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The Effect of Differential Modulation of the N-methyl-D-aspartate Receptor on Growth and intracellular Calcium Ion Concentration in Normal Human Oral Keratinocytes

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Authors

Kim, Insoo; Paik, Ki-Suk; Chang, Mi-Sook; Lee, Seung-Pyo

Issue Date
2010-12
Publisher
KOREAN TISSUE ENGINEERING REGENERATIVE MEDICINE SOC
Citation
TISSUE ENGINEERING AND REGENERATIVE MEDICINE; Vol.7(5), pp. 489-500
Keywords
N-methyl-D-aspartateintracellular calcium ionarachidonic acidnitric oxide synthase inhibitornormal human oral keratinocyte
Abstract
The purpose of this study was to examine the interaction between the agents [N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOS-I)] and cultured normal human oral keratinocytes (NHOK) in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. We also examined whether AA and NOS-I could protect NHOK from glutamate cytotoxicity and the change of intracellular calcium ion concentration. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. Cell viability was measured by the MTT assay and DNA synthesis by the BrdU assay. The microscopic features of NHOK were observed and changes in intracellular calcium ion concentration were measured. NMDA and NNA induced an initial increase in cell number. Low concentration of AA induced an increase in cell number while high concentrations of AA induced a decrease in cell number. The decrease in cell number induced by NMDA at the seventh day was abolished by the addition of low concentrations of AA or NOS inhibitors. Low concentrations of AA or NMDA with high concentrations of AA significantly increased the DNA synthesis rate at four hours. NMDA and AA both induced an increase in intracellular calcium ion concentration, and AA enhanced NMDA-induced intracellular calcium ion in a concentration-dependent manner. NMDA-induced NHOK death was associated with intracellular calcium ion change and the promotion of cell differentiation. Low concentrations of AA protected NHOK cells from NMDA-induced death.
ISSN
1738-2696
Language
English
URI
https://hdl.handle.net/10371/81335
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