Innovative microphysiological systems for long-term real-time imaging in biomedicine

The discovery and development of new drugs is a complex and highly regulated process that, since its dawn, is afflicted by an intrinsic and fundamental problem: how to maintain cells or tissues alive out of human body in such a way they could behave like in vivo. So far, identification and selection of pharmaceutical compounds have occurred through standardized protocols based on traditional in vitro cultures, animal models and, lastly, human trials. This well-established operating structure consented to advance people’s health conditions all over the world. However, conventional in vitro cultures and animal experimentations have limited predictive capabilities in detecting therapeutic efficacy and toxicity of candidate compounds. Due to that, drug discovery and development has been for a long time recording a reduced yield characterized by a small number of new molecules yearly delivered on the market and elevated production costs. Concomitantly, the pressure from unmet medical needs is rising together with ethical controversies related to animal use during preclinical trials. This negative and persistent situation is driving drug research towards an inevitable change of paradigm. Currently, one of the more discussed solutions that might be adopted to invert the trend of drug R&D and provide patients with better therapies is represented by microphysiological systems (MPSs), also noted as Organ on Chip (OoC) devices. This recent strategy tackles the primal issue mentioned above, which is at the very basis of our capabilities i) to understand patho-physiological processes and ii) screen more effective and specific drugs based on their efficacy and toxicity. MPSs reform traditional in vitro cultures providing biological samples with artificial microenvironments aimed to induce physiologicalresponses, for example, to xenobiotics. MPSs exploit cutting edge technologies from the advancements of different scientific fields (microfluidics and biomaterials firsts in line) not just to recreate in vivo-like environments but also to acquire quantitative data and detailed biomedical information. They can integrate sensors to monitor the microenvironmental parameters and molecules concentrations (with the perspective of introducing feedback loops to control those variables) and be compatible with the most advanced imaging methods. In the context of this industrial doctoral thesis and as a contributor of the EU-funded project ITN-DeLIVER (grant agreement n°766181), two MPS were conceived, realized, and validated at Cherry Biotech SAS. Then, the two systems were preliminary tested at the laboratories of two project’s partners: the University of Tromsø and the University of Birmingham. The tests were conducted using cells and tissues deeply involved in hepatic patho-physiology, namely liver sinusoidal endothelial cells (LSECs). Inside the body, these cells are subjected to peculiar conditions of mechanical stress and molecular gradients that were replicated implementing the MPS platforms object of this thesis work. Further, as LSECs have nanoscale structures, the MPS were adapted to enable super resolution imaging that, so far, is the only approach allowing the observation of live cells under the resolution limit imposed by light diffraction. Among the innovative features of those novel MPS devices, it can be highlighted: i) an unprecedented adaptability to standard cell CUs, which facilitates the implementation of advanced tools in research laboratories respect to other commercially available products; ii) an unprecedent integration of microfluidic and chip-based super resolution imaging (SRI), which bolsters the transition to advanced microscopy techniques that might reveal biological phenomena so far undetectable; iii) an unprecedented versatility of those MPS that, when combined with other traditional instrumentation such as perfusion pumps and microfluidic temperature controllers, could work as all in-one CUs enabling gas-content-controlled perfusion, precise incubator-free thermalization and 6 sample live-imaging. The preliminary results from biological experiments carried out in collaboration with the partners of ITN-DeLIVER consortium confirmed the feasibility in adopting those innovative MPS devices, also indicating improvable aspects. The experimental outcomes together with a patent (already granted in France, EU and US) and a literature-based benchmark against competitors’ systems available in the market suggest that the MPS treated in this thesis have the potential to promote the adoption of advanced tools for in vitro cultures in life-science and pharmaceutical research.

Level thesis:degree_level

thesis.doctoral

Grantor dc:publisher

Universität Bielefeld

Year

2024

Author dc:creator

• Boninsegna, Matteo

Repository record source_url

https://pub.uni-bielefeld.de/record/2988168

OAI identifier oai:identifier

oai:pub.uni-bielefeld.de:2988168