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Hands and Eyes on Single-cells: Silver Nanowire Endoscopy for Intracellular Delivery and Sensing

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Category
Ph D Defense
Date
2021-01-21 13:00
Venue
KU Leuven, Universiteitshal, Promotiezaal, 01.46 - Naamsestraat 22
3000 Leuven, België

Promovendus/a: Monica Ricci

Promotor(en): Prof. dr. Hiroshi Ujii, Prof. dr. Maurilio Sampaolesi, Prof. dr. Susana Rocha, Mevrouw Beatrice Fortuni

The heterogeneous essence of life can be observed at nearly all biological scales and levels, yet it becomes evident when looking at all the different species that evolution has created. But heterogeneity is much further deep into living systems, where even cells from "apparently identical" populations are in fact heterogeneous, owning a wide range of different characteristics, likewise individuals within one species exhibit unique properties. Is every day more clear that observing, and manipulating, single cells plays a significant role in shaping our essential understanding of biological phenomena and diseases. The detection of tumour cells and cancer treatment, gene therapy and regenerative medicine, and drug discovery are only a few examples. In this framework, extremely beneficial is the ability to address single-living cells with a high spatiotemporal resolution, while retaining the cell in its native, healthy, state. The ultimate goal is to observe cellular responses to the introduction of exogenous materials, as well as to unravel biochemical events happening in living cells.

One way to interrogate single-living cells is through a technique called silver nanowire endoscopy. Silver nanowires can be inserted, and retracted, into, and from, single living cells in the same way as a needle can be manually inserted into ravioli. Endoscopy derives from the Greek words "endo", meaning "within", and the verb "skopein", meaning "to view", and to observe inside something is precisely what silver nanowires do. Silver nanowires make their way into the cell in a minimally invasive fashion by virtue of their 1D configuration and small sizes compared to the cell dimensions. These features make them an ideal candidate for delivery exogenous material into the cells.

The first focus of this thesis is, therefore, the development of an intracellular delivery system based on silver nanowires. The key challenge here is to find the best strategy to load the material onto the nanowire and to release it into the cell quickly and with high efficiency. The main focus of this part is, therefore, the optimisation of the loading and releasing strategy. This challenge is tackled by using "smart" polymers, named after their smart responses to external stimuli, such as changes in temperature, pH or light. The study was centred on the delivery of genetic material, being it highly applied in gene therapy, as well as in cancer treatment.

The peculiar optical properties of silver nanowires enable us to perform surface-enhanced Raman scattering (SERS) spectroscopy analyses. SERS is a powerful technique in which the interaction between light and nanostructures enable to observe molecules and obtain information about their conformation, structure, and interactions with surrounding items. Simply put, SERS outcomes can be seen as a molecule's identity card.

The thesis hereafter, therefore, focuses on the development of a reliable platform for high-sensitive intracellular sensing, based on SERS spectroscopy. The first key step is a further improvement of the SERS performances of the silver nanowire endoscopic probes. This is important because it enables more accessible operational protocols, which translate in better versatility. Moreover, the better SERS performances allow for application to real case studies, as shown by the solid proof of concept provided. Finally, we step into the real world by studying the intracellular dynamics of anticancer drugs, being this aspect fundamental in envisioning new, highly efficient, chemotherapeutics.

The journey of this thesis is concluded by perspectives and directions for future studies, with the final goal of developing a "2 in 1" delivery-sensing platform, ready to be applied across the nanomedicine realm.
 
 

All Dates

  • 2021-01-21 13:00

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