scholarly journals Evaluation and Implementation of Biocompatible Methods for the Cross-linking of Plasma Proteins

2021 ◽  
Vol 7 (2) ◽  
pp. 187-190
Author(s):  
Tom Bode ◽  
Kai Höltje ◽  
Sara Leal-Marin ◽  
Marc Müller ◽  
Birgit Glasmacher
Keyword(s):  
Plasma Proteins ◽  
Water Solubility ◽  
Patient Specific ◽  
Cross Linking ◽  
Reaction Products ◽  
Cross Link ◽  
Autologous Plasma ◽  
Amino Group Content ◽  
Primary Amino

Abstract Autologous plasma proteins can be used to fabricate patient specific cardiovascular implants but need to be cross-linked to increase their mechanical strength and reduce water solubility. Glutaraldehyde is the state-of-the-art solution but its reaction products have been shown to be cytotoxic and pro-inflammatory. In this work, it has been shown, that cross-linking of plasma proteins with biocompatible alternatives to glutaraldehyde is possible. This was achieved by identifying four candidate substances (thrombin, transglutaminase, genipin, EDC) from current literature and investigating their ability to cross-link porcine plasma proteins in vitro. The degree of crosslinking was examined using calorimetric (DSC) and spectroscopic (FTIR, Raman) methods, mapping the influence of cross-linking on the denaturation temperature and primary amino-group content of the proteins. It could be shown that thrombin, genipin and EDC are able to cross-link plasma proteins to a satisfactory degree and thus represent useful alternatives to glutaraldehyde. Transglutaminase, on the other hand, could not sufficiently cross-link the plasma proteins and was therefore ruled out as an alternative.

scholarly journals Cross-linking modifications of HDL apoproteins by oxidized phospholipids: structural characterization, in vivo detection, and functional implications

2020 ◽  
Vol 295 (7) ◽  
pp. 1973-1984
Author(s):  
Detao Gao ◽  
Mohammad Z. Ashraf ◽  
Lifang Zhang ◽  
Niladri Kar ◽  
Tatiana V. Byzova ◽  
...  
Keyword(s):  
Density Lipoprotein ◽  
Cross Linking ◽  
Oxidized Phospholipids ◽  
Cross Link ◽  
In Vivo Detection ◽  
Lysine Residues ◽  
Cross Links ◽  
Human Atheroma

Apolipoprotein A-I (apoA-I) is cross-linked and dysfunctional in human atheroma. Although multiple mechanisms of apoA-I cross-linking have been demonstrated in vitro, the in vivo mechanisms of cross-linking are not well-established. We have recently demonstrated the highly selective and efficient modification of high-density lipoprotein (HDL) apoproteins by endogenous oxidized phospholipids (oxPLs), including γ-ketoalkenal phospholipids. In the current study, we report that γ-ketoalkenal phospholipids effectively cross-link apoproteins in HDL. We further demonstrate that cross-linking impairs the cholesterol efflux mediated by apoA-I or HDL3 in vitro and in vivo. Using LC-MS/MS analysis, we analyzed the pattern of apoprotein cross-linking in isolated human HDL either by synthetic γ-ketoalkenal phospholipids or by oxPLs generated during HDL oxidation in plasma by the physiologically relevant MPO-H2O2-NO2− system. We found that five histidine residues in helices 5–8 of apoA-I are preferably cross-linked by oxPLs, forming stable pyrrole adducts with lysine residues in the helices 3–4 of another apoA-I or in the central domain of apoA-II. We also identified cross-links of apoA-I and apoA-II with two minor HDL apoproteins, apoA-IV and apoE. We detected a similar pattern of apoprotein cross-linking in oxidized murine HDL. We further detected oxPL cross-link adducts of HDL apoproteins in plasma and aorta of hyperlipidemic LDLR−/− mice, including cross-link adducts of apoA-I His-165–apoA-I Lys-93, apoA-I His-154–apoA-I Lys-105, apoA-I His-154–apoA-IV Lys-149, and apoA-II Lys-30–apoE His-227. These findings suggest an important mechanism that contributes to the loss of HDL's atheroprotective function in vivo.

scholarly journals A SEX-LINKED DEFECT IN THE CROSS-LINKING OF COLLAGEN AND ELASTIN ASSOCIATED WITH THE MOTTLED LOCUS IN MICE

1974 ◽  
Vol 139 (1) ◽  
pp. 180-192 ◽  
Author(s):  
David W. Rowe ◽  
Ermona B. McGoodwin ◽  
George R. Martin ◽  
Michael D. Sussman ◽  
Douglas Grahn ◽  
...  
Keyword(s):  
Coat Color ◽  
Cross Linking ◽  
Genetic Abnormality ◽  
Cross Link ◽  
Skin Collagen ◽  
The Cross ◽  
Tissue Abnormalities ◽  
Link Formation ◽  
Experimental Lathyrism

A genetic abnormality in collagen and elastin cross-linking resembling experimental lathyrism has been identified in mice. The defect is an X-linked trait, attributed to the mottled locus which also influences coat color. The affected mice have aneurysms of the aorta and its branches, weak skin, and bone deformities in a spectrum of severity varying with the alleles at the mottled locus. A defect in the cross-linking of collagen was demonstrated in the skin of the affected animals by a marked increase in collagen extractability and a reduced proportion of cross-linked components in the extracted collagen. A decrease in lysine-derived aldehyde levels was found in both skin collagen and aortic elastin similar to that found in lathyritic tissue. Furthermore the in vitro formation of lysine-derived aldehyde was reduced. Thus the cause of the connective tissue abnormalities in these mice appears to be a defect in cross-link formation due to an impairment in aldehyde formation.

scholarly journals In Vitro Assembly of Sindbis Virus Core-Like Particles from Cross-Linked Dimers of Truncated and Mutant Capsid Proteins

2001 ◽  
Vol 75 (6) ◽  
pp. 2810-2817 ◽  
Author(s):  
Timothy L. Tellinghuisen ◽  
Rushika Perera ◽  
Richard J. Kuhn
Keyword(s):  
Capsid Protein ◽  
Sindbis Virus ◽  
Terminal Region ◽  
Full Length ◽  
Capsid Proteins ◽  
Size Difference ◽  
Cross Linking ◽  
Cross Link ◽  
Amino Terminal

ABSTRACT A nucleic acid-bound capsid protein dimer was previously identified using a Sindbis virus in vitro nucleocapsid assembly system and cross-linking reagents. Cross-link mapping, in combination with a model of the nucleocapsid core, suggested that this dimer contained one monomer from each of two adjacent capsomeres. This intercapsomere dimer is believed to be the initial intermediate in the nucleocapsid core assembly mechanism. This paper presents the purification of cross-linked dimers of a truncated capsid protein and the partial purification of cross-linked dimers of a full-length assembly-defective mutant. The assembly of core-like particles from these cross-linked capsid protein dimers is demonstrated. Core-like particles generated from cross-linked full-length mutant CP(19-264)L52D were examined by electron microscopy and appeared to have a morphology similar to that of wild-type in vitro-assembled core-like particles, although a slight size difference was often visible. Truncated cross-linked CP(81-264) dimers generated core-like particles as well. These core-like particles could subsequently be disassembled when reversible cross-linking reagents were used to form the dimers. The ability of the covalent intercapsomere cross-link to rescue capsid proteins with assembly defects or truncations in the amino-terminal region of the capsid protein supports the previous model of assembly and suggests a possible role for the amino-terminal region of the protein.

scholarly journals STRUCTURAL, FUNCTIONAL PROPERTIES AND IN VITRO DIGESTIBILIGY OF MAIZE STARCH UNDER HEAT-MOISTURE AND ATMOSPHERIC-COLD PLASMA TREATMENTS

2018 ◽  
Vol 56 (6) ◽  
pp. 671 ◽  
Author(s):  
Linh Thùy Nguyễn ◽  
Sơn Khánh Trịnh
Keyword(s):  
Water Solubility ◽  
Argon Plasma ◽  
Cross Linking ◽  
In Vitro Digestibility ◽  
Maize Starch ◽  
Raw Starch ◽  
Plasma Treatments ◽  
Water Absorbance ◽  
Swelling Factor

Maize starch is one of an important material which widely using in food applications and other industries. However, natural properties of raw starch can not be suitable for processed foods. So, the modification of starch is very important. In this study, heat-moisture and atmospheric cold argon-plasma treatments were applied in maize starch; then, structural, fuctional properties and digestibility of modified starch was investigated. Raw starch was heated at 20, 25 and 30% of moisture content. Subsequently, sample was treated under argon-plasma environment at fixed paramentes (137,5 V; 1.0 A for 10 min). Sample was investigated degree of cross-linking, degree of relative crystallinity (DRC), degree of hydrolysis using alpha-amylase, in vitro digestibility, changes in the hydration properties such as water absorbance index, swelling factor and water solubility index. Results show that degree of cross-linking, DRC, resistant starch of samples significantly increase under heat-moisture and plasma treatments; especially, sample of 20% heat-moisture contains 3-folded to 10-folded increase comparing to raw starch base on with or without pre-boiling process. Furthermore, water absorbance index and swelling factor decrease but water solubility index increase under plasma treatment.

A multicomponent complex is required for the AAUAAA-dependent cross-linking of a 64-kilodalton protein to polyadenylation substrates

1990 ◽  
Vol 10 (3) ◽  
pp. 1244-1248 ◽  
Author(s):  
J Wilusz ◽  
T Shenk ◽  
Y Takagaki ◽  
J L Manley
Keyword(s):  
Uv Light ◽  
Cross Linking ◽  
Cross Link ◽  
Multicomponent Complex ◽  
Specificity Factor ◽  
The Individual ◽  
64 Kda Protein ◽  
Rna Interaction ◽  
Cleavage Stimulation Factor

A 64-kilodalton (kDa) polypeptide that is cross-linked by UV light specifically to polyadenylation substrate RNAs containing a functional AAUAAA element has been identified previously. Fractionated HeLa nuclear components that can be combined to regenerate efficient and accurate polyadenylation in vitro have now been screened for the presence of the 64-kDa protein. None of the individual components contained an activity which could generate the 64-kDa species upon UV cross-linking in the presence of substrate RNA. It was necessary to mix two components, cleavage stimulation factor and specificity factor, to reconstitute 64-kDa protein-RNA cross-linking. The addition of cleavage factors to this mixture very efficiently reconstituted the AAUAAA-specific 64-kDa protein-RNA interaction. The 64-kDa protein, therefore, is present in highly purified, reconstituted polyadenylation reactions. However, it is necessary to form a multicomponent complex to efficiently cross-link the protein to a substrate RNA.

scholarly journals Transglutaminase-catalyzed matrix cross-linking in differentiating cartilage: identification of osteonectin as a major glutaminyl substrate.

1995 ◽  
Vol 129 (3) ◽  
pp. 881-892 ◽  
Author(s):  
D Aeschlimann ◽  
O Kaupp ◽  
M Paulsson
Keyword(s):  
Tissue Transglutaminase ◽  
Physiological Mechanism ◽  
Cross Linking ◽  
Endochondral Bone Formation ◽  
Cross Link ◽  
Endochondral Bone ◽  
Tracheal Cartilage ◽  
Matrix Stabilization ◽  
The Matrix

The expression of tissue transglutaminase in skeletal tissues is strictly regulated and correlates with chondrocyte differentiation and cartilage calcification in endochondral bone formation and in maturation of tracheal cartilage (Aeschlimann, D., A. Wetterwald, H. Fleisch, and M. Paulsson. 1993. J. Cell Biol. 120:1461-1470). We now demonstrate the transglutaminase reaction product, the gamma-glutamyl-epsilon-lysine cross-link, in the matrix of hypertrophic cartilage using a novel cross-link specific antibody. Incorporation of the synthetic transglutaminase substrate monodansylcadaverine (amine donor) in cultured tracheal explants reveals enzyme activity in the pericellular matrix of hypertrophic chondrocytes in the central, calcifying areas of the horseshoe-shaped cartilages. One predominant glutaminyl substrate (amine acceptor) in the chondrocyte matrix is osteonectin as revealed by incorporation of the dansyl label in culture. Indeed, nonreducible osteonectin-containing complexes of approximately 65, 90, and 175 kD can be extracted from mature tracheal cartilage. In vitro cross-linking of osteonectin by tissue transglutaminase gives similar products of approximately 90 and 175 kD, indicating that the complexes in cartilage represent osteonectin oligomers. The demonstration of extracellular transglutaminase activity in differentiating cartilage, i.e., cross-linking of osteonectin in situ, shows that tissue transglutaminase-catalyzed cross-linking is a physiological mechanism for cartilage matrix stabilization.

scholarly journals Tissue fixation by osmium tetroxide. A possible role for proteins.

1979 ◽  
Vol 27 (5) ◽  
pp. 997-999 ◽  
Author(s):  
A J Nielson ◽  
W P Griffith
Keyword(s):  
Normal Tissue ◽  
Osmium Tetroxide ◽  
Chemical Nature ◽  
Side Chain ◽  
Cross Linking ◽  
Tissue Fixation ◽  
Reaction Products ◽  
Cholesteryl Acetate ◽  
Unsaturated Lipids

The osmiophilia, under the conditions of normal tissue fixation, of the histidine, lysine, tryptophan, cysteine and methionine side chain of proteins is suggested by in vitro studies on blocked amino acids representative of such protein side chains, and the chemical nature of the reaction products elucidated. The chemical feasibility of inter- or intramolecular cross-linking of protein by OsO4 at these and other sites is demonstrated, as in the cross-linking of protein with unsaturated lipids such as methyl oleate, methyl linoleate and linolenate, and cholesteryl acetate. The relevance of these results to the process of tissue fixation by OsO4 is discussed.

scholarly journals Non-enzymic glycation of fibrous collagen: reaction products of glucose and ribose

1995 ◽  
Vol 305 (2) ◽  
pp. 385-390 ◽  
Author(s):  
A J Bailey ◽  
T J Sims ◽  
N C Avery ◽  
E P Halligan
Keyword(s):  
Physical Properties ◽  
High Molecular Mass ◽  
Diabetic Patients ◽  
Reaction Products ◽  
Cross Link ◽  
Dermal Tissue ◽  
Total Fluorescence ◽  
Collagen Molecules ◽  
Cross Links

Non-enzymic glycation of collagen involves a series of complex reactions ultimately leading to the formation of intermolecular cross-links resulting in changes in its physical properties. During analysis for the fluorescent cross-link pentosidine we identified the presence of an additional component (Cmpd K) in both glucose and ribose incubations. Cmpd K was formed more quickly than pentosidine in glucose incubations and more slowly than pentosidine in ribose incubations. Cmpd K represented 45% of the total fluorescence compared with 15% for pentosidine in glucose incubations and 25% of the total fluorescence compared with 30% for pentosidine in the ribose incubations. Cmpd K is not an artefact of in vitro incubations, as it was shown to be present in dermal tissue from diabetic patients. Subsequent high-resolution h.p.l.c. analysis of glucose-incubated collagen revealed Cmpd K comprise two components (K1 and K2). Further, a similar analysis of Cmpd K from the ribose incubations revealed two different components (K3 and K4). These differences indicate alternative mechanisms for the reactions of glucose and ribose with collagen. The amounts of these fluorescent components and the pentosidine cross-link determined for both glucose and ribose glycation were found to be far too low (about one pentosidine molecules per 200 collagen molecules after 6 months incubation with glucose) to account for the extensive cross-linking responsible for the changes in physical properties, suggesting that a further additional series of cross-links are formed. We have analysed the non-fluorescent high-molecular-mass components and identified a new component that increases with time of in vitro incubation and is present in the skin of diabetic patients. This component is present in sufficient quantities (estimated at one cross-link per two collagen molecules) to account for the changes in physical properties occurring in vitro.

scholarly journals Structural Mechanism for Alteration of Collagen Gel Mechanics by Glutaraldehyde Crosslinking

2007 ◽  
Author(s):  
Preethi L. Chandran ◽  
Dezba G. Coughlin ◽  
David C. Paik ◽  
Jeffrey W. Holmes
Keyword(s):  
Collagen Gel ◽  
Cross Linking ◽  
Collagen Gels ◽  
Cross Link ◽  
Structural Mechanism ◽  
Micro Scale ◽  
Macro Scale ◽  
Link Density ◽  
Cross Link Density

Cross-linking of fibrous collagenous tissues occurs during late-stage wound healing and during aging. The attendant changes in micro-scale kinematics and macro-scale mechanics are not well understood. In this study we used glutaraldehyde as a model cross-linking agent, and in vitro reconstituted collagen gel as a model collagenous tissue. Collagen gels are in vitro assembled hydrated networks of collagen fibrils. Glutaraldehyde is a commonly used cross-linking agent for bioprosthetic tissues and is chemically well-characterized. Glutaraldehyde cross-linking is known to decrease the deformability of arterial valves, but the micro-scale mechanism of its action is not known. In this study, collagen gels with anisotropic fibril orientation were subjected to increasing cross-link density, and the concurrent change in biaxial mechanical properties was monitored. The extent of cross-linking is determined by biochemical analysis. Structural modeling of the biaxial mechanics of a fibrous microstructure was performed for three potential cross-link mechanisms. The trends in the simulated mechanics for increasing cross-link density were compared against that of the experimental data.

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