DIT PhD Project
Supervisor name & contact details:
Hugh Byrne
hugh.byrne@dit.ie
0872169501
Alan Casey
alan.casey@dit.ie
+35314027932
Research Centre:
Focas Research Institute
Research Centre website:
http://www.dit.ie/focas/
http://www.dit.ie/nanolab
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.
Additionally, the Nanolab research centre has unparalleled expertise in state of the art nano
material characterisation and the analysis of the interaction of Nanomaterials with biological
systems, both mammalian and aquatic (fresh and marine water) systems. Its researchers explore
standards and methods for the characterisation of nanomaterials with respect to physical,
chemical and biological properties including the toxicity and biocompatibility of a variety of
nanomaterials such as carbonaceous, polymeric, metallic and composite nano material systems.
The centre’s emphasis is on establishing structure activity relationships governing particle uptake,
trafficking, fate and organism response. Model systems are employed to improve fundamental
understanding, to validate current and develop new biological testing protocols for
nanomaterials, while real life exposure scenarios are explored to assess risk.
Nanolab is active in promoting awareness of the impacts of nanotechnology among stakeholders
through national and international workshops as well as in pedagogical research for the
advancement of education in nano sciences.
Title & brief description of PhD project (suitable for publication on web):
Impact of nanoparticles on differentiation and maturation of primary cell lines
Recent years have seen a growth in the use of nanomaterials in all areas of industry. The
Department of Enterprise, Trade and Innovation estimated that 10% of ~€150 billion goods and
services exported from Ireland in 2008 were enabled by nanoscience and nanotechnologies. This
increased usage raises issues regarding the health and environmental risks associated with
engineered nanomaterials. Of particular concern is the potential of these particles to disrupt
placental and foetal development. Therefore it is important to examine the effects of
nanoparticles on stem cells and stem cell differentiation.
This project will investigate the effects of commercially available nanoparticles to primary
osteoblasts (Promocell, UK) and an ostoblast-like cell line, U20S (Health Protection Agency, UK)
by examining uptake, toxicity and differentiation. Polystyrene nanoparticles are chosen as models
as they have been shown to be uptaken by cells and trafficked internally. They can be
fluorescently labelled, and so their interactions can be easily visualised. Neutral nanoparticles are
nontoxic to cells, while their aminated counterparts cause oxidative stress leading to apoptosis5.
Primary osteoblasts will initially be monitored, but due to high costs and limited passaging, the
osteoblast-like cell line U20S will be examined in parallel. Investigations will employ classical
techniques of cytotoxicity, immunocytochemistry, fluorescence microscopy, flow cytometry and
spectroscopic imaging.
Using polystyrene nanoparticles (http://www.kisker-biotech.com/shop/categorie-106.htm) of
varying size and surface chemistry as models, this project will:
- Assess in vitro toxicity of polystyrene nanoparticles on undifferentiated primary and cell line
osteoblasts
- Examine the in vitro toxicity of these nanoparticles to differentiated osteoblasts
- Elucidate whether the polystyrene nanoparticles affect the normal mechanisms of osteoblast
differentiation in vitro
Ciência sem Fronteiras / Science Without Borders Priority Area:
Health and Biomedical Sciences