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Cellular and Molecular Bioengineering

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https://www.readbyqxmd.com/read/28163790/hypertrophy-changes-3d-shape-of-hipsc-cardiomyocytes-implications-for-cellular-maturation-in-regenerative-medicine
#1
Cassady E Rupert, Heidi H Chang, Kareen L K Coulombe
Advances in the use of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes for heart regeneration and in vitro disease models demand a greater understanding of how these cells grow and mature in 3-dimensional space. In this study, we developed an analysis methodology of single cardiomyocytes plated on 2D surfaces to assess their 3D myofilament volume and its z-height distribution, or shape, upon hypertrophic stimulation via phenylephrine (PE) treatment or long-term culture ("aging"). Cardiomyocytes were fixed and labeled with α-actinin for confocal microscopy imaging to obtain z-stacks for 3D myofilament volume analysis...
February 2017: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28138348/cholesterol-enrichment-impairs-capacitative-calcium-entry-enos-phosphorylation-shear-stress-induced-no-production
#2
Allison M Andrews, Tenderano T Muzorewa, Kelly A Zaccheo, Donald G Buerk, Dov Jaron, Kenneth A Barbee
Endothelial dysfunction, characterized by decreased production or availability of nitric oxide (NO), is widely believed to be the hallmark of early-stage atherosclerosis. In addition, hypercholesterolemia is considered a major risk factor for development of atherosclerosis and is associated with impaired flow-induced dilation. However, the mechanism by which elevated cholesterol levels leads to decreased production of NO is unclear. NO is released in response to shear stress and agonist-evoked changes in intracellular calcium...
February 2017: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28239428/improvement-in-electrotransfection-of-cells-using-carbon-based-electrodes
#3
Chun-Chi Chang, Mao Mao, Yang Liu, Mina Wu, Tuan Vo-Dinh, Fan Yuan
Electrotransfection has been widely used as a versatile, non-viral method for gene delivery. However, electrotransfection efficiency (eTE) is still low and unstable, compared to viral methods. To understand potential mechanisms of the unstable eTE, we investigated effects of electrode materials on eTE and viability of mammalian cells. Data from the study showed that commonly used metal electrodes generated a significant amount of particles during application of pulsed electric field, which could cause precipitation of plasmid DNA from solutions, thereby reducing eTE...
December 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28220141/adhesive-peptide-sequences-regulate-valve-interstitial-cell-adhesion-phenotype-and-extracellular-matrix-deposition
#4
Yan Wu, K Jane Grande-Allen, Jennifer L West
Knowledge of how extracellular matrix (ECM) binding impacts valve interstitial cells (VICs) is critical not only to better understanding the etiology of valvular diseases but also to constructing living valve substitutes that can grow and remodel. Use of ECM-mimicking adhesive peptides with specific affinity to different receptors provides insights into adhesion-mediated cell signaling and downstream outcomes. Expression of adhesion receptors by VICs was assessed by flow cytometry and used to guide the choice of peptides studied...
December 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28083075/biorheology-of-platelet-activation-in-the-bloodstream-distal-to-thrombus-formation
#5
Jevgenia Zilberman-Rudenko, Asako Itakura, Jeevan Maddala, Sandra M Baker-Groberg, Ralf Vetter, Erik I Tucker, András Gruber, Christoph Gerdes, Owen J T McCarty
Thrombus growth at the site of vascular injury is mediated by the sequential events of platelet recruitment, activation and aggregation concomitant with the initiation of the coagulation cascade, resulting in local thrombin generation and fibrin formation. While the biorheology of a localized thrombus formation has been well studied, it is unclear whether local sites of thrombin generation propagate platelet activation within the bloodstream. In order to study the physical biology of platelet activation downstream of sites of thrombus formation, we developed a platform to measure platelet activation and microaggregate formation in the bloodstream...
