Glycated Collagen Impairs Endothelial Cell Response to Cyclic Stretch

Figueroa, Dannielle Solomon; Kemeny, Steven Frank; Clyne, Alisa Morss

doi:10.1007/s12195-011-0176-9

Cited by 11 publications

(23 citation statements)

References 77 publications

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“…Collagen glycation was vali dated by increased fluorescence intensity and decreased separation via sodium dodecyl sulfate-polycrylamide gel electrophoresis (SDS-PAGE) [31]. In brief, for SDS-PAGE the samples were digested with 7.5 u/ml collagenase prior to separation.…”

Section: Cs Applicationmentioning

confidence: 99%

“…Native or glycated collagen was covalently bonded onto polydimethylsiloxane (PDMS, Sylgard 184, Dow Coming) as previously described [31,34]. In brief, plasma treated PDMS was incubated with 10% 3-aminopropyltriethoxysilane in ethanol, heated to 60 °C for 60 min, incubated with 3% glutaraldehyde in PBS for 20 min, and finally coated with native or glycated collagen (100 /tg/ml) for 3h.…”

Section: Cs Applicationmentioning

confidence: 99%

“…A custom-built stretching device [31] was used to apply 10% linear CS, 0.5 Hz to PAEC on both native and glycated collagen coated membranes in supplemented medium for 0-24 h. In the device, PDMS membranes were chemically bonded on both ends to polystyrene clamps, which were then attached to a fixed cross bar on one end and a moveable crossbar on the other end. The moveable crossbar was attached to a DC motor via a linear slider (Specialty Motions), which allowed smooth translation from rota tional to linear motion.…”

Section: Cs Applicationmentioning

confidence: 99%

“…Samples were then incubated with a goat antimouse AlexaFluor 488 secondary antibody (Invitrogen, 1:200 dilution, 1 h at room temperature) and imaged using an Olympus 1X81 inverted confocal microscope with a 60 x or lOOx oil-immersion objective. Images were taken at four loca tions in the central region, which was previously validated to experience uniform strain [31]. Fibronectin fiber alignment was quantified using an edge detection method developed in our labo ratory [35].…”

Section: Fibronectin and Actin Fiber Alignment Quantificationmentioning

confidence: 99%

“…We recently showed that glycated collagen inhibits endothelial cell actin alignment and FAK activation in response to shear stress and cyclic strain [31,32]. Since matrix remodeling depends on actin tension, we hypothesized that ECM protein remodeling would decrease in endothelial cells exposed to CS on glycated collagen.…”

Section: Introductionmentioning

confidence: 99%

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Glycated Collagen Decreased Endothelial Cell Fibronectin Alignment in Response to Cyclic Stretch Via Interruption of Actin Alignment

Figueroa

Kemeny

Clyne

2014

Journal of Biomechanical Engineering

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Hyperglycemia is a defining characteristic of diabetes, and uncontrolled blood glucose in diabetes is associated with accelerated cardiovascular disease. Chronic hyperglycemia glycates extracellular matrix (ECM) collagen, which can lead to endothelial cell dysfunction. In healthy conditions, endothelial cells respond to mechanical stimuli such as cyclic stretch (CS) by aligning their actin cytoskeleton. Other cell types, specifically fibroblasts, align their ECM in response to CS. We previously demonstrated that glycated collagen inhibits endothelial cell actin alignment in response to CS. The aim of this study was to determine the effect of glycated collagen on ECM remodeling and protein alignment in response to stretch. Porcine aortic endothelial cells (PAEC) seeded on native or glycated collagen coated elastic substrates were exposed to 10% CS. Cells on native collagen aligned subcellular fibronectin fibers in response to stretch, whereas cells on glycated collagen did not. The loss of fibronectin alignment was due to inhibited actin alignment in response to CS, since fibronectin alignment did not occur in cells on native collagen when actin alignment was inhibited with cytochalasin. Further, while ECM protein content did not change in cells on native or glycated collagen in response to CS, degradation activity decreased in cells on glycated collagen. Matrix metalloproteinase 2 (MMP-2) and membrane-associated type 1 matrix metalloproteinase (MT1-MMP) protein levels decreased, and therefore MMP-2 activity also decreased. These MMP changes may relate to c-Jun N-terminal kinase (Jnk) phosphorylation inhibition with CS, which has previously been linked to focal adhesion kinase (FAK). These data demonstrate the importance of endothelial cell actin tension in remodeling and aligning matrix proteins in response to mechanical stimuli, which is critical to vascular remodeling in health and disease.

