The aim of this article is to compare the distribution and levels of xenobiotic metabolism between ex vivo human skin and Labskin. This comparison is performed to identify whether it is possible to use Labskin as a prediction tool for xenobiotic metabolism in human skin via proteomic and substrate-based mass spectrometry imaging (SB-MSI) analysis.
- Ex vivo skin was procured from the Human Tissue Bank at the University of Bradford.
- Proteomics: After homogenizing the samples and treatment with detergent, the debris was removed. The crude fraction was centrifuged and the cytosolic fraction was collected and digested to make it ready for analysis through label-free quantification and identification of peptides.
- SB-MSI: SB-MSI probes (for example, methyl paraben) were used for treating the surface of the samples for 48 hours. The samples were snapped frozen, sectioned to a size of 12 µm, and coated with MALDI matrix to render them ready for analysis.
Proteomic analysis revealed consistent expression of various xenobiotic metabolizing enzymes (XME) between ex vivo human skin and Labskin. The XME profile between ex vivo human skin and Labskin was acceptable (see Table 1).
Table 1. Expression of soluble phase I and II XMEs in Labskin and ex vivo human skin. The variation in color highlights the similarities in XME expression between Labskin and ex vivo skin (fmol/µg, ND = not detected).
SB-MSI identified esterase activity for the metabolism of methyl paraben into p-hydroxybenzoic acid (see Figure 1).
Figure 1. (A) Diagram showing metabolism reaction of SB-MSI probe methyl paraben with esterase to form p-hydroxybenzoic acid. (B) MSI image of Labskin treated with methyl paraben highlighting esterase activity.
Label-free proteomic analysis pointed out the comparable distribution of XMEs in ex vivo human skin and Labskin. In Labskin, the esterase activity site was determined with the help of an XME probe (methyl paraben).
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