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Effects of fluid shear stress on mRNA expression of carbonic anhydrase II in polarized rat osteoclasts
Author(s) -
Zhang Qinghong,
Liang Xing,
Zhu Baoming,
Dong Qiang,
Xu Ling,
Xia Lu,
Hu Jian,
Fu Jun,
Liu Mengtao
Publication year - 2006
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2006.05.002
Subject(s) - osteoclast , acid phosphatase , staining , chemistry , cytoplasm , carbonic anhydrase ii , resorption , carbonic anhydrase , messenger rna , bone resorption , vacuole , microbiology and biotechnology , biology , biochemistry , in vitro , enzyme , endocrinology , gene , genetics
The present study was designed to determine the effects of fluid shear stress on the mRNA expression of carbonic anhydrase II (CAII) in polarized rat osteoclasts. Cellular morphology of the polarized osteoclasts generated by a mechanical anatomical technique was examined by tartrate‐resistant acid phosphatase (TRAP) staining and the osteoclastic resorption of dentine slices. The polarized osteoclasts were then stress‐loaded by using a flow shear stress device newly developed by the osteoclast research group (patent number 200420034438; China), at 9 dyne/cm 2 for various time periods [0 (control group), 15, 30, 60, and 120 min], or at various stress levels [0 (control), 0.9, 2.9, 8.7, and 26.3 dyne/cm 2 ] for 30 min. The mRNA expression of CAII was quantified using real‐time fluorescent quantitative PCR (RT‐PCR) and the data were analyzed with SPSS 12.0 software. The polarized osteoclasts were larger than regular monocytes (about 30 μm diameter) with irregular configuration, and the majority of polarized osteoclasts appeared to be spherical and had approximately 2–20 nuclei. The TRAP positive polarized osteoclasts showed asymmetrical red staining in the cytoplasm, and had many filaments and vacuoles. These cells formed resorptive pits in dentine slices. The levels of CAII mRNA expression were shown to be time‐dependent, with the E+5 copy numbers being 7.88 ± 0.09, 11.14 ± 0.12, 15.83 ± 0.18, 1.94 ± 0.02, and 1.37 ± 0.01 in cells treated at 9 dyne/cm 2 for 0, 15, 30, 60 and 120 min, respectively ( P < 0.05). The levels of CAII mRNA expression (E+5 copy numbers) in cells treated with the stress levels of 0, 0.9, 2.9, 8.7 and 26.3 dyne/cm 2 were 7.97 ± 0.201, 11.26 ± 0.688, 15.94 ± 0.201, 31.88 ± 1.496, and 45.08 ± 2.639, respectively ( P < 0.05). These results indicate that there is a relationship between the fluid shear stress and the mRNA expression of CAII in polarized rat osteoclasts.