutrient sampling at a polluted reef in Panama (Image: Daniel Wangpraseurt)

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World’s Largest 3D Printed Coral Reef Now Resides in the Maldives

Coral reefs are important not only to the millions of species that live in them, but to humans as well. Reefs protect coastlines from storm surges during violent storms, and they also offer a great deal of income from tourism and fishing. Coral reefs are in quite a bit of trouble at the moment, though. 50 percent of the world’s reefs have died in the last 30 years, and 90 percent of the surviving reefs may die off in the next century if action isn’t taken to save them. The reefs are suffering from numerous threats – climate change, overfishing, damage from boats, and other human-caused damage.

Humans may be responsible for threatening the coral reefs, but many humans are also taking responsibility for saving and restoring them. 3D printing has been found to be an effective way of creating artificial coral reefs, which are then dropped into the ocean where it’s hoped, free-floating coral polyps will attach themselves to them and grow into new coral structures. That coral will then attract the fish and other species that make their homes in reefs, and a new, natural reef will grow from the artificial seed material.

Several reefs have already been 3D printed and sunk into the ocean, and now Reef Design Lab, an Australian organization dedicated to the creation of artificial reefs, has created what, it turns out, is the largest 3D printed artificial coral reef so far. 3D molds of the reef structures were 3D printed over 24 hours, then used to cast the structures in ceramic, which is similar to the calcium carbonate found in actual coral reefs. The 3D printed molds were printed in Australia and shipped to the Maldives, filled with concrete, assembled at Summer Island and then dropped into the ocean, where they now rest seven meters below the surface.

Live coral was then transplanted into the artificial reef, where it will hopefully grow and colonize the artificial structure, forming a new, living reef.

“3D printing technology helps us to develop more innovative ways of protecting coral reefs. The technology allows us to mimic the complexity of natural reef structures, so we can design artificial reefs that closely resemble those found in nature,” said Alex Goad of Reef Design Lab. “We hope this will be a more effective way of growing and restoring corals.”

The creation of artificial reefs isn’t new – humans have utilized sunken ships and concrete blocks to attract coral in the past. 3D printing, however, allows for the natural structure of coral reefs to be imitated, which attracts more marine life. In this case, Goad used CAD to design replicas of naturally-occurring coral reef structures before 3D printing them. The complex shapes of natural reefs are difficult to create in any other way, so 3D printing has become a valuable asset in the creation of these reefs.

It will take several years for coral and the other species that make reefs their home to populate the artificial structures, so we won’t know for some time if these 3D printed experiments are truly working. They show a lot of promise, though, and for the 25% of marine species that depend on coral, help can’t come soon enough. Scientists are hoping that the Summer Island reef will be covered in coral within the next two years, and if the project succeeds, more will likely follow.

“Projects like the 3D printed reef are popular among guests, who like that we protect our environment,” said Mari Shareef, Manager of the nearby Summer Island Resort. “And it’s not only for the guests. Our staff, most of whom are Maldivian, want to protect their environment. “Ultimately, we want to help promote a culture of environmental stewardship, not just at Summer Island, but across the Maldives.”

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[Source: Verdict/Images: Reef Design Lab]

Team Effort Uses 3D Printing to Restore Coral Reefs

[Image: SECORE: Paul Selvaggio]

Coral reefs are the most diverse ecosystems on Earth, with thousands of animal and plant species living in their colorful ocean-floor habitats. These reefs are in quite a bit of trouble currently, however. In the past 30 years, 50 percent of the world’s coral reefs have died and if changes aren’t made to slow the progression of climate change and curb other human-caused damage to the reefs, 90 percent of them may die in the next century. Coral reefs aren’t just vital to the plants and animals that call them home, but to humans as well – they provide a lot of income through tourism and fishing, as well as protecting coastlines during violent storms.

Saving them, therefore, is critical, and involves some human intervention at this point. Coral are sessile animals, meaning that they take root like plants but capture their food from the ocean water. Coral polyps root themselves in ocean rocks, gradually reproducing and growing until they form the lush, brightly colored reefs that people travel thousands of miles to see. It’s a slow process, though – coral reefs grow by centimeters each year, taking thousands of years to become large and thriving. Right now, coral reefs don’t have thousands of years, so they need our help.

