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AN IN VIVO BIOENGINEERED CHIP AS A SMART INTRAVITAL MULTIPHOTON IMAGING WINDOW FOR NEW VALIDATION PROTOCOLS OF BIOMATERIALS

Periodic Reporting for period 2 - IN2SIGHT (AN IN VIVO BIOENGINEERED CHIP AS A SMART INTRAVITAL MULTIPHOTON IMAGING WINDOW FOR NEW VALIDATION PROTOCOLS OF BIOMATERIALS)

Reporting period: 2022-03-01 to 2023-08-31

Routine clinical use of biomaterials must comply with the biocompatibility tests stated by the ISO10993 EU norms that require a wide use of laboratory animals with an excessive economic burden, particularly for SMEs, and with heavy ethical costs for the society.
IN2SIGHT wants to renovate the way the tests are made in order to reduce the number of animals used and to increase the statistical reliability of the analyses.

The objectives are to:
1. develop a new technology, based on non-linear optical microscopy, capable to observe the immune reaction to the implant of a biomaterial for weeks, during the evolution of the possible reaction, without the need to sacrifice the animal;
2. formulate a novel statistical approach to analyze the data relative to the immune reaction;
3. make the IN2SIGHT technology known among the stakeholders and initiate the process for an update of the norms through contacts with the relevant regulatory bodies in Europe and USA;
3. make the society aware of the possibility to reduce the number of animals in the tests and of alternative methods for the biomaterial and drugs testing;
4. develop an innovative numerical simulation method for the immune reaction to the implant of a biomaterial based on agent based techniques with the aim to enhance our interpretation of the immune reaction observed experimentally.
The main results of the project in the first 30 months are:
1. development of protocols for the fabrication of the device by means of 2-photon polymerization and nano imprint lithography;
2 fabrication of microlenses for in vivo imanging of biological samples;
3. assessment of any possible immune response of the implant of the In2Sight device;
4. In-vivo imaging on implanted In2Sight devices
5. contacts with regulatory bodies for the presentation of the projet.
6. activation and use of social media channels: twitter and linkedin.

Part of the above results have been published (6 publications open access and 2 proceedings) and presented to congresses as talks (26 talks ) or posters (9 posters).
The Consortium has been present at the following events:
- Eusaat Congress 2022;
- SPIE Optical Metrology 2023;
- EOS- European optical society annual meeting;
- 28th Congress of the European Society of Biomechanics 2023 (ESB2023);
- Tissue Engineering and Regenerative Medicine International Society (TERMIS) 2023;
- CLEO2023, Muniche June 2023;
- EOSAM2023, Porto Sept 2023;
- FOM2022, Porto April 2022;
- 3R’ Italian center congress, Milano, September 2023;
- Icton, Bucharest, July 2023;
- Photonica 2023, Belgrade, August 2023;
- Optica (OSA) Imaging Congress 2023, Boston .
- FisMat 23 Congress, Milano, 4-8 September 2023.

Additional information can be found on the project web site: www.in2sight.eu
The consortium R&D actions has already produced innovative results in the fields of the laser micro-structuring of resins based on silica and Zirconium, the test of their biocompatibility and of their rigidity for long term implants.

Moreover, additional improvement beyond the state of the art are in the field of the optical setup for in-vivo imaging. These encompass the micro-fabrication with resins engineered for high index of refraction and low auto-fluorescence. The major leap forward in R&D of the second period of the project is the possibility to use the IN2SIGHT device to obtain in vivo images when implanted. The Consortium has started the exploitation by filing the first patent (May 2022).

We are proceeding also in the formulation of a modular and effective framework for the simulation of the immune reaction in a confined, well defined environment constituted by the micro-structured niche that is the exact replica of the one actually embedded in the IN2SIGHT device.

The economic impact of the project for the first year is primarily in the R&D activities and comes from the integration of the technologies developed in the laser fabrication facilities of the consortium to the level of microfabrication through nano-imprinting lithography. From the societal point of view, our impact can be judged from our effort to communicate (at congresses and workshops in the field of regulatory actions, beyond the field of photonics research, se for example our presence at the EUSAAT 2022 conference) and to disseminate the results of the project. We keep constantly updated the web site for the project and make use of the twitter and linkedin pages. The IN2SIGHT consortium has also organized workshops and dedicated sessions at international congresses.
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