Fluid-induced, shear stress-regulated extracellular matrix and matrix metalloproteinase genes expression on human annulus fibrosus cells

Abstract

BackgroundMechanical loading plays an important role in the regulation of extracellular matrix (ECM) homeostasis as well as pathogenesis of intervertebral disc (IVD) degeneration. The human annulus fibrosus (hAF) in the IVD is subjected to contact shear stress during body motion. However, the effects of shear stress on hAF cells remain unclear. This aim of the study was to investigate the expression of the ECM (COLI, COLIII and aggrecan) and matrix metalloproteinase (MMP-1, MMP-3 and ADAMTS-4) genes in hAF cells following fluid-induced shear stress in a custom-fabricated bio-microfluidic device.MethodshAF cells were harvested from degenerated disc tissues in routine spine surgery, staged by magnetic resonance imaging, expanded in monolayers and then seeded onto the bio-microfluidic device. The experimental groups were subjected to 1 and 10 dyne/cm2 shear stress for 4 h, and no shear stress was applied to the control group. We used real time polymerase chain reaction for gene expression.ResultsShear stress of 1 dyne/cm2 exerted an anabolic effect on COLI and COLIII genes and catabolic effects on the aggrecan gene, while 10 dyne/cm2 had an anabolic effect on the COLI gene and a catabolic effect on COLIII and aggrecan genes. The COLI gene was upregulated in a stress-dependent manner. Expression of MMP-1 was significantly higher in the 10 dyne/cm2 group compared to the control group (P < 0.05), but was similar in the control and 1 dyne/cm2 groups. Expression of MMP-3 and ADAMTS-4 were similar in all three groups.ConclusionTaken together, hAF cells responded to shear stress. The findings help us understand and clarify the effects of shear stress on IVD degeneration as well as the development of a new therapeutic strategy for IVD degeneration.

Genetic dimension of intervertebral disc degeneration: polymorphism of matrix metalloproteinase 1 and 3 in the north iranian population

“…However, in high concentrations it is known as the most important cause of intervertebral disc degeneration. It has also been shown that MMP-1 expression is associated with the degradation of the matrix in disc degeneration (8). Promoter polymorphisms have been identified for both MMP-1 and MMP-3.…”

“…Studies have found a number of matrix metalloproteinase (MMPs) (including MMP type 1,28,32,23,33,10,8,25,7,16) associated with disc degeneration (33). These proteases are the most important catabolic enzymes in the destruction of intervertebral discs (4,21).…”

Effects of the NF‑κB/p53 signaling pathway on intervertebral disc nucleus pulposus degeneration

“…The most widely accepted factor responsible for causing low back pain is discogenic low back pain, which is characterized by the chronic process of progressive intervertebral disc degeneration (idd) (2). idd is primarily characterized by a small number of active cells in the nucleus pulposus of the intervertebral disc, the breakdown of extracellular matrix and the emergence of proinflammatory mediators in nucleus pulposus cells (3). Ye et al (4) demonstrated that short stature homeobox 2 expression levels were significantly decreased in human idd tissues, and the expression levels of metallopeptidases with thrombospondin type 1 motifs (adaMTSs) and matrix metallopeptidases (MMPs) were increased in idd tissues and cells (5,6).…”

The Effect of Drag Force and Flow Rate on Mesenchymal Stem Cells in Packed-Bed Perfusion Bioreactor

“…However, the expression of death-associated protein kinase (DAPK) was also reported to be regulated by the mechanical stimulus affecting cytokine-activated apoptosis [29]. In a microfluidic culture system, the effect of shear stress on human annulus fibrosus cells was evaluated [30]. As stated, a shear stress of 0.1 N/m 2 caused an anabolic effect on collagen type-1 and -3, while the catabolic effects on the aggrecan were reported.…”