Abstract P447: Layer-by-layer Fabrication Of Thick Cardiac Muscle Patches With High Viability And Maturity

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Lu Wang ◽  
Jianyi Zhang
Keyword(s):  
Cardiac Muscle ◽  
Calcium Handling ◽  
Tunel Staining ◽  
Global Alignment ◽  
Layer By Layer ◽  
Intercalated Discs ◽  
Transmission Electron ◽  
High Viability ◽  
A Cell ◽  
Induced Pluripotent

Conventionally manufactured human cardiac muscle patches (hCMPs) typically rely on diffusion for oxygen and nutrient delivery to the cells and, consequently, the thickness of the hCMP is limited to 100-200 μm. Here, we tested our hypothesis that our layer-by-layer (lbl) manufacturing protocol could increase hCMP thickness without compromising the viability of individual cell populations. Human induced-pluripotent stem cells were differentiated into cardiomyocytes (CMs) or endothelial cells (ECs); then, layers composed of CMs or ECs were fabricated by mixing a cell-containing fibrinogen solution with thrombin and pouring the mixture into a mold, and the layers were stacked into a CM-EC-CM “sandwich”. hCMP viability was assessed via TUNEL staining, vascularization via expression of the EC marker CD144, and maturation via immunofluorescence and qPCR assessments of the expression of contractile (cTnT, cTnI), connective (Cx43, N-cad), and calcium-handling (RyR2, SERCA2, BIN1, Kir2.1) proteins. Sarcomere length was evaluated by staining for α-sarcomeric actinin, and hCMP ultrastructure was examined via transmission electron microscopy. The lbl-manufactured hCMP was 10 mm x 10 mm and 1.8 mm thick after fabrication. Minimal cells in the lbl-hCMP were apoptotic or necrotic. Compared to assessments in a single-layered patch containing the same number and proportion of cells, the lbl-hCMP was vascularized with higher levels of contractile, connective, and calcium-handling protein expression. Contractile proteins were also better aligned in the lbl-hCMP (deviation from global alignment 12.3 ± 3° vs. 24.3° ± 1.9°, p<0.05). The lbl-hCMP contained longer sarcomeres (2.05 ± 0.015 vs. 1.74 ± 0.026 um, p<0.05) with well-ordered I-bands, A-bands, M-lines, Z-lines, desmosomes, and intercalated discs. Thus, our lbl-manufacturing technique produced thicker and more mature patches that can be potentially clinically applicable for hearts with postinfarction LV remodeling.

Nuclear Reactions in Deuterium Titanium

1990 ◽  
Vol 48 (2) ◽  
pp. 134-135
Author(s):  
D.M. Vanderwalker
Keyword(s):  
Nuclear Reactions ◽  
Ion Beam ◽  
Exothermic Reaction ◽  
Pure Titanium ◽  
Fundamental Interest ◽  
Transmission Electron ◽  
Iodide Titanium ◽  
A Cell ◽  
After Hours ◽  
Quantity Of Heat

There is a fundamental interest in electrochemical fusion of deuterium in palladium and titanium since its supposed discovery by Fleischmann and Pons. Their calorimetric experiments reveal that a large quantity of heat is released by Pd after hours in a cell, suggesting fusion occurs. They cannot explain fusion by force arguments, nor can it be an exothermic reaction on the formation of deuterides because a smaller quantity of heat is released. This study examines reactions of deuterium in titanium.Both iodide titanium and 99% pure titanium samples were encapsulated in vacuum tubes, annealed for 2h at 800 °C. The Ti foils were charged with deuterium in a D2SO4 D2O solution at a potential of .45V with respect to a calomel reference junction. Samples were ion beam thinned for transmission electron microscopy. The TEM was performed on the JEOL 200CX.The structure of D charged titanium is α-Ti with hexagonal and fee deuterides.

Transmission Electron Microscopy characterization of epitaxial III-V semiconductor thin films grown on Si(100) by ion-assisted deposition

1990 ◽  
Vol 48 (4) ◽  
pp. 686-687
Author(s):  
L. Hultman ◽  
C.-H. Choi ◽  
R. Kaspi ◽  
R. Ai ◽  
S.A. Barnett
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Ion Irradiation ◽  
High Energy ◽  
Nucleation Mechanism ◽  
Layer By Layer ◽  
Extended Defect ◽  
Island Formation ◽  
Si Substrates ◽  
Transmission Electron

