Periodic Reporting for period 1 - HumanINK (Human based bioinks to engineer physiologically relevant tissues)
Berichtszeitraum: 2022-11-01 bis 2024-04-30
Bioprinting techniques hold a great potential to develop innovative strategies for integrating living cells into natural or synthetic biomaterials, utilized as bioinks, to address key biomedical and healthcare challenges that hinder advancements in the medical, pharmaceutical and food industries. HumanINK aims to develop and validate human protein-based bioinks to create robust humanized 3D environments with unprecedented biofunctionality for cell culture, with the goal of fully recapitulate the native microenvironment of a variety of human tissues and organs. With this intent, human-derived proteins modified with photo-responsive groups will be tested and validated as bioinks to develop geometrically controlled macro-scale 3D soft hydrogels with customizable mechanical properties. The protein-rich content of these materials provides cell-anchoring moieties to support cell growth and bioinstructive cues to regulate cell function, guiding the process of tissue morphogenesis. The innovative aspect of this platform lies in the human origin of the biomaterial proposed 3D bioprinting, offering a physiologically-relevant and easy-to-use approach to recreating native tissues in vitro, thereby accelerating drug discovery and clinical purposes. In the HumanINK, we propose to optimize the printability, robustness, reproducibility, and scalability of the human-based bioinks. Additionally, the biological response of multiple human cell types will be investigated and the bioinks will be benchmarked with the main competitors in the market.
Overall, our proposed technology aims to enhance the probability of successful drug development while simultaneously reducing the cost and time of development and supporting animal welfare, reducing the need for animal testing. Leveraging the unique properties of our products, HumanINK represents a valuable opportunity to develop bioinstructive materials for tissue engineering and accurate disease models that can effectively bridge the gap between fundamental research and drug validation. Such approach is expected to withstand a high and broad market potential among pharmaceutical companies, clinical institutions, or academic research.
Overall, our proposed technology aims to enhance the probability of successful drug development while simultaneously reducing the cost and time of development and supporting animal welfare, reducing the need for animal testing. Leveraging the unique properties of our products, HumanINK represents a valuable opportunity to develop bioinstructive materials for tissue engineering and accurate disease models that can effectively bridge the gap between fundamental research and drug validation. Such approach is expected to withstand a high and broad market potential among pharmaceutical companies, clinical institutions, or academic research.
In the HumanINK project, human methacryloyl platelet lysates (PLMA) and decellularized extracellular matrix (dECM) derived from placental tissues have been combined to create a human bioink with complementary biochemical properties. As newly human biomaterials purposed as great alternatives for the biomaterials of animal origin, the lipidomic profile of human adipose-derived stem cells (hASCs) cultured on PLMA hydrogels was analyzed, revealing a closer correlation to freshly isolated cells than the Matrigel-cultured ones. Pursuing the biological validation of the developed materials for disease modelling, PLMA was explored for the development of complex co-culture tumor models, demonstrating excellent suitability to support native cell functionality. Moreover, a novel placenta-derived biomaterial, the methacryloyl chorionic membrane (CMMA), was produced and tested for its biocompatibility and angio-vasculogenic competence.
Taking into account the exceptional biocompatibility of these protein-rich materials, their potential as bioink was addressed in the HumanINK project through various innovative approaches. To improve the viscoelasticity and shear thinning behavior of platelet lysates (PL)-based solutions, the available amine groups on PL proteins were exploited for coupling with carboxyl groups in PLMA, by leveraging carbodiimide chemistry. The creation of a pre-gel resulted in inks with controlled viscosities and elasticities, which enabled the fabrication of 3D printed multilayered constructs with high shape-fidelity. These PL-based ink scaffolds showcased mechanical robustness and the ability to support hACSs culture.
