Development and characterization of a tissue-engineered human oral mucosa equivalent produced in a serum-free culture system
Development and characterization of a tissue-engineered human oral mucosa equivalent produced in a serum-free culture system
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DOI：
10.1177/00220345000790030301
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发表时间：
2000-03-01
影响因子：
7.6
通讯作者：
Feinberg, SE
中科院分区：
文献类型：
--
作者：
Izumi, K;Terashi, H;Feinberg, SE
颌面外科医生面临的一个问题是缺乏足够的自体口腔粘膜来重建口腔。中厚或口腔粘膜移植需要一次以上的外科手术，并可能导致供区并发症。皮肤的附属器结构和角质化模式与口腔粘膜不同。在这项研究中，我们成功地在体外组装了一种由表皮和真皮成分组成的人类口腔粘膜等效物，在一种定义的、缺乏必需脂肪酸的、无血清的、没有饲养层的培养液中，可以用于人类口腔内移植。将自体口腔角质形成细胞种植到身体真皮--异体真皮上。口腔黏膜在气液界面培养2周。所产生的等价物有一个分层良好的角化旁上皮层，类似于天然的口腔角化粘膜。分化标志物微丝蛋白和细胞角蛋白10/13的表达提示早期角化状态。增殖细胞核抗原(增殖细胞核抗原)和Ki-67等增殖标记物的存在提示肿瘤处于过度增殖状态。等价物的脂肪酸组成与体外培养的口腔角质形成细胞相似，但与体内天然组织的脂肪酸组成不同，分别为18：2和20：4，16：1和18：1。相应的角质形成细胞似乎比天然角化粘膜处于更活跃和增殖的状态。口腔黏膜上细胞群体的动态特性可能有利于口腔内移植手术和角质形成细胞的转基因。
A problem maxillofacial surgeons face is a lack of sufficient autogenous oral mucosa for reconstruction of the oral cavity. Split-thickness or oral mucosa grafts require more than one surgical procedure and can result in donor site morbidity. Skin has disadvantages of adnexal structures and a different keratinization pattern than oral mucosa. In this study, we successfully assembled, ex vivo, a human oral mucosa equivalent, consisting of epidermal and dermal components, in a defined, essential-fatty-acid-deficient, serum-free culture medium without a feeder layer, that could be used for intra-oral grafting in humans. Autogenous oral keratinocytes were seeded onto a cadaveric dermis, AlloDerm(TM). The oral mucosa equivalent was cultured at an air-liquid interface for 2 wks. The resulting equivalent had a well-stratified parakeratinized epithelial layer similar to native oral keratinized mucosa. Expression of differentiation markers, filaggrin and cytokeratin 10/13, suggested a premature keratinized state. The presence of proliferation markers, proliferating cell nuclear antigen (PCNA) and Ki-67, suggested a state of hyperproliferation. Fatty acid composition of the equivalent was similar to that of in vitro cultured oral keratinocytes but differed from the that of in vivo native tissue, showing a lower content of 18:2 and 20:4, and a higher content of 16:1 and 18:1 fatty acids, respectively. The keratinocytes of the equivalent appeared to be in a more active and proliferative state than native keratinized mucosa. The dynamic nature of the cell population on the oral mucosa equivalent may be beneficial for intra-oral grafting procedures and for transfection of the keratinocytes.