SKIN MICROBIOME
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Figure 1: Image of Labskin-S construct in culture
of an adult acne sufferer (a 26 year-old black female volunteer). A validated sampling method was used whereby a sterile stainless-steel ring was place against the cheek and 1mL of GS25 buffer (PH neutral saline solution with 0.1% mild detergent) added to the ring. The skin was gently agitated with a sterile
Perspex wand and the GS25 buffer containing the microbiome sample recovered to a sterile tube for transport. The microbiome sample was transferred to the York laboratories were the microbiome sample was recovered and transferred onto the
surface of Labskin-S constructs. The whole microbiome was cultured but
specific investigation was placed on Cutibacteria spp. As the bacteria responsible for an acne presentation when out of balance with the other constituents of the skin microbiome.
Experimental protocol Labskin-S constructs, consisting of a 1.1cm 2 surface area, were cultured to maturation with a 12-day air-liquid-interface to provide a dry top surface for microbiome and product
Colonise Labskin with Microbiome sample Uncolonised Labskin Negative Labskin Positive Labskin Test Item Labskin
Figure 2: Photograph of cheek area of the volunteer providing the skin microbiome sample
applications. Each Labskin-S construct was colonized with an equal amount of microbiome sample, except for the uncolonized controls. All incubations were performed at 37°C/ 5% CO2
.
The microbiome sample was allowed to establish on the surface of Labskin-S for 48 hours prior to application of the products. An uncolonized control (no microbiome) was used. A negative sample of Dulbecco’s phosphate buffered saline was used as this would have no impact on the Cutibacteria present in the microbiome sample. A known over-the-counter anti-acne
product containing 5% benzoyl peroxide was used as the positive control. Finally, the product that the volunteer was using to treat her acne used. This contained 2% succinic acid, 2% colloidal sulphur and 1% salicylic acid. The Labskin-S was incubated for a further 48 hours post application of the treatment groups.
Assessment Visual The constructs were imaged by light photography at each of the protocol stages.
Apply dPBS
Apply Positive control
Apply Test Item
Microbiology Selective agar media for Cutibacteria spp. was used to perform microbiology enumeration. Eight-millimetre punch biopsies were used to take full thickness samples though the Labskin-S constructs. The microbiome sample was recovered using GS25 buffer and serial dilutions plated out onto selective agar.11
Sampling for:
Sampling for: IL-1α
Microbial enumeration Histology IL-1α
Figure 3: Protocol overview showing the timeline of the experimental procedure
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Immunological The culture media from each construct well was assessed for the presence of Il-1α using a commercially available ELISA kit (RnD Systems). Il-1a is a pro-inflammatory cytokine and is used as a key marker for irritation in the skin. Keratinocyte cells are able to release
November 2025 PERSONAL CARE
Incubate for˜72h
Incubate for 48h
Incubate for 48h
Incubate for 48h
Incubate for 48h
Incubate for 48h
Incubate for 48h
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