Department of Plastic and Reconstructive Surgery, Erasmus MC, Rotterdam, The Netherlands.
Cells Tissues Organs. 2012;195(3):185-96. doi: 10.1159/000324918. Epub 2011 Apr 14.
The aim of this study was to create and characterize a tissue-engineered mucosal equivalent (TEM) that closely resembles native mucosa. TEM consists of human primary keratinocytes and fibroblasts isolated from biopsies taken from healthy donors and seeded onto a de-epidermized dermis and cultured for 14 days at the air/liquid interface. The structure of TEM was examined and compared with native nonkeratinizing oral mucosa (NNOM). The various components of the newly formed epidermal layer, basement membrane and underlying connective tissue were analyzed using immunohistochemistry. The mucosal substitute presented in this study showed a mature stratified squamous epithelium that was similar to that of native oral mucosa, as demonstrated by K19, desmoglein-3 and involucrin staining. In addition, the expression of basement membrane components collagen type IV, laminin-5 and integrin α6 and β4 in TEM proved to be consistent with native oral mucosa. The expression of PAS, Ki67, K10 and K13, however, appeared to be different in TEM compared to NNOM. Nevertheless, the similarities with native oral mucosa makes TEM a promising tool for studying the biology of mucosal pathologies such as oral mucositis or fibrosis as well as the development of new therapies.
本研究旨在构建和表征一种组织工程化的黏膜等效物（TEM），使其尽可能地类似于天然黏膜。TEM 由从健康供体活检中分离出的人原代角质形成细胞和成纤维细胞组成，接种到去表皮真皮上，并在气/液界面培养 14 天。我们对 TEM 的结构进行了检查，并与天然非角化口腔黏膜（NNOM）进行了比较。使用免疫组织化学分析了新形成的表皮层、基底膜和下方结缔组织的各种成分。本研究中提出的黏膜替代物呈现出成熟的复层鳞状上皮，与天然口腔黏膜相似，这一点通过 K19、桥粒芯糖蛋白 3 和兜甲蛋白染色得到证实。此外，TEM 中基底膜成分 IV 型胶原、层粘连蛋白 5 和整合素 α6 和 β4 的表达与天然口腔黏膜一致。然而，与 NNOM 相比，TEM 中 PAS、Ki67、K10 和 K13 的表达似乎有所不同。尽管如此，TEM 与天然口腔黏膜的相似性使其成为研究口腔黏膜炎或纤维化等黏膜病理生物学以及开发新疗法的有前途的工具。
