Nitric Oxide Response to Shear Stress by Human Bone Cell Cultures Is Endothelial Nitric Oxide Synthase Dependent

Bone cells, in particular osteocytes, are extremely sensitive to shear stress, a phenomenon that may be related to mechanical adaptation of bone. In this study we examined whether human primary bone cells produce NO in response to fluid shear stress and established by RT/PCR which NOS isoforms were...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1998-09, Vol.250 (1), p.108-114
Main Authors: Klein-Nulend, J., Helfrich, M.H., Sterck, J.G.H., MacPherson, H., Joldersma, M., Ralston, S.H., Semeins, C.M., Burger, E.H.
Format: Article
Language:English
Subjects:
Adaptation, Physiological
Adolescent
Adult
Aged
Aged, 80 and over
Base Sequence
Bone and Bones - cytology
Bone and Bones - metabolism
Bone and Bones - physiology
bone cells
Cells, Cultured
Child
DNA Primers
ecNOS
fluid shear stress
Humans
mechanical stress
Middle Aged
nitric oxide
Nitric Oxide - metabolism
nitric oxide synthase
Nitric Oxide Synthase - genetics
Nitric Oxide Synthase - metabolism
Nitric Oxide Synthase Type III
Physical Stimulation
Polymerase Chain Reaction
RNA, Messenger - genetics
Stress, Physiological - enzymology
Stress, Physiological - metabolism
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Summary:Bone cells, in particular osteocytes, are extremely sensitive to shear stress, a phenomenon that may be related to mechanical adaptation of bone. In this study we examined whether human primary bone cells produce NO in response to fluid shear stress and established by RT/PCR which NOS isoforms were expressed before and after application of shear stress. One hour pulsating fluid flow (PFF; 0.7 ± 0.02 Pa, 5 Hz) caused a rapid (within 5 min) 2 to 4-fold increase in NO production. NO release was only transiently increased during the first 15 min of exposure to PFF, and remained at control levels during a 1-24 hr postincubation period. In both control and PFF-treated cells, mRNA was easily detected for ecNOS, but not nNOS, and only minimal amounts iNOS were found. mRNA levels for ecNOS increased 2-fold at 1 hr after 1 hr PFF treatment. These results suggest that the rapid production of NO by human bone cells in response to fluid flow results from activation of ecNOS. PFF also leads to an increase in ecNOS mRNA which is likely related to the shear stress responsive element in the promoter of ecNOS.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1998.9270