Purpura Valeria, Bondioli Elena, Graziano Antonio, Trovato Letizia, Melandri Davide, Ghetti Martina, Marchesini Andrea, Cusella De Angelis Maria Gabriella, Benedetti Laura, Ceccarelli Gabriele, Riccio Michele
Burns Centre and Emilia Romagna Regional Skin Bank.
Human Brain Wave srl;
J Vis Exp. 2016 Mar 4(109):e53579. doi: 10.3791/53579.
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.
在生物技术领域已开发出几种新方法，以便在手术期间获得自体细胞悬液，从而为急慢性皮肤损伤提供一步治疗。此外，由创伤、糖尿病、感染和其他原因导致的慢性和急性伤口的处理仍然具有挑战性。在本研究中，我们描述了一种从单个患者的皮肤组织创建自体微移植体的新方法及其临床应用。此外，还对来自皮肤、去表皮真皮（Ded）以及多器官和/或多组织供体的真皮的皮肤组织进行了体外生物学特性分析。所有组织均通过这一新方案进行解离，使我们能够获得有活力的微移植体。特别是，我们报告称，这种创新方案能够创建由自体微移植体和胶原海绵组成的生物复合物，随时可应用于皮肤损伤。还报告了自体生物复合物在腿部损伤上的临床应用，显示出再上皮化过程和损伤部位柔软度均有所改善。此外，我们的体外模型表明，使用该系统进行机械解离后，细胞活力在长达七（7）天的培养时间内得以维持。通过流式细胞术分析，我们还观察到，解离获得的细胞群由多种细胞类型组成，包括间充质干细胞，由于其高再生潜力，这些细胞在组织再生和修复过程中发挥着关键作用。最后，我们在体外证明，当在琼脂培养皿中培养时，该程序可保持微移植体的无菌状态。总之，我们得出结论，这种新的再生方法可能成为临床医生的一种有前景的工具，可一步获得有活力、无菌且随时可用的微移植体，这些微移植体可单独应用或与最常见的生物支架联合应用。
