[Detection of reactive oxygen species of human dental pulp cells by flow cytometry].
OBJECTIVE
To establish a method for detection of reactive oxygen species(ROS) of human dental pulp cells(HDPCs) by flow cytometry.


METHODS
HDPCs were obtained using tissue explant technique in vitro. The subcultured cells were exposed to peroxide oxygen(H2O2) of different concentrations from 50 μmol/L to 400 μmol/L for 30 minutes, then incubated with two different concentrations of 2',7'-dichlorofluorescein diacetate (DCFH-DA), which were 10 μmol/L and 20 μmol/L for 20 minutes at 37 degrees centigrade in dark. The fluorescence intensities of intracellular dichlorofluorescein(DCF) were detected by flow cytometry. Statistical analysis was carried out by SPSS 13.0 software software.


RESULTS
The positive rate varied with different concentrations of detectors. The fluorescence intensities remained insignificant difference among samples incubated with the same concentration of detector and H(2)O(2),and increased by rising of the incubating concentration of H(2)O(2).


CONCLUSIONS
The detector with concentration of 20 μmol/L shows higher detector loading rate(positive rate). The intracellular ROS level changes as the H(2)O(2) treatment concentration rising from 50 to 400 μmol/L. The application of flow cytometry to measure the ROS in HDPCs is simple, reliable and stable.
Department of General Dentistry, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China. wutiantian_happy@126.com
Tian-tian
TT
Wu
Rong
R
Du
Li-fen
LF
Li
Ya-qin
YQ
Zhu
D003782:Dental Pulp; D005434:Flow Cytometry; D005452:Fluoresceins; D006801:Humans; D006861:Hydrogen Peroxide; D017382:Reactive Oxygen Species
D005452:Fluoresceins; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; D006861:Hydrogen Peroxide