Development of a 3D oral mucosa model for in vitro studies

Abstract

Several studies have been developed to evaluate the oral mucosa repair process. So far, most of the in vitro studies were performed using two dimensional cell culture methods, where cells are seeded in a monolayer model. These studies have provided great contributions about cell behavior at normal conditions and in response to different stimuli. However, the two dimensional cell culture models show some limitations, such as poor cell interactions to extracellular matrix and the interactions of different cell lines. To overcome these limitations, the three dimensional (3D) cell culture models were developed, to better simulate the in vivo conditions of cells interactions and phenotypes expression. To date, the development and standardization of a 3D oral mucosa model containing stratified epithelium and cellularized connective tissue would not only improve the comprehension of cells interactions, but also the elucidation of the etiopathogenesis of several oral conditions, such as oral mucositis, improving also the treatment strategies and protocols. The aim of this study will be to develop a 3D oral mucosa equivalent model (EVPOME), using an AlloDerm acellular matrix, for in vitro studies. The development of this model, that shows similar morphology and phenotype as oral mucosa will enlarge the use of 3D oral mucosa model for elucidation of several oral conditions and treatment protocols. Oral keratinocytes and fibroblasts will be obtained from healthy young individuals and 3D cell culture model will be obtained using the AlloDerm scaffold for fibroblasts cell culture, followed by the keratinocyte cell culture. Cell viability, cell culture morphology and cell proliferation will be evaluated by MTT Assay, H&E staining and PCNA immunohistochemistry, respectively. (AU)