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Feasibility of a porcine oral mucosa equivalent: A preclinical study

机译：猪口腔粘膜当量的可行性：一项临床前研究

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Oral tissue engineering aims to treat and fill tissue deficits caused by congenital defects, facial trauma, or malignant lesion surgery, as well as to study the biology of oral mucosa. The Food and Drug Administration (FDA) and the European Medicines Agency (EMA) require a large animal model to evaluate cell-based devices, including tissue-engineered oral mucosa, prior to initiating human clinical studies. Porcine oral mucosa is non-keratinized and resembles that of humans more closely than any other animal in terms of structure and composition; however, there have not been any reports on the reconstruction of a porcine oral mucosa equivalent, probably due to the difficulty to culture porcine fibroblasts. In this study, we demonstrate the feasibility of a 3D porcine oral mucosa equivalent based on a collagen-GAG-chitosan scaffold, as well as reconstructed porcine epithelium by using an amniotic membrane as support, or without any support in form of epithelial cell sheets by using thermoresponsive culture plates. Explants technique was used for the isolation of the porcine fibroblasts and a modified fibroblast medium containing 20% fetal calf serum was used for their culture. The histological and transmission electron microscopic analyses of the resulting porcine oral mucosa models showed the presence of non-keratinized epithelia expressing keratin 13, the major differentiation marker of non-keratinized oral mucosa, in all models, and the presence of newly synthesized collagen fibers in the lamina propria equivalent of the full-thickness model, indicating the functionality of porcine fibroblasts.
机译：口腔组织工程旨在治疗和填补由先天性缺陷，面部创伤或恶性病变手术引起的组织缺陷，以及研究口腔粘膜的生物学特性。在开始人类临床研究之前，美国食品药品监督管理局（FDA）和欧洲药品管理局（EMA）需要大型动物模型来评估基于细胞的设备，包括组织工程化的口腔粘膜。猪的口腔粘膜是非角质化的，在结构和组成方面比任何其他动物都更接近人类。然而，尚无关于重建猪口腔粘膜等同物的报道，可能是由于培养猪成纤维细胞的困难。在这项研究中，我们证明了以胶原蛋白-GAG-壳聚糖支架为基础的3D猪口腔粘膜等效物的可行性，以及通过使用羊膜作为支持物或不使用任何形式的上皮细胞片的支持物来重建猪上皮的可行性。使用热响应培养板。使用外植体技术分离猪成纤维细胞，并使用含有20％胎牛血清的改良成纤维细胞培养基进行培养。对所得猪口腔粘膜模型的组织学和透射电镜分析表明，在所有模型中均存在表达角蛋白13的非角化上皮细胞，这是所有模型中非角化口腔粘膜的主要分化标志物，并且存在新合成的胶原纤维。全层模型的固有层当量，表明猪成纤维细胞的功能。

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