December 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27818713/mechanism-of-enhanced-cellular-uptake-and-cytosolic-retention-of-mk2-inhibitory-peptide-nano-polyplexes
#6
Kameron V Kilchrist, Brian C Evans, Colleen M Brophy, Craig L Duvall
Electrostatic complexation of a cationic MAPKAP kinase 2 inhibitory (MK2i) peptide with the anionic, pH-responsive polymer poly(propylacrylic acid) (PPAA) yields MK2i nano-polyplexes (MK2i-NPs) that significantly increase peptide uptake and intracellular retention. This study focused on elucidating the mechanism of MK2i-NP cellular uptake and intracellular trafficking in vascular smooth muscle cells. Small molecule inhibition of various endocytic pathways showed that MK2i-NP cellular uptake involves both macropinocytosis and clathrin mediated endocytosis, whereas the free peptide exclusively utilizes clathrin mediated endocytosis for cell entry...
September 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27800035/oncogene-knockdown-via-active-loading-of-small-rnas-into-extracellular-vesicles-by-sonication
#7
Tek N Lamichhane, Anjana Jeyaram, Divya B Patel, Babita Parajuli, Natalie K Livingston, Navein Arumugasaamy, John S Schardt, Steven M Jay
Extracellular vesicles (EVs), including exosomes and microvesicles, have emerged as promising drug delivery vehicles for small RNAs (siRNA and miRNA) due to their natural role in intercellular RNA transport. However, the application of EVs for therapeutic RNA delivery may be limited by loading approaches that can induce cargo aggregation or degradation. Here, we report the use of sonication as a means to actively load functional small RNAs into EVs. Conditions under which EVs could be loaded with small RNAs with minimal detectable aggregation were identified, and EVs loaded with therapeutic siRNA via sonication were observed to be taken up by recipient cells and capable of target mRNA knockdown leading to reduced protein expression...
September 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27795742/mechanisms-of-reduced-astrocyte-surface-coverage-in-cortical-neuron-glia-co-cultures-on-nanoporous-gold-surfaces
#8
Christopher A R Chapman, Hao Chen, Marianna Stamou, Pamela J Lein, Erkin Seker
Nanoporous gold (np-Au) is a promising multifunctional material for neural electrodes. We have previously shown that np-Au nanotopography reduces astrocyte surface coverage (linked to undesirable gliosis) while maintaining high neuronal coverage in a cortical primary neuron-glia co-culture model as long as two weeks in vitro. Here, we investigate the potential influence of secreted soluble factors from cells grown on np-Au on the cell type-specific surface coverage of cells grown on conventional tissue culture plastic and test the hypothesis that secretion of factors is responsible for inhibiting astrocyte coverage on np-Au...
September 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27752289/mechanical-properties-of-the-tumor-stromal-microenvironment-probed-in-vitro-and-ex-vivo-by-in-situ-calibrated-optical-trap-based-active-microrheology
#9
Jack R Staunton, Wilfred Vieira, King Leung Fung, Ross Lake, Alexus Devine, Kandice Tanner
One of the hallmarks of the malignant transformation of epithelial tissue is the modulation of stromal components of the microenvironment. In particular, aberrant extracellular matrix (ECM) remodeling and stiffening enhances tumor growth and survival and promotes metastasis. Type I collagen is one of the major ECM components. It serves as a scaffold protein in the stroma contributing to the tissue's mechanical properties, imparting tensile strength and rigidity to tissues such as those of the skin, tendons, and lungs...
