Periodic Reporting for period 1 - OSTEONET (In vitro 3D cell models of healthy and OSTEOpathological ageing bone tissue for implantation and drug testing in a multidisciplinary NETwork)
Okres sprawozdawczy: 2023-01-01 do 2024-12-31
The project OSTEONET aims at developing a lab-scale model of human bone tissue, both mimicking the healthy and the osteoporotic bone, which can be used for basic physiology or pathology studies as well as serve as a model for pre-clinical drug-candidate screening. To reach this ambitious goal, expertise is needed in various fields of science and technology, such as biomedical engineering, cellular biology, fluid dynamics and microfluidics, electronics, AI.
Indeed, nowadays there is a lack of lab-scale models of human tissues for all kinds of molecular medicine applications and animals (i.e. in vivo models) or 2D-cultured cells (so-called in vitro) are used instead. Both available in vivo and in vitro models are unsuitable for addressing specific quests: the former implies the usage of animal models, thereby more simple organisms as compared to the human body in addition to the ethical issues, the latter overlooking the complex three-dimensional architecture of human tissues thus, being unable to reliably model certain mechanisms, such as intercellular contacts.
Moreover, human derived samples (e.g. from healthy donors or patients willing to help the research) are not easy to get in general, due to sample isolation, culture and ethics related issues too.
To meet the urge of shifting the models-paradigm in biomedical research, the Consortium is developing an automated 3D-cell culture system, taking advantage of the bioreactor developed by Cellex (namely, BioAxFlow or BAF) that allows for 3D-cell culture upon scaffolds, i.e. mechanical supports similar to bone architecture, tailor-made for resembling either the healthy or the pathological condition (i.e. with different percentages of porosity: more porous scaffolds mimicking osteoarthritis and less porous mimicking the healthy condition). Hand-in-hand with a thorough choice of the most suitable cell line(s) that need to be used, computational modelling is helping by improving the performances of the cell culture process occurring within BAF (e.g. by predicting the fluid-dynamical behaviour of cell culture media) and software development, as well as AI, are being used for augmenting knowledge related to human bone tissue architecture and to transfer it to the scaffolds that are continuously being generated.
Indeed, nowadays there is a lack of lab-scale models of human tissues for all kinds of molecular medicine applications and animals (i.e. in vivo models) or 2D-cultured cells (so-called in vitro) are used instead. Both available in vivo and in vitro models are unsuitable for addressing specific quests: the former implies the usage of animal models, thereby more simple organisms as compared to the human body in addition to the ethical issues, the latter overlooking the complex three-dimensional architecture of human tissues thus, being unable to reliably model certain mechanisms, such as intercellular contacts.
Moreover, human derived samples (e.g. from healthy donors or patients willing to help the research) are not easy to get in general, due to sample isolation, culture and ethics related issues too.
To meet the urge of shifting the models-paradigm in biomedical research, the Consortium is developing an automated 3D-cell culture system, taking advantage of the bioreactor developed by Cellex (namely, BioAxFlow or BAF) that allows for 3D-cell culture upon scaffolds, i.e. mechanical supports similar to bone architecture, tailor-made for resembling either the healthy or the pathological condition (i.e. with different percentages of porosity: more porous scaffolds mimicking osteoarthritis and less porous mimicking the healthy condition). Hand-in-hand with a thorough choice of the most suitable cell line(s) that need to be used, computational modelling is helping by improving the performances of the cell culture process occurring within BAF (e.g. by predicting the fluid-dynamical behaviour of cell culture media) and software development, as well as AI, are being used for augmenting knowledge related to human bone tissue architecture and to transfer it to the scaffolds that are continuously being generated.
The Consortium has, so-far, tested the bioreactor BAF with different cell lines to mimic the bone tissue, namely the human dental pulp stem cells (hDPSCs), the osteosarcoma cell line SAOS-2 and the mesenchymal stem cells (MSCs), all of which are cells which are either precursor of the bone cells (i.e. can differentiate into osteoblasts or osteoclasts) or are more easily proliferating thus, populating the scaffolds that have been produced. Additionally, with the help of software developers working with AI, a comprehensive pool of scaffolds like the bone tissue architecture have been generated. This implies exploring several geometries of the natural bone trabecular architectures that are populated by bone tissue-cells to get to scaffolds-geometries enabling the desired cells to adhere and proliferate.
To let such scaffolds be as close as possible to the healthy and pathological human bone tissue, a technique called micro computational tomography (µCT) has been used to scan real bone tissue samples to assess the exact geometries. With the help of software developers and AI, we are trying to get to an automatically calculated library of different geometries that can be used to generate the scaffolds thus, overcoming the need of using µCT. Moreover, different materials and 3D-printing techniques are being used, including the 3D bio-printing, which uses bioinks, i.e. inks of bioinspired materials, as well as the LCM technology, enabling the generation of β-tricalcium phosphate- made scaffolds.
Additionally, cell culture protocols for long term cryopreservation of constructs (i.e. bone-derived cells grown onto different scaffolds) are being optimized to reach the goal of having tissue-engineering intervention material for human treatment, in the future.
To let such scaffolds be as close as possible to the healthy and pathological human bone tissue, a technique called micro computational tomography (µCT) has been used to scan real bone tissue samples to assess the exact geometries. With the help of software developers and AI, we are trying to get to an automatically calculated library of different geometries that can be used to generate the scaffolds thus, overcoming the need of using µCT. Moreover, different materials and 3D-printing techniques are being used, including the 3D bio-printing, which uses bioinks, i.e. inks of bioinspired materials, as well as the LCM technology, enabling the generation of β-tricalcium phosphate- made scaffolds.
Additionally, cell culture protocols for long term cryopreservation of constructs (i.e. bone-derived cells grown onto different scaffolds) are being optimized to reach the goal of having tissue-engineering intervention material for human treatment, in the future.
The integration of expertise on innovative techniques in scaffold manufacturing, cell culture, and long-term tissue preservation together with mathematical modelling, advanced imaging and image analysis techniques, bioinformatics, artificial intelligence and machine learning applied to the drug-candidate discovery process will lead to a deep share of knowledge giving rise to important market opportunities in the field of pharmaceutics, tissue engineering and regenerative medicine, and to innovative protocols for more effective management of bone dysfunctions in the elderly.
Through effective coordination, knowledge sharing, and practical hands-on and theoretical training of professionals with different backgrounds, the OSTEONET project is also aimed at making inexperienced personnel aware of the advantages and limits of in vitro dynamic culture of 3D models, at making them able to exploit the culture features for pharmacological or basic studies and to correctly analyse the experiment outcome.
This way also inexperienced personnel participate in the development and translation to clinics and research labs of such innovative and reliable models of healthy and ageing bone. The real breakthrough is to make accessible to a vast cohort of potential users the innovative technology developed within OSTEONET, as the only way to successfully personalize therapies and enable better preventive care for the elderly.
Through effective coordination, knowledge sharing, and practical hands-on and theoretical training of professionals with different backgrounds, the OSTEONET project is also aimed at making inexperienced personnel aware of the advantages and limits of in vitro dynamic culture of 3D models, at making them able to exploit the culture features for pharmacological or basic studies and to correctly analyse the experiment outcome.
This way also inexperienced personnel participate in the development and translation to clinics and research labs of such innovative and reliable models of healthy and ageing bone. The real breakthrough is to make accessible to a vast cohort of potential users the innovative technology developed within OSTEONET, as the only way to successfully personalize therapies and enable better preventive care for the elderly.