In vitro cultured autologous pre-confluent oral keratinocytes for experimental prefabrication of oral mucosa.

摘要: The reconstruction of large defects after head and neck cancer resection often requires composite tissue transfer to replace a combination bone, muscle mucosa. Thus, engineering techniques may be useful for oral mucosal reconstructive surgery prefabricate on the flap in vivo, instead using conventional skin-bearing flaps. aim this study was investigate whether autogenous pre-confluent keratinocytes (PCOK) cultured vitro can create coverage single grafting procedure. In 30 Wistar rats, with small piece mucosa (2 mm x 5 mm), were isolated then seeded hydrophilic PTFE membrane (n = 50) serum-free culture condition. After 48 h, membrane, together PCOK, transplanted onto gracilis fabricate vivo. wound bed closed primarily until time examination. Biopsies carried out 1, 2, 3, 4 weeks, respectively, transplantation evaluated immunohistochemically (AE1/AE3 anti-pancytokeratin, cytokeratin 5/6, collagen IV, laminin, lectin-specific labeling N-acetylglucosamine oligomeres endothelial cells) relation following criteria: (1) graft acceptance; (2) inflammatory signs; (3) structural changes keratinocyte lining; (4) expression basement components; (5) vascularization. Ninety-one percent grafts showed uniform epithelial layers. mean number reconstructed cell layers 1.7, 2.0, 1.85 2.7 at 3 respectively (P 0.342). Collagen laminin capillaries developed between neoepithelium underlying muscular layer. Only two specimens signs infection 2 weeks transplantation. conclusion, experiment demonstrated that PCOK vivo achieve multi-layered short period time. This technique alternative tool oropharyngeal is also worth considering further clinical studies.