September 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28163791/mechanobiology-of-chromatin-and-the-nuclear-interior
#10
Stephen T Spagnol, Travis J Armiger, Kris Noel Dahl
The view of the cell nucleus has evolved from an isolated, static organelle to a dynamic structure integrated with other mechanical elements of the cell. Both dynamics and integration appear to contribute to a mechanical regulation of genome expression. Here, we review physical structures inside the nucleus at different length scales and the dynamic reorganization modulated by cellular forces. First, we discuss nuclear organization focusing on self-assembly and disassembly of DNA structures and various nuclear bodies...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27990180/the-effect-of-rgd-peptide-on-2d-and-miniaturized-3d-culture-of-hepm-cells-mscs-and-adscs-with-alginate-hydrogel
#11
Jenna Dumbleton, Pranay Agarwal, Haishui Huang, Nathaniel Hogrebe, Renzhi Han, Keith J Gooch, Xiaoming He
Advancements in tissue engineering require the development of new technologies to study cell behavior in vitro. This study focuses on stem cell behavior within various miniaturized three-dimensional (3D) culture conditions of alginate biomaterials modified with the Arg-Gly-Asp (RGD) peptide known for its role in cell adhesion/attachment. Human embryonic palatal mesenchyme (HEPM) cells, bone marrow derived mesenchymal stem cells (MSCs), and human adipose derived stem cells (ADSCs) were cultured on a flat hydrogel of different concentrations of alginate-RGD, and in the miniaturized 3D core of microcapsules with either a 2% alginate or 2% alginate-RGD shell...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27570565/nuclear-lamins-in-cancer
#12
Jerome Irianto, Charlotte R Pfeifer, Irena L Ivanovska, Joe Swift, Dennis E Discher
Dysmorphic nuclei are commonly seen in cancers and provide strong motivation for studying the main structural proteins of nuclei, the lamins, in cancer. Past studies have also demonstrated the significance of microenvironment mechanics to cancer progression, which is extremely interesting because the lamina was recently shown to be mechanosensitive. Here, we review current knowledge relating cancer progression to lamina biophysics. Lamin levels can constrain cancer cell migration in 3D and thereby impede tumor growth, and lamins can also protect a cancer cell's genome...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27499815/lincing-defective-nuclear-cytoskeletal-coupling-and-dyt1-dystonia
#13
Cosmo A Saunders, G W Gant Luxton
Mechanical forces generated by nuclear-cytoskeletal coupling through the LINC (linker of nucleoskeleton and cytoskeleton) complex, an evolutionarily conserved molecular bridge in the nuclear envelope (NE), are critical for the execution of wholesale nuclear positioning events in migrating and dividing cells, chromosome dynamics during meiosis, and mechanotransduction. LINC complexes consist of outer (KASH (Klarsicht, ANC-1, and Syne homology)) and inner (SUN (Sad1, UNC-84)) nuclear membrane proteins. KASH proteins interact with the cytoskeleton in the cytoplasm and SUN proteins in the perinuclear space of the NE...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27453760/a-case-for-the-nuclear-membrane-as-a-mechanotransducer
#14
Balázs Enyedi, Philipp Niethammer
The cell nucleus is becoming increasingly recognized as a mechanosensitive organelle. Most research on nuclear mechanosignaling focuses on the nuclear lamina and coupled actin structures. In this commentary, we discuss the possibility that the nuclear membrane senses and transduces mechanical signals similar to the plasma membrane. We briefly summarize possible (i) pathophysiological sources of nuclear membrane tension, (ii) features that render nuclear membranes particularly suited for mechanotransduction, and (iii) molecular sensing mechanisms...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27330571/an-unresolved-linc-in-the-nuclear-envelope
#15
Mehdi Torbati, Tanmay P Lele, Ashutosh Agrawal
The nuclear envelope segregates the nucleoplasm from the cytoplasm and is a key feature of eukaryotic cells. Nuclear envelope architecture is comprised of two concentric membrane shells which fuse at multiple sites and yet maintain a uniform separation of 30-50 nm over the rest of the membrane. Studies have revealed the roles for numerous nuclear proteins in forming and maintaining the architecture of the nuclear envelope. However, there is a lack of consensus on the fundamental forces and physical mechanisms that establish the geometry...
