Heparin and heparansulfate modulate cell-matrix interactions of fibroblasts and endothelial cells in vitro and interact with specific binding sites

1993 ◽  
Vol 6 (1) ◽  
pp. 5
Author(s):  
Th. Schaefer ◽  
M. Roux ◽  
H.W. Stuhlsatz ◽  
Th. Krieg ◽  
H. Smola
Keyword(s):  
Endothelial Cells ◽  
Binding Sites ◽  
Specific Binding ◽  
Cell Matrix ◽  
Specific Binding Sites ◽  
Matrix Interactions ◽  
Cell Matrix Interactions

scholarly journals Synthetic extracellular matrices with tailored adhesiveness and degradability support lumen formation during angiogenic sprouting

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jifeng Liu ◽  
Hongyan Long ◽  
Dagmar Zeuschner ◽  
Andreas F. B. Räder ◽  
William J. Polacheck ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Material Properties ◽  
In Vitro Model ◽  
Matrix Remodeling ◽  
Lumen Formation ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Vitro Model ◽  
Cell Matrix Interactions

AbstractA major deficit in tissue engineering strategies is the lack of materials that promote angiogenesis, wherein endothelial cells from the host vasculature invade the implanted matrix to form new blood vessels. To determine the material properties that regulate angiogenesis, we have developed a microfluidic in vitro model in which chemokine-guided endothelial cell sprouting into a tunable hydrogel is followed by the formation of perfusable lumens. We show that long, perfusable tubes only develop if hydrogel adhesiveness and degradability are fine-tuned to support the initial collective invasion of endothelial cells and, at the same time, allow for matrix remodeling to permit the opening of lumens. These studies provide a better understanding of how cell-matrix interactions regulate angiogenesis and, therefore, constitute an important step towards optimal design criteria for tissue-engineered materials that require vascularization.

Glycosaminoglycans modulate cell-matrix interactions of human fibroblasts and endothelial cells in vitro

1996 ◽  
Vol 109 (2) ◽  
pp. 479-488
Author(s):  
T. Schaefer ◽  
M. Roux ◽  
H.W. Stuhlsatz ◽  
R. Herken ◽  
B. Coulomb ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Heparan Sulfate ◽  
Interleukin 6 ◽  
Binding Sites ◽  
Human Fibroblasts ◽  
Collagen I ◽  
Collagen Gels ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions

Contact of various cells with extracellular matrix molecules modulates their cellular functions and phenotype. Most investigations have employed dishes coated with purified matrix constituents or plain collagen I lattices omitting the effects of other important matrix components such as proteoglycans. In this study we analyze the effect of purified glycosaminoglycans (GAGs) on human fibroblasts and human umbilical vein endothelial cells (HUVEC) embedded within collagen I/III lattices. HUVEC contracted collagen I/III gels far less efficiently than fibroblasts and addition of heparan sulfate and heparin almost completely inhibited contraction. In collagen gels HUVEC down-regulated collagenase mRNA while increasing collagen I, IV mRNA expression. Addition of heparin and heparan sulfate reversed the collagen IV mRNA induction whereas hyaluronic acid and chondroitin sulfate enhanced fibronectin and collagenase transcripts. Fibroblasts readily contracted collagen gels, and mRNA levels for fibronectin, collagenase and interleukin-6 were stimulated. Gel contraction was mostly unaffected by the different glycosaminoglycans. Fibroblasts responded to the addition of dermatan sulfate, heparan sulfate and heparin with a decrease in fibronectin, collagenase and interleukin-6 mRNA. Binding studies revealed saturable binding sites on fibroblasts and HUVEC for 35S-labelled heparin, demonstrating specificity for heparin and heparan sulfate over other GAGs in competition experiments. This study implies that glycosaminoglycans participate in cell-matrix interactions by effectively modulating the cellular phenotype via high affinity binding sites.

Atrial natriuretic factor binding sites in the jejunum

1989 ◽  
Vol 256 (2) ◽  
pp. G436-G441 ◽  
Author(s):  
C. Bianchi ◽  
G. Thibault ◽  
A. De Lean ◽  
J. Genest ◽  
M. Cantin
Keyword(s):  
Binding Sites ◽  
Atrial Natriuretic Factor ◽  
Specific Binding ◽  
Rat Tissues ◽  
Rat Jejunum ◽  
Specific Binding Sites ◽  
Natriuretic Factor ◽  
Atrial Natriuretic

We have studied the localization and the characterization of atrial natriuretic factor (ANF) binding sites by radioautographic techniques. Quantitative in vitro radioautography with a computerized microdensitometer demonstrated the presence of high-affinity, low-capacity 125I-ANF-(99-126) binding sites (Kd, 48 pM; Bmax, 63 fmol/mg protein) mainly in the villi of 20-microns slide-mounted transverse sections of the rat jejunum. Competition curves showed 50% inhibitory concentrations of 55 and 1,560 pM for ANF-(99-126) and ANF-(103-123), respectively. In vivo electron microscope radioautography showed that 80% of the silver grains were localized on the lamina propria fibroblast-like cells, 18% on mature enterocytes, and 2% on capillaries. Bradykinin and adrenocorticotropin did not compete with ANF binding. These results demonstrate that ANF binding sites in the rat jejunum possess the pharmacological characteristics of functional ANF receptors encountered in other rat tissues, and ultrastructural radioautographs show their cellular distribution. Taken together, these results demonstrate the presence and the localization of specific binding sites for ANF in the jejunal villi of the rat small intestine.

scholarly journals Imaging and Analysis of Cellular Locations in Three-Dimensional Tissue Models

2019 ◽  
Vol 25 (3) ◽  
pp. 753-761 ◽  
Author(s):  
Warren Colomb ◽  
Matthew Osmond ◽  
Charles Durfee ◽  
Melissa D. Krebs ◽  
Susanta K. Sarkar
Keyword(s):  
Three Dimensional ◽  
Human Mesenchymal Stem Cells ◽  
Basic Research ◽  
Light Sheet ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Tissue Models ◽  
Alginate Matrix

AbstractThe absence of quantitative in vitro cell–extracellular matrix models represents an important bottleneck for basic research and human health. Randomness of cellular distributions provides an opportunity for the development of a quantitative in vitro model. However, quantification of the randomness of random cell distributions is still lacking. In this paper, we have imaged cellular distributions in an alginate matrix using a multiview light sheet microscope and developed quantification metrics of randomness by modeling it as a Poisson process, a process that has constant probability of occurring in space or time. We imaged fluorescently labeled human mesenchymal stem cells embedded in an alginate matrix of thickness greater than 5 mm with $\sim\! {\rm 2}{\rm. 9} \pm {\rm 0}{\rm. 4}\,\mu {\rm m}$ axial resolution, the mean full width at half maximum of the axial intensity profiles of fluorescent particles. Simulated randomness agrees well with the experiments. Quantification of distributions and validation by simulations will enable quantitative study of cell–matrix interactions in tissue models.

scholarly journals Peptide gels of fully-defined composition and mechanics for probing cell-cell and cell-matrix interactions in vitro

2020 ◽  
Vol 85-86 ◽  
pp. 15-33 ◽  
Author(s):  
J.C. Ashworth ◽  
J.L. Thompson ◽  
J.R. James ◽  
C.E. Slater ◽  
S. Pijuan-Galitó ◽  
...  
Keyword(s):  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Cell Cell

Autoradiographic detection of kinin receptors in the human medulla of control, hypertensive, and diabetic donors

2002 ◽  
Vol 80 (4) ◽  
pp. 249-257 ◽  
Author(s):  
Hudson de Sousa Buck ◽  
Brice Ongali ◽  
Gaétan Thibault ◽  
Charles J Lindsey ◽  
Réjean Couture
Keyword(s):  
Receptor Binding ◽  
Binding Sites ◽  
Specific Binding ◽  
Inferior Olivary Nucleus ◽  
Binding Studies ◽  
Specific Binding Sites ◽  
Kinin Receptors ◽  
Medullary Nuclei ◽  
Inferior Olivary

Kinins have been elected to the status of central neuromediators. Their effects are mediated through the activation of two G-protein-coupled receptors, denoted B1 and B2. Functional and binding studies suggested that B1 and B2 receptors are upregulated in the medulla and spinal cord of hypertensive and diabetic rats. The aim of this study was to localize and quantify kinin receptors in post-mortem human medulla obtained from normotensive, hypertensive, and diabetic subjects, using in vitro receptor autoradiography with the radioligands [125I]HPP-HOE140 (B2 receptor) and [125I]HPP[des-Arg10]-HOE140 (B1 receptor). Data showed specific binding sites for B2 receptor (0.4–1.5 fmol/mg tissue) in 11 medullary nuclei from 4 control specimens (paratrigeminal > ambiguus > cuneate, gelatinous layer of the caudal spinal trigeminal nucleus > caudal and interpolar spinal trigeminal, external cuneate, solitary tract > hypoglossal > gracile > inferior olivary nuclei). Increased density of B2 receptor binding sites was observed in seven medullary nuclei of four hypertensive specimens (paratrigeminal > external cuneate > interpolar and caudal spinal trigeminal, gracile, inferior olivary > hypoglossal nuclei). B2 receptor binding sites were seemingly increased in the same medullary nuclei of two diabetic specimens. Specific binding sites for B1 receptor (1.05 and 1.36 fmol/mg tissue) were seen only in the inferior olivary nucleus in two out of the ten studied specimens. The present results support a putative role for kinins in the regulation of autonomic, nociceptive, and motor functions at the level of the human medulla. Evidence is also provided that B2 receptors are upregulated in medullary cardiovascular centers of subjects afflicted of cardiovascular diseases.Key words: bradykinin, hypertension, diabetes, human brain.

scholarly journals An Inducible Model for Unraveling the Effects of Advanced Glycation End-Products in Collagen with Age and Diabetes

2020 ◽  
Author(s):  
Austin G. Gouldin ◽  
Jennifer L. Puetzer
Keyword(s):  
Cell Viability ◽  
Blue Light ◽  
Advanced Glycation End Products ◽  
Advanced Glycation ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Glycation End Products ◽  
End Products ◽  
Cell Matrix Interactions

AbstractIn connective tissues there is a clear link between increasing age and degeneration. It is believed advanced glycation end-products (AGEs) play a central role in this degeneration. AGEs are sugar induced non-enzymatic crosslinks which accumulate in collagen with age and diabetes, altering tissue mechanics and cellular function. Despite ample correlative evidence linking collagen glycation to degeneration, little is known how AGEs impact cell-matrix interactions, limiting therapeutic options. One reason for this limited understanding is AGEs are typically induced in vitro using high concentrations of ribose which decrease cell viability and make it impossible to investigate cell-matrix interactions. The objective of this study was to develop a system to trigger AGE accumulation while maintaining cell viability. Using cell-seeded high density collagen gels, we investigated the effect of two different systems for AGE induction, ribose at low concentrations (30, 100, and 200 mM) over 15 days of culture and riboflavin (0.25 mM and 0.75mM) induced with blue light for 40 seconds. We found ribose and riboflavin with blue light are capable of producing a wide range of AGE crosslinks which match and/or exceed reported human AGE levels for various tissues, ages, and diseases, without affecting cell viability and metabolism. Interestingly, a single 40 second treatment of riboflavin and blue light produced similar levels of AGEs as 3 days of 100 mM ribose treatment and matched aged mouse tendon AGE levels. This riboflavin treatment option is an exciting means to trigger AGE crosslinks on demand in vivo or in vitro without impacting cell metabolism or viability and holds great promise for further unraveling the mechanism of AGEs in age and diabetes related tissue degeneration.

Understanding and Regulating Cell-Matrix Interactions Using Hydrogels of Designable Mechanical Properties

2021 ◽  
Vol 17 (2) ◽  
pp. 149-168
Author(s):  
Jiapeng Yang ◽  
Yu Zhang ◽  
Meng Qin ◽  
Wei Cheng ◽  
Wei Wang ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Mechanical Response ◽  
Research Direction ◽  
Mechanical Environment ◽  
Cell Functions ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Sense And Respond ◽  
Cell Matrix Interactions

Similar to natural tissues, hydrogels contain abundant water, so they are considered as promising biomaterials for studying the influence of the mechanical properties of extracellular matrices (ECM) on various cell functions. In recent years, the growing research on cellular mechanical response has revealed that many cell functions, including cell spreading, migration, tumorigenesis and differentiation, are related to the mechanical properties of ECM. Therefore, how cells sense and respond to the extracellular mechanical environment has gained considerable attention. In these studies, hydrogels are widely used as the in vitro model system. Hydrogels of tunable stiffness, viscoelasticity, degradability, plasticity, and dynamical properties have been engineered to reveal how cells respond to specific mechanical features. In this review, we summarize recent process in this research direction and specifically focus on the influence of the mechanical properties of the ECM on cell functions, how cells sense and respond to the extracellular mechanical environment, and approaches to adjusting the stiffness of hydrogels.

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