Hi guys. So far, I have enough assays to show that its necrosis without apoptosis by ZnO np in SH-SY5Y cells. But I'm having trouble writing about it since many papers claim that its mainly apoptosis and apoptosis + necrosis. There hasn't been any that I found to directly say that ZnO np in that cell model or other models induces necrosis only. Any ideas how to defend this claim?
Please advise. Finishing my research this year. Thanks
If you have results on the inflammatory markers and apoptotic markers, then maybe you can talk about necrosis is more likely to occur in your studies supported by elevated inflammatory markers (which is associated with necrosis) and insignificant rises/normal level in the apoptotic markers, for examples, caspases levels. In term of cell morphology, you can support your conclusion by stating absence of apoptotic features like no blebbing, no apoptotic bodies seen etc..
I believe there are many papers which mention about the association of nanoparticles and inflammation in different systems, either to be in vitro or in vivo like C. elegans etc. So you can link presence of inflammation is likely to related to necrosis instead of apoptosis as the latter is rarely to be associated with inflammation.. Then you can discuss on possible ways on how ZnO nanoparticles cause necrosis but not apoptosis..
Run other apoptotic and necrotic markers as well. Try to do a titer experiment with lower doses of ZnO with your cell lines. Sometimes higher concentration of NPs induce necrotic cell death. But the markers will help you in making your point.
I agree, that you should do dose responses and time kinetics curves. May be at earlier time points and lower doses apoptosis is induced and later necrosis follows, then its not easy to detect apoptosis. Also its good to have a positive control for apoptosis to see your assay is working and also good to use more than one assay.
Best of Luck!
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