III-V semiconductor films nucleate by the Stranski-Krastanov (SK) mechanism on Si substrates. Many of the extended defects present in the films are believed to result from the island formation and coalescence stage of SK growth. We have recently shown that low (-30 eV) energy, high flux (4 ions per deposited atom), Ar ion irradiation during nucleation of III-V semiconductors on Si substrates prolongs the 1ayer-by-layer stage of SK nucleation, leading to a decrease in extended defect densities. Furthermore, the epitaxial temperature was reduced by >100°C due to ion irradiation. The effect of ion bombardment on the nucleation mechanism was explained as being due to ion-induced dissociation of three-dimensional islands and ion-enhanced surface diffusion.For the case of InAs grown at 380°C on Si(100) (11% lattice mismatch), where island formation is expected after ≤ 1 monolayer (ML) during molecular beam epitaxy (MBE), in-situ reflection high-energy electron diffraction (RHEED) showed that 28 eV Ar ion irradiation prolonged the layer-by-layer stage of SK nucleation up to 10 ML. Otherion energies maintained layer-by-layer growth to lesser thicknesses. The ion-induced change in nucleation mechanism resulted in smoother surfaces and improved the crystalline perfection of thicker films as shown by transmission electron microscopy and X-ray rocking curve studies.

scholarly journals CdSe QD Biosynthesis in Yeast Using Tryptone-Enriched Media and Their Conjugation with a Peptide Hecate for Bacterial Detection and Killing

Nanomaterials ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 1463 ◽  
Author(s):  
Vishma Pratap Sur ◽  
Marketa Kominkova ◽  
Zaneta Buchtova ◽  
Kristyna Dolezelikova ◽  
Ondrej Zitka ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Synthesis Process ◽  
Synthesis Methods ◽  
Bacterial Cells ◽  
Cdse Qds ◽  
Yeast Saccharomyces Cerevisiae ◽  
Shape Distribution ◽  
Transmission Electron ◽  
A Cell ◽  
Physical And Chemical

The physical and chemical synthesis methods of quantum dots (QDs) are generally unfavorable for biological applications. To overcome this limitation, the development of a novel “green” route to produce highly-fluorescent CdSe QDs constitutes a promising substitute approach. In the present work, CdSe QDs were biosynthesized in yeast Saccharomyces cerevisiae using a novel method, where we showed for the first time that the concentration of tryptone highly affects the synthesis process. The optimum concentration of tryptone was found to be 25 g/L for the highest yield. Different methods were used to optimize the QD extraction from yeast, and the best method was found to be by denaturation at 80 °C along with an ultrasound needle. Multiple physical characterizations including transmission electron microscopy (TEM), dynamic light scattering (DLS), energy-dispersive X-ray spectroscopy (EDX), and spectrophotometry confirmed the optical features size and shape distribution of the QDs. We showed that the novel conjugate of the CdSe QDs and a cell-penetrating peptide (hecate) can detect bacterial cells very efficiently under a fluorescent microscope. The conjugate also showed strong antibacterial activity against vancomycin-resistant Staphylococcus aureus (VRSA), methicillin-resistant Staphylococcus aureus (MRSA), and Escherichia coli, which may help us to cope with the problem of rising antibiotic resistance.

scholarly journals Mitochondria and Pharmacologic Cardiac Conditioning—At the Heart of Ischemic Injury

2021 ◽  
Vol 22 (6) ◽  
pp. 3224
Author(s):  
Christopher Lotz ◽  
Johannes Herrmann ◽  
Quirin Notz ◽  
Patrick Meybohm ◽  
Franz Kehl
Keyword(s):  
Cell Death ◽  
Mitochondrial Permeability Transition ◽  
Permeability Transition ◽  
Research Field ◽  
Calcium Handling ◽  
Control Mechanisms ◽  
Intrinsic Resistance ◽  
Atp Production ◽  
Cardiac Conditioning ◽  
A Cell

Pharmacologic cardiac conditioning increases the intrinsic resistance against ischemia and reperfusion (I/R) injury. The cardiac conditioning response is mediated via complex signaling networks. These networks have been an intriguing research field for decades, largely advancing our knowledge on cardiac signaling beyond the conditioning response. The centerpieces of this system are the mitochondria, a dynamic organelle, almost acting as a cell within the cell. Mitochondria comprise a plethora of functions at the crossroads of cell death or survival. These include the maintenance of aerobic ATP production and redox signaling, closely entwined with mitochondrial calcium handling and mitochondrial permeability transition. Moreover, mitochondria host pathways of programmed cell death impact the inflammatory response and contain their own mechanisms of fusion and fission (division). These act as quality control mechanisms in cellular ageing, release of pro-apoptotic factors and mitophagy. Furthermore, recently identified mechanisms of mitochondrial regeneration can increase the capacity for oxidative phosphorylation, decrease oxidative stress and might help to beneficially impact myocardial remodeling, as well as invigorate the heart against subsequent ischemic insults. The current review highlights different pathways and unresolved questions surrounding mitochondria in myocardial I/R injury and pharmacological cardiac conditioning.

