A multishear microfluidic device for quantitative analysis of calcium dynamics in osteoblasts

Kou, Songzi; Pan, Leiting; Noort, Danny van; Meng, Guixian; Wang, Xian; Sun, Hai‐Ying; Xu, Jingyi; Lee, Imshik

doi:10.1016/j.bbrc.2011.04.044

Cited by 57 publications

(50 citation statements)

References 35 publications

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“…Latency in building up calcium responses was first described to occur within 1 min of shear stress on bovine ECs (3), in which IP 3 was found to precede and peak after 20 -30 s of flow in human umbilical vein endothelial cells (4,27). In more recent studies, a significant lag between the onset of fluid-flow and [Ca 2ϩ ] i increase was also observed in intercalated, renal, and bone cells subjected to shear stress (20,22,33,34,41). Although likely to be a function of the nature of the force applied, this lag time differs between cell types, but the peak occurrence was typically found to be between 10 and 30 s after stimulus.…”

Section: Discussionmentioning

confidence: 95%

Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

Melchior

Frangos

2012

American Journal of Physiology-Cell Physiology

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Disturbed flow patterns, including reversal in flow direction, are key factors in the development of dysfunctional endothelial cells (ECs) and atherosclerotic lesions. An almost immediate response of ECs to fluid shear stress is the increase in cytosolic calcium concentration ([Ca(2+)](i)). Whether the source of [Ca(2+)](i) is extracellular, released from Ca(2+) intracellular stores, or both is still undefined, though it is likely dependent on the nature of forces involved. We have previously shown that a change in flow direction (retrograde flow) on a flow-adapted endothelial monolayer induces the remodeling of the cell-cell junction along with a dramatic [Ca(2+)](i) burst compared with cells exposed to unidirectional or orthograde flow. The heterotrimeric G protein-α q and 11 subunit (Gα(q/11)) is a likely candidate in effecting shear-induced increases in [Ca(2+)](i) since its expression is enriched at the junction and has been previously shown to be activated within seconds after onset of flow. In flow-adapted human ECs, we have investigated to what extent the Gα(q/11) pathway mediates calcium dynamics after reversal in flow direction. We observed that the elapsed time to peak [Ca(2+)](i) response to a 10 dyn/cm(2) retrograde shear stress was increased by 11 s in cells silenced with small interfering RNA directed against Gα(q/11). A similar lag in [Ca(2+)](i) transient was observed after cells were treated with the phospholipase C (PLC)-βγ inhibitor, U-73122, or the phosphatidylinositol-specific PLC inhibitor, edelfosine, compared with controls. Lower levels of inositol 1,4,5-trisphosphate accumulation seconds after the onset of flow correlated with the increased lag in [Ca(2+)](i) responses observed with the different treatments. In addition, inhibition of the inositol 1,4,5-trisphosphate receptor entirely abrogated flow-induced [Ca(2+)](i). Taken together, our results identify the Gα(q/11)-PLC pathway as the initial trigger for retrograde flow-induced endoplasmic reticulum calcium store release, thereby offering a novel approach to regulating EC dysfunctions in regions subjected to the reversal of blood flow.

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Section: Discussionmentioning

confidence: 95%

Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

Melchior

Frangos

2012

American Journal of Physiology-Cell Physiology

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“…mRNA expression of osteogenic genes Runx2, ALP, Col I, and osteocalcin was also increased under low FSS (Yu et al, 2014). Similar to high FSS experiments, PPFC were often used to provide the required stimuli in low FSS applications (Kou et al, 2011; Xing et al, 2014). In addition, rockers and orbital shakers were also used to generate FSS in this range (Liegibel et al, 2004; Aisha et al, 2015).…”

Section: Bone Mechanotransductionmentioning

confidence: 99%

In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

Wittkowske

Reilly

Lacroix

et al. 2016

Front. Bioeng. Biotechnol.

237

225

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This review describes the role of bone cells and their surrounding matrix in maintaining bone strength through the process of bone remodeling. Subsequently, this work focusses on how bone formation is guided by mechanical forces and fluid shear stress in particular. It has been demonstrated that mechanical stimulation is an important regulator of bone metabolism. Shear stress generated by interstitial fluid flow in the lacunar-canalicular network influences maintenance and healing of bone tissue. Fluid flow is primarily caused by compressive loading of bone as a result of physical activity. Changes in loading, e.g., due to extended periods of bed rest or microgravity in space are associated with altered bone remodeling and formation in vivo. In vitro, it has been reported that bone cells respond to fluid shear stress by releasing osteogenic signaling factors, such as nitric oxide, and prostaglandins. This work focusses on the application of in vitro models to study the effects of fluid flow on bone cell signaling, collagen deposition, and matrix mineralization. Particular attention is given to in vitro set-ups, which allow long-term cell culture and the application of low fluid shear stress. In addition, this review explores what mechanisms influence the orientation of collagen fibers, which determine the anisotropic properties of bone. A better understanding of these mechanisms could facilitate the design of improved tissue-engineered bone implants or more effective bone disease models.

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“…Moreover, polydimethylsiloxane (PDMS) channels make it possible to observe dynamic changes of cell morphology in situ [15, 16]. Some previous studies have shown the responses of calcium signals and the changes of morphology and cytoskeleton in osteoblasts using microfluidic devices [6, 17]. …”

Section: Introductionmentioning

confidence: 99%

Impact of flow shear stress on morphology of osteoblast-like IDG-SW3 cells

Duan

Ren

et al. 2017

J Bone Miner Metab

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This study constructed an in situ cell culture, real-time observation system based originally on a micro-fluidic channel, and reported the morphological changes of late osteoblast-like IDG-SW3 cells in response to flow shear stress (FSS). The effects of high (1.2 Pa) and low (0.3 Pa) magnitudes of unidirectional FSS and three concentrations of extracellular Type I collagen (0.1, 0.5, and 1 mg/mL) coating on cell morphology were investigated. IDGSW3 cells were cultured in polydimethylsiloxane microfluidic channels. Cell images were recorded real-time under microscope at intervals of 1 min. Cell morphology was characterized by five parameters: cellular area, cell elongation index, cellular alignment, cellular process length, and number of cellular process per cell. Immunofluorescence assay was used to detect stress fiber distribution and vinculin expression. The results showed that 1.2 Pa, but not 0.3 Pa of FSS induced a significant morphological change in late osteoblast-like IDG-SW3 cells, which may be caused by the alteration of cellular adhesion with matrix in response to FSS. Moreover, the amount of collagen matrix, alignment of fiber stress and expression of vinculin were closely correlated with the morphological changes of IDG-SW3 cells. This study suggests that osteoblasts are very responsive to the magnitudes of FSS, and extracellular collagen matrix and focal adhesion are directly involved in the morphological changes adaptive to FSS.

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A multishear microfluidic device for quantitative analysis of calcium dynamics in osteoblasts

Cited by 57 publications

References 35 publications

Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

Impact of flow shear stress on morphology of osteoblast-like IDG-SW3 cells

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A multishear microfluidic device for quantitative analysis of calcium dynamics in osteoblasts

Cited by 57 publications

References 35 publications

Gαq/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

Gαq/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

Impact of flow shear stress on morphology of osteoblast-like IDG-SW3 cells

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Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells

Gα_q/11-mediated intracellular calcium responses to retrograde flow in endothelial cells