Fukahori Mioko, Chitose Shun-Ichi, Sato Kiminori, Sueyoshi Shintaro, Kurita Takashi, Umeno Hirohito, Monden Yu, Yamakawa Ryoji
Department of Otolaryngology-Head and Neck Surgery, Kurume University School of Medicine, Kurume, Fukuoka, Japan.
Department of Ophthalmology, Kurume University School of Medicine, Kurume, Fukuoka, Japan.
PLoS One. 2016 Jan 5;11(1):e0146151. doi: 10.1371/journal.pone.0146151. eCollection 2016.
Scarred vocal folds result in irregular vibrations during phonation due to stiffness of the vocal fold mucosa. To date, a completely satisfactory corrective procedure has yet to be achieved. We hypothesize that a potential treatment option for this disease is to replace scarred vocal folds with organotypic mucosa. The purpose of this study is to regenerate vocal fold mucosa using a tissue-engineered structure with autologous oral mucosal cells.
Animal experiment using eight beagles (including three controls).
A 3 mm by 3 mm specimen of canine oral mucosa was surgically excised and divided into epithelial and subepithelial tissues. Epithelial cells and fibroblasts were isolated and cultured separately. The proliferated epithelial cells were co-cultured on oriented collagen gels containing the proliferated fibroblasts for an additional two weeks. The organotypic cultured tissues were transplanted to the mucosa-deficient vocal folds. Two months after transplantation, vocal fold vibrations and morphological characteristics were observed.
A tissue-engineered vocal fold mucosa, consisting of stratified epithelium and lamina propria, was successfully fabricated to closely resemble the normal layered vocal fold mucosa. Laryngeal stroboscopy revealed regular but slightly small mucosal waves at the transplanted site. Immunohistochemically, stratified epithelium expressed cytokeratin, and the distributed cells in the lamina propria expressed vimentin. Elastic Van Gieson staining revealed a decreased number of elastic fibers in the lamina propria of the transplanted site.
The fabricated mucosa with autologous oral mucosal cells successfully restored the vocal fold mucosa. This reconstruction technique could offer substantial clinical advantages for treating intractable diseases such as scarring of the vocal folds.
瘢痕化的声带由于声带黏膜僵硬，在发声时会导致不规则振动。迄今为止，尚未实现完全令人满意的矫正手术。我们假设，针对这种疾病的一种潜在治疗选择是用器官型黏膜替代瘢痕化的声带。本研究的目的是使用具有自体口腔黏膜细胞的组织工程结构来再生声带黏膜。
使用八只比格犬（包括三只对照犬）进行动物实验。
手术切除3mm×3mm的犬口腔黏膜标本，并将其分为上皮组织和上皮下组织。分别分离并培养上皮细胞和成纤维细胞。将增殖的上皮细胞与增殖的成纤维细胞共同培养在定向胶原凝胶上，再培养两周。将器官型培养组织移植到黏膜缺损的声带处。移植两个月后，观察声带振动和形态特征。
成功构建了由复层上皮和固有层组成的组织工程化声带黏膜，其与正常分层的声带黏膜非常相似。喉动态镜检查显示移植部位有规则但略小的黏膜波。免疫组织化学检测显示，复层上皮表达细胞角蛋白，固有层中分布的细胞表达波形蛋白。弹性范吉森染色显示移植部位固有层中的弹性纤维数量减少。
用自体口腔黏膜细胞构建的黏膜成功修复了声带黏膜。这种重建技术可为治疗声带瘢痕等难治性疾病提供显著的临床优势。
