Fabrication of tissue engineered myo-mucosal flap by grafting the complex of autologous oral keratinocytes and platelet rich plasma (prp) in a rat model

ABSTRACT

BACKGROUNDS: To overcome limited amount of autogenous mucosa for the reconstruction of various mucosal defect including oral mucosal defect, tissue engineered mucosa has been recently introduced. However, introduced conventional technique of tissue engineered mucosa still have serious pitfalls such as long fabrication time, fragility of the reconstructed mucosa, and complexity of the technique. AIM OF THE STUDY: To examine whether the complex of preconfluent autologous keratinocytes and autologous PRP (Platelet rich plasma) can reconstruct oral mucosa on the muscular flap with easier and faster way compared to conventional mucosal tissue engineering technique. MATERIALS AND METHODS: One day before the operation, oral mucosa (3mm in diameter) were taken and treated for extraction of oral keratinocytes according to the routine manner. The day of operation, oral keratinocytes were prepared in the laboratory and then moved to the operating theater. Autologous PRP was also prepared and then mixed with oral keratinocytes just before grafting on the prepared muscular flap. After keratinocyte-PRP complex was seated, then a sterilized rubber sheet was placed on the graft and the elevated skin flap was replaced and sutured. Biopsies were proceeded at 3,5,7,14 and 21 days. Tissue samples were evaluated clinically, histologically, and immunohistochemically. RESULTS: All of the oral keratinocyte-PRP complexes were successfully grafted on the recipient sites (100%). On 3 days after the operation, 1-2 continuous epithelial layer and many inflammatory cells were observed. On 5 days after the operation, increase of layers of keratinocyte was observed with less inflammatory response. Thickness of the layers was gradually increased from 7 to 21 days after the operation. Cytokeratin confirms epithelium in every specimen. CONCLUSIONS: Preconfluent graft of autogenous oral keratinocytes mixed with autogenous PRP have successfully reconstructed myo-mucosal flap. This technique could be a useful alternative for oral mucosal reconstruction in the near future.