Comparative cell adhesion properties of cysteine extended peptide architectures

AbstractIn the current study the in vitro outcome of a degradable magnesium alloy (AZ91D) and standard titanium modified by nanostructured-hydroxyapatite (n-HA) coatings concerning cell adhesion and osteogenic differentiation was investigated by direct cell culture. The n-HA modification was prepared via radio-frequency magnetron sputtering deposition and proven by field emission scanning electron microscopy and X-ray powder diffraction patterns revealing a homogenous surface coating. Human mesenchymal stem cell (hMSCs) adhesion was examined after one and 14 days displaying an enhanced initial cell adhesion on the n-HA modified samples. The osteogenic lineage commitment of the cells was determined by alkaline phosphatase (ALP) quantification. On day one n-HA coated AZ91D exhibited a comparable ALP expression to standard tissue culture polystyrene samples. However, after 14 days solely little DNA and ALP amounts were measurable on n-HA coated AZ91D due to the lack of adherent cells. Titanium displayed excellent cell adhesion properties and ALP was detectable after 14 days. An increased pH of the culture was measured for AZ91D as well as for n-HA coated AZ91D. We conclude that n-HA modification improves initial cell attachment on AZ91D within the first 24 h. However, the effect does not persist for 14 days in in vitro conditions.

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A Conserved NXIP Motif Is Required for Cell Adhesion Properties of the Syndecan-4 Ectodomain

Journal of Biological Chemistry ◽

10.1074/jbc.m605553200 ◽

2006 ◽

Vol 281 (43) ◽

pp. 32156-32163 ◽

Cited By ~ 33

Author(s):

James R. Whiteford ◽

John R. Couchman

Keyword(s):

Cell Adhesion ◽

Cell Attachment ◽

Focal Adhesions ◽

Cell Types ◽

Binding Activity ◽

Cytoskeletal Reorganization ◽

Cell Binding ◽

Adhesion Properties ◽

Support Cell ◽

Β1 Integrins

Syndecans are cell surface proteoglycans involved in cell adhesion and motility. Syndecan-4 is an important component of focal adhesions and is involved in cytoskeletal reorganization. Previous work has shown that the syndecan-4 ectodomain can support cell attachment. Here, three vertebrate syndecan-4 ectodomains were compared, including that of the zebrafish, and we have demonstrated that the cell binding activity of the syndecan-4 ectodomain is conserved. Cell adhesion to the syndecan-4 ectodomain appears to be a characteristic of mesenchymal cells. Comparison of syndecan-4 ectodomain sequences led to the identification of three conserved regions of sequence, of which the NXIP motif is important for cell binding activity. We have shown that cell adhesion to the syndecan-4 ectodomain involves β1 integrins in several cell types.

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Effect of Calcium Chloride Hydrothermal Treatment of Titanium on Protein, Cellular, and Bacterial Adhesion Properties

Journal of Clinical Medicine ◽

10.3390/jcm9082627 ◽

2020 ◽

Vol 9 (8) ◽

pp. 2627 ◽

Cited By ~ 1

Author(s):

Takuya Haraguchi ◽

Yasunori Ayukawa ◽

Yukie Shibata ◽

Toru Takeshita ◽

Ikiru Atsuta ◽

...

Keyword(s):

Cell Adhesion ◽

Epithelial Cell ◽

Calcium Chloride ◽

Hydrothermal Treatment ◽

Bacterial Adhesion ◽

Chloride Solution ◽

Cell Attachment ◽

Calcium Chloride Solution ◽

Adhesion Properties ◽

Dental Implant Surface

Topographical modification of the dental implant surface is one of the main topics for the improvement of the material, however, the roughened surface has some risks for peri-implantitis. A hydrothermal treatment (HT) of titanium with calcium chloride solution was reported to improve osseointegration and soft tissue sealing without changing the surface topography; however, its mechanism is unclear. We herewith investigated the interaction between extracellular matrix (ECM) protein and HT titanium. Furthermore, we also clarified the bacterial interaction. We employed two kinds of HT, HT with water (DW-HT) and HT with calcium chloride solution (Ca-HT). As a result, the adsorptions of both laminin-332 and osteopontin onto the Ca-HT surface were enhanced. In contrast, the adsorption of albumin, which was reported to have no cell adhesion capacity, was not influenced by Ca-HT. Osteoblast adhesion onto Ca-HT was also enhanced. Although Ca-HT was reported to enhance both epithelial cell attachment strength and in vivo peri-implant epithelial bonding, the number of epithelial cell attachment was not increased even after HT. Ca-HT had no impact in the adhesion of Streptococcus gordonii. These results suggest that Ca-HT enhances cell adhesion onto titanium without increasing bacterial adhesion, and the improvement of ECM protein adsorption is supposed to contribute to cell adhesion.

