Application of in vitro and zebrafish models to investigate nanomaterial mediated pulmonary fibrosis

"This is a PhD research project in Biology (specialising in Toxicology)
Objectives:
Monoculture of cells
1. Investigate the cytotoxicity o f nanomaterials (NMs) to alveolar epithelial cells, fibroblasts and macrophages in vitro using alamar blue and lactate dehydrogenase (LDH) assays
a. Identify what studies already exists for the NMs of interest (to avoid duplication of research)
i. NMs: CNTs, Ag, ZnO, CuO
ii. Include fibrosis positive controls (e.g. bleomycin, quartz)
b. Compare toxicity when NMs are exposed in an aqueous suspension vs ALI dry cloud
2. Investigate collagen deposition (microscopy), cytokine (e.g. TGF-B) production (ELISA) and matrix metalloproteinase in monocultures of cells following single exposure
3. Investigate toxicity of NMs following single and repeated exposure (up to 3 weeks) in monocultures e.g.
a. Cytotoxicity
b. Cytokine production
c. Collagen deposition
d. Matrix metalloproteinase content / activity
e. Cell morphology (light / electron microscopy)
3D co-culture of cells
1. Investigate cytotoxicity of nanomaterials to a co-culture of epithelial cells, fibroblasts and macrophages in vitro following single and repeated exposure
a. Endpoints as above
Zebrafish
1. Investigate collagen deposition and inflammatory cell accumulation in transgenic zebrafish following single and repeated exposure
-exposure via gills or microinjection into swimbladder (as surrogates for lung)
-use non-protected life stages where possible (can use later life stages if required)"