I am dealing with support-mediated drug delivery systems, and in particular I have some cancer cells adherent on a collagen surface. I loaded a hydrophilic drug on collagen by physisorption, then seeded cells, but I could not detect any drug inside the cells after 24h, even if I clearly saw (by fluorescence) cells attached on the drug-rich areas at the interface, and a drug release from the surface over time.
Has someone any suggestion, or can give me some explanations more about the molecular transport processes across the cell membrane on the support side?
I don't deal with live cells, but I do use alpha-hemolysin to transport single molecules at 2 nm or less across lipid bilayers. Perhaps there is some kind of protein nanopore out there that can be used.
For example, lets say your drug is around 1.8 nm (for convenience, there are lots of biological nanopore proteins out there of different sizes and function, and I would be happy to provide further detail if this idea sounds intriguing). You could use a patch pipette method detailed in my paper's supplemental ( Article Ligand-Induced Structural Changes of Thiolate-Capped Gold Na...
) to apply a single alpha-hemolysin protein complex onto the surface. You could then release the drug into the vicinity giving it some time to enter the cell (this takes only a second), then spike in something like streptavidin to seal up the pore to avoid having the cell lose salts.This is just my immediate thought and probably would take a little trial and error to find the right protein/method. Not to mention the difficulties of doing this in vivo. Hopefully you get something useful out of this answer.
Hi Vittorio Ferrara
you can use a carrier such as liposome or micelle to transfer a small molecule into the cell
Dear Bobby Cox, I find your suggestion to use a pore-forming protein a very good idea. They generally work well in solution, then I might try to translate this approach on a solid interface.
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