*Pole of Morphology, Institute of Experimental and Clinical Research, Université catholique de Louvain, Brussels, Belgium †Pole of Experimental Surgery and Transplantation, Institute of Experimental and Clinical Research, Université catholique de Louvain, Brussels, Belgium ‡Department of Plastic and Reconstructive Surgery, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium §Department for Biomedical Research, University of Bern, Bern, Switzerland ¶Department of Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston-Salem, NC ||Pole of Pharmacology and Therapeutic, Institute of Experimental and Clinical Research, Université catholique de Louvain, Brussels, Belgium **Department of Medical Imaging, Cliniques Universitaires Saint-Luc, Université catholique de Louvain, Brussels, Belgium ††Division of Cell Matrix Biology and Regenerative Medicine, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK ‡‡Department of Plastic, Reconstructive and Hand Surgery, Inselspital, University Hospital of Bern, Bern, Switzerland.
Ann Surg. 2017 Nov;266(5):754-764. doi: 10.1097/SLA.0000000000002396.
During the last decade, face allotransplantation has been shown to be a revolutionary reconstructive procedure for severe disfigurements. However, offer to patients remains limited due to lifelong immunosuppression. To move forward in the field, a new pathway in tissue engineering is proposed.
Our previously reported technique of matrix production of a porcine auricular subunit graft has been translated to a human face model.
5 partial and 1 total face grafts were procured from human fresh cadavers. After arterial cannulation, the specimens were perfused using a combined detergent/polar solvent decellularization protocol. Preservation of vascular patency was assessed by imaging, cell and antigen removal by DNA quantification and histology. The main extracellular matrix proteins and associated cytokines were evaluated. Lip scaffolds were cultivated with dermal, muscle progenitor and endothelial cells, either on discs or in a bioreactor.
Decellularization was successful in all facial grafts within 12 days revealing acellular scaffolds with full preservation of innate morphology. Imaging demonstrated a preservation of the entire vascular tree patency. Removal of cells and antigens was confirmed by reduction of DNA and antigen markers negativation. Microscopic evaluation revealed preservation of tissue structures as well as of major proteins. Seeded cells were viable and well distributed within all scaffolds.
Complex acellular facial scaffolds were obtained, preserving simultaneously a cell-friendly extracellular matrix and a perfusable vascular tree. This step will enable further engineering of postmortem facial grafts, thereby offering new perspectives in composite tissue allotransplantation.
在过去的十年中，面部同种异体移植已被证明是一种革命性的严重毁容重建手术。然而，由于需要终身免疫抑制，患者的选择仍然有限。为了在该领域取得进展，提出了组织工程的新途径。
我们之前报道的猪耳廓亚单位移植物基质生产技术已被转化为人类面部模型。
从人类新鲜尸体中采集了 5 个部分和 1 个全脸移植物。在动脉插管后，使用联合去污剂/极性溶剂脱细胞方案对标本进行灌注。通过成像评估血管通畅性的保留情况，通过 DNA 定量和组织学评估细胞和抗原的去除情况。评估了主要细胞外基质蛋白及其相关细胞因子。将真皮、肌肉祖细胞和内皮细胞培养在盘状或生物反应器中的脂肪支架上。
所有面部移植物在 12 天内均成功脱细胞，显示出具有完整形态保留的无细胞支架。成像显示整个血管树的通畅性得以保留。通过减少 DNA 和抗原标志物的阴性化，证实了细胞和抗原的去除。显微镜评估显示保留了组织结构以及主要蛋白质。种子细胞在所有支架中均存活且分布均匀。
获得了复杂的无细胞面部支架，同时保留了细胞友好的细胞外基质和可灌注的血管树。这一步将使死后面部移植物的进一步工程化成为可能，从而为复合组织同种异体移植提供新的前景。
