Utilizing a Living Skin Equivalent to Examine Wound Healing

Objective

To develop methods to reproduce wounding of Labskin and to measure mass spectra directly from the tissue during the wound healing process using Matrix Assisted Laser Desorption Ionization Mass Spectrometry Imaging (MALDI-MSI).

Methods

  • Controlled depth penetration with a scalpel blade was used to wound each Labskin sample. The sample was then assessed every 24 h for 5 days.
  • For histology, samples were formalin fixed paraffin embedded (FFPE) and, in preparation for mass spectrometry imaging, samples were embedded in 20% gelatin and flash frozen.
  • FFPE tissue samples were sectioned (5 μm) and stained with haematoxylin and eosin (H&E).
  • Fresh frozen samples were sectioned (10 μm), sprayed with MALDI matrix and analysis was carried out to look for lipids.

Results

The migration of keratinocytes into the wound site immediately post wounding in an initial wound healing response is mimicked with this model.

Photographic image of wounding Labskin with scalpel blade

Figure 1. Photographic image of wounding Labskin with scalpel blade

MSI image and corresponding H&E image of Labskin 3 days post wounding. MSI image of two distinct ion species in the epidermis (green = m/z 721.4) and dermis (red = m/z 725.4)

Figure 2. MSI image and corresponding H&E image of Labskin 3 days post wounding. MSI image of two distinct ion species in the epidermis (green = m/z 721.4) and dermis (red = m/z 725.4)

Summary

This demonstrates that it is possible to achieve reproducible wounding with Labskin allowing for the wound healing process to be studied through a range of techniques including MALDI-MSI. As such, the activity of ingredients and formulations in the wound healing process can be assessed using Labskin, which can be benchmarked against products of recognized activity.

Labskin can be used within the same experimental design to evaluate several endpoints including cytokine responses (i.e. IL-1a, IL-6, IL-8, PGE2, TNFa, IL-10 etc.), histological changes, wound repair and photoreactivity in addition to skin commensal and pathogenic microorganisms.

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 4:42 AM

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