An in-vivo engineered chip as a smart intravital multiphoton imaging window for new validation protocols of biomaterials

Duration 1/03/2021 – 28/02/2025- Funded by EU Horizon 2020-FET OPEN – G.A. 964481

PI: Prof. Giberto Chirico, Bicocca University, Milano, Italy – Official website: in2sight.eu

VIDEO with project results (relevant publications are linked at the bottom of this page)

Project abstract

Routine clinical use of biomaterials requires the reduction of the economical and ethical costs of biocompatibility tests (ISO10993 EU norm) which are unsustainable for small-medium industries and for the society. In this project we foster an unprecedented breakthrough in in-vivo optical imaging that will radically renew the biocompatibility tests of biomaterials.
A micro-structured chip, built by two-photon laser polymerization (2PP), will be implanted in lab animals, host a biomaterial and contain micro-features that guide the spontaneous regeneration of vascularized tissue within a thin gap (0.15mm) in contact with the biomaterial and act as beacons to correct the optical aberrations. The same chip carries a micro-lenses array for in-situ multi-spot imaging, with no need of external high numerical aperture objectives, dramatically improving light penetration in tissue. This chip will recast our thinking of deep tissue in-vivo imaging: the mice carry their own imaging optics, thus reducing substantially image aberration issues allowing unprecedented quantitative and longitudinal analyses of the host inflammatory response to the implant, without sacrificing the mice at each time step.
This project will allow unique quantification of the immune reaction to biomaterials at the cellular level (scientific impact), reduce (at least threefold) the number of used animals (societal impact) and the costs of biomaterial discovery (economical impact), and will Refine and Reduce protocols for biocompatibility on a single revolutionary device (regulatory impact). We open here a new visionary path for in-vivo imaging with high Replacement potential in oncological pharmaceutics and immune-therapies. 3 academic units, 2 public research institute and 2 SMEs ensure a highly inter-sectorial/interdisciplinary approach encompassing non-linear intravital imaging, bioengineering design, 2PP material science, biocompatibility protocols design and numerical simulations of immune response.

Publications

2023

Conci C, Sironi L, Jacchetti E, Panzeri D, Inverso D, Martinez Vasquez R, Osellame R, Collini M, Cerullo G, Chirico G, Raimondi MT. In vivo label-free tissue histology through a microstructured imaging window. APL Bioeng. 8, 016102 (2024). https://doi.org/10.1063/5.0165411. Repository link: https://re.public.polimi.it/handle/11311/1258496

Marini, M., Nardini, A., Martínez Vázquez, R., Conci, C., Bouzin, M., Collini, M., Osellame, R., Cerullo, G., Kariman, B. S., Farsari, M., Kabouraki, E., Raimondi, M. T., Chirico, G., Microlenses Fabricated by Two-Photon Laser Polymerization for Cell Imaging with Non-Linear Excitation Microscopy. Adv. Funct. Mater. 2023, 2213926. https://doi.org/10.1002/adfm.202213926

2022

Conci C, Jacchetti E, Sironi L, Gentili L, Cerullo G, Osellame R, Chirico G, Raimondi MT. A miniaturized imaging window to quantify intravital tissue regeneration within a 3D micro scaffold in longitudinal studies. Adv. Optical Mater. 2022, 2101103.  https://doi.org/10.1002/adom.202101103

Claudio Conci, Emanuela Jacchetti, Laura Sironi, Lorenzo Gentili, Giulio Cerullo, Rebeca Martinez, Roberto Osellame, Mario Marini, Margaux Bouzin, Maddalena Collini, Laura D’Alfonso, Elmina Kabouraki, Maria Farsari, Anthi Ranella, Nikos Kehagias, Giuseppe Chirico, and Manuela T. Raimondi. A miniaturized chip for 3D optical imaging of tissue regeneration in vivo, Proc. SPIE 12144, Biomedical Spectroscopy, Microscopy, and Imaging II, 121440D (27 May 2022); https://doi.org/10.1117/12.2629824

Musi CA, Colnaghi L, Giani A, Priori EC, Marchini G, Tironi M, Conci C, Cerullo G, Osellame R, Raimondi MT, Remuzzi A, Borsello T. Effect of 3D Synthetic Microscaffold Nichoid on the Morphology of Cultured Hippocampal Neurons and Astrocytes. Cells. 2022 Jun 23;11(13):2008. doi: 10.3390/cells11132008