In vitro assessment of shampoos eye stinging potential using Transient Receptor Potential Vanilloid Type 1 (TRPV1) Frédéric AMARAL L’Oréal Research and.

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In vitro assessment of shampoos eye stinging potential using Transient Receptor Potential Vanilloid Type 1 (TRPV1) Frédéric AMARAL L’Oréal Research and Innovation , Aulnay, France

Presentation Outline Role of TRPV1 in nociception Objectives of the study/experimental design Results Conclusions and next steps

Role of TRPV1 in nociception (1) Receptor type: calcium permeable ion channel belonging to the TRP (Transient Receptor Potential) family Location: C-fibers, CNS but also in skin, cornea and mucosal tissue Activation: heat, acidic conditions, Capsaicin (CP) calcium flux from extracellular sources Release of neurotransmiters, neuropeptides induction of pain, burning, pruritus

Role of TRPV1 in nociception (2) Need for an in vitro assay for identifying the stinging potential of personal care products (ingredients and formulae)

Objectives of the Study/experimental design To demonstrate the PoC with the « Nociocular » in vitro assay (based on SH-SY5Y overexpressing TRPV1) Collaboration with the University of Stockholm (A Forsby) Read out of the assay: calcium influx (fura2-AM) Test articles used: TRPV1 agonist: capsaicine (CP) for sensitivity TRPV1 antagonist: capsazepine (CPZ) for specificity A selection of 10 coded shampoos (adults and children/baby)

Results: in vitro assay data 3 categories of profiles have been identified Case C induction of Ca2+ flux No impact of CPZ TRPV1-mediated effect? Case A No induction of Ca2+ flux Case B induction of Ca2+ flux Decreased response with CPZ TRPV1-mediated effect

Results: in vitro/in vivo correlation Good correlation, especially for extreme classes Good trend for the moderate class except for formula G

Conclusions We have demontrated that: Next steps: The « Nociocular » assay could be used to evaluate eye stinging personal care products The assay was applicable to complex water-soluble formula For some formulae, the response was TRPV1-mediated The in vitro responses correlated well with clinical data Next steps: To evaluate a more diverse set of formulae and ingredients To confirm the robustness of the assay To refine the discomfort classes proposed To develop a model for non water-soluble formulae To sick for additional assays to get a better understanding of the responses observed

University of Stockholm Acknowledgments L’OREAL Linda Bourouf Reine Note Thomas Delanne Gladys Ouédraogo Sophie Loisel-Joubert José Cotovio Jean-Roch Meunier University of Stockholm Hanegraaf Maaike Anna Forsby