Jank Bernhard J, Goverman Jeremy, Guyette Jacques P, Charest Jon M, Randolph Mark, Gaudette Glenn R, Gershlak Joshua R, Purschke Martin, Javorsky Emilia, Nazarian Rosalynn M, Leonard David A, Cetrulo Curtis L, Austen William G, Ott Harald C
1 Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Medical School , Boston, Massachusetts.
2 Divison of Burns, Department of Surgery, Massachusetts General Hospital, Harvard Medical School , Boston, Massachusetts.
Tissue Eng Part A. 2017 Jul;23(13-14):696-707. doi: 10.1089/ten.TEA.2016.0487. Epub 2017 Jun 19.
Full-thickness skin loss is a challenging problem due to limited reconstructive options, demanding 75 million surgical procedures annually in the United States. Autologous skin grafting is the gold standard treatment, but results in donor-site morbidity and poor aesthetics. Numerous skin substitutes are available on the market to date, however, none truly functions as full-thickness skin due to lack of a vascular network. The creation of an autologous full-thickness skin analogue with a vascular pedicle would result in a paradigm shift in the management of wounds and in reconstruction of full-thickness skin defects. To create a clinically relevant foundation, we generated an acellular skin flap scaffold (SFS) with a perfusable vascular pedicle of clinically relevant size by perfusion decellularization of porcine fasciocutaneous flaps. We then analyzed the yielded SFS for mechanical properties, biocompatibility, and regenerative potential in vitro and in vivo. Furthermore, we assessed the immunological response using an in vivo model. Finally, we recellularized the vascular compartment of an SFS and reconnected it to a recipient's blood supply to test for perfusability. Perfusion decellularization removed all cellular components with preservation of native extracellular matrix composition and architecture. Biaxial testing revealed preserved mechanical properties. Immunologic response and biocompatibility assessed via implantation and compared with native xenogenic skin and commercially available dermal substitutes revealed rapid neovascularization and complete tissue integration. Composition of infiltrating immune cells showed no evidence of allorejection and resembled the inflammatory phase of wound healing. Implantation into full-thickness skin defects demonstrated good tissue integration and skin regeneration without cicatrization. We have developed a protocol for the generation of an SFS of clinically relevant size, containing a vascular pedicle, which can be utilized for perfusion decellularization and, ultimately, anastomosis to the recipient vascular system after precellularization. The observed favorable immunological response and good tissue integration indicate the substantial regenerative potential of this platform.
全层皮肤缺损是一个具有挑战性的问题，因为重建选择有限，在美国每年需要进行7500万例外科手术。自体皮肤移植是金标准治疗方法，但会导致供区发病和美学效果不佳。迄今为止，市场上有许多皮肤替代品，但由于缺乏血管网络，没有一种能真正起到全层皮肤的作用。创建一个带有血管蒂的自体全层皮肤类似物将导致伤口处理和全层皮肤缺损重建的范式转变。为了建立一个临床相关的基础，我们通过对猪筋膜皮瓣进行灌注去细胞化，生成了一个具有临床相关大小的可灌注血管蒂的无细胞皮瓣支架（SFS）。然后，我们在体外和体内分析了所产生的SFS的力学性能、生物相容性和再生潜力。此外，我们使用体内模型评估了免疫反应。最后，我们对SFS的血管腔进行了再细胞化，并将其重新连接到受体的血液供应中以测试灌注能力。灌注去细胞化去除了所有细胞成分，同时保留了天然细胞外基质的组成和结构。双轴测试显示力学性能得以保留。通过植入评估免疫反应和生物相容性，并与天然异种皮肤和市售真皮替代品进行比较，结果显示血管快速新生和组织完全整合。浸润免疫细胞的组成没有显示出同种异体排斥的迹象，并且类似于伤口愈合的炎症阶段。植入全层皮肤缺损处显示出良好的组织整合和皮肤再生，没有瘢痕形成。我们已经开发了一种方案，用于生成具有临床相关大小、包含血管蒂的SFS，该方案可用于灌注去细胞化，并最终在预细胞化后与受体血管系统进行吻合。观察到的良好免疫反应和良好的组织整合表明该平台具有巨大的再生潜力。
