ECIS, Cellular Adhesion and Migration in Keratinocytes

Background: The development of materials with bioregenerative properties is critically important for vital pulp therapies and regenerative endodontic procedures. The aim of this study was to evaluate the cytocompatibility and cytotoxicity of a new endodontic biomaterial, PulpGuard, in comparison with two other biomaterials widely used in endodontic procedures, ProRoot Mineral Trioxide Aggregate (MTA) and Biodentine. Methods: Apical papilla cells (APCs) were isolated from third molars with incomplete rhizogenesis from patients with orthodontic indication for dental extraction. Cultured APCs were incubated for 24, 48, or 72 h with different dilutions of eluates prepared from the three materials. Cellular viability, mobility, and proliferation were assessed in vitro using the Alamar Blue assay and a wound-healing test. The cells were also cultured in direct contact with the surface of each material. These were then analyzed via Scanning Electron Microscopy (SEM), and the surface chemical composition was determined by Energy-Dispersive Spectroscopy (EDS). Results: Cells incubated in the presence of eluates extracted from ProRoot MTA and PulpGuard presented rates of viability comparable to those of control cells; in contrast, undiluted Biodentine eluates induced a significant reduction of cellular viability. The wound-healing assay revealed that eluates from ProRoot MTA and PulpGuard allowed for unhindered cellular migration and proliferation. Cellular adhesion was observed on the surface of all materials tested. Consistent with their disclosed composition, EDS analysis found high relative abundance of calcium in Biodentine and ProRoot MTA and high abundance of silicon in PulpGuard. Significant amounts of zinc and calcium were also present in PulpGuard discs. Concerning solubility, Biodentine and ProRoot MTA presented mild weight loss after eluate extraction, while PulpGuard discs showed significant water uptake. Conclusions: PulpGuard displayed a good in vitro cytocompatibility profile and did not significantly affect the proliferation and migration rates of APCs. Cells cultured in the presence of PulpGuard eluates displayed a similar profile to those cultured with eluates from the widely used endodontic cement ProRoot MTA.

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Natural Compounds as Target Biomolecules in Cellular Adhesion and Migration: From Biomolecular Stimulation to Label-Free Discovery and Bioactivity-Based Isolation

Biomedicines ◽

10.3390/biomedicines9121781 ◽

2021 ◽

Vol 9 (12) ◽

pp. 1781

Author(s):

Beatrix Péter ◽

Imre Boldizsár ◽

Gábor M. Kovács ◽

Anna Erdei ◽

Zsuzsa Bajtay ◽

...

Keyword(s):

Natural Compounds ◽

Cellular Adhesion ◽

Bioactive Metabolites ◽

Label Free ◽

Antiproliferative Effects ◽

Receptor Sites ◽

Measuring Techniques ◽

Surface Receptor ◽

And Migration ◽

Active Substances

Plants and fungi can be used for medical applications because of their accumulation of special bioactive metabolites. These substances might be beneficial to human health, exerting also anti-inflammatory and anticancer (antiproliferative) effects. We propose that they are mediated by influencing cellular adhesion and migration via various signaling pathways and by directly inactivating key cell adhesion surface receptor sites. The evidence for this proposition is reviewed (by summarizing the natural metabolites and their effects influencing cellular adhesion and migration), along with the classical measuring techniques used to gain such evidence. We systematize existing knowledge concerning the mechanisms of how natural metabolites affect adhesion and movement, and their role in gene expression as well. We conclude by highlighting the possibilities to screen natural compounds faster and more easily by applying new label-free methods, which also enable a far greater degree of quantification than the conventional methods used hitherto. We have systematically classified recent studies regarding the effects of natural compounds on cellular adhesion and movement, characterizing the active substances according to their organismal origin (plants, animals or fungi). Finally, we also summarize the results of recent studies and experiments on SARS-CoV-2 treatments by natural extracts affecting mainly the adhesion and entry of the virus.

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Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells

The Journal of Cell Biology ◽

10.1083/jcb.201002119 ◽

2010 ◽

Vol 190 (4) ◽

pp. 675-691 ◽

Cited By ~ 186

Author(s):

Mark T. Howes ◽

Matthew Kirkham ◽

James Riches ◽

Katia Cortese ◽

Piers J. Walser ◽

...

Keyword(s):

High Capacity ◽

Leading Edge ◽

Cellular Adhesion ◽

Endocytic Pathway ◽

Membrane Turnover ◽

Major Pathway ◽

Endocytic Sorting ◽

And Migration ◽

Sorting Station ◽

Migrating Cells

Although the importance of clathrin- and caveolin-independent endocytic pathways has recently emerged, key aspects of these routes remain unknown. Using quantitative ultrastructural approaches, we show that clathrin-independent carriers (CLICs) account for approximately three times the volume internalized by the clathrin-mediated endocytic pathway, forming the major pathway involved in uptake of fluid and bulk membrane in fibroblasts. Electron tomographic analysis of the 3D morphology of the earliest carriers shows that they are multidomain organelles that form a complex sorting station as they mature. Proteomic analysis provides direct links between CLICs, cellular adhesion turnover, and migration. Consistent with this, CLIC-mediated endocytosis of key cargo proteins, CD44 and Thy-1, is polarized at the leading edge of migrating fibroblasts, while transient ablation of CLICs impairs their ability to migrate. These studies provide the first quantitative ultrastructural analysis and molecular characterization of the major endocytic pathway in fibroblasts, a pathway that provides rapid membrane turnover at the leading edge of migrating cells.

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Fabrication of Single and Multi-Layer Fibrous Biomaterial Scaffolds for Tissue Engineering

Volume 2: Biomedical and Biotechnology Engineering ◽

10.1115/imece2008-67964 ◽

2008 ◽

Author(s):

Amrinder S. Nain ◽

Eric Miller ◽

Metin Sitti ◽

Phil Campbell ◽

Cristina Amon

Keyword(s):

Tissue Engineering ◽

Fiber Diameter ◽

Living Cells ◽

Cellular Adhesion ◽

C2c12 Cells ◽

Fiber Axis ◽

Cellular Interactions ◽

Fiber Spacing ◽

Biomaterial Scaffolds ◽

And Migration

For regenerative medicine applications, we need to expand our understanding of the mechanisms by which nature assembles and functionalizes specialized complex tissues to form a complete organism. The first step towards this goal involves understanding the underlying complex mechanisms of highly organized behavior spanning not only diverse scientific fields, but also nano, micro and macro length-scales. For example, an engineered fibrous biomaterial scaffold possessing the hierarchal spatial properties of a native extracellular matrix (ECM) can serve as a building block upon which living cells are seeded for repair or regeneration. The hierarchical nature of ECM along with the inherent topological constraints of fiber diameter, fiber spacing, multi-layer configurations provide different pathways for living cells to adapt and conform to the surrounding environment. Our previously developed Spinneret based Tunable Engineered Parameters (STEP) technique to deposit biomaterial scaffolds in aligned configurