scholarly journals Effects of particulates and lipids on the hydraulic conductivity of Matrigel

2008 ◽  
Vol 105 (2) ◽  
pp. 621-628 ◽  
Author(s):  
William J. McCarty ◽  
Melissa F. Chimento ◽  
Christine A. Curcio ◽  
Mark Johnson
Keyword(s):  
Extracellular Matrix ◽  
Hydraulic Conductivity ◽  
Weight Fraction ◽  
Low Density Lipoproteins ◽  
Blood Vessel Wall ◽  
Connective Tissues ◽  
Rigid Particles ◽  
Ldl Particles ◽  
Transmission Electron ◽  
Theoretical Predictions

The hydraulic conductivity of a connective tissue is determined both by the fine ultrastructure of the extracellular matrix and the effects of larger particles in the interstitial space. In this study, we explored this relationship by examining the effects of 30- or 90-nm-diameter latex nanospheres or low-density lipoproteins (LDL) on the hydraulic conductivity of Matrigel, a basement membrane matrix. The hydraulic conductivity of Matrigel with latex nanospheres or LDL particles added at 4.8% weight fraction was measured and compared with the hydraulic conductivity of Matrigel alone. The LDL-derived lipids in the gel were visualized by transmission electron microscopy and were seen to have aggregated into particles up to 500 nm in size. The addition of these materials to the medium markedly decreased its hydraulic conductivity, with the LDL-derived lipids having a much larger effect than did the latex nanospheres. Debye-Brinkman theory was used to predict the effect of addition of particles to the hydraulic conductivity of the medium. The theoretical predictions matched well with the results from adding latex nanospheres to the medium. However, LDL decreased hydraulic conductivity much more than was predicted by the theory. The validation of the theoretical model for rigid particles embedded in extracellular matrix suggests that it could be used to make predictions about the influence of particulates (e.g., collagen, elastin, cells) on the hydraulic conductivity of the fine filamentous matrix (the proteoglycans) in connective tissues. In addition, the larger-than-predicted effects of lipidlike particles on hydraulic conductivity may magnify the pathology associated with lipid accumulation, such as in Bruch's membrane of the retina during macular degeneration and the blood vessel wall in atherosclerosis.

scholarly journals Hemicentin-1 is an essential extracellular matrix component of the dermal–epidermal and myotendinous junctions

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Daniela Welcker ◽  
Cornelia Stein ◽  
Natalia Martins Feitosa ◽  
Joy Armistead ◽  
Jin-Li Zhang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Developmental Stages ◽  
Cell Types ◽  
Connective Tissues ◽  
Functional Behavior ◽  
Transmission Electron ◽  
Cellular Microenvironments ◽  
And Function ◽  
Myotendinous Junctions

AbstractThe extracellular matrix architecture is composed of supramolecular fibrillar networks that define tissue specific cellular microenvironments. Hemicentins (Hmcn1 and Hmcn2) are ancient and very large members (> 600 kDa) of the fibulin family, whose short members are known to guide proper morphology and functional behavior of specialized cell types predominantly in elastic tissues. However, the tissue distribution and function of Hemicentins within the cellular microenvironment of connective tissues has remained largely unknown. Performing in situ hybridization and immunofluorescence analyses, we found that mouse Hmcn1 and Hmcn2 show a complementary distribution throughout different tissues and developmental stages. In postnatal dermal–epidermal junctions (DEJ) and myotendinous junctions (MTJ), Hmcn1 is primarily produced by mesenchymal cells (fibroblasts, tenocytes), Hmcn2 by cells of epithelial origin (keratinocytes, myocytes). Hmcn1−/− mice are viable and show no overt phenotypes in tissue tensile strength and locomotion tests. However, transmission electron microscopy revealed ultrastructural basement membrane (BM) alterations at the DEJ and MTJ of Hmcn1−/− mice, pointing to a thus far unknown role of Hmcn1 for BM and connective tissue boundary integrity.

Specific Hydraulic Conductivity of Corneal Stroma as Seen by Quick-Freeze/Deep-Etch

2000 ◽  
Vol 123 (2) ◽  
pp. 154-161 ◽  
Author(s):  
Darryl Overby ◽  
Jeffrey Ruberti ◽  
Haiyan Gong ◽  
Thomas F. Freddo ◽  
Mark Johnson
Keyword(s):  
Extracellular Matrix ◽  
Hydraulic Conductivity ◽  
Flow Resistance ◽  
Corneal Stroma ◽  
High Flow ◽  
Depth Of Field ◽  
Resistance Characteristic ◽  
Connective Tissues ◽  
Electron Microscopic ◽  
Microscopic Techniques

Previous studies of the hydraulic conductivity of connective tissues have failed to show a correspondence between ultrastructure and specific hydraulic conductivity. We used the technique of quick-freeze/deep-etch to examine the ultrastructure of the corneal stroma and then utilized morphometric studies to compute the specific hydraulic conductivity of the corneal stroma. Our studies demonstrated ultrastructural elements of the extracellular matrix of the corneal stroma that are not seen using conventional electron microscopic techniques. Furthermore, we found that these structures may be responsible for generating the high flow resistance characteristic of connective tissues. From analysis of micrographs corrected for depth-of-field effects, we used Carmen-Kozeny theory to bound a morphometrically determined specific hydraulic conductivity of the corneal stroma between 0.46×10−14 and 10.3×10−14 cm2. These bounds encompass experimentally measured values in the literature of 0.5×10−14 to 2×10−14 cm2. The largest source of uncertainty was due to the depth-of-field estimates that ranged from 15 to 51 nm; a better estimate would substantially reduce the uncertainty of these morphometrically determined values.

scholarly journals Hemicentin-1 is An Essential Extracellular Matrix Component of the Dermal-Epidermal and Myotendinous Junctions

2021 ◽  
Author(s):  
Daniela Welcker ◽  
Cornelia Stein ◽  
Natalia Martins Feitosa ◽  
Joy Armistead ◽  
Jin-Li Zhang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Developmental Stages ◽  
Cell Types ◽  
Connective Tissues ◽  
Functional Behavior ◽  
Transmission Electron ◽  
Cellular Microenvironments ◽  
Ecm Architecture ◽  
And Function ◽  
Myotendinous Junctions

Abstract The extracellular matrix (ECM) architecture is composed of supramolecular fibrillar networks that define tissue specific cellular microenvironments. Hemicentins (Hmcn1 and Hmcn2) are ancient and very large members (>600 kDa) of the fibulin family, whose short members are known to guide proper morphology and functional behavior of specialized cell types predominantly in elastic tissues. However, the tissue distribution and function of Hemicentins within the cellular microenvironment of connective tissues has remained largely unknown. Performing in situ hybridization and immunofluorescence analyses, we found that Hmcn1 and Hmcn2 show a complementary distribution throughout different tissues and developmental stages. In postnatal dermal-epidermal junctions (DEJ) and myotendinous junctions (MTJ), Hmcn1 is primarily produced by mesenchymal cells (fibroblasts, tenocytes), Hmcn2 by cells of epithelial origin (keratinocytes, myocytes). Hmcn1-/-mice are viable and show no overt phenotypes in tissue tensile strength and locomotion tests. However, transmission electron microscopy revealed ultrastructural basement membrane (BM) alterations at the DEJ and MTJ of Hmcn1-/-mice, pointing to a thus far unknown role of Hmcn1 for BM and connective tissue boundary integrity.

X-ray microanalysis of thin specimens in the transmission electron microscope at voltages up to 1000 Kv.

1978 ◽  
Vol 36 (1) ◽  
pp. 540-541
Author(s):  
G. Cliff ◽  
M.J. Nasir ◽  
G.W. Lorimer ◽  
N. Ridley
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Weight Fraction ◽  
Present Series ◽  
Constant Independent ◽  
X Ray ◽  
Transmission Electron ◽  
Series Of Experiments ◽  
Image Position ◽  
X Ray Absorption

In a specimen which is transmission thin to 100 kV electrons - a sample in which X-ray absorption is so insignificant that it can be neglected and where fluorescence effects can generally be ignored (1,2) - a ratio of characteristic X-ray intensities, I1/I2 can be converted into a weight fraction ratio, C1/C2, using the equationwhere k12 is, at a given voltage, a constant independent of composition or thickness, k12 values can be determined experimentally from thin standards (3) or calculated (4,6). Both experimental and calculated k12 values have been obtained for K(11<Z>19),kα(Z>19) and some Lα radiation (3,6) at 100 kV. The object of the present series of experiments was to experimentally determine k12 values at voltages between 200 and 1000 kV and to compare these with calculated values.The experiments were carried out on an AEI-EM7 HVEM fitted with an energy dispersive X-ray detector.

A crystallographic analysis of the CoSi/Si(111) interface using Transmission Electron Microscopy

1990 ◽  
Vol 48 (4) ◽  
pp. 574-575
Author(s):  
A.C. Daykin ◽  
C.J. Kiely ◽  
R.C. Pond ◽  
J.L. Batstone
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Defect Structure ◽  
Room Temperature ◽  
Theoretical Method ◽  
Crystallographic Analysis ◽  
Si Substrate ◽  
Transmission Electron ◽  
Theoretical Predictions

When CoSi2 is grown onto a Si(111) surface it can form in two distinct orientations. A-type CoSi2 has the same orientation as the Si substrate and B-type is rotated by 180° degrees about the [111] surface normal.One method of producing epitaxial CoSi2 is to deposit Co at room temperature and anneal to 650°C.If greater than 10Å of Co is deposited then both A and B-type CoSi2 form via a number of intermediate silicides .The literature suggests that the co-existence of A and B-type CoSi2 is in some way linked to these intermediate silicides analogous to the NiSi2/Si(111) system. The phase which forms prior to complete CoSi2 formation is CoSi. This paper is a crystallographic analysis of the CoSi2/Si(l11) bicrystal using a theoretical method developed by Pond. Transmission electron microscopy (TEM) has been used to verify the theoretical predictions and to characterise the defect structure at the interface.

Asymmetric Cracking in Znse/ZnSxSel−x Superlattices Grown by Molecular Bean Epitaxy

1987 ◽  
Vol 102 ◽  
Author(s):  
Richard J. Dalby ◽  
John Petruzzello
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Thermal Expansion ◽  
Molecular Beam Epitaxy ◽  
Molecular Beam ◽  
Lattice Mismatch ◽  
Sulfur Concentration ◽  
Transmission Electron ◽  
Theoretical Predictions ◽  
Superlattice Layer

ABSTRACTOptical and transmission electron microscopy have been used to study cracks appearing in ZnSe/ZnSxSe1−x (x ∼ 0.38) superlattices grown by Molecular Beam Epitaxy. It Is shown that when a fracture occurs it is confined, in most cases, to the superlattice and propagates along <011> cleavage directions in these <001> oriented epilayers. Cracks were not observed in all superlattices and their onset is discussed in relation to sulfur concentration, overall superlattice height, individual superlattice layer thicknesses, and stress, tensile or compressive, due to lattice mismatch and thermal expansion differences between buffer layer and superlattice. It was found that by adjusting the controllable parameters, cracks in the superlattices could be eliminated. Orientation and density of these features have been related to asynnmetric cracking associated with the zincblende structure of these II-VI materials. Experimental results are shown to be in agreement with theoretical predictions of critical heights for the onset of cracking.

scholarly journals Genetically modified human type II collagen for N- and C-terminal covalent tagging

2018 ◽  
Vol 96 (2) ◽  
pp. 204-211
Author(s):  
Andrew Wieczorek ◽  
Clara K. Chan ◽  
Suzana Kovacic ◽  
Cindy Li ◽  
Thomas Dierks ◽  
...  
Keyword(s):  
Developmental Disorders ◽  
Connective Tissue Diseases ◽  
Sulfhydryl Group ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Structural Protein ◽  
Connective Tissues ◽  
Human Type ◽  
C Terminus ◽  
Transmission Electron

Collagen is the predominant structural protein in vertebrates, where it contributes to connective tissues and the ECM; it is also widely used in biomaterials and tissue engineering. Dysfunction of this protein and its processing can lead to a wide variety of developmental disorders and connective tissue diseases. Recombinantly engineering the protein is challenging due to post-translational modifications generally required for its stability and secretion from cells. Introducing end labels into the protein is problematic, because the N- and C-termini of the physiologically relevant tropocollagen lie internal to the initially flanking N- and C-propeptide sequences. Here, we introduce mutations into human type II procollagen in a manner that addresses these concerns and purify the recombinant protein from a stably transfected HT1080 human fibrosarcoma cell line. Our approach introduces chemically addressable groups into the N- and C-telopeptide termini of tropocollagen. Simultaneous overexpression of formylglycine generating enzyme (FGE) allows the endogenous production of an aldehyde tag in a defined, substituted sequence in the N terminus of the mutated collagen, whereas the C-terminus of each chain presents a sulfhydryl group from an introduced cysteine. These modifications are designed to enable specific covalent end-labelling of collagen. We find that the doubly mutated protein folds and is secreted from cells. Higher order assembly into well-ordered collagen fibrils is demonstrated through transmission electron microscopy. Chemical tagging of thiols is successful; however, background from endogenous aldehydes present in wild-type collagen has thus far obscured the desired specific N-terminal labelling. Strategies to overcome this challenge are proposed.

scholarly journals LTBP1 promotes fibrillin incorporation into the extracellular matrix

2021 ◽  
Author(s):  
Matthias Przyklenk ◽  
Veronika Georgieva ◽  
Fabian Metzen ◽  
Sebastian Mostert ◽  
Birgit Kobbe ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Wide Spectrum ◽  
Extracellular Matrix Protein ◽  
Connective Tissues ◽  
Pathogenic Variants ◽  
Human Pathology ◽  
Fibrillin 1

LTBP1 is a large extracellular matrix protein and an associated ligand of fibrillin-microfibrils. Knowledge of LTBP1 functions is largely limited to its role in targeting and sequestering TGFβ growth factors within the extracellular matrix, thereby regulating their bioavailability. However, the recent description of a wide spectrum of phenotypes in multiple tissues in patients harboring LTBP1 pathogenic variants suggests a multifaceted role of the protein in the homeostasis of connective tissues. To better understand the human pathology caused by LTBP1 deficiency it is important to investigate its functional role in extracellular matrix formation. In this study, we show that LTBP1 coordinates the incorporation of fibrillin-1 and -2 into the extracellular matrix in vitro. We also demonstrate that this function is differentially exerted by the two isoforms, the short and long forms of LTBP1. Thereby our findings uncover a novel TGFβ-independent LTBP1 function potentially contributing to the development of connective tissue disorders.

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