Wound Healing Society

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The Effect Of An Autologous Keratinocyte Spray On Human Skin Wound Healing Transcriptome
Kristo Nuutila1, Antti Siltanen2, Matti Korhonen3, Johanna Nystedt3, Jussi Valtonen4, Andrew Lindford4, Jyrki Vuola4, Esko Kankuri2.
1Brigham and Women's Hospital, Boston, MA, USA, 2University of Helsinki, Helsinki, Finland, 3Finnish Red Cross Blood Service, Helsinki, Finland, 4Helsinki Burn Center, Helsinki, Finland.

Background: Most of the molecular-level wound-healing data have been obtained from various animal models. This may lead to misinformation and affect the translation of results to patient care. To overcome this issue, we have utilized the split-thickness skin graft donor site as a clinical wound model. The donor site wound offers an excellent opportunity for studying the mechanisms of wound healing because they are standardized, non-infected and clean. Importantly, serial small biopsy samples can be harvested without major ethical issues. The purpose of this study was to investigate the effect of an autologous keratinocyte cell therapy on wound healing transcriptome. Methods: Small skin biopsies were collected from burn patients undergoing split-thickness skin grafting, in Helsinki Burn Center. The biopsies were immediately delivered to Finnish Red Cross Blood Service where keratinocytes were isolated under GMP conditions. During the same operation, the cell suspension was delivered back to the burn center and sprayed on the donor site wound. Biopsies were collected on the intact skin and on days 7, 14 and 21 postoperatively with a 3-mm biopsy punch. Non-treated donor site wounds acted as controls. The harvested samples were processed for human full genome microarray and subsequently gene ontology enrichment analyses were performed. Results: The results demonstrated significant differences in gene profiles between different time points and treatments. E.g. when comparing the day 14 treated and non-treated donor sites the analyses showed that in total 19 gene pathways were significantly upregulated in cell spray treated wounds. These pathways included keratinization, keratinocyte differentiation, skin development, epidermis development, epidermal cell differentiation, cornification ja establishment of skin barrier, demonstrating on a molecular level that the cell therapy is promoting healing. Conclusions: This study provides the first objective molecular level information on the effect of cell therapy on human wound healing over time.

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