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Cellular and Molecular Bioengineering

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https://www.readbyqxmd.com/read/27818713/mechanism-of-enhanced-cellular-uptake-and-cytosolic-retention-of-mk2-inhibitory-peptide-nano-polyplexes
#1
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
#2
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
#3
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
#4
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/27570565/nuclear-lamins-in-cancer
#5
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
#6
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
#7
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
#8
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/27114743/effects-of-intermittent-and-incremental-cyclic-stretch-on-erk-signaling-and-collagen-production-in-engineered-tissue
#9
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
#10
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
#11
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
#12
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
https://www.readbyqxmd.com/read/27042236/poly-ethylene-glycol-hydrogel-scaffolds-containing-cell-adhesive-and-protease-sensitive-peptides-support-microvessel-formation-by-endothelial-progenitor-cells
#13
Erica B Peters, Nicolas Christoforou, Kam W Leong, George A Truskey, Jennifer L West
The development of stable, functional microvessels remains an important obstacle to overcome for tissue engineered organs and treatment of ischemia. Endothelial progenitor cells (EPCs) are a promising cell source for vascular tissue engineering as they are readily obtainable and carry the potential to differentiate towards all endothelial phenotypes. The aim of this study was to investigate the ability of human umbilical cord blood-derived EPCs to form vessel-like structures within a tissue engineering scaffold material, a cell-adhesive and proteolytically degradable poly(ethylene glycol) (PEG) hydrogel...
March 1, 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/26958093/local-influence-of-cell-viability-on-stretch-induced-permeability-of-alveolar-epithelial-cell-monolayers
#14
M J Song, C I Davis, G G Lawrence, S S Margulies
Ventilator induced lung injury (VILI), often attributed to over-distension of the alveolar epithelial cell layer, can trigger loss of barrier function. Alveolar epithelial cell monolayers can be used as an idealized in vitro model of the pulmonary epithelium, with cell death and tight junction disruption and permeability employed to estimate stretch-induced changes in barrier function. We adapted a method published for vascular endothelial permeability, compare its sensitivity with our previously published method, and determine the relationship between breeches in barrier properties after stretch and regions of cell death After 4-5 days in culture, primary rat alveolar epithelial cells seeded on plasma treated polydimethylsiloxane membrane coated with biotin-labeled fibronectin, or fibronectin alone were stretched in the presence of FITC-tagged streptavidin (biotin-labeled membrane) or BODIPY-ouabain...
March 1, 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/26951764/modeling-the-progression-of-epithelial-leak-caused-by-overdistension
#15
Katharine L Hamlington, Baoshun Ma, Bradford J Smith, Jason H T Bates
Mechanical ventilation is necessary for treatment of the acute respiratory distress syndrome but leads to overdistension of the open regions of the lung and produces further damage. Although we know that the excessive stresses and strains disrupt the alveolar epithelium, we know little about the relationship between epithelial strain and epithelial leak. We have developed a computational model of an epithelial monolayer to simulate leak progression due to overdistension and to explain previous experimental findings in mice with ventilator-induced lung injury...
March 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27547269/targeted-programming-of-the-lymph-node-environment-causes-evolution-of-local-and-systemic-immunity
#16
James I Andorko, Joshua M Gammon, Lisa H Tostanoski, Qin Zeng, Christopher M Jewell
Biomaterial vaccines offer cargo protection, targeting, and co-delivery of signals to immune organs such as lymph nodes (LNs), tissues that coordinate adaptive immunity. Understanding how individual vaccine components impact immune response has been difficult owing to the systemic nature of delivery. Direct intra-lymph node (i.LN.) injection offers a unique opportunity to dissect how the doses, kinetics, and combinations of signals reaching LNs influence the LN environment. Here, i.LN. injection was used as a tool to study the local and systemic responses to vaccines comprised of soluble antigen and degradable polymer particles encapsulating toll-like receptor agonists as adjuvants...
2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27547268/predictive-model-of-lymphocyte-specific-protein-tyrosine-kinase-lck-autoregulation
#17
Jennifer A Rohrs, Pin Wang, Stacey D Finley
Lymphocyte-specific protein tyrosine kinase (LCK) is a key activator of T cells; however, little is known about the specific autoregulatory mechanisms that control its activity. We have constructed a model of LCK autophosphorylation and phosphorylation by the regulating kinase CSK. The model was fit to existing experimental data in the literature that presents an in vitro reconstituted membrane system, which provides more physiologically relevant kinetic measurements than traditional solution-based systems...
2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/27335609/roles-of-cross-membrane-transport-and-signaling-in-the-maintenance-of-cellular-homeostasis
#18
Inchul Cho, Mark R Jackson, Joe Swift
Organelles allow specialized functions within cells to be localized, contained and independently regulated. This separation is oftentimes achieved by selectively permeable membranes, which enable control of molecular transport, signaling between compartments and containment of stress-inducing factors. Here we consider the role of a number of membrane systems within the cell: the plasma membrane, that of the endoplasmic reticulum, and then focusing on the nucleus, depository for chromatin and regulatory centre of the cell...
2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/26900408/heading-in-the-right-direction-understanding-cellular-orientation-responses-to-complex-biophysical-environments
#19
Chiara Tamiello, Antonetta B C Buskermolen, Frank P T Baaijens, Jos L V Broers, Carlijn V C Bouten
The aim of cardiovascular regeneration is to mimic the biological and mechanical functioning of tissues. For this it is crucial to recapitulate the in vivo cellular organization, which is the result of controlled cellular orientation. Cellular orientation response stems from the interaction between the cell and its complex biophysical environment. Environmental biophysical cues are continuously detected and transduced to the nucleus through entwined mechanotransduction pathways. Next to the biochemical cascades invoked by the mechanical stimuli, the structural mechanotransduction pathway made of focal adhesions and the actin cytoskeleton can quickly transduce the biophysical signals directly to the nucleus...
2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/26900407/fibronectin-mechanobiology-regulates-tumorigenesis
#20
Karin Wang, Bo Ri Seo, Claudia Fischbach, Delphine Gourdon
Fibronectin (Fn) is an essential extracellular matrix (ECM) glycoprotein involved in both physiological and pathological processes. The structure-function relationship of Fn has been and is still being studied, as changes in its molecular structure are integral in regulating (or dysregulating) its biological activities via its cell, matrix component, and growth factor binding sites. Fn comprises three types of repeating modules; among them, FnIII modules are mechanically unstable domains that may be extended/unfolded upon cell traction and either uncover cryptic binding sites or disrupt otherwise exposed binding sites...
2016: Cellular and Molecular Bioengineering
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