Building on the recent advancements on granular materials, supramolecular granular materials were prepared from the fragmentation of bulk hydrogels made of human decellularized amniotic membrane (dAM), combined with the proto-responsive acryloyl b-cyclodextrin. Due to the non-covalent interactions between the cyclodextrin and the proteins, the extrudable granular materials exhibited self-healing and self-curing abilities, forming cohesive and stable structures. Using a similar strategy, PLMA microparticles and methacryloyl hAM (AMMA) were combined to produce a jammed microgel ink. In this case, AMMA was incorporated as a photocrosslinkable interstitial matrix that resides between microgels. The jammed ink demonstrated good extrudability and shape fidelity, enabling the production of compartmentalized bulk structures with improved nutrient availability for encapsulated cells.
The outcomes of the HumanINK project highlight the potential of human-based bioinks as a promising technology for creating more physiologically relevant platforms for tissue engineering and more precise disease models. We envision these advancements bridging the gap between fundamental research and drug validation, moving towards an animal-free drug development process.
Taking into account the exceptional biocompatibility of these protein-rich materials, their potential as bioink was addressed in the HumanINK project through various innovative approaches. To improve the viscoelasticity and shear thinning behavior of platelet lysates (PL)-based solutions, the available amine groups on PL proteins were exploited for coupling with carboxyl groups in PLMA, by leveraging carbodiimide chemistry. The creation of a pre-gel resulted in inks with controlled viscosities and elasticities, which enabled the fabrication of 3D printed multilayered constructs with high shape-fidelity. These PL-based ink scaffolds showcased mechanical robustness and the ability to support hACSs culture.
Building on the recent advancements on granular materials, supramolecular granular materials were prepared from the fragmentation of bulk hydrogels made of human decellularized amniotic membrane (dAM), combined with the proto-responsive acryloyl b-cyclodextrin. Due to the non-covalent interactions between the cyclodextrin and the proteins, the extrudable granular materials exhibited self-healing and self-curing abilities, forming cohesive and stable structures. Using a similar strategy, PLMA microparticles and methacryloyl hAM (AMMA) were combined to produce a jammed microgel ink. In this case, AMMA was incorporated as a photocrosslinkable interstitial matrix that resides between microgels. The jammed ink demonstrated good extrudability and shape fidelity, enabling the production of compartmentalized bulk structures with improved nutrient availability for encapsulated cells.
The outcomes of the HumanINK project highlight the potential of human-based bioinks as a promising technology for creating more physiologically relevant platforms for tissue engineering and more precise disease models. We envision these advancements bridging the gap between fundamental research and drug validation, moving towards an animal-free drug development process.
The project successfully demonstrated the development of bioinks derived from chemically modified platelet lysate and placenta proteins. These bioinks have shown promising results in terms of biocompatibility and support for cell growth, indicating a significant potential impact in the fields of tissue engineering, in vitro testing, disease modeling, and regenerative medicine. By offering an innovative solution for creating human protein-based structures that can mimic the microenvironment of human tissues, this project advances capabilities in 3D bioprinting, potentially leading to more effective and personalized in vitro diagnostics and medical treatments.
Additionally, using expired blood units and placenta-derived proteins, which are often considered medical waste, aligns with sustainable practices and could reduce reliance on animal-based materials.
To ensure further uptake and success, several key needs must be addressed. Firstly, optimizing printability fidelity and construct stability is crucial for ensuring consistent and precise fabrication of constructs. Furthermore, establishing reliable sources of placentas is essential for large-scale production and the standardization of bioink components.
We have submitted several patents and are preparing a new one on the use of the whole placenta. To facilitate a successful product launch and implementation, understanding and navigating access to markets, along with strategic plans for commercialization, are essential.
Additionally, using expired blood units and placenta-derived proteins, which are often considered medical waste, aligns with sustainable practices and could reduce reliance on animal-based materials.
To ensure further uptake and success, several key needs must be addressed. Firstly, optimizing printability fidelity and construct stability is crucial for ensuring consistent and precise fabrication of constructs. Furthermore, establishing reliable sources of placentas is essential for large-scale production and the standardization of bioink components.
We have submitted several patents and are preparing a new one on the use of the whole placenta. To facilitate a successful product launch and implementation, understanding and navigating access to markets, along with strategic plans for commercialization, are essential.