Radiation Effects on a 3-Dimensional Model of Oral Mucositis

We studied the radiation effect on cell culture model of oral mucositis. The musositis model (tissues) used is an organotypic model which consisted of 3 dimensional (3-D) cell cultures of primary human oral keratinocytes grown on top of primary oral fibroblasts and resembles oral mucosal (tissues). The tissues were purchased from MatTek corporation (Ashland, MA) and were grown on top of microporous membrane and supplemented with MatTek serum free media. These tissues were irradiated with 12 and 2 Gy. Six hours after the irradiation the tissues were cut in half. Half of the tissue was used to extract total RNA (RNeasy Plus Mini kit, Qiagen, Germantown, MD) and the other half was used to evaluate the histology and the apoptosis. We had the following groups: Non-Irradiated Tissue, Irradiated Tissue with 12 Gy and Irradiated Tissues with 2 Gy. Human Prirmary 3-D Cultures of Human Keratinocytes grown on top of Human Fibroblasts were irradiated with 12 or 2 Gy and the effect of the radiation was studied 6 hours after the irradiation using histology, apoptosis assays and gene expression microarrays. There are 3 samples the control (non irradiated) is NTNI. Each sample was done in triplicate.