Periodic Reporting for period 2 - SEE (MAPPING THE SKIN-IMMUNE INTERACTIONS OF NOVEL 2D MATERIALS: MXENES)
Reporting period: 2023-07-01 to 2024-06-30
In conclusion, MXenes were highly biocompatible at both the skin and immune levels. A versatile multiplexed label-free single-cell detection strategy based on CyTOF and ion beam imaging by time-of-flight (MIBI-TOF) was developed for MXenes. This strategy, Label-free sINgle-cell tracKing of 2D matErials by mass cytometry and MIBI-TOF Design (LINKED), enables nanomaterial detection and simultaneous measurement of multiple cell and tissue features. In addition MXene ability to promote wound healing was observed. These findings pave the way for further research and development for the applications of MXenes in the field of nanomedicine.
Overview of the results and their exploitation and dissemination:
Within the project, different main interesting results were identified, which have been selected and prioritized due to their high potential to be exploited – meaning to make use and derive benefits- downstream the value chain of MXenes, and act as an important input to policy.
i) Considering the critical unmet need to detect and image 2D materials within single cells and tissues while surveying a high degree of information from single samples, the project proposed a versatile multiplexed label-free single-cell detection strategy based on CyTOF and MIBI-TOF. This strategy, Label-free sINgle-cell tracKing of 2D matErials by mass cytometry and MIBI-TOF Design (LINKED), enabled nanomaterial detection and simultaneous measurement of multiple cell and tissue features. The label-free detection of 2D materials by mass cytometry at the single-cell level, on multiple cell subpopulations, and in multiple organs simultaneously, is expected to enable exciting new opportunities in biomedicine. This method was extended to other nanomaterials sharing similar properties, thus paving the way for new discoveries in nanomedicine and materials science.
ii) Within the project, MXenes with different characteristics were developed and the material physicochemical properties were correlated to their biological impact with a particular focus on immune cells, including macrophages and peripheral blood mononuclear cells and human skin.
iii) Detecting nanomaterial uptake from the skin is challenging. Thanks to the LINKED approach developed within the project, it was possible to evaluate the absorption of nanomaterials at the skin level. The advanced approach and analysis methodology can be applied to other kinds of nanostructures and nanomaterials with similar properties.
Exploitation mechanisms and activities to maximize the expected impact focus on identifying end-users to ensure impact and uptake of the results:
1. Several companies interested in the SEE project results have been identified. These have been targeted by different communication and dissemination actions and networking/clustering activities.
2. Results on the immune impact were obtained in advance of the scheduled time, resulting in a provisional patent application (U.S. Provisional Application No. 63/374,460) was submitted.
3. The safe use of MXenes will be beneficial not only for their biomedical applications, but also for other sectors where MXenes are emerging as promising candidates, from energy storage, catalysis, and nanoelectronics, Artificial Intelligence, to communications and sensing.
4. Understanding the effects and customizing the design of MXenes with tunable properties is expected to paw the way for a wide variety of new opportunities and discoveries for MXene technologies in several fields.
5. It will accelerate the rapid and safe adoption of newly developed materials, also facilitating general public awareness and trust in 2D material advances.
6. Can potentially contribute to the efforts of regulatory agencies in developing specific guidelines for nanomaterial safety assessment.
7. In addition, by exploring the biological impact of MXenes and shade light on their classification, the project is expected to facilitate the application-oriented design of MXenes making it possible to scale up MXene synthesis well past laboratory scales, and potentially providing a path toward the direct commercialization of MXene and MXene-based technologies.
8. The Fellow was selected among the 10 finalist teams of Start Cup Padova, a competition for the development of a star-up, proposing MXenes for biomedical applications.
Within the project, different main interesting results were identified, which have been selected and prioritized due to their high potential to be exploited – meaning to make use and derive benefits- downstream the value chain of MXenes, and act as an important input to policy.
i) Considering the critical unmet need to detect and image 2D materials within single cells and tissues while surveying a high degree of information from single samples, the project proposed a versatile multiplexed label-free single-cell detection strategy based on CyTOF and MIBI-TOF. This strategy, Label-free sINgle-cell tracKing of 2D matErials by mass cytometry and MIBI-TOF Design (LINKED), enabled nanomaterial detection and simultaneous measurement of multiple cell and tissue features. The label-free detection of 2D materials by mass cytometry at the single-cell level, on multiple cell subpopulations, and in multiple organs simultaneously, is expected to enable exciting new opportunities in biomedicine. This method was extended to other nanomaterials sharing similar properties, thus paving the way for new discoveries in nanomedicine and materials science.
ii) Within the project, MXenes with different characteristics were developed and the material physicochemical properties were correlated to their biological impact with a particular focus on immune cells, including macrophages and peripheral blood mononuclear cells and human skin.
iii) Detecting nanomaterial uptake from the skin is challenging. Thanks to the LINKED approach developed within the project, it was possible to evaluate the absorption of nanomaterials at the skin level. The advanced approach and analysis methodology can be applied to other kinds of nanostructures and nanomaterials with similar properties.
Exploitation mechanisms and activities to maximize the expected impact focus on identifying end-users to ensure impact and uptake of the results:
1. Several companies interested in the SEE project results have been identified. These have been targeted by different communication and dissemination actions and networking/clustering activities.