Several organizations have been trying to help coral by 3D printing artificial reefs and sinking them in the ocean in hopes of attracting free-floating coral polyps to embed themselves and begin reproducing. An organization called SECORE International (Sexual Coral Reproduction) is also using 3D printing, but taking a more hands-on, aggressive approach. SECORE is a nonprofit global network of scientists, public aquarium professionals and local stakeholders working to protect and restore coral reefs. Along with its partners, which include the California Academy of Sciences (CAS) and the Nature Conservancy, SECORE is developing restoration processes that leverage the natural reproductive habits of coral.

3D printed seeding units. [Image: SECORE/Valérie Chamberland]

Certain coral species naturally broadcast egg and sperm cells, which are collected by SECORE, fertilized, and then raised in tanks until they become freely swimming larvae. Those larvae are then introduced to 3D printed “seeding units” that resemble places on natural reefs where coral would attach. Once the coral have embedded themselves, the seeding units are planted on reef areas in need of restoration.

It’s an effective approach, but a costly one, unfortunately.

“One of the ways SECORE is aiming to reduce these costs is by designing seeding units that do not need to be manually attached to the reef, but rather can be sown from a boat or other method, similar to how a farmer would sow seeds in a field,” said SECORE Project and Workshop Manager Aric Bickel.

3D printing is another way to keep costs down, as well as to rapidly produce the seeding units. SECORE aims to produce a million of the units by 2021, and hundreds of thousands of units annually by then. Phase One of the project is taking place in the Caribbean, with research and training hubs in Mexico, Curaçao and the Bahamas.

“3D printing allows us to do a bit of rapid prototyping. We were looking at several different materials, and 3D printing allows us to print a variety of materials,” Bickel said. “It also saves the cost of having to make molds or castings which, particularly for the initial prototypes, would be a significant amount of money invested.”

A diver with a tray of the seeding units [Image: SECORE/Benjamin Mueller]

CAS is one of SECORE’s primary funding providers, and because SECORE is a small team with limited engineering capabilities, CAS turned to the Autodesk Foundation, with which it looked into various design firms for help with the development of the seeding units.

“In collaboration with the Foundation, we reached out to several design firms,” Bickel said. “Emerging Objects seemed like they would be the best folks to help us out with this next design phase and hopefully with the iterative design phases as we go forward.”

One of the main challenges SECORE has been having is finding the best material and design combination for the seeding units. Not just any shape can be used – the units need to be able to wedge themselves into the reefs without manual assistance. The material is an issue, too. SECORE had been using rough cement for the seeding units, but that material worked a little too well – in addition to attracting corals, it also attracted quite a few competing organisms.

“One issue was with competition from other species on the units themselves,” said Bickel. “What the trials showed is that a slicker surface will cut down on that potential competition. The needle that you have to thread here is having a surface that’s rough enough for corals to settle on and to attach to but smooth enough that it’s not a good location for other organisms such as sponges and algae to attach to.”

Several years of trials and experiments revealed ceramic to be a good potential material for the seeding units. Emerging Objects has plenty of experience in the experimental use of 3D printed ceramic, but needed to be able to 3D print the material on a large scale, so the company reached out to Boston Ceramics for help.

“Boston Ceramics is one of the few companies we’re aware of in the world that can potentially meet some of the demands for the number of substrates we’ll be using,” said Bickel.

The team used Autodesk Netfabb to design the original shape, a tetrapod, for the seeding units, and has been experimenting with other designs that are better suited to landing and wedging themselves in the surfaces of the reefs and protecting the larvae. One of those designs looks like a ninja throwing star.

[Image: SECORE/Valérie Chamberland]

“The question we posed to our working group was, ‘Can you give us your best impression of what promotes coral larvae to grow, and what’s going to allow them to survive in the ocean as they grow up in these early life stages?’” said Bickel.

The SECORE project is not one of immediate gratification. The organization grows its corals from embryos in small conglomerations of cells, and depending on the species, it can take several years for the corals to become sexually mature. In earlier life stages, however, the coral can still provide habitats for fish and other species.

This elkhorn coral was outplanted by SECORE five years ago. Since then, it has grown into a mature colony, which now spawns with other elkhorn colonies in the waters of Curaçao. [Image: SECORE/Paul Selvaggio]

“It’s definitely an investment in the future,” Bickel said. “I think that with really complicated ecosystems, we’re talking many years before you start seeing comparable structure return to areas that are being restored. The main focus at the moment is, can we improve our methods and our technologies to upscale this type of restoration to the levels needed to counteract the decline?”

SECORE isn’t the only organization working to do so, and the hope is that with enough of them putting effort into restoring coral reefs, the damage can be mitigated and even reversed.

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[Source: Autodesk]