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- Title
THE INFLUENCE OF CA<sup>2+</sup> IONOPHOR IONOMYCIN ON THE MITOCHONDRIAL TRANSITION PERMEABILITY PORE IN NORMAL AND DRUG TREATED GINGIVAL FIBROBLASTS.
- Authors
Goriuc, Ancuta; Petreus, T.; Peptenariu, D.; Iancu, Roxana; Costuleanu, M.
- Abstract
Apoptosis represent a form of programmed cell death, characterized by well-defined morphological and biochemical attributes. Until present time, few data regarding Ca2+ involvement in normal gingival fibroblasts apoptosis are available. This paper purpose is to emphasize the effect of ionomycin on normal gingival fibroblasts and also on fibroblasts treated by CsA, nifedipine, phenytoin, by using flow cytometry methods. Gingival fibroblasts were achieved from 6 week old male rats, 150 - 170 g weight, by gingival explants and grown up in specific culture medium consisting of DMEM, 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 mg/ml streptomycin in an atmosphere containing 5% CO2 at 37°C. Medium was supplemented with CsA treatment (1μg/ml), nifedipine (3mM) and phenytoin (2,5mM). We have used a control group that received no treatment. Following these steps, flow cytometry using FL1 settings 623 V, 505 V, FL2, 10,000 events, 488 nm laser was performed. Data were processed using FlowJo 7.6.1 software.Statistical data were analyzed by One Way ANOVA method (completed by Student-Newman-Keuls method). Results were considered statistically significant for a p value <0,05 and were expressed as mean value (S.E.M). CsA, nifedipine and phenytoin treated fibroblasts were periodically examined and recorded by a image acquisition system attached to the inverted microscope. We have observed and recorded morphological changes and major count changes between cells treated by CsA, nifedipine and phenytoin and untreated cells. Our results show major morphological differences between treated and untreated fibroblasts. Calcein together with CoCl2 appears to be a viable mode; to evaluate Mitochondrial transient permeability pore opens following intracellular and intramitochondrial Ca2+ concentrations variation. Mitochondrial calcein load for normal and treated fibroblasts does not show significant differences.
- Subjects
APOPTOSIS; CELL death; FIBROBLASTS; NIFEDIPINE; PHENYTOIN; CYTOMETRY; NEWMAN-Keuls method (Statistics)
- Publication
Annals of the Romanian Society for Cell Biology, 2010, Vol 15, Issue 2, p341
- ISSN
2067-3019
- Publication type
Article