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Quantifying bio-nano interactions of nanoparticles through microfluidic live cell Raman spectroscopy

Periodic Reporting for period 1 - qBioNano (Quantifying bio-nano interactions of nanoparticles through microfluidic live cell Raman spectroscopy)

Reporting period: 2017-08-21 to 2019-08-20

Bio-nano science is an emerging area of research at the interface of engineering and medicine. The ultimate aim is to provide new treatments for diseases. One key challenge in this area is understanding how nanoparticles interacts with cells and tissues: “bio-nano interactions”. The objective of this MSCA Individual Fellowship and the qBionano project was to develop new methods to facilitate quantitative bio-nano studies. The MSCA fellow described some of these challenges in a popular science article published in ‘The Conversation’: https://theconversation.com/how-diseases-can-be-targeted-using-nanotechnology-and-why-its-difficult-82434

By investigating a range of emerging technologies, the qBionano project has made substantial progress in the areas of nanomaterial preparation and bio-nano analysis. Nanoparticles and microparticles with tailored properties were engineered. Raman spectroscopy was employed to examine the interaction of these materials with biological environments. And finally, microfluidic devices were investigated to explore new avenues to research the interactions of nanomaterials with biological cells.

Outcomes of the project include an increasing understanding of how these types of systems should be designed. The project proposal envisioned “Raman active” particles and during the project, particle design evolved from being based around conventional Raman spectroscopy to also include particles incorporating elements enabling surface-enhanced Raman spectroscopy which can greatly increase sensitivity. In addition, thanks to the highly collaborative environment and strong network of the host group, there was an opportunity to pursue a complementary approach based on split fusion proteins. During the qBionano project, the MSCA fellow has co-supervised MSc and PhD candidates, and some of them are continuing this work. The comparative study (comparing Raman approaches and split fusion protein approaches for the study of bio-nano interactions) is showing great promise and has provided a wealth of data and insights so far. The results from this study are intended for publication in a peer-reviewed journal.

Secondly, an opportunity to further engage with the broader objective of this project (i.e. to facilitate quantitative bio-nano studies) occurred during project. The outcome was the development of the MIRIBEL guidelines which were published in September 2018 (Faria & Björnmalm et al., Nature Nanotechnology, open access link http://hdl.handle.net/11343/216237 ). The development of these guidelines received a lot of attention from the broader community (for some examples see the editorial of the July 2019 issue of Nature Nanotechnology). A follow-up publication, co-authored by the MSCA fellow, was further published open access in the Journal of Controlled Release (open access link https://doi.org/10.1016/j.jconrel.2019.06.027 ).

Overall, the qBionano project has led to a range of new insights in the area of bio-nano interactions, and contributed substantially to the overall goal of facilitating quantitative bio-nano studies. By being placed in the highly interdisciplinary Stevens Group at Imperial College London, with access to extensive networks and collaborators, the MSCA fellow has gained extensive new scientific and professional expertise and experience. Additionally, the MSCA fellow has had the opportunity to pursue the objectives of the qBionano project from several directions, which has greatly amplified the outcomes and impact. For the host group, the qBionano project has laid a solid foundation for continuing work in this area, with several projects in the Stevens Group having benefitted from these efforts. Ongoing work in the Stevens Group is building on the outcomes from qBionano, thus ensuring its continuing impact.
The following summarizes the main research tasks and results from this fellowship:

1. To prepare the nanomaterials, alkyne-modified poly(methacrylic acid) was synthesized and particles were prepared following the layer-by-layer assembly strategy. Nanomaterials were characterized using a suite of methods, including optical microscopy, electron microscopy, and dynamic light scattering analysis.
2. Following particle preparation, Raman spectroscopy experiments were performed. The signal achieved for this new type of nanomaterial was deemed to be quite low, a potential challenge identified already in the original project proposal. The extensive expertise and collaboration network in the host group was leveraged to address this challenge, and two complementary strategies were incorporated into the project. One approach uses materials with surface-enhanced Raman spectroscopy (SERS) properties and the other strategy leverages fusion protein constructs used to study nanoscale interactions.
3. A microfluidic device was designed to explore cell behavior and particle behavior in flow environments. The compatibility of standard microfluidic devices (using cover glasses meant for fluorescence microscopy as the bottom layer) was determined to be suboptimal for Raman spectroscopy. New microfluidic devices were therefore envisioned and designed with the help of collaborators. This next generation device was also specifically developed to be able to incorporate complex biological tissue constructs.
4. The qBionano project additionally contributed to the development of the MIRIBEL guidelines. This effort aligns well with the central objective of the qBionano project (i.e. to facilitate quantitative bio-nano studies).

The key findings from this project were disseminated in several ways. Examples include presenting at international research conferences such as the Drug Carriers in Medicine and Biology Gordon Research Conference in Vermont, USA in 2018; and the International Nanomedicine Conference in Sydney, Australia in 2019. The MSCA fellow has also given guest lectures, including at RWTH Aachen in Germany, Zhejiang University in China, Shandong University in China, and at the University of Melbourne in Australia. Several publications were also authored and co-authored by the MSCA fellow. Two further publications are in preparation for submission to high-quality journals (all publications acknowledge MSCA funding and follow all relevant open access policies). The MSCA fellow has also published a popular science article aimed for the public, and participated in several science outreach events, including at the Science Museum in London.
The results from this Marie Skłodowska Curie Actions project have advanced the state-of-the-art in several areas. Firstly, several new nanomaterials and particle types have been developed suitable for exploring bio-nano interactions in general and intracellular interactions in specific. Secondly, the use of Raman spectroscopy in this area remains a nascent field, and new insights from the qBionano project is helping to advance this area. Thirdly, the design and development of a new generation of microfluidic devices is expected to advance this field greatly. Finally, the development of the MIRIBEL guidelines has already had a substantial impact on the bio-nano science community. This project has thus had scientific impact, as well as impacted on the broader community. In the longer term, this project is helping to advance our knowledge towards the development of new technologies and strategies for the treatment of diseases.
Bio-nano studies of engineered nanomaterials