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Neutrophil Extracellular Traps as an Adhesion Substrate for Different Tumor Cells Expressing RGD-Binding Integrins

International Journal of Molecular Sciences ◽

10.3390/ijms19082350 ◽

2018 ◽

Vol 19 (8) ◽

pp. 2350 ◽

Cited By ~ 5

Author(s):

Marcello Monti ◽

Viviana De Rosa ◽

Francesca Iommelli ◽

Maria Vincenza Carriero ◽

Cristina Terlizzi ◽

...

Keyword(s):

Cell Adhesion ◽

Cancer Cells ◽

Cell Lines ◽

Defense Mechanism ◽

Cell Attachment ◽

Thrombus Formation ◽

Neutrophil Extracellular Traps ◽

Adhesion Properties ◽

Extracellular Traps ◽

Relevant Role

Neutrophil extracellular traps (NETs), in addition to their function as a host defense mechanism, play a relevant role in thrombus formation and metastatic dissemination of cancer cells. Here we screened different cancer cell lines endogenously expressing a variety of integrins for their ability to bind to NETs. To this end, we used NETs isolated from neutrophil-like cells as a substrate for adhesion assays of HT1080, U-87 MG, H1975, DU 145, PC-3 and A-431 cells. Levels of α5, αIIb, αv, β1, β3 and β5 chains were determined by western blot analysis in all cell lines and levels of whole integrins on the plasma membrane were assessed by fluorescence-activated cell sorting (FACS) analysis. We found that high levels of α5β1, αvβ3 and αvβ5 enhance cell adhesion to NETs, whereas low expression of α5β1 prevents cell attachment to NETs. Excess of cyclic RGD peptide inhibited cell adhesion to NETs by competing with fibronectin within NETs. The maximal reduction of such adhesion was similar to that obtained by DNase 1 treatment causing DNA degradation. Our findings indicate that NETs from neutrophil-like cells may be used as a substrate for large screening of the adhesion properties of cancer cells expressing a variety of RGD-binding integrins.

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Smart Hybrid Mussel Adhesive Materials for Cell and Tissue Engineering

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.47-50.861 ◽

2008 ◽

Vol 47-50 ◽

pp. 861-864

Author(s):

Dong Soo Hwang ◽

Seong Hye Lim ◽

Hyung Joon Cha

Keyword(s):

Tissue Engineering ◽

Cell Adhesion ◽

Cell Attachment ◽

Attachment Site ◽

Peptide Sequence ◽

Adhesion Properties ◽

Adhesion Ability ◽

A Cell ◽

Mussel Adhesive ◽

Adhesive Materials

Mussel foot protein is widely known for candidate of strong adhesive materials. Six times fp-1 decapeptide repeats and fp-5 are chosen among the various type of foot protein and fused into (MAP) fp-151. Through various cell-adhesion analyses, we previously demonstrated that fp- 151 has the potential to be used as a cell or tissue bioadhesive. In the present study, to improve the cell-adhesion properties of fp-151, it was designed a new cell-adhesive protein, fp-151-RGD, which is a fusion with the GRGDSP residues, a RGD peptide sequence that has previously been identified at the cell-attachment site of fibronectin, at the C-terminus of fp-151. Also, recombinant fp-151- RGD maintained the advantages associated with fp-151, such as a high production yield in Escherichia coli and simple purification, it showed superior spreading ability, which is important for cell proliferation under serum-free conditions, as well as better cell-adhesion ability compared with other commercially produced cell-adhesion materials such as poly-L-lysine (PLL) and the naturally extracted MAP mixture Cell-Tak. The excellent adhesion and spreading abilities of fp- 151-RGD might be due to the fact that it utilizes three types of cell-binding mechanisms: DOPA adhesion of Cell-Tak, cationic binding force of PLL, and RGD sequence-mediated adhesion of fibronectin. Therefore, the new recombinant fp-151-RGD is suitable for use as a cell-adhesion material in cell culture or tissue engineering, and in any other area where efficient cell adhesion is required.

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Microgravity Induces Transient EMT in Human Keratinocytes by Early Down-Regulation of E-Cadherin and Cell-Adhesion Remodeling

Applied Sciences ◽

10.3390/app11010110 ◽

2020 ◽

Vol 11 (1) ◽

pp. 110

Author(s):

Giulia Ricci ◽

Alessandra Cucina ◽

Sara Proietti ◽

Simona Dinicola ◽

Francesca Ferranti ◽

...

Keyword(s):

Cell Adhesion ◽

Human Keratinocytes ◽

Short Exposure ◽

Migration And Invasion ◽

Hacat Cells ◽

Invasive Phenotype ◽

Adhesion Properties ◽

Mesenchymal Transition ◽

Cell Matrix ◽

E Cadherin

Changes in cell–matrix and cell-to-cell adhesion patterns are dramatically fostered by the microgravity exposure of living cells. The modification of adhesion properties could promote the emergence of a migrating and invasive phenotype. We previously demonstrated that short exposure to the simulated microgravity of human keratinocytes (HaCaT) promotes an early epithelial–mesenchymal transition (EMT). Herein, we developed this investigation to verify if the cells maintain the acquired invasive phenotype after an extended period of weightlessness exposure. We also evaluated cells’ capability in recovering epithelial characteristics when seeded again into a normal gravitational field after short microgravity exposure. We evaluated the ultra-structural junctional features of HaCaT cells by Transmission Electron Microscopy and the distribution pattern of vinculin and E-cadherin by confocal microscopy, observing a rearrangement in cell–cell and cell–matrix interactions. These results are mirrored by data provided by migration and invasion biological assay. Overall, our studies demonstrate that after extended periods of microgravity, HaCaT cells recover an epithelial phenotype by re-establishing E-cadherin-based junctions and cytoskeleton remodeling, both being instrumental in promoting a mesenchymal–epithelial transition (MET). Those findings suggest that cytoskeletal changes noticed during the first weightlessness period have a transitory character, given that they are later reversed and followed by adaptive modifications through which cells miss the acquired mesenchymal phenotype.

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Integrins: An Important Link between Angiogenesis, Inflammation and Eye Diseases

Cells ◽

10.3390/cells10071703 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1703

Author(s):

Małgorzata Mrugacz ◽

Anna Bryl ◽

Mariusz Falkowski ◽

Katarzyna Zorena

Keyword(s):

Cell Adhesion ◽

Tissue Repair ◽

Normal Development ◽

Cell Attachment ◽

Biological Activities ◽

Integrin Receptors ◽

Cytokine Activation ◽

Eye Disorders ◽

Membrane Bound ◽

Cell To Cell Adhesion

Integrins belong to a group of cell adhesion molecules (CAMs) which is a large group of membrane-bound proteins. They are responsible for cell attachment to the extracellular matrix (ECM) and signal transduction from the ECM to the cells. Integrins take part in many other biological activities, such as extravasation, cell-to-cell adhesion, migration, cytokine activation and release, and act as receptors for some viruses, including severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2). They play a pivotal role in cell proliferation, migration, apoptosis, tissue repair and are involved in the processes that are crucial to infection, inflammation and angiogenesis. Integrins have an important part in normal development and tissue homeostasis, and also in the development of pathological processes in the eye. This review presents the available evidence from human and animal research into integrin structure, classification, function and their role in inflammation, infection and angiogenesis in ocular diseases. Integrin receptors and ligands are clinically interesting and may be promising as new therapeutic targets in the treatment of some eye disorders.