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Section: Cs Applicationmentioning

confidence: 99%

Section: Cs Applicationmentioning

confidence: 99%

Section: Cs Applicationmentioning

confidence: 99%

Section: Fibronectin and Actin Fiber Alignment Quantificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Glycated Collagen Decreased Endothelial Cell Fibronectin Alignment in Response to Cyclic Stretch Via Interruption of Actin Alignment

Figueroa

Kemeny

Clyne

2014

Journal of Biomechanical Engineering

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Tuning of Collagen Scaffold Properties Modulates Embedded Endothelial Cell Regulatory Phenotype in Repair of Vascular Injuries In Vivo

Unterman

Freiman

Beckerman

et al. 2015

Adv Healthcare Materials

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Perivascularly implantated matrix embedded endothelial cells (MEECs) are potent regulators of inflammation and intimal hyperplasia following vascular injuries. ECs in porous collagen scaffolds adopt a reparative phenotype with significant therapeutic potential. Although the biological effects of MEECs are increasingly understood, less work has studied the tuning of scaffold properties to control cell-substrate interactions and subsequent biological outcomes. We hypothesized that modulating scaffold degradation would change EC phenotype. Scaffolds with differential degradation rates were prepared by crosslinking and pre-degradation. Vascular injury increased degradation and the presence of MEECs retarded injury-mediated degradation. MEECs responded to differential scaffold properties with altered cell viability in vivo, suppression of smooth muscle cell (SMC) proliferation in vitro, and changes in IL-6 and MMP-9 expression. When implanted perivascularly to a murine carotid wire injury model, tuned scaffolds changed MEEC effects on vascular repair and inflammation. Live animal imaging allowed for real-time tracking of cell viability, inflammation, and scaffold degradation in vivo, affording an unprecedented understanding of interactions between cells, substrate, and tissue. MEEC-treated injuries exhibited improved endothelialization and reduced SMC hyperplasia over 14 days. These data demonstrate the potent role material design plays in tuning MEEC efficacy in vivo, with implications for the design of clinical therapies.

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Pre‐glycation impairs gelation of high concentration collagen solutions

Diamantides¹,

Slyker

Martin

et al. 2022

J Biomedical Materials Res

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There remains a need for stiffer collagen hydrogels for tissue engineering and disease modeling applications. Pre‐glycation, or glycation of collagen in solution prior to gelation, has been shown to increase the mechanics of collagen hydrogels while maintaining high viability of encapsulated cells. The stiffness of glycated collagen gels can be increased by increasing the collagen concentration, sugar concentration, and glycation time. However, previous studies on pre‐glycation of collagen have used low collagen concentrations and/or low sugar concentrations and have not investigated the effect of glycation time. Therefore, the objective of this study was to determine the effects of pre‐glycation with high sugar concentrations (up to 500 mM) and extended glycation times (up to 21 days) on high concentration collagen (8 mg/ml). The addition of sugar to the collagen and the formation of advanced glycation end products (AGEs) were quantified. The ability to gel successfully and rheological properties were determined and correlated with biochemical characterizations. Successful collagen gelation and rheological properties of pre‐glycated collagen were found to be strongly dependent on the ratio of added sugars to added AGEs with high ratios impairing gelation and low ratios resulting in optimal storage moduli. There is likely a competing effect during pre‐glycation of the formation of AGEs resulting in crosslinking of collagen and the formation of Amadori intermediates acting to increase collagen solubility. Overall, this study shows that collagen glycation can be optimized by increasing the formation of AGEs while maintaining a low ratio of added sugar to added AGEs.

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Glycated Collagen Impairs Endothelial Cell Response to Cyclic Stretch

Cited by 11 publications

References 77 publications

Glycated Collagen Decreased Endothelial Cell Fibronectin Alignment in Response to Cyclic Stretch Via Interruption of Actin Alignment

Glycated Collagen Decreased Endothelial Cell Fibronectin Alignment in Response to Cyclic Stretch Via Interruption of Actin Alignment

Tuning of Collagen Scaffold Properties Modulates Embedded Endothelial Cell Regulatory Phenotype in Repair of Vascular Injuries In Vivo

Pre‐glycation impairs gelation of high concentration collagen solutions

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