scholarly journals Altered Electrical, Biomolecular, and Immunologic Phenotypes in a Novel Patient-Derived Stem Cell Model of Desmoglein-2 Mutant ARVC

2021 ◽  
Vol 10 (14) ◽  
pp. 3061
Author(s):  
Robert N. Hawthorne ◽  
Adriana Blazeski ◽  
Justin Lowenthal ◽  
Suraj Kannan ◽  
Roald Teuben ◽  
...  
Keyword(s):  
Stem Cell ◽  
Structural Changes ◽  
Cell Model ◽  
Arrhythmogenic Right Ventricular Cardiomyopathy ◽  
Induced Pluripotent Stem Cell ◽  
Cytokine Expression ◽  
Calcium Handling ◽  
Disease Manifestation ◽  
Time To Peak ◽  
Induced Pluripotent

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a progressive heart condition which causes fibro-fatty myocardial scarring, ventricular arrhythmias, and sudden cardiac death. Most cases of ARVC can be linked to pathogenic mutations in the cardiac desmosome, but the pathophysiology is not well understood, particularly in early phases when arrhythmias can develop prior to structural changes. Here, we created a novel human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) model of ARVC from a patient with a c.2358delA variant in desmoglein-2 (DSG2). These DSG2-mutant (DSG2Mut) hiPSC-CMs were compared against two wildtype hiPSC-CM lines via immunostaining, RT-qPCR, Western blot, RNA-Seq, cytokine expression and optical mapping. Mutant cells expressed reduced DSG2 mRNA and had altered localization of desmoglein-2 protein alongside thinner, more disorganized myofibrils. No major changes in other desmosomal proteins were noted. There was increased pro-inflammatory cytokine expression that may be linked to canonical and non-canonical NFκB signaling. Action potentials in DSG2Mut CMs were shorter with increased upstroke heterogeneity, while time-to-peak calcium and calcium decay rate were reduced. These were accompanied by changes in ion channel and calcium handling gene expression. Lastly, suppressing DSG2 in control lines via siRNA allowed partial recapitulation of electrical anomalies noted in DSG2Mut cells. In conclusion, the aberrant cytoskeletal organization, cytokine expression, and electrophysiology found DSG2Mut hiPSC-CMs could underlie early mechanisms of disease manifestation in ARVC patients.

Increased cell death in osteopontin-deficient cardiac fibroblasts occurs by a caspase-3-independent pathway

2004 ◽  
Vol 287 (4) ◽  
pp. H1730-H1739 ◽  
Author(s):  
Ron Zohar ◽  
Baoqian Zhu ◽  
Peter Liu ◽  
Jaro Sodek ◽  
C. A. McCulloch
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Cardiac Fibroblasts ◽  
Chromatin Condensation ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Wild Type ◽  
Nuclear Fragmentation ◽  
A Cell

Reperfusion-induced oxidative injury to the myocardium promotes activation and proliferation of cardiac fibroblasts and repair by scar formation. Osteopontin (OPN) is a proinflammatory cytokine that is upregulated after reperfusion. To determine whether OPN enhances fibroblast survival after exposure to oxidants, cardiac fibroblasts from wild-type (WT) or OPN-null (OPN−/−) mice were treated in vitro with H2O2to model reperfusion injury. Within 1 h, membrane permeability to propidium iodide (PI) was increased from 5 to 60% in OPN−/−cells but was increased to only 20% in WT cells. In contrast, after 1–8 h of treatment with H2O2, the percent of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-stained cells was more than twofold higher in WT than OPN−/−cells. Electron microscopy of WT cells treated with H2O2showed chromatin condensation, nuclear fragmentation, and cytoplasmic and nuclear shrinkage, which are consistent with apoptosis. In contrast, H2O2-treated OPN−/−cardiac fibroblasts exhibited cell and nuclear swelling and membrane disruption that are indicative of cell necrosis. Treatment of OPN−/−and WT cells with a cell-permeable caspase-3 inhibitor reduced the percentage of TUNEL staining by more than fourfold in WT cells but decreased staining in OPN−/−cells by ∼30%. Although the percentage of PI-permeable WT cells was reduced threefold, the percent of PI-permeable OPN−/−cells was not altered. Restoration of OPN expression in OPN−/−fibroblasts reduced the percentage of PI-permeable cells but not TUNEL staining after H2O2treatment. Thus H2O2-induced cell death in OPN-deficient cardiac fibroblasts is mediated by a caspase-3-independent, necrotic pathway. We suggest that the increased expression of OPN in the myocardium after reperfusion may promote fibrosis by protecting cardiac fibroblasts from cell death.

