In-Vitro Development of A Mucocutaneous Junction for Lip Reconstruction

Publication date: Available online 13 April 2016
Source:Journal of Oral and Maxillofacial Surgery
Author(s): Gurkan Rasit Bayar, Shiuhyang Kuo, Cynthia L. Marcelo, Stephen E. Feinberg
PurposePresentation of a simple and reproducible technique to create, in vitro, a construct containing a mucocutaneous junction (MCJ) with a transitional zone (vermillion) for fabrication of a microvascular prelaminated flap for use in lip reconstruction.Material and MethodsCultured primary human skin keratinocytes and oral mucosal epithelial cells at premixed ratios of 50% skin-50% oral, 25% skin-75% oral, and 75% skin-25% oral were grown on an AlloDerm® dermal equivalent to create a MCJ equivalent with a lip or transitional zone (vermillion) utilizing a novel three dimensional (3D) culture device with a barrier to separate co-cultured skin and oral cells. The three different cell ratios were compared by staining for the following specific differentiation markers; filaggrin, cytokeratin 10 (CK10), CK19, and small proline-rich protein 3 (SPR3) to define the different areas of skin and mucosal keratinocytes.ResultsImmunohistochemical results showed that MCJ equivalents seeded with premixed cells were similar to the differentiation patterns of tissue-engineered 3D cultures using 100% of oral mucosal epithelial cells or skin keratinocytes. The engineered MCJ equivalent constructs, grown in the 3D device specifically constructed with a cell-free gap at the barrier site, formed a transitional zone (vermillion) at the barrier site of intermingling of the skin and oral keratinocytes. The results showed different and unique expression patterns of filaggrin, CK10, 19, and SPR3 by those cells migrating into the gap which were similar to that seen in human lip tissue. This pattern was not seen in MCJ equivalents created using pre-mixed skin and oral cells.ConclusionUsing a device to separately co-culture human oral and skin keratinocytes, to allow the cells to migrate into a cell-free zone resulted in phenotypic expression closer to what is seen in native tissue, in comparison to premixing the skin and oral cells prior to seeding.



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