Rabbit tendon cells produce MMP-3 in response to fluid flow without significant calcium transients.
Forces applied to tendon during movement cause cellular deformation, as well as fluid movement. The goal of this study was to test the hypothesis that rabbit tendon fibroblasts detect and respond to fluid-induced shear stress. Cells were isolated from the paratenon of the rabbit Achilles tendon and then subjected to fluid flow at 1 dyn/cm(2) for 6h in a specially designed multi-slide flow device. The application of fluid flow led to an increased expression of the collagenase-1 (MMP-1), stromelysin-1 (MMP-3), cyclooxygenase II (COX-2) and interleukin-1beta (IL-1beta) genes. The release of proMMP-3 into the medium exhibited a dose-response with the level of fluid shear stress. However, not all cells aligned in the direction of flow. In other experiments, the same cells were incubated with the calcium-reactive dye FURA-2 AM, then subjected to laminar fluid flow in a parallel plate flow chamber. The cells did not significantly increase intracellular calcium concentration when exposed to fluid shear stress levels of up to 25 dyn/cm(2). These results show that gene expression in rabbit tendon cells is sensitive to fluid flow, but that signal transduction is not dependent on intracellular calcium transients. The upregulation of the MMP-1, MMP-3 and COX-2 genes shows that fluid flow could be an important mechanical stimulus for tendon remodelling or injury.
Archambault
Joanne M
JM
Faculty of Kinesiology, Human Performance Laboratory, University of Calgary, 2500 University Drive, NW Calgary, Alberta, Canada T2N 1N4.
Elfervig-Wall
Michelle K
MK
Tsuzaki
Mari
M
Herzog
Walter
W
Banes
Albert J
AJ
D000125:Achilles Tendon / Q000166:cytology / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001696:Biomechanical Phenomena; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D003470:Culture Media; D018076:DNA, Complementary / Q000302:isolation & purification; D003627:Data Interpretation, Statistical; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D016257:Fura-2 / Q000494:pharmacology; D005784:Gene Amplification; D005786:Gene Expression Regulation; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000235:genetics* / Q000378:metabolism*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013314:Stress, Mechanical; D013708:Tendon Injuries / Q000378:metabolism / Q000503:physiopathology; D013710:Tendons / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D015854:Up-Regulation
D003470:Culture Media; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D007375:Interleukin-1; D012313:RNA; D011451:Prostaglandin-Endoperoxide Synthases; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1; D002118:Calcium; D016257:Fura-2