Skimune™ Technologies for sensitization and potency assessment   Unique tools for the Cosmetic and Chemical Industries. www.alcyomics.com  

Skin Explant Technology Skimune TM technology Blood Donor Immature Mo-DC´s Mature DC´s + compound + T Cells Sensitized T cells + Skin biopsy - Response + Response

Histopathological evaluation Grade I Grade II Grade III Grade IV in vitro tissue damage can predict in vivo tissue damage, tissue damage graded I-IV (Lerner, K.G., et al., Transplant Proc, 1974. 6(4): p. 367-7) Grade 0 = normal skin Grade I = mild vacuolization of epidermal cells Grade II = diffuse vacuolization of basal cells with scattered dyskeratotic bodies Grade III = sub-epidermal cleft formation Grade IV = complete epidermal separation

SkimuneTM assay What it models: Damaged skin The sensitization and elicitation phases of the skin sensitization process. Adapted from (http://www.kao.com/jp/en/corp_rd/skills_01_04.html)) The SkimuneTM model models the in vivo acute contact dermatitis (ACD) model

95% specificity and 95% sensitivity, concordance 95% Alcyomics Assay evaluation SkimuneTM – LLNA The SkimuneTM assay was evaluated against the mouse local lymph node assay (LLNA), which is the European gold standard for sensitization testing Number of positive LLNA chemicals = 20 SkimuneTM prediction of positive chemicals = 19 Number of negative LLNA chemicals = 20 SkimuneTM prediction of negative chemicals = 19 95% specificity and 95% sensitivity, concordance 95%

SkimuneTM technologies detect human reactions Reactions to Nickel Sulphate ( NiSO4) cannot be picked up using mouse models of toxicology-mouse physiology is not human physiology !!

Potency assessment Potency assessment of cinnamic alcohol and oxazalone. Skin explant results show histopathological damage in skin in response to 0.1µM, 0.05µM and 0.01µM concentration of each chemical. For Cinnamic alcohol 0.1µM gave a grade III positive response, 0.05µM and 0.01µM gave a grade I negative response. Oxazalone gave a grade III positive response at 0.1µM and 0.05µM and a grade I negative response at 0.01µM. Results show oxazalone is a more potent chemical than cinnamic alcohol

Case study 1- Cosmeceuticals Determine the effect of repeated applications of product X in skin Positive control Product X Day2 Product X Day3 Negative control Result: Repeated application of Product X over a 3 day period showed an increase in damage. Results show a grade II/III positive response on day 2 and a grade III positive response on day 3. Results indicate that product X has the potential to cause an adverse response.

Case study 2- Cosmeceuticals Determine in-vitro responses to wheat protein products Medium Grade I Compound 1 Grade I Compound 2Grade III Two wheat protein products were tested for immunogenic responses. Compound 2 gave a positive response showing a high degree of histological damage indicating that is likely to cause an adverse reaction. Compound 1 showed no damage response in skin histology

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