Tissue engineering of human oral mucosa on different scaffolds for clinical applications

In oral and maxillofacial surgery, there is a great demand for oral mucosa for intraoral grafting after tumor dissections or traumas. In this study, three different scaffolds were tested for suitability as an underlay for a three-dimensional reconstruction of human autologous oral mucosa for clinical applications. For this purpose, primary human oral fibroblasts and keratinocytes were cultured and increased in vitro. They were seeded on matrices: TissuFoil E (TFE) Dermal Regeneration Template Single Layer (DRT) and Vicyl to create a three-dimensional reconstruction of oral mucosa. First only fibroblasts were seeded on scaffolds to obtain a dermal equivalent after seven days. The seeding was analyzed and characterized using electron-microscopy and immunostaining. In the experiments that followed, keratinocytes were seeded on dermal equivalents to develop organotypic co-cultures. They were cultured for seven and fourteen days to get full-thickness oral mucosa substitutes. Frozen sections of the probes were stained against CD90 (fibroblasts) and cytokeratin 14 (keratinocytes) with fluorescent dyestuffs. The results showed that all of the matrices were biocompatible because cells were able to grow on them. In oral mucosa substitutes on Vicryl, no natural stratification of the cells could be shown, but there was good biostability. TFE provided the best cell growth with good stratification and natural arrangement of epithelial and dermal components, but bad biostability of the membrane. The combination of good biostability and good cell growth with natural stratification could be seen only on the DRT-construct. For this reason, we consider DRT to be the most suitable for its intended use.