June 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28042345/biomimetic-microstructure-morphology-in-electrospun-fiber-mats-is-critical-for-maintaining-healthy-cardiomyocyte-phenotype
#16
Rutwik Rath, Jung Bok Lee, Truc-Linh Tran, Sean F Lenihan, Cristi L Galindo, Yan Ru Su, Tarek Absi, Leon M Bellan, Douglas B Sawyer, Hak-Joon Sung
Despite recent advances in biomimetic substrates, there is still only limited understanding of how the extracellular matrix (ECM) functions in the maintenance of cardiomyocyte (CM) phenotype. In this study, we designed electrospun substrates inspired by morphologic features of non-failing and failing human heart ECM, and examined how these substrates regulate phenotypes of adult and neonatal rat ventricular CMs (ARVM and NRVM, respectively). We found that poly(ε-caprolactone) fiber substrates designed to mimic the organized ECM of a non-failing human heart maintained healthy CM phenotype (evidenced by cell morphology, organized actin/myomesin bands and expression of β-MYH7 and SCN5A...
March 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27114743/effects-of-intermittent-and-incremental-cyclic-stretch-on-erk-signaling-and-collagen-production-in-engineered-tissue
#17
Jillian B Schmidt, Kelley Chen, Robert T Tranquillo
Intermittent cyclic stretching and incrementally increasing strain amplitude cyclic stretching were explored to overcome the reported adaptation of fibroblasts in response to constant amplitude cyclic stretching, with the goals of accelerating collagen production and understanding the underlying cell signaling. The effects of constant amplitude, intermittent, and incremental cyclic stretching regimens were investigated for dermal fibroblasts entrapped in a fibrin gel by monitoring the extracellular signal-regulated kinase (ERK1/2) and p38 pathways, collagen transcription, and finally the deposited collagen protein...
March 1, 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27110295/physical-and-chemical-signals-that-promote-vascularization-of-capillary-scale-channels
#18
Raleigh M Linville, Nelson F Boland, Gil Covarrubias, Gavrielle M Price, Joe Tien
Proper vascularization remains critical to the clinical application of engineered tissues. To engineer microvessels in vitro, we and others have delivered endothelial cells through preformed channels into patterned extracellular matrix-based gels. This approach has been limited by the size of endothelial cells in suspension, and results in plugging of channels below ~30 μm in diameter. Here, we examine physical and chemical signals that can augment direct seeding, with the aim of rapidly vascularizing capillary-scale channels...
March 1, 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27087859/neurogenic-potential-of-engineered-mesenchymal-stem-cells-overexpressing-vegf
#19
Alan J Man, Gregory Kujawski, Travis S Burns, Elaine N Miller, Fernando A Fierro, J Kent Leach, Peter Bannerman
Numerous signaling molecules are altered following nerve injury, serving as a blueprint for drug delivery approaches that promote nerve repair. However, challenges with achieving the appropriate temporal duration of recombinant protein delivery have limited the therapeutic success of this approach. Genetic engineering of mesenchymal stem cells (MSCs) to enhance the secretion of proangiogenic molecules such as vascular endothelial growth factor (VEGF) may provide an alternative. We hypothesized that the administration of VEGF-expressing human MSCs would stimulate neurite outgrowth and proliferation of cell-types involved in neural repair...
March 1, 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27042237/effect-of-pseudopod-extensions-on-neutrophil-hemodynamic-transport-near-a-wall
#20
Anne D Rocheleau, Weiwei Wang, Michael R King
During inflammation, circulating neutrophils roll on, and eventually tether to, the endothelial lining of blood vessels, allowing them to exit the bloodstream and enter the surrounding tissue to target pathogens. This process is mediated by the selectin family of adhesion proteins expressed by endothelial cells. Interestingly, only 10% of activated, migrating neutrophils transmigrate into the extravascular space; the other 90% detach from the wall and rejoin the blood flow. Neutrophils extrude pseudopods during the adhesion cascade; however, the transport behavior of this unique cell geometry has not been previously addressed...
March 1, 2016: Cellular and Molecular Bioengineering
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