Odontoblast cell culture – Characterisation of cells from the dentin-pulp interface

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As an epithelial layer at the dentin-pulp interface odontoblasts express unique physiological properties. These cells build dentin as a hard tissue, they are part of the cellular immune system, and as mechanosensitive cells they are involved in nociception. Odontoblasts are target cells of restorative dental materials and essential for biocompatibility studies as well. Unfortunately, it is current scientific consensus that primary odontoblasts are non-dividing cells and thus inculturable. In preliminary studies we isolated primary cells from the dentin-pulp interface of several third molars of young patients, expanded and characterized the morphology. It is aim of the current proposal to clarify the identity of these cells as primary odontoblasts. They major difficulty is the lack of a unique and specific marker for this cell type. Thus the isolated primary cultures will be characterized by a pattern of various properties in comparison with cells of different origin including pulp cells, cells from the periodontal ligament, dental papilla cells, osteoblasts, skin fibroblasts, HeLa, and lung epithelial cells. We described the clearly different morphology of the cells in a preliminary work. Here we continue with the analysis of the expression of proteins and genes such as DSPP, DMP1, NCAM, nestin, pannexin 2 runx2, osteocalcin or colA1. Immunophenotyping following current guidelines for the identification of mesenchymal stem cells will be performed by cytometry, and mineralization will be investigated as well. The expression of these parameters will allow for the evaluation of the identity of the cells isolated from the dentin-pulp interface. If it was possible to keep primary odontoblasts in culture we could investigate cellular mechanisms behind odontoblast functions. These findings would be a major step forward in the development of strategies for the therapy of carious lesions, clinically relevant options of pharmacological support of these therapies, and local tissues regeneration.