Stimulation of nitric oxide mechanotransduction in single osteoblasts using atomic force microscopy

McGarry, James G.; Maguire, Pierce; Campbell, Veronica A.; O’Connell, Brian; Prendergast, Patrick J.; Jarvis, Suzanne P.

doi:10.1002/jor.20515

Cited by 21 publications

(17 citation statements)

References 41 publications

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“…Accordingly, the continued increase of fluorescence during prolonged exposure (i.e., > 60 s) to constant shear stress may not mean that NOS activity also gradually increases during this period, but rather may reflect an intracellular accumulation of fluorescent DAF derivatives together with continued NOS activity at an elevated but steady level. Similar patterns of DAF-related fluorescence with induced NOS activity have been reported for other cell types [23-25]. …”

Section: Discussionsupporting

confidence: 80%

Shear stress activation of nitric oxide synthase and increased nitric oxide levels in human red blood cells

et al. 2011

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Red blood cells (RBC) play an important role in the balance between generation and scavenging of nitric oxide (NO) and hence its local bioavailability and influence on vasomotor control. Previous studies have reported increased NO levels in RBC suspensions subsequent to exposure to shear forces; the present study was designed to further investigate changes in intracellular NO concentration and possible mechanisms involved for RBC exposed to well-controlled shear forces. Attached human RBC were subjected to shear stresses up to 0.1 Pa in a parallel-plate flow channel; fluorescent methods were used to monitor changes in intracellular NO and calcium concentrations. Intracellular NO concentration, estimated by the fluorescence level of 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM), increased sharply within 30 s following the application of shear stress between 0.013 to 0.1 Pa. This increase was only partially prevented by the absence of L-arginine and by the presence of L-N-acetyl-methyl-arginine (L-NAME), strongly suggesting that this response was in part related to the activation of NO-synthase (NOS) enzyme. The increase in intracellular NO concentration under shear stress was also inhibited by calcium chelation in the suspending medium, indicating the role of calcium entry for NOS activation. Increases of intracellular calcium concentrations under the same shearing conditions were demonstrated by monitoring Fluo-3/AM fluorescence in RBC exposed to shear stress. Serine 1177 phosphorylated NOS protein, the activated form of the enzyme determined by immunohistochemistry, was found to be significantly increased following the exposure of RBC to 0.1 Pa shear stress for 1 min. These data confirm that RBC possess a NOS enzyme that is actively synthesizing NO and activated by effective shear forces. The data also suggest that there may be additional (e.g., non-enzymatic) NO generating mechanisms in RBC that are also enhanced under shear stress.

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Section: Discussionsupporting

confidence: 80%

Shear stress activation of nitric oxide synthase and increased nitric oxide levels in human red blood cells

et al. 2011

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“…In addition to the direct toxicity, NO could also influence many physiological processes from DNA transcription and replication to protein synthesis and secretion (Pavanelli et al 2010). NO production is demonstrated to be involved in mechanotransduction of mechanically stimulated bone cells (McGarry et al 2008) and endothelial cells (Rizzo et al 1998), and regulated by the effects of simulated microgravity (Random Positioning Machine) in endothelial cells (Grenon et al 2013) and macrophages (Hsieh et al 2005). NO is synthesized by a class of NADPH-dependent NO synthases (NOSs) and regulated by arginase (Shatanawi et al 2015).…”

Section: Discussionmentioning

confidence: 99%

Effects of Simulated Microgravity on Functions of Neutrophil-like HL-60 Cells

Wang

Zhang

et al. 2015

Microgravity Sci. Technol.

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Altered gravity, especially microgravity affects cellular functions of immune cells and can result in immune dysfunction for long-term, manned spaceflight and space exploration. The underlying mechanism, however, of sensing and responding to the gravity alteration is poorly understood. Here, a rotary cell culture system (RCCS) bioreactor was used to elucidate the effects of simulated microgravity on polymorphonuclear neutrophils (PMN)-like HL-60 cells. Alteration of cell morphology, up-regulation of (nitric oxide) NO production, enhancement of interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein 1 (MCP-1) secretion, and diversity of cellular adhesion molecule expression were observed for the cells cultured in RCCS, leading to the up-regulated inflammatory immune responses and host defense. It was also indicated that such alterations in biological responses of PMNs mediated the reduced rolling velocity and decreased adhesion of PMNlike HL-60 cells on endothelial cells under shear flow. This work furthers the understandings in the effects and mechanism of microgravity on PMN functions, which are Chengzhi Wang and Ning Li, both authors contributed equally to this work. Electronic supplementary material

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“…A single osteocyte can disseminate a mechanical stimulus to its surrounding osteocytes via extracellular soluble signaling factors like nitric oxide (Vatsa et al, 2007). A wide variety of devices have been tested for mechanical stimulation of cells and tissues in vitro, namely of osteocytes and osteoblasts (Appleford et al, 2007;Brown, 2000;Lewandowska-Szumiel et al, 2007;McGarry et al, 2008;Tanaka, 1999), although many of these systems are difficult to adapt to an in vivo device. Cell responses depend upon the strain, load and frequency of the stimulus; dynamic, short loading exerts the strongest bone adaptation response, and bone cells tend to accommodate to a routine, so the stimulus must vary in order to elicit a same level of response; a stochastic bone cells response in vitro and in vivo has been reported (Bacabac et al, 2006;Bakker et al, 2001;Burr et al, 2002;Cullen et al, 2001;Hsieh and Turner, 2001;Robling et al, 2001;Tanaka et al, 2003aTanaka et al, , 2003bTurner et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

Polymeric piezoelectric actuator substrate for osteoblast mechanical stimulation

Frias

Reis

Silva

et al. 2010

Journal of Biomechanics

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a b s t r a c tBone mass distribution and structure are dependent on mechanical stress and adaptive response at cellular and tissue levels. Mechanical stimulation of bone induces new bone formation in vivo and increases the metabolic activity and gene expression of osteoblasts in culture. A wide variety of devices have been tested for mechanical stimulation of cells and tissues in vitro. The aim of this work was to experimentally validate the possibility to use piezoelectric materials as a mean of mechanical stimulation of bone cells, by converse piezoelectric effect. To estimate the magnitude and the distribution of strain, finite numerical models were applied and the results were complemented with the optical tests (Electronic Speckle Pattern Interferometric Process). In this work, osteoblasts were grown on the surface of a piezoelectric material, both in static and dynamic conditions at low frequencies, and total protein, cell viability and nitric oxide measurement comparisons are presented.

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Stimulation of nitric oxide mechanotransduction in single osteoblasts using atomic force microscopy

Cited by 21 publications

References 41 publications

Shear stress activation of nitric oxide synthase and increased nitric oxide levels in human red blood cells

Shear stress activation of nitric oxide synthase and increased nitric oxide levels in human red blood cells

Effects of Simulated Microgravity on Functions of Neutrophil-like HL-60 Cells

Polymeric piezoelectric actuator substrate for osteoblast mechanical stimulation

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