Radiation effects on a 3D cell culture of primary human keratinocytes and the protective effect of N-acetyl-cysteine (NAC) and the Traditional Chinese Medicine Qingre-Liyan decoction (QYD). Homo sapiens

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. 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 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; Irradiated Tissue with 12 Gy pretreated with N-acetylcysteine (NAC); Irradiated Tissue with 12 Gy pretreated with the Traditional Chinese Medicine Qingre-Liyan decoction (QYD); and Irradiated Tissue with 12 Gy pretreated with 50:50 w/w NAC-QYD. Overall design: Group 1: Human Prirmary 3-D Cultures of Human Keratinocytes were irradiated with 12 Gy; Group 2: Human Prirmary 3-D Cultures of Human Keratinocytes were pretereated with N-acetylcysteine (NAC) and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with the Qingre-Liyan Decoction (QYD) and then irradiated with 12 Gy; Human Prirmary 3-D Cultures of Human Keratinocytes were pretreated with a 50:50 w/w mixture of NAC-QYD and then irradiated with 12 Gy. Each sample was done in triplicate.