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目的利用4种不同支架材料构建复合式口腔黏膜,并比较其组织结构特点。方法体外培养人口腔黏膜的成纤维细胞和角质形成细胞,在4种支架材料中加入成纤维细胞,培养7d后,在支架表面加入角质形成细胞,培养4d后,移至气-液界面继续培养7d。苏木精-伊红染色镜下观察构建的复合式口腔黏膜的组织形态学特点。结果 4种支架均可构建形成复层上皮。其中,上皮层与脱细胞真皮基质材料(de-epidermised dermis,DED)结合紧密,形成的人工黏膜有明显的上皮钉突。不同于以往报道,上皮层与Alloderm结合并不十分紧密。以胶原凝胶为基质形成的人工口腔黏膜最厚,有明显分层。以胶原海绵-胶原凝胶为基质形成的复层上皮在部分区域长入至胶原海绵的空隙中。结论以DED和胶原凝胶为支架构建的口腔黏膜更接近于天然结构,而后者脆性较大,限制了其临床应用的可能。
Objective To construct composite oral mucosa by using four kinds of scaffolds and compare their histological features. Methods Human oral mucosa fibroblasts and keratinocytes were cultured in vitro. Four kinds of scaffolds were added with fibroblasts. After cultured for 7 days, keratinocytes were added to the scaffolds. After cultured for 4 days, the cells were transferred to gas-liquid interface and cultured continuously 7d. Morphological characteristics of the compound oral mucosa observed under hematoxylin-eosin staining. Results All four kinds of scaffolds could be constructed to form stratified epithelium. Among them, the epithelial layer and acellular dermal matrix material (de-epidermised dermis, DED) closely, the formation of artificial mucosa obvious epithelial nail. Unlike previous reports, the epithelial layer and Alloderm combination is not very close. The artificial oral mucosa formed by collagen gel is the thickest with obvious stratification. The collagen-sponge-collagen gel-based stratified epithelium grows in some areas into the collagen sponge voids. Conclusion Oral mucosa constructed with DED and collagen gel is closer to the natural structure, while the latter is more brittle, which limits the possibility of its clinical application.