Measuring the Stiffness of &lt;em&gt;Ex Vivo&lt;/em&gt; Mouse Aortas Using Atomic Force Microscopy

Bae, Yongho; Liu, Shulin; Byfield, Fitzroy J.; Janmey, Paul A.; Assoian, Richard K.

doi:10.3791/54630

Cited by 18 publications

(17 citation statements)

References 32 publications

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“…AFM Measurement for Arterial Stiffness : The biomechanical properties of arteries from ApoE −/− mice were measured on the aortic arch ex vivo using AFM (Dimension Icon, Bruker) as described previously 26b,46. Briefly, mice were anesthetized and perfused with PBS for 10 min after treatment.…”

Section: Methodsmentioning

confidence: 99%

Surface Engineered Polymersomes for Enhanced Modulation of Dendritic Cells During Cardiovascular Immunotherapy

Zhang

Sangji

et al. 2019

Adv Funct Materials

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The principle cause of cardiovascular disease (CVD) is atherosclerosis, a chronic inflammatory condition characterized by immunologically complex fatty lesions within the intima of arterial vessel walls. Dendritic cells (DCs) are key regulators of atherosclerotic inflammation, with mature DCs generating pro-inflammatory signals within vascular lesions and tolerogenic DCs eliciting atheroprotective cytokine profiles and regulatory T-cell (Treg) activation. Here, the surface chemistry and morphology of synthetic nanocarriers composed of poly(ethylene glycol)-bpoly(propylene sulfide) copolymers to enhance the targeted modulation of DCs by transporting the anti-inflammatory agent 1,25-dihydroxyvitamin D3-(aVD) and ApoB-100-derived antigenic peptide P210 are engineered. Polymersomes decorated with an optimized surface display and density for a lipid construct of the P-D2 peptide, which binds CD11c on the DC surface, significantly enhance the cytosolic delivery and resulting immunomodulatory capacity of aVD in vitro. Weekly lowdose intravenous administration of DC-targeted, aVD-loaded polymersomes significantly inhibit atherosclerotic lesion development in high-fat-diet-fed ApoE −/− mice. The results validate the key role of DC immunomodulation during aVD-dependent inhibition of atherosclerosis and demonstrate the therapeutic enhancement and dosage lowering capability of cell-targeted nanotherapy in the treatment of CVD.

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Section: Methodsmentioning

confidence: 99%

Surface Engineered Polymersomes for Enhanced Modulation of Dendritic Cells During Cardiovascular Immunotherapy

Zhang

Sangji

et al. 2019

Adv Funct Materials

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“…AFM was used to measure intracellular stiffness as previously described [1, 2, 30]. Briefly, to measure intracellular stiffness, MEFs cultured on soft and stiff polyacrylamide hydrogels in phenol red-free DMEM with 10% FBS were indented with a silicon nitride cantilever (Bruker; spring constant, 0.06 N/m) with a conical AFM tip (40 nm in diameter).…”

Section: Methods Detailsmentioning

confidence: 99%

Mechanosensitive Expression of Lamellipodin Governs Cell Cycle Progression and Intracellular Stiffness

Brazzo

Lee

Bae

2020

Preprint

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Cells exhibit pathological behaviors in response to increased extracellular matrix (ECM) stiffness, including accelerated cell proliferation and migration, which are correlated with increased intracellular stiffness and tension. The biomechanical signal transduction of ECM stiffness into relevant molecular signals and resultant cellular processes is mediated through multiple proteins associated with the actin cytoskeleton in lamellipodia. However, the molecular mechanisms by which lamellipodial dynamics regulate cellular responses to ECM stiffening remain unclear. Previous work described that lamellipodin, a phosphoinositide- and actin filament-binding protein that is known mostly for controlling cell migration, promotes ECM stiffness-mediated early cell cycle progression, revealing a potential commonality between the mechanisms controlling stiffness-dependent cell migration and those controlling cell proliferation. However, i) whether and how ECM stiffness affects the levels of lamellipodin expression and ii) whether stiffness-mediated lamellipodin expression is required throughout cell cycle progression and for intracellular stiffness have not been explored. Here, we show that the levels of lamellipodin expression in cells are significantly increased by a stiff ECM and that this stiffness-mediated lamellipodin upregulation persistently stimulates cell cycle progression and intracellular stiffness throughout the cell cycle, from the early G1 phase to M phase. Finally, we show that both Rac activation and intracellular stiffening are required for the mechanosensitive induction of lamellipodin. More specifically, inhibiting Rac1 activation in cells on stiff ECM reduces the levels of lamellipodin expression, and this effect is reversed by the overexpression of activated Rac1 in cells on soft ECM. We thus propose that lamellipodin is a critical molecular lynchpin in the control of mechanosensitive cell cycle progression and intracellular stiffness.

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“…Moreover, ECM stiffness is required for capillary maintenance and is related to vascular pathology [ 132 ]. Data on blood vessel stiffness largely differ depending on the method used, and can vary by several orders of magnitude [ 133 , 134 , 135 ], also depending on whether the whole vessel or only the subendothelial layers were analyzed [ 136 , 137 ]. It clearly differs between arterial and venous ECs, between species and between material of different age [ 138 , 139 ].…”

Section: Tgfβ/bmp Signaling Crosstalk At Focal Adhesions With Extrmentioning

confidence: 99%

It Takes Two to Tango: Endothelial TGFβ/BMP Signaling Crosstalk with Mechanobiology

Hiepen

Mendez

Knaus

2020

Cells

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Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-beta (TGFβ) superfamily of cytokines. While some ligand members are potent inducers of angiogenesis, others promote vascular homeostasis. However, the precise understanding of the molecular mechanisms underlying these functions is still a growing research field. In bone, the tissue in which BMPs were first discovered, crosstalk of TGFβ/BMP signaling with mechanobiology is well understood. Likewise, the endothelium represents a tissue that is constantly exposed to multiple mechanical triggers, such as wall shear stress, elicited by blood flow or strain, and tension from the surrounding cells and to the extracellular matrix. To integrate mechanical stimuli, the cytoskeleton plays a pivotal role in the transduction of these forces in endothelial cells. Importantly, mechanical forces integrate on several levels of the TGFβ/BMP pathway, such as receptors and SMADs, but also global cell-architecture and nuclear chromatin re-organization. Here, we summarize the current literature on crosstalk mechanisms between biochemical cues elicited by TGFβ/BMP growth factors and mechanical cues, as shear stress or matrix stiffness that collectively orchestrate endothelial function. We focus on the different subcellular compartments in which the forces are sensed and integrated into the TGFβ/BMP growth factor signaling.

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Measuring the Stiffness of <em>Ex Vivo</em> Mouse Aortas Using Atomic Force Microscopy

Cited by 18 publications

References 32 publications

Surface Engineered Polymersomes for Enhanced Modulation of Dendritic Cells During Cardiovascular Immunotherapy

Surface Engineered Polymersomes for Enhanced Modulation of Dendritic Cells During Cardiovascular Immunotherapy

Mechanosensitive Expression of Lamellipodin Governs Cell Cycle Progression and Intracellular Stiffness

It Takes Two to Tango: Endothelial TGFβ/BMP Signaling Crosstalk with Mechanobiology

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