3D Printing of Ultra-fideLity tissues using Space for anti-ageing solutions on Earth

Project Information

PULSE

Grant agreement ID: 101099346

DOI

10.3030/101099346

EC signature date 19 January 2023

Start date 1 April 2023

End date 31 March 2028

Funded under

• The European Innovation Council (EIC)

Total cost

€ 4 597 578,75

EU contribution

€ 4 597 578,75

Coordinated by

UNIVERSITEIT MAASTRICHT
Netherlands

Periodic Reporting for period 2 - PULSE (3D Printing of Ultra-fideLity tissues using Space for anti-ageing solutions on Earth)

Reporting period: 2024-04-01 to 2025-07-31

Summary of the context and overall objectives of the project

PULSE project is poised to develop paradigm-changing bioprinting technology for applications in space and on Earth. By combining magnetic and acoustic levitation into an innovative bioprinting platform, PULSE’s device should be capable of achieving unparalleled spatiotemporal control of cell deposition. This new technology facilitates the precise manipulation of biological materials, enabling the creation of highly sophisticated and realistic organoids that closely mimic the complexity of the corresponding human organs.

Bioprinting in Space is one of the novel promising and perspective research directions in the rapidly emerging field of biofabrication. There are several advantages of bioprinting in Space. First, under the conditions of microgravity, it is possible to bioprint constructs employing more fluidic channels and, thus, more biocompatible bio-inks. Second, microgravity conditions enable 3D bioprinting of tissue and organ constructs of more complex geometries with voids, cavities, and tunnels. Third, a novel scaffold-free, label-free, and nozzle-free technology based on multi-levitation principles can be implemented under the condition of microgravity. The ideal Space bioprinters must be safe, automated, compact, and user friendly.

Systematic exploration of 3D bioprinting in Space will advance biofabrication and bioprinting technology per se. Vice versa 3D bioprinted tissues could be used to study pathophysiological biological phenomena when exposed to microgravity and cosmic radiation that will be useful on Earth to understand ageing conditioning of tissues, and in space for the crew of deep space manned missions. As a proof of concept study, in PULSE will use this newly developed bioprinting technology to create cardiac 3D in vitro models able to better mimic cardiac physiology compared to organoids. We will use such models to study cardiac ageing and test the efficacy of anti-inflammatory/anti-oxidative drugs with anti-ageing potential.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

In the second reporting period of the project, we have further defined the operating principles of the PULSE device. We have refined a mathematical and numerical model of the system, which provides insights on the required process parameters to perform magneto-acoustic levitation. The refined model provides a better understanding of how cellular spheroids would be first brought together by magnetic levitation and then shaped into the final tissue construct by acoustic levitation. Accordingly, the first prototype capable of both magnetic and acoustic levitation within a closed bioassembly chamber was designed, manufactured and assembled for ground experiments. The first prototype of the PULSE magneto-acoustic levitation device has gone through a number of iterations to provide a system that could work on ground (Earth), yet being qualified also for operating on the ISS. The design is currently being finalized to enter into testing and validation phase for space mission in 2026. In parallel, we have generated an adapted design to be launched for a parabolic flight in October 2025. The parabolic flight serves as an intermediate experimental testbed for the device under conditions that simulate the closest possible the launch to ISS in 2027. Currently, the ground experiments include polystyrene beads for the validation of the levitation process as well as cell spheroids and organoids for optimizing process parameters of the PULSE device.

Hydrogel selection has been completed, identifying hydrogels specific compositions as best candidates for all cells. In particular, the selected hydrogels showed to support the culture of the cardiovascular cells and cardioids, supporting vascularization.
We have further characterized endothelial cells, cardiomyocytes, cardiofibroblasts, and cardioids in culture conditions emulating some of the challenging microenvironment conditions that cells will experience during transport and launching to the ISS. Results showed that cells are robust to maintain viability and functionality in refrigerated conditions with specific culture media conditions.

We have developed a complete framework as preparatory work on primary and commercial human cardiovascular cell lines to identify a suitable platform and model for testing under simulated microgravity conditions and to elucidate possible effects on target cells. We validated the space condition simulator platform and obtained preliminary results of the effect of radiotherapy radiation and microgravity on heart organoids.

We have further refined the requirements for launching of the PULSE device to the ISS. We have established the initial system engineering framework for the qualification and performance verification of the PULSE bioprinter. The document defines model philosophy, verification subjects, verification approach, verification strategy, and verification tools. The verification campaign will include several types of tests. Qualification and environmental tests, including random vibration, EMC, DC magnetic field, micro-g disturbances, audible noise, leak test, and proof pressure tests. Interface tests will be conducted using the PULSE device and the ground model of the ICE Cubes Facility, ensuring mechanical and electrical fit, as well as compatibility with communication, software, and thermal interfaces. Functional tests will be conducted to verify functional and performance requirements. The functional test campaign will be complemented by extensive breadboard testing for each subsystem.

Finally, we are building the PULSE stakeholder network, engaging a wide range of partners to build confidence on PULSE technology. To do so, we have set up an advisory board with key representatives from several sectors close to the PULSE innovation, and we started to contact ESA groups involved in the follow-up of the technology in space. We are also reaching out to a vast network of students and young generation scientists connected to space medicine and tissue engineering via the organization of a summer school. Next steps will involve building relationships with pharma companies interested in cardiac organoids, ESA, and the ISS National Laboratory.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

One of the key applications of PULSE’s innovative bioprinting technology is the creation of in vitro 3D human heart models, which are essential tools for studying the effects of space and radiation on the human cardiovascular system. These advanced heart models will provide invaluable insights into cardiac physiology and pathology, facilitating the development of preventive and therapeutic solutions for astronauts embarking on long-term space missions and cancer patients undergoing radiation therapy. Bioprinted organoids that closely replicate the complexity of human organs have the potential to reduce the reliance on animal experimentation and provide a more accurate and efficient platform to study disease mechanisms and evaluate drug responses.

Illustration of space radiation affecting a representation of the project’s cardiac model.

Illustration of combination of magnetic and acoustic levitation.

Conceptual illustration of the project's multi-levitation bioprinter assembling cardiac tissue.