Characterizing Xenobiotic Metabolizing Enzymes Using Labskin

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.

Method

  • 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.

Results

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).

(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.

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.

Summary

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).

About Labskin

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.

Whether you need rapid, focused analysis or flexible, tailor-made research programs we can help you develop and validate skincare ingredients and products which really work.

Our skin model is a 3D human skin equivalent that incorporates vital biological components to model normal skin function.

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).

“An ideal platform for basic or applied skin research, testing compounds or formulated products for the cosmetic, pharmaceutical and chemical sectors.”


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Last updated: Oct 28, 2019 at 4:43 AM

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