Hi, I want to start my cell culture to run a MTT assay , how can I sterilize my polymer?
Dear Ghazal!
Pleae You look at this publication: (attached)
Article Sterilized chitosan-based composite hydrogels: Physicochemic...
2.3 | Fabrication of hydrogels
Finally, hydrogels were sterilized in phosphatebuffered saline (PBS) using a steam autoclave SE 510 (Yamato Scientific, Tokyo, Japan) at 121C and 103.4 KPa for 15 min
Article The Potential of Stimuli-Responsive Nanogels in Drug and Act...
3.2.Sterilization of Nanogels
The sterilization methods currently used for pharmaceutical products are dry and moist heat, chemical cold sterilization that employs ethylene oxide and other gases, filtration, and radiation. Sterilization of the final formulation is the most employed method. Nevertheless, if the pharmaceutical product cannot be sterilized during fabrication, aseptic processing will be required with the special complications derived from handling and transfer of materials in a sterile environment [89].
Among the variety of sterilization methods, membrane filtration is a useful procedure for sterilization of light or heat sensible nanosystems without adverse effects for the nanosystem properties [90]. However, in the case of particles with a broad size distribution, sterilization by filtration might not be the most adequate method because membrane pore size cannot be tailored for filtration particles of different sizes [91]. In addition, rigid or inflexible particles can lead to clogging of the sterilization membranes [92]. Despite the flexibility of nanogel particles, solid content loss can occur when passing through the standard filtration sterilization membranes (0.22 µm pore size).
4.1. Cytocompatibility Assays in Nanogels
Polymeric NGs synthesis implies using monomers, initiators, and surfactants that can have toxic characteristics. To provide information about their biocompatibility and the most safe and effective concentrations, “in vitro” studies are necessary .Cell viability is analyzed using tests such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, AlamarBlue, and trypan blue analysis. The MTT test, one of the most employed techniques, is a colorimetric test based on the selective ability of viable cells to reduce MTT into an insoluble compound, formazan.
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