2. Results on the immune impact were obtained in advance of the scheduled time, resulting in a provisional patent application (U.S. Provisional Application No. 63/374,460) was submitted.
3. The safe use of MXenes will be beneficial not only for their biomedical applications, but also for other sectors where MXenes are emerging as promising candidates, from energy storage, catalysis, and nanoelectronics, Artificial Intelligence, to communications and sensing.
4. Understanding the effects and customizing the design of MXenes with tunable properties is expected to paw the way for a wide variety of new opportunities and discoveries for MXene technologies in several fields.
5. It will accelerate the rapid and safe adoption of newly developed materials, also facilitating general public awareness and trust in 2D material advances.
6. Can potentially contribute to the efforts of regulatory agencies in developing specific guidelines for nanomaterial safety assessment.
7. In addition, by exploring the biological impact of MXenes and shade light on their classification, the project is expected to facilitate the application-oriented design of MXenes making it possible to scale up MXene synthesis well past laboratory scales, and potentially providing a path toward the direct commercialization of MXene and MXene-based technologies.
8. The Fellow was selected among the 10 finalist teams of Start Cup Padova, a competition for the development of a star-up, proposing MXenes for biomedical applications.
i) The effects of a panel of MXenes (Ti3C2, V2C, Ta4C3, etc.) with different physical and chemical properties (size, lateral dimension, chemical structure, etc) were evaluated on immune cells.Results were obtained even beyond the envisaged objectives, by developing a versatile multiplexed label-free single-cell detection strategy based on CyTOF and ion beam imaging by time-of-flight (MIBI-TOF) for MXenes. This strategy, Label-free sINgle-cell tracKing of 2D matErials by mass cytometry and MIBI-TOF Design (LINKED), enables nanomaterial detection and simultaneous measurement of multiple cell and tissue features, enabling new opportunities in biomedicine (a provisional patent application was submitted (U.S. Provisional Application No. 63/374,460).
ii) The biocompatibility of MXene on the skin was demonstrated. To evaluate the material capability to penetrate the skin, CyTOF technology demonstrated the ability to detect and quantify MXene at the single-cell level in primary skin cells (NHEK) and immortalized human keratinocytes (HaCaT). In addition MXene ability to promote wound healing was observed. These findings pave the way for further research and development in the field of MXenes for skin applications.
iii) The results demonstrating the biocompatibility of MXene with human macrophages are highly significant. The material did not reduce cell viability nor compromise cellular metabolism, indicating that MXene is safe for use in biological systems. This biocompatibility is crucial for potential biomedical applications, such as drug delivery, tissue engineering, and biosensing, where macrophages play a vital role in immune response and tissue homeostasis. The ability of MXene to maintain macrophage health and functionality opens up new avenues for its integration into medical devices and therapeutic strategies, ensuring that the immune system remains uncompromised while benefiting from the advanced properties of MXene materials. These findings pave the way for further research and development in the field of biomedicine, leveraging MXene's unique characteristics for innovative healthcare solutions.
iv) The Skin irritation induced by MXene was evaluated on human skin model, following the skin Irritation Test-42bis, a test compliant with the OECD Test Guideline (TG) No. 439 for testing of chemicals. These findings are significant as they suggest that MXene does not compromise the viability of human epidermal keratinocytes. This lack of cytotoxicity is crucial for potential biomedical and cosmetic applications, where maintaining skin health is paramount. Furthermore, the use of the skin models provides a robust in vitro platform for assessing skin irritation, offering insights that are highly relevant to human skin physiology. In summary, MXene has been shown to be non-irritating, indicating its safety for topical applications. This conclusion supports the further development and potential use of MXenes in products intended for direct skin contact.
ii) The biocompatibility of MXene on the skin was demonstrated. To evaluate the material capability to penetrate the skin, CyTOF technology demonstrated the ability to detect and quantify MXene at the single-cell level in primary skin cells (NHEK) and immortalized human keratinocytes (HaCaT). In addition MXene ability to promote wound healing was observed. These findings pave the way for further research and development in the field of MXenes for skin applications.
iii) The results demonstrating the biocompatibility of MXene with human macrophages are highly significant. The material did not reduce cell viability nor compromise cellular metabolism, indicating that MXene is safe for use in biological systems. This biocompatibility is crucial for potential biomedical applications, such as drug delivery, tissue engineering, and biosensing, where macrophages play a vital role in immune response and tissue homeostasis. The ability of MXene to maintain macrophage health and functionality opens up new avenues for its integration into medical devices and therapeutic strategies, ensuring that the immune system remains uncompromised while benefiting from the advanced properties of MXene materials. These findings pave the way for further research and development in the field of biomedicine, leveraging MXene's unique characteristics for innovative healthcare solutions.
iv) The Skin irritation induced by MXene was evaluated on human skin model, following the skin Irritation Test-42bis, a test compliant with the OECD Test Guideline (TG) No. 439 for testing of chemicals. These findings are significant as they suggest that MXene does not compromise the viability of human epidermal keratinocytes. This lack of cytotoxicity is crucial for potential biomedical and cosmetic applications, where maintaining skin health is paramount. Furthermore, the use of the skin models provides a robust in vitro platform for assessing skin irritation, offering insights that are highly relevant to human skin physiology. In summary, MXene has been shown to be non-irritating, indicating its safety for topical applications. This conclusion supports the further development and potential use of MXenes in products intended for direct skin contact.