组织工程人口腔黏膜的制备及异体移植的临床应用

目的观察自行制备的复层组织工程人口腔黏膜移植后生长情况.方法取3月龄患儿唇裂术中切取的多余口腔黏膜组织,分离成纤维细胞与上皮细胞,分别接种于聚乳酸/聚羟基乙酸共聚物(polylactic/glycolic acid copolymer,PLGA)胶原复合膜上培养,后将其移至气液面进行复合,制成复层组织工程口腔黏膜.7例行口腔内良性肿瘤切除后遗留黏膜缺损的患者,缺损范围为1.8 cm×1.6 cm～3.2 cm×2.6 cm.采用组织工程口腔黏膜修复,术后观察其生长情况.1例志愿者在移植术后18和30 d分别取移植部位黏膜行组织学观察.结果制备的组织工程口腔黏膜生长良好,具有上皮层和上皮下层双层结构,之间为PLGA膜;上皮层5～6层细胞,角蛋白染色阳性,上皮下层3～7层细胞,角蛋白染色阴性.7例患者移植修复术后10 d,组织工程口腔黏膜与创面生长良好,颜色较正常黏膜深;18 d后仍可辨出移植区与正常黏膜;30 d后无法区别界限,创面均Ⅰ期愈合,无明显瘢痕组织.组织学观察:18 d创面上皮层及下方肉芽组织生长良好,毛细血管增生有上皮钉突形成,成纤维细胞卵圆形;30 d胶原化更明显,移植区结构与周边正常组织相似.结论应用组织工程口腔黏膜异体移植修复后,黏膜生长良好,但愈合组织上皮的来源还需进一步研究.

GROWTH OF COMPOUND LAYER TISSURE ENGINEERED ORAL MUCOSA AND ITS CLINICAL APPLICATION IN HETERO-TRANSPLANTATION

Objective To investigate the growth of the tissue engineered mucosa after its hetero-transplantation. Methods The epithelial cells and fibroblasts were isolated from a postoperative tissue of the 3-month patient with labial cleft. The epithelial cells and fibroblasts were separately seeded on the polylactic/glycolic acid copolymer membrane, and then they were exposed to the air-liquid interface. Seven volunteer patients, whose traumatic beds were repaired with the tissue engineered oral mucosa. The biopsy tissue from one of the seven patients was observed under light microscope 18 and 30 days after transplantation, respectively. Results The tissue engineered oral mucosa having 5-6 layers anti-cytoeratin staining positively cells in the epithelial layer and 3-7 layers anti-Cytoeratin staining negatively cells in the subepithelial layer grew well after the hetero-transplantation. No difference could be found between the transplanting and normal areas. At 18 days, the epithelial layer and lamina propria grew well and the fibroblasts were found; at 30 days, collagen was obviously observed. The structures in both the transplanting and the normal areas were similar. Conclusion The tissue engineered oral mucosa can grow well after the hetero-transplantation.