3 replies to this topic

[Jakare]

Hi everyone, I am considering the possibility of testing the anticancer properties of C60OO using cell culture for my 3rd year project. After some initial talks with some teachers, the problem of its solubility arises. C60OO is obviously not water soluble and It can be difficult to dissolve it on DMEM as an example. Is there any easy way to dissolve it?

The type of cell is another problem. I have been told I would probably use a glioblastoma cell line but I am not sure that C60OO does something to pre-existing cancers (or does it?). What I was originally planning was to treat a healthy cell line with a carcinogen and see If C60OO prevents cancers at any rate or if it produces apoptosis on cells that become cancer cells.

Thoughts? Thanks in advance.

[niner]

Jakare, it's great that you're thinking of looking at this! C60-oo starts out as a triglyceride c60 adduct. When we consume it orally, one of the early steps in its digestion is lipase-mediated hydrolysis, forming the fatty acid c60 adduct that is the presumed active species. I don't know if cells can absorb a triglyceride and perform the hydrolysis themselves. One way to get around this would be to make a version of c60 that is attached to ethyl oleate. It's likely that ethyl oleate would get into the cell and the ethyl group would be removed by some generic esterase. One of our members has made some- it's quite easy. You'd probably need some DMSO to get it into solution. Another possibility would be to treat the c60-oo with a lipase first. If you were to hydrolyze either the oleate or the olive oil product, you'd get a soap, so it shouldn't be too hard to get it into solution. You could hydrolyze either of them chemically by heating in a basic solution (classic soap-making), although I would be concerned about what sorts of chemistry might occur on the c60 in a process like this. C60 is happiest when it's protected from light and oxygen. Making up a batch of ethyl oleate-c60 and using it with DMSO might be the easiest.

[Jakare]

Thank you niner! Awaiting an answer from the module leader. Let's hope he shares my enthusiasm although I have been told they are not keen on spending any, even reasonable, amount of money on just a 3rd year project...
For what you said it seems like using a lipase would be the best option as all the metabolites will be available to the cells. But logistics rule so lets hope I will be asking you soon for the protocol to make that ethyl oleate-c60...
Thanks again!

[hav]

Thank you niner! Awaiting an answer from the module leader. Let's hope he shares my enthusiasm although I have been told they are not keen on spending any, even reasonable, amount of money on just a 3rd year project...
For what you said it seems like using a lipase would be the best option as all the metabolites will be available to the cells. But logistics rule so lets hope I will be asking you soon for the protocol to make that ethyl oleate-c60...
Thanks again!

Hi, Jakare. Best wishes on your c60 research idea going forward. I've tested mixing c60 in ethyl oleate and found it does dissolve more readily than in olive oil, I was able to dissolve c60 in ethyl oleate at just under 2 mg per ml. I followed the Baati method of mixing with a magnetic stirrer for 2 weeks at room temperature in the dark, filtered afterwards, but skipped the centrifuge step.

At the risk of complicating your choice of methods, when reading up on how lipophilic/hydrophobic drugs are tested with cell cultures, i came across a variation of the approach suggested by niner:

Development of a liposomal nanodelivery system for nevirapine

And this paper on fullerene liposomes:

Liposome Formulation of Fullerene-Based Molecular Diagnostic and Therapeutic Agents

Howard

Edited by hav, 18 March 2014 - 10:01 PM.