title: Tissue Characterization after a New Disaggregation Method for Skin Micro-Grafts Generation.
abstract: Several new methods have been developed in the field of biotechnology to obtain autologous cellular suspensions during surgery, in order to provide one step treatments for acute and chronic skin lesions. Moreover, the management of chronic but also acute wounds resulting from trauma, diabetes, infections and other causes, remains challenging. In this study we describe a new method to create autologous micro-grafts from cutaneous tissue of a single patient and their clinical application. Moreover, in vitro biological characterization of cutaneous tissue derived from skin, de-epidermized dermis (Ded) and dermis of multi-organ and/or multi-tissue donors was also performed. All tissues were disaggregated by this new protocol, allowing us to obtain viable micro-grafts. In particular, we reported that this innovative protocol is able to create bio-complexes composed by autologous micro-grafts and collagen sponges ready to be applied on skin lesions. The clinical application of autologous bio-complexes on a leg lesion was also reported, showing an improvement of both re-epitalization process and softness of the lesion. Additionally, our in vitro model showed that cell viability after mechanical disaggregation with this system is maintained over time for up to seven (7) days of culture. We also observed, by flow cytometry analysis, that the pool of cells obtained from disaggregation is composed of several cell types, including mesenchymal stem cells, that exert a key role in the processes of tissue regeneration and repair, for their high regenerative potential. Finally, we demonstrated in vitro that this procedure maintains the sterility of micro-grafts when cultured in Agar dishes. In summary, we conclude that this new regenerative approach can be a promising tool for clinicians to obtain in one step viable, sterile and ready to use micro-grafts that can be applied alone or in combination with most common biological scaffolds.
authors.affiliation: Burns Centre and Emilia Romagna Regional Skin Bank.
authors.forename: Valeria
authors.initials: V
authors.lastname: Purpura
authors.affiliation: Burns Centre and Emilia Romagna Regional Skin Bank.
authors.forename: Elena
authors.initials: E
authors.lastname: Bondioli
authors.affiliation: Human Brain Wave srl; lab@hbwsrl.com.
authors.forename: Antonio
authors.initials: A
authors.lastname: Graziano
authors.affiliation: Human Brain Wave srl.
authors.forename: Letizia
authors.initials: L
authors.lastname: Trovato
authors.affiliation: Burns Centre and Emilia Romagna Regional Skin Bank.
authors.forename: Davide
authors.initials: D
authors.lastname: Melandri
authors.affiliation: Burns Centre and Emilia Romagna Regional Skin Bank.
authors.forename: Martina
authors.initials: M
authors.lastname: Ghetti
authors.affiliation: Plastic and Reconstructive Surgery, AOU "Ospedali Riuniti"
authors.forename: Andrea
authors.initials: A
authors.lastname: Marchesini
authors.affiliation: Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia; C.H.T Centre for Health Technologies, University of Pavia.
authors.forename: Maria Gabriella
authors.initials: MG
authors.lastname: Cusella De Angelis
authors.affiliation: Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia; C.H.T Centre for Health Technologies, University of Pavia.
authors.forename: Laura
authors.initials: L
authors.lastname: Benedetti
authors.affiliation: Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia; C.H.T Centre for Health Technologies, University of Pavia.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ceccarelli
authors.affiliation: Plastic and Reconstructive Surgery, AOU "Ospedali Riuniti"
authors.forename: Michele
authors.initials: M
authors.lastname: Riccio
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology; D012038:Regeneration; D016038:Skin Transplantation / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*; D014945:Wound Healing