Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium; Department of Plastic and Reconstructive Surgery, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10, B-1200 Brussels, Belgium.
Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium.
Acta Biomater. 2018 Jun;73:339-354. doi: 10.1016/j.actbio.2018.04.009. Epub 2018 Apr 11.
Human ear reconstruction is recognized as the emblematic enterprise in tissue engineering. Up to now, it has failed to reach human applications requiring appropriate tissue complexity along with an accessible vascular tree. We hereby propose a new method to process human auricles in order to provide a poorly immunogenic, complex and vascularized ear graft scaffold.
12 human ears with their vascular pedicles were procured. Perfusion-decellularization was applied using a SDS/polar solvent protocol. Cell and antigen removal was examined by histology and DNA was quantified. Preservation of the extracellular matrix (ECM) was assessed by conventional and 3D-histology, proteins and cytokines quantifications. Biocompatibility was assessed by implantation in rats for up to 60 days. Adipose-derived stem cells seeding was conducted on scaffold samples and with human aortic endothelial cells whole graft seeding in a perfusion-bioreactor.
Histology confirmed cell and antigen clearance. DNA reduction was 97.3%. ECM structure and composition were preserved. Implanted scaffolds were tolerated in vivo, with acceptable inflammation, remodeling, and anti-donor antibody formation. Seeding experiments demonstrated cell engraftment and viability.
Vascularized and complex auricular scaffolds can be obtained from human source to provide a platform for further functional auricular tissue engineered constructs, hence providing an ideal road to the vascularized composite tissue engineering approach.
The ear is emblematic in the biofabrication of tissues and organs. Current regenerative medicine strategies, with matrix from donor tissues or 3D-printed, didn't reach any application for reconstruction, because critically missing a vascular tree for perfusion and transplantation. We previously described the production of vascularized and cell-compatible scaffolds, from porcine ear grafts. In this study, we ---- applied findings directly to human auricles harvested from postmortem donors, providing a perfusable matrix that retains the ear's original complexity and hosts new viable cells after seeding. This approach unlocks the ability to achieve an auricular tissue engineering approach, associated with possible clinical translation.
人类耳朵重建被认为是组织工程学中的标志性工程。到目前为止，它还没有达到人类应用的要求，需要适当的组织复杂性和可接近的血管树。为此，我们提出了一种新的方法来处理人耳，以提供一种免疫原性低、复杂且血管化的耳朵移植物支架。
从 12 个带有血管蒂的人耳中获取样本。使用 SDS/极性溶剂方案进行灌注去细胞化处理。通过组织学检查和 DNA 定量来检查细胞和抗原的去除情况。通过常规和 3D 组织学、蛋白质和细胞因子定量来评估细胞外基质（ECM）的保留情况。将支架植入大鼠体内 60 天，评估其生物相容性。对支架样本进行脂肪源性干细胞接种，并在灌注生物反应器中对整个移植物进行人主动脉内皮细胞接种。
组织学检查证实了细胞和抗原的清除。DNA 减少了 97.3%。ECM 结构和组成得以保留。植入的支架在体内可耐受，炎症、重塑和抗供体抗体形成可接受。接种实验表明细胞定植和活力。
可以从人源获得血管化和复杂的耳廓支架，为进一步构建功能性耳廓组织工程化构建体提供平台，从而为血管化复合组织工程方法提供理想的途径。
耳朵是组织和器官生物制造的标志。目前的再生医学策略，使用来自供体组织的基质或 3D 打印，都没有达到任何重建应用，因为严重缺乏用于灌注和移植的血管树。我们之前描述了从猪耳朵移植物中生产血管化和细胞相容的支架。在这项研究中，我们----直接应用从死后供体中获取的人耳的研究结果，提供了一种可灌注的基质，保留了耳朵的原始复杂性，并在接种后容纳新的有活力的细胞。这种方法为实现耳廓组织工程方法提供了可能性，也可能实现临床转化。
