• Models close to the actual conditions of use of the products to be tested.

• The skin conserves a normal morphology, structure and metabolism.

• Several application modes are possible: topically or solubilised in the survival medium, single or chronic application. Possibility of making injections, laser treatments, etc.

• Possibility of reproducing external interactions: exposure to UV or pollutants, chemical attacks, wound, sensitisation, hair removal, etc.

• Carrying out histological analyses

Our laboratory has a battery of stainings and immunomarkers that enable different cellular and tissue markers to be visualised. The marked parameters may be quantified by image analysis software.

• Carrying out biological analyses

On whole skin, separate epidermises, homogenates, extracts or in the culture medium. All types of extractions, separations, chromatograms and assays may be envisaged.

• Carrying out genomic studies (in partnership with the Genex Laboratory)

Large-scale screening and analysis of genomic data. Identification of the effects of the products on human skin thanks to the study of the genic expression (DNA chip, miRNA chip, qPCR) correlated with the protein expression of the genes of interest.

• Thickness of the stratum corneum
• Exfoliation

• Filaggrin (NMF precursor)
• Membrane Transglutaminase – TGM (formation and cohesion of the stratum corneum)
• Tight junctions

– ZO-1

– Occludin-1

– Claudin-1

• Kallikreins
• Hyaluronic acid
• Small proline-rich proteins – SPRRs (corneal envelope assembly)
• Ceramides
• Neutral lipids
• Terminal differentiation of keratinocytes

– Loricrin

– Involucrin

– Caspase 14

• Serine proteases inhibitor – LEKTI (desquamation regulation)
• Aquaporins (protein regulating the hydric flow between the dermis and the stratum corneum)

BIO-EC Laboratory may, using this device, evaluate the kinetics of the tissue penetration and distribution of your product, the skin modifications caused by your product across various parameters, the hydration rate in the stratum corneum and the epidermis, the distribution of the NMF components, of the cutaneous lipids, the carotenoids, etc.

Thanks to our partnership with BIC (Bordeaux Imaging Center), it is possible to complement our histological studies with TEM observations, such as the evaluation of the quality of lipid bilayers.

With the help of this devic BIO-EC Laboratory can carry out topographic studies on the surface of explants with an accuracy evaluated at a tenth of a nanometre. This enables various parameters to be analysed such as the depth and width of wrinkles and micro-lines and the roughness of the skin in order to evaluate a filmogenic effect.

Visualisation of melanin by Fontana-Masson.

• Tyrosinase-Related Protein-1 and 2 (melanin synthesis)
• Tyrosinase (melanin synthesis)
• Melan-A (synthesis of melanosomes)
• MelanoCortin-1 Receptor (migration of melanosomes)
• Premelanosome protein – PMEL17 (formation of melanosomes)

Screening test to evidence melanin produced by melanocytes: DOPA oxidase test

Test carried out on separate epidermis.

Observation of the appearance of Sun Burn Cells (SBCs).

• DNA protection after UVB irradiation

  • p53
  • Caspase-3
  • Thymine dimers
  • H2AX

• Evaluation of the oxidative stress after UVA irradiations

  • Oxidative-Stress response-1 – OXSR-1
  • 8-hydroxy-2-deoxyguanosine – 8-OHdG
  • Hème oxygénase – HO-1 (mitochondrial enzyme)

• Markers evaluated after infrared ray irradiation

  • Tropoelastin (elastin precursor)
  • Metalloproteinases – MMPs (remodelling of the extra-cellular matrix)
  • Fibrillin (principal constituent of the extra-cellular matrix microfibrils)

• Markers evaluated after irradiation by visible light (blue light)

  • MMP-1 (protease involved in the remodelling of the ECM)
  • Involucrin (protein involved in the stratum corneum organisation and connections)
  • Sirtuin 1 – SIRT1 (protein involved in the cellular renewal process)
  • NF-E2-related factor – Nrf2 (anti-oxidant)

• MDA (malonedialdehyde acid), lipid peroxidation marker.

• Cytokines

In the culture medium:

• Fatty acids
• Glycerol
• Triglycerides

• Masson’s Trichrome Staining
• Measurement of the average size of the adipocytes

This marking enables demonstrating the conversion of white adipocytes into smaller, more easily eliminated brown adipocytes.

General morphology

After topical application of the product.

Immunomarking

Langerhans cells: CD1a and langerin.

General morphology

Determination of the irritant character in compliance with GLP (OECD method 439)

• Localised burn caused by a strong dose of UV in the centre of the explant
• Burn by application of chemical products
• Mechanical injury to the explant

Evaluation of the re-epithelialisation speed (structure of the growth bud).

• Basal and supra-basal keratins – Stratification
• Fibronectin – Adhesion
• Integrin β4 – Adhesion
• Laminin-5 – Dermo-epidermal junction
• Perlecan – Dermo-epidermal junction and epidermal differentiation
• PECAM-1 – Micro-vascularisation
• αSMA – Papillary dermis below the growth bud

Impact of the products on the cutaneous microbiome bacteria (commensal or pathogenic), influence of the product on their adhesion (formation of biofilms, proliferation, etc.).

• Β defensins 2, 3 (peptides possessing an antimicrobial activity)
• Cathelicidin LL 37 (peptide possessing an anti-microbial activity)
• Toll like receptor 2 -TLR-2 (receptor linked to the production of cytokines and cellular activation)

• Cytokines (IL8, IL6, TNFα…)

Process exclusive to BIO-EC Laboratory of exposure of explants to atmospheric pollutants:

– Hydrocarbons,

– Heavy metals,

– Diesel particles,

– Ozone,

– Cigarette smoke, etc…

This device enables pollutants to be vaporised into cloud form and to be put into contact only with the surface of the skin to resemble physiological conditions as closely as possible.

• Aromatic hydrocarbons receptor – AhR (activation of cytochrome and detoxification enzyme)
• NF-E2-related factor – Nrf2 (anti-oxidant)
• Metallothionein – MT-1H 5 (stress resistance and detoxification enzyme)
• Hème oxygenase – HO-1 (mitochondrial enzyme)

• MDA (malonedialdehyde acid), lipid peroxidation marker.

• NF-E2-related factor – Nrf2 (anti-oxidant)
• OXSR-1 protein (stress response enzyme)
• 8-hydroxy-2-deoxyguanosine -8-OHdG (DNA transcription regulation molecule)
• Mitochondrial aconitase (protection from oxidative stress)
• Metallothionein – MT-1H (anti-oxidant)
• TNFα (glycoprotein stress marker)
• Interleukin 1 alpha – IL1α (inflammatory response cytokine)

• MDA (malonedialdehyde acid), lipid peroxidation marker
• Assays of interleukins (IL-1α, 8, 6), of the TNFα and other proteins involved in the inflammatory response.

• General morphology of the sebaceous glands using Masson’s Trichrome.
• Staining of the sebum.

• Interleukins (IL)