Naslov
Standardization of the method for cigarette smoke extract preparation and its toxicity towards polymorphonuclear leukocytes

Autori
Somborac Bačura, Anita ; Rumora, Lada ; Popović-Grle, Sanja ; Žanić Grubišić, Tihana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Clinical Chemistry and Laboratory Medicine / Plebani, Mario - : Walter de Gruyter, 2015

Skup
15th EFLM Postgraduate Course in Clinical Chemistry and Laboratory Medicine: ''How to assess the quality of your method?''

Mjesto i datum
Zagreb, Hrvatska, 24.10.2015. - 25.10.2015

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
cigarette smoke extract ; method standardization ; poymorphonuclear leukocytes ; toxicity

Sažetak
Background: Cigarette smoke is known as oxidative stress causing agent. For laboratory studies of its toxic effects on different cell models cigarette smoke extract (CSE) is used. The aim of this study was to evaluate the method for CSE preparation and to examine cytotoxic effect of CSE on polymorphonuclear leukocytes (PMN). Materials and Methods: 100 % CSE was freshly prepared by bubbling the smoke from two Kentucky 3R4F research-reference cigarettes through the 25 mL of cell culture medium with the use of vacuum pump. Standardization of CSE preparation was performed by measurement of the absorbance of 1.25 - 100 % CSE at 280 and 320 nm using the UV/VIS spectrophotometer (Cecil Aquarius CE 7200). PMN were isolated from blood of healthy volunteer and exposed to 4 % CSE or cell culture medium (control). After incubation for different time periods (1, 4, 20 or 22 h), cytotoxicity of CSE was examined by measurement of LDH activity (Herbos Dijagnostika reagent ; Trace 30 analyzer) in PMN supernatants. Statistical analysis was performed using the SigmaStat program. Results: Mean absorbance of 100 % CSE at 320 nm was 1.717 ± 0.386 (N = 6 ; CV = 22.5 %), while it was too high for measurement at 280 nm. Variations in mean absorbances for diluted CSE solutions were slightly higher than for 100 % CSE at 320 nm. The catalytic activities of LDH measured in PMN supernatants exposed to 4 % CSE were not statistically different when compared to untreated PMN for all tested time periods (P = 0.086). Conclusions: In this work the method for CSE preparation was evaluated and no cytotoxic effects of low dose CSE on PMN was found. Standardization of CSE preparation is the basis for further research of the role of cigarette smoke in pathogenesis of diseases caused by cigarette smoking.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti, Farmacija

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