2022-11-014 - COAXIAL (3D-MUSCLE) - BIOPRINTING KIT FOR SKELETAL MUSCLE TISSUE

COAXIAL (3D-MUSCLE)

2022-11-014

Innovative 3D bioprinting system to fabricate skeletal muscle tissues with biomimetic design in terms of structure and functionality. Our coaxial method allows to 3D-bioprint muscle fibres that remain individual and not merge with each other

Our project aligns with ESG values by promoting sustainability and ethical responsibility. Environmental: It reduces reliance on animal testing and minimizes material waste through more efficient 3D bioprinting methods. Social: The technology offers new treatments for muscle injuries and supports cruelty-free research in cosmetics and pharmaceuticals. Governance: By adhering to ethical standards and providing nonanimal alternatives for drug testing, the project enhances regulatory compliance and supports responsible innovation in biomedical fields.

Our project has the potential to create a transformative impact across multiple dimensions: Drug Development: our biomimetic tissue models offer a more accurate and ethical alternative to animal testing, accelerating drug discovery and reducing costs in pharmaceutical and cosmetic industries. Sustainability: by reducing animal testing and material waste, the technology supports more environmentally friendly research practices, promoting sustainability in biotech and pharmaceuticals. Market Growth: Filling gaps in the current 3D bioprinting market will drive further innovation and business opportunities. Regenerative Medicine: the production of biocompatible, functional tissue implants opens new possibilities for personalized medicine and faster recovery times for patients.

Muscles in human body are subject to several disorders such as ageing, traumatic injury or diseases called myopathies, among others. Hence, skeletal muscles are of high relevance for several industries like: cosmetics, since sustained contractions of facial muscles cause wrinkles; biomedicine, since diseases like muscle dystrophies are very heterogeneous and generally understudied; and tissue engineering and regenerative medicine, since implants like muscle grafts could help heal people with serious muscle damage. However, up until recently, most of the research involving skeletal muscle tissue was done in a 2D environment because its three-dimensional structure was hard to be replicated. Recent advances in 3D bioprinting have allowed the fabrication of bioengineered muscle models to mimic the complex hierarchical organization of native tissues. However, it is currently not possible to bioprint several muscle fibres and assemble them in individual fascicles. So, there is an urgent need for a method to obtain thin and individual myofibers that accurately replicate the structure of human muscle tissue for research and tissue replacement purposes. We have developed an innovative 3D bioprinting system for obtaining individual free-form width-controlled biological fibers, suitable to fabricate skeletal muscle tissues with biomimetic design in terms of structure and functionality. Our technology also includes a bioink formulation, the hybrid biomimetic structure comprising one or more of said individual fibers and the methods of manufacturing such structure.

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Our asset can be marketed as a kit for 3D bioprinting (bioink + bioprinting methodology) to print skeletal muscle tissue for bioengineering research applications. The asset will be licensed to companies in the field of 3D-Bioprinting manufacturing, with marketing and sales capabilities to bring our product to the market. Additionally, it could be exploited by a company (third party licensee or a spin-off of our research center) focused on regenerative therapies to develop bioimplants for muscular disorders.

We anticipate developing several products from our 3D bioprinting coaxial technology for various applications: • A 3D-bioprinting kit for producing cell tissues, particularly skeletal muscle, which is missing from the portfolios of leading 3D-bioprinting companies. • A drug discovery platform for cosmetics and pharmaceuticals to study compounds on 3D skeletal muscle models. With the FDA no longer requiring animal tests and animal testing banned in EU cosmetics, nonanimal methods will grow in importance. • Biocompatible implants for regenerative medicine, allowing functional tissue implants for patients with muscle injuries or myopathies.

Global 3D cell culture market: USD 2.5 billion in 2024, expected to reach USD 6.2 billion by 2032 (CAGR of 12.1%; source: FortuneBusinessInsights). Additional markets relevant for the project: • Cosmetics: market size USD 360 billion in 2024, expected USD 420 in 2033 (Source: Business Research Insights). • Tissue engineering and regenerative medicine: market size USD 18.3 billion in 2024, expected USD 43 in 2030 (source: GrandViewResearch market analysis).

The technology has been developed and validated at lab scale. No specific funding needs exist before licensing the technology to a player in the 3D bioprinting field. If a further validation of our platform for drug development applications were required, such a project shall be peformed in collaboration with an interested industrial partner with an estimated cost of 80,000€ (6-months duration).

Licensees in the field of 3D bioprinting or industrial partners for co-development.

Achieved: • Full validation for skeletal muscle tissue (fibre morphology versus printing parameters, cell proliferation and differentiation), demonstrating improved cell differentiation and greater force outputs. • Testing for specific bioink formulations and drug applications. • Characterisation of different protocols to obtain high quality fibres with a wide combination of materials. • Successful validation of a force measurement platform based on our technology for drug testing. Next milestones: • Optimize our printing method to obtain more sophisticated tissues • Scale-up and industrialization • Validate the versatility of our method by fabricating fiber-like tissues other than skeletal muscle

• Excellent replication of highly structured skeletal muscle tissue: - Individual fibres are printed with a controlled width - No significant fusion of adjacent fibres • Versatile system, compatible with a wide range of cell-laden hydrogels (even without support baths). • The solution is based on coaxial nozzles that can be used with different 3D printing systems. • Our method does not rely on the use of sodium alginate, as most coaxial approaches do, which is not well tolerated by certain tissues such as skeletal muscle.

- While growing, the demand for non-animal testing methods for drug development and its full adoption from the pharma sector is still in its early stages. - Potential lack of consistency and standardization of 3D tissue model. - High cost of implementing 3D cell culture technologies. - Complex development for bioimplants for tissue replacement.

2018-2020: design and characterization of 3D bioprinted actuartors based on skeletal muscle tissue; 2020: IP protection; 2020-2022: drug testing validation in collaboration with the industry; 2022-2024: full validation and optimization of our 3D bioprinted tissue model; 2025: Tech transfer and business development effort for achieving licensing agreement; 2026-2027: scale-up, industrialization and first commercialization.

Our system was protected by a patent application (priority year: 2020; PCT publication n. WO2022003203A1), currently in national phases (Europe, US), pending to be granted.

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Know-how on 3D bioprinting, soft-robotics and biofabrication of bioactuators.

The quality management strategy for our 3D bioprinting technology will ensure precision, consistency, and scalability. Process Optimization: printing parameters (e.g., fiber morphology, cell proliferation, and tissue functionality) shall be refined and continuously monitored to ensure each batch meets set standards. Scale-up and Industrialization: as production will be scaled up by the future licensee, SOPs and automated systems will be required to monitor material consistency, bioink quality, and environmental factors to ensure uniformity across larger volumes. Validation of Versatility: for new tissue applications, quality benchmarks shall be established and biomimetic structures, functionality, and biocompatibility shall be validated, maintaining versatility without compromising quality. All processes will have to adhere to industry standards and regulations.

Patents: WO 2022003203

Our coaxial method allows to 3D-bioprint muscle fibres that remain individual and not merge with each other, mimicking the higher-level structure of muscle tissue. Bioactuators were obtained with our method and applied as force measurement platform for drug testing. Main advantages of our method are: the fine thickness control of the bioprinted fibers; their contraction forces that are 3 times higher than conventional printing; the high versatility in terms of hydrogel composition.