Natural products have many healing effects on the skin with minimal or no adverse effects. In this study, we analyzed the regenerative properties of a waste product (hydrolate) derived from Helichrysum italicum (HH) on scratch-tested skin cell populations seeded on a fluidic culture system. Helichrysum italicum has always been recognized in the traditional medicine of Mediterranean countries for its wide pharmacological activities. We recreated skin physiology with a bioreactor that mimics skin stem cell (SSCs) and fibroblast (HFF1) communication as in vivo skin layers. Dynamic culture models represent an essential instrument for recreating and preserving the complex multicellular organization and interactions of the cellular microenvironment. Both cell types were exposed to two different concentrations of HH after the scratch assay and were compared to untreated control cells. Collagen is the constituent of many wound care products that act directly on the damaged wound environment. We analyzed the role played by HH in stimulating collagen production during tissue repair, both in static and dynamic culture conditions, by a confocal microscopic analysis. In addition, we performed a gene expression analysis that revealed the activation of a molecular program of stemness in treated skin stem cells. Altogether, our results indicate a future translational application of this natural extract to support skin regeneration and define a new protocol to recreate a dynamic process of healing.

Center for Developmental Biology and Reprogramming CEDEBIOR Department of Biomedical Sciences University of Sassari Viale San Pietro 43 B 07100 Sassari Italy

Department of Biomedical Sciences University of Sassari Viale San Pietro 43 B 07100 Sassari Italy

Department of Life and Environmental Sciences University of Cagliari University Campus 09042 Cagliari Italy

Department of Medical Surgical and Experimental Sciences University of Sassari 07100 Sassari Italy

Laboratory of Molecular Biology and Stem Cell Engineering Eldor Lab Istituto Nazionale di Biostrutture e Biosistemi 40128 Bologna Italy

R and D Laboratory Center InoCure s r o Politickych veziu 935 13 Nové Mesto Praha 1 110 00 Prague Czech Republic

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