To compare sensitizing and non-sensitizing materials using an IL-18 / MTT based in vitro sensitization assay to determine Labskin’s response as an in vitro skin model.
- Labskin Maintenance Medium (4.5 mL) was added to each well. The 12 well plate configuration was used to conduct the experiment.
- Known sensitizing materials (4-Nitrobenzyl bromide [NBB] and Eugenol) along with known non-sensitizing material (Lactic Acid) were used to dose tissues (52 μL).
- A 12 well plate was used to incubate tissues with 1 mL of MTT (1 mg/mL) for 3.5 hours.
- In each 12 well plate insert, Isopropanol (2 mL) was placed and a further 2 mL Isopropanol was added to the top of the tissues. Extraction occurred overnight.
- The next day spectrophotometer (OD 540-690) was used to read 200 μL aliquots for viability once removed. ELISA (MBL) was used to analyze the media for IL-18 secretion into the culture media.
As expected, there was no increase in IL-18 with the non-sensitizer Lactic Acid in comparison to vehicle control (AOO [Acetone: Olive OIL]).
However, where a response should have been seen, there was no increase in IL-18 for the sensitizing material Eugenol in comparison to the vehicle control. Fold increases in IL-18 for NBB was detected.
Figure 1. IL-18 Secretion detected in culture media
In conclusion, this experiment demonstrated that Labskin full thickness in vitro skin model is able to respond to sensitizing/allergic materials.
At Labskin we deliver human skin microbiology services to support your product R&D activities in the cosmetic, personal care, medical device and pharmaceutical sectors. With our sector experience and use of technology, you will be accessing industry-focused services supported by world-leading skin science expertise.
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Developed over 12 years with more than 30 scientific journal publications, it is made from young keratinocytes (human skin cells) and adult fibroblasts (metabolically-active, collagen-producing human skin cells).
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