I am synthesizing chitosan nanoparticle by ionic gelation. The pH of chitosan solution while synthesizing nanoparticles is 4.6. The pH of treatment media is 7.4.
You should decrease the medium pH to 6.5. Encapsulation of BSA can also be useful if the first approach does not work.
Maybe you can add a tensioactive like poloxamer at low concentration, I know that the poloxamer can be in touch with cells and not kill them. There are different types of poloxamer that can help you in order to avoid aggregation.
As mentioned above, pH of the medium for unmodified chitosan nanoparticles has to be 6.5 or lower. If you are using trimethylated chitosan, consider using HEPES instead of phophate buffered saline. sometimes it also helps to use 5 mM buffers instead of 10 mM. Lastly, perform a pH stability study of your nps in different medium/pH before treating them with cell cultures.
Hi Amit,
Have you thought of purifying your nanosuspension before putting it on cells ? From previous experience, it helps a lot . You can, for example, perform either rigorous dialysis: 10 mL against 1.0 L of ultrapure water, 3 x 24 h, MWCO of dialysis membrane: 2.5 x that of your 'bigger' molecules. You can also use tangential filtration to wash your nanosupension. If it doesn't improve sufficiently your situation, you could add a small amount of tensioactive (poloxamers work well) in your purified nanosuspension. Of course, you'll have to modify consequently your controls, but it may be worth it.
Good luck with your work !
Hello Mr. Yadav, You should send me more detaile information about the ionic gelation proccess and medium treatment parameters (gelation reagent used etc.) .
May be you can try with the capping agents that alter medium zetapotential your nanopaticles are positive use positive capping agents.other wise negative capping agents is preferable.
This article may help you for your research
Article FORMULATION AND CHARACTERIZATION OF EPIGALLOCATECHIN GALLATE...
Pharm Res 15(9):1326–1331
This will help you. This suggests adding monovalent salt to the solvent which screens out to the electrostatic repulsion between the positively charged amine groups on the chitosan backbone. This will increase the flexibility of the polymer chains in the solution and thus increase their stability.
Valérie Gaëlle Roullin Thank you for your suggestions.
Which poloxamers would you reccommend?
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