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EC number: -
CAS number: -
The dermal irritation potential was assessed by applying EXP0700332 (10
µL) to the exposed surface of three EpiSkin reconstructed human
epidermis (RhE) units for 15 min. The surface area of the EpiSkin was
0.38 cm^2, therefore the application rate was 26.3 µL/cm^2. After the 15
min exposure period, the surface of the EpiSkin was rinsed with
Dulbecco’s phosphate-buffered saline (PBS; 25 mL). Initial rinsing with
PBS did not remove all of the applied dose from the EpiSkin surface.
Therefore, cotton swabs soaked in PBS were used to gently remove any
remaining test item from the surface of the EpiSkin units. The EpiSkin
was then incubated in maintenance media (2 mL) for a recovery period of
42.5 hours in a humidified incubator set to maintain temperature and CO2
levels of 37 degrees Celsius and 5%, respectively. Following incubation,
the EpiSkin units were transferred to assay medium (2 mL) containing MTT
(0.3 mg/mL) and returned to the incubator (set to maintain temperature
and CO2 levels of 37 degrees Celsius and 5%, respectively) for 3 hours.
Biopsies of the EpiSkin membranes were removed, added to acidified
isopropanol and refrigerated for ca 68 h in order to extract the
formazan. Formazan production (cell viability) was assessed by measuring
the optical density of the extracts at a wavelength of 550 nm. Three
replicates of the positive control, aqueous sodium dodecyl sulphate
(SDS) solution (5%, w/v), and the negative control, PBS, were tested in
parallel to demonstrate the efficacy of the assay. The viability of each
individual EpiSkin tissue was calculated as a percentage of the mean
negative control viability (defined as 100%). Exposure to EXP0700332
resulted in a mean EpiSkin viability of 99.90% +/- 9.42% of the negative
control value. Exposure to the positive control, aqueous SDS solution
(5%, w/v), resulted in a mean EpiSkin viability of 10.62% +/- 3.54% of
the negative control value.
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