Download Development of 3D in vitro tissue models for the analysis of

DIT PhD Project
Supervisor name & contact details:
Hugh Byrne
[email protected]
Research Centre:
Focas Research Institute
Research Centre website:
http://www.dit.ie/focas/
Supervisor’s List of Publications
http://arrow.dit.ie/do/search/?q=author_lnam
e%3A%22Byrne%22%20author_fname%3A%22
Hugh%22&start=0&context=490738
Brief summary of research centre activity:
The Focas Research Institute has extensive experience in the field of microscopy and imaging. The
institute is equipped with cutting edge instrumentation, from conventional confocal fluorescence
microscope to Raman and Infrared imaging systems (http://www.dit.ie/focas/facilities/). In
recent years, research in Focas has greatly contributed to a better understanding of the potential
of vibrational spectroscopy for biological applications. The wide range of application covers
studies going for single cells to tissue analysis. On one hand, Infrared spectroscopy is particularly
suitable for the analysis of large tissue area and has been proven suitable for diagnostic
applications and the specific identification of pathological samples in the case of cervical and
lung. On the other hand, the high spatial resolution when working with Raman spectroscopy has
allowed to access information at the subcellular but also sub-nuclear level. The mapping of single
cells in vitro can deliver crucial information regarding cell-drug interaction interaction but can
also help to the localization and tracking of nano-particles inside the cells without requiring any
labeling. The use of more suitable models to represent in vivo conditions such as collagen gels
have been demonstrated to be perfectly suitable for Raman spectroscopy giving access to the
study of live cells in a 3D environment. Additionally, Raman spectroscopy is a powerfull tool for
the investigation of tissue samples. Recent work using Raman spectroscopic mapping to identify
and chemically characterise the dermal and stratum corneum layers in samples and distinguish
further substructures of the epidermis it has been possible to identify DNA damage, loss of lipidic
signatures and changes in protein signatures and track trends in these changes according to the
exposure times of the samples to solar radiations. Raman spectroscopy can identify a significant
degree of biochemical damage at stages where minimal loss in tissue viability is observed,
suggesting the modality holds great potential for early disease diagnosis applied to skin cancer.
Title: Development of 3D in vitro tissue models for the analysis of solar radiation damage of
skin
The objectives of this work are to develop novel in vitro skin models for the evaluation of the
impact of solar radiation at a molecular level using Raman spectroscopy. The project will address
such research questions as to how does solar radiation affect skin barrier function on a molecular
level, whether artificial skin models can be developed for the investigation of skin function and
whether Raman spectroscopy can be established as a routine, rapid screening technique for
evaluation of skin function and dysfunction. New regulations against human and animal testing
(EU Directive-2010/63/EU and US Public Law 106-545, 2010, 106th Congress) necessitate the
development of suitable in vitro models for skin research. Therefore, for the conduction of in
vitro studies, several substitutes have been developed, commonly called artificial skin or skin
recombinant, mimicking the 3D organization of the human skin. Although the techniques of cell
culture, and for instance skin culture, have evolved considerably, the study of the skin
composition remains challenging, due to invasiveness of some approaches. For example,
techniques such as HPLC and GCMS are routinely used in analytical chemistry but require
chemical extraction for specific molecular charaterisation. This project aims to provide a new
alternative for the analysis of skin samples in vitro, providing a specific molecular fingerprint of
the sample without interfering with the complex organization of the skin in sub-layers. Raman
spectroscopic microscopy will be employed for the investigation of UV induced damaging of 3D
skin models and alteration of the skin function barrier. The main advantage of this approach is
the absence of extraction or sample processing before analysis allowing to preserve the
molecular organization intact during the recording of the data. Using Raman spectroscopy to
perform Z chemical profiling to access the molecular composition of the underlying sub-layers of
the epidermis, the skin differentiation will be investigated. The objective of the proposed study is
an in depth understanding at the molecular level how solar radiation can penetrate the epidermis
affecting the keratinocyte maturation and the events or factors conducting to an alteration of the
skin permeability thus a loss of the skin barrier function.
Ciência sem Fronteiras / Science Without Borders Priority Area:
Health and Biomedical Sciences