scholarly journals Propagation of Action Potentials and the Structure of the Nexus in Cardiac Muscle

1965 ◽  
Vol 48 (5) ◽  
pp. 797-823 ◽  
Author(s):  
L. Barr ◽  
M. M. Dewey ◽  
W. Berger
Keyword(s):  
Guinea Pig ◽  
Cardiac Muscle ◽  
Action Potentials ◽  
Structural Changes ◽  
Sucrose Solution ◽  
Electrical Coupling ◽  
Intercalated Discs ◽  
Sucrose Gap ◽  
Specialized Structure ◽  
Frog Atrium

The hypothesis that the nexus is a specialized structure allowing current flow between cell interiors is corroborated by concomitant structural changes of the nexus and changes of electrical coupling between cells due to soaking in solutions of abnormal tonicity. Fusiform frog atrial fibers are interconnected by nexuses. The nexuses, desmosomes, and regions of myofibrillar attachment of this muscle are not associated in a manner similar to intercalated discs of guinea pig cardiac muscle. Indeed, nexuses occur wherever cell membranes are closely apposed. Action potentials of frog atrial bundles detected extracellularly across a sucrose gap change from monophasic to diphasic when the gap is shunted by a resistor. This indicates that action potentials are transmitted across the gap when sufficient excitatory current is allowed to flow across the gap. When the sucrose solution in the gap is made hypertonic, propagation past the gap is blocked and the resistance between the cells in the gap increases. Electron micrographs demonstrate that the nexuses of frog atrium and guinea pig ventricle are ruptured by hypertonic solutions.

scholarly journals Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices.

1984 ◽  
Vol 99 (4) ◽  
pp. 1398-1404 ◽  
Author(s):  
C Decker ◽  
R Greggs ◽  
K Duggan ◽  
J Stubbs ◽  
A Horwitz
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Cardiac Fibroblasts ◽  
Cell Types ◽  
Extracellular Matrices ◽  
Common Denominator ◽  
Cell Matrix ◽  
Skeletal Myoblasts ◽  
A Cell ◽  
Cell Matrix Adhesion

Neff et al. (1982, J. Cell Biol., 95:654-666) have described a monoclonal antibody, CSAT, directed against a cell surface antigen that participates in the adhesion of skeletal muscle to extracellular matrices. We used the same antibody to compare and parse the determinants of adhesion and morphology on myogenic and fibrogenic cells. We report here that the antigen is present on skeletal and cardiac muscle and on tendon, skeletal, dermal, and cardiac fibroblasts; however, its contribution to their morphology and adhesion is different. The antibody produces large alterations in the morphology and adhesion of skeletal myoblasts and tendon fibroblasts; in contrast, its effects on the cardiac fibroblasts are not readily detected. The effects of CSAT on the other cell types, i.e., dermal and skeletal fibroblasts, cardiac muscle, 5-bromodeoxyuridine-treated skeletal muscle, lie between these extremes. The effects of CSAT on the skeletal myoblasts depends on the calcium concentration in the growth medium and on the culture age. We interpret these differential responses to CSAT as revealing differences in the adhesion of the various cells to extracellular matrices. This interpretation is supported by parallel studies using quantitative assays of cell-matrix adhesion. The likely origin of these adhesive differences is the progressive display of different kinds of adhesion-related molecules and their organizational complexes on increasingly adhesive cells. The antigen to which CSAT is directed is present on all of the above cells and thus appears to be a lowest common denominator of their adhesion to extracellular matrices.

Formation of Sic, Si3N4 and SiO2 by High-Dose Ion Implantation and Laser Annealing

1980 ◽  
Vol 1 ◽  
Author(s):  
S.W. Chiang ◽  
Y.S. Liu ◽  
R.F. Reihl
Keyword(s):  
Ion Implantation ◽  
Polycrystalline Silicon ◽  
Laser Annealing ◽  
Pulsed Laser ◽  
High Dose ◽  
Pulsed Laser Annealing ◽  
Transmission Electron ◽  
A Cell ◽  
Ft Ir ◽  
Ft Ir Spectroscopy

ABSTRACTHigh-dose ion implantation (1017 ions-cm−2) of C+, N+, and O+ at 50 KeV into silicon followed by pulsed laser annealing at 1.06 μm was studied. Formation of SiC, Si3N4, and SiO2 has been observed and investigated using Transmission Electron Microscopy (TEM) and Differential Fourier-Transform Infrared (FT-IR) Spectroscopy. Furthermore, in N+-implanted and laser-annealed silicon samples, we have observed a cell-like structure which has been identified to be spheroidal polycrystalline silicon formed by the rapid laser irradiation.

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