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Redistribution and dysfunction of integrins in cultured renal epithelial cells exposed to oxidative stress

AJP Renal Physiology ◽

10.1152/ajprenal.1993.264.1.f149 ◽

1993 ◽

Vol 264 (1) ◽

pp. F149-F157 ◽

Cited By ~ 14

Author(s):

J. Gailit ◽

D. Colflesh ◽

I. Rabiner ◽

J. Simone ◽

M. S. Goligorsky

Keyword(s):

Oxidative Stress ◽

Cell Adhesion ◽

Epithelial Cell ◽

Epithelial Cells ◽

Cell Surface ◽

Apical Cell ◽

Flow Cytometric Analysis ◽

Adhesion Properties ◽

Renal Tubular Cells ◽

Renal Tubular

Tubular obstruction by detached renal tubular epithelial cells is a major cause of oliguria in acute renal failure. Viable renal tubular cells can be recovered from urine of patients with acute tubular necrosis, suggesting a possible defect in cell adhesion to the basement membrane. To study this process of epithelial cell desquamation in vitro, we investigated the effect of nonlethal oxidative stress on the integrin adhesion receptors of the primate kidney epithelial cell line BS-C-1. Morphological and functional studies of cell adhesion properties included the following: interference reflection microscopy, intravital confocal microscopy and immunocytochemistry, flow cytometric analysis of integrin receptor abundance, and cell-matrix attachment assay. High levels of the integrin subunits alpha 3, alpha v, and beta 1 were detected on the cell surface by fluorescence-activated cell sorting (FACS) analysis, as well as lower levels of alpha 1, alpha 2, alpha 4, alpha 5, alpha 6, and beta 3. Exposure of BS-C-1 cells to nonlethal oxidative stress resulted in the disruption of focal contacts, disappearance of talin from the basal cell surface, and in the redistribution of integrin alpha 3-subunits from predominantly basal location to the apical cell surface. As measured in a quantitative cell attachment assay, oxidative stress decreased BS-C-1 cell adhesion to type IV collagen, laminin, fibronectin, and vitronectin. Defective adhesion was not associated with a loss of alpha 3-, alpha 4-, or alpha v-integrin subunits from the cell surface.(ABSTRACT TRUNCATED AT 250 WORDS)

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Receptor functions for the integrin VLA-3: fibronectin, collagen, and laminin binding are differentially influenced by Arg-Gly-Asp peptide and by divalent cations.

The Journal of Cell Biology ◽

10.1083/jcb.112.1.169 ◽

1991 ◽

Vol 112 (1) ◽

pp. 169-181 ◽

Cited By ~ 260

Author(s):

M J Elices ◽

L A Urry ◽

M E Hemler

Keyword(s):

Cell Adhesion ◽

Divalent Cations ◽

Small Cell Lung Carcinoma ◽

Rgd Peptide ◽

Dependent Manner ◽

Cell Lung Carcinoma ◽

Ligand Affinity ◽

Rgd Site ◽

Inhibition Studies ◽

Laminin Binding

The capability of the integrin VLA-3 to function as a receptor for collagen (Coll), laminin (Lm), and fibronectin (Fn) was addressed using both whole cell adhesion assays and ligand affinity columns. Analysis of VLA-3-mediated cell adhesion was facilitated by the use of a small cell lung carcinoma line (NCI-H69), which expresses VLA-3 but few other integrins. While VLA-3 interaction with Fn was often low or undetectable in cells having both VLA-3 and VLA-5, NCI-H69 cells readily attached to Fn in a VLA-3-dependent manner. Both Arg-Gly-Asp (RGD) peptide inhibition studies, and Fn fragment affinity columns suggested that VLA-3, like VLA-5, may bind to the RGD site in human Fn. However, unlike Fn, both Coll and Lm supported VLA-3-mediated adhesion that was not inhibited by RGD peptide, and was totally unaffected by the presence of VLA-5. In addition, VLA-3-mediated binding to Fn was low in the presence of Ca++, but was increased 6.6-fold with Mg++, and 30-fold in the presence of Mn++. In contrast, binding to Coll was increased only 1.2-fold with Mg++, and 1.7-fold in Mn++, as compared to the level seen with Ca++. Together, these experiments indicate that VLA-3 can bind Coll, Lm, and Fn, and also show that (a) VLA-3 can recognize both RGD-dependent and RGD-independent ligands, and (b) different VLA-3 ligands have distinctly dissimilar divalent cation sensitivities.

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