1. Outline – the need for single cell measures of nanotoxicity
1. There is more than one way for a cell to die...
2. Necrosis" vs. "Apoptosis"
3. There are other forms of "toxicity"
4. Some other challenges in measuring toxicity of nanomaterials
2. Necrosis vs. Apoptosis mechanisms
1. Necrosis is unplanned "cell injury"
2. Apoptosis is planned "programmed cell death"
3. Why it is important to distinguish between necrosis and apoptosis
3. Single cell assays for necrosis and apoptosis
1. Dye exclusion assays for necrosis
2. TUNEL assays for late apoptosis
3. Annexin V assays for early apoptosis
4. COMET assays for DNA damage and repair
5. Light scatter assays
4. Nanotoxicity in vivo – some additional challenges
1. Single cell nanotoxicity, plus....
2. Accumulations of nanoparticles can change toxicity locally to tissues and organs
3. Filtration issues of nanoparticles – size matters – toxicity to liver and lung

• Chana, W-H, Nion-Shiao, N-H, Pin-Zhen Lu, P-Z. CdSe quantum dots induce apoptosis in human neuroblastoma cells via mitochondrial-dependent pathways and inhibition of survival signals. Toxicol. Lett. (2006), doi:10.1016/j.toxlet.2006.09.007
• Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F. Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry. 1997 Jan 1;27(1):1-20.
• Kirchner,C. Liedl, T., Kudera, S., Pellegrino,T., Munoz Javier, A., Hermann E. Gaub,H.E., Stolzle,S., N. Fertig, Parak, W.P., Cytotoxicity of Colloidal CdSe and CdSe/ZnS Nanoparticles. Nano Lett., Vol. 5, No. 2, 331-338, 2005.
• Oberdörster,G., Oberdörster, E. Oberdörster, J. Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles. Environmental Health Perspectives 113(7): 2005
• Ryman-Rasmussen, J.P., Riviere, J.E.,Monteiro-Riviere, N.A. Surface Coatings Determine Cytotoxicity and Irritation Potential of Quantum Dot Nanoparticles in Epidermal Keratinocytes. Journal of Investigative Dermatology. 10 August 2006; doi:10.1038/sj.jid.5700508
• Shiohara, A., Hoshino, A., Hanaki, K., Suzuki, K., Yamamoto, K. On the cytotoxicity caused by quantum dots. Microbiol. Immunol. 48(9): 669-675, 2004.

Researchers should cite this work as follows:

• James Leary (2007), "BME 695N Lecture 17: Assessing nanotoxicity at the single cell level," http://nanohub.org/resources/3497.

1. biomedical engineering 1
2. cells 1
3. course lecture 1
4. drug delivery 1
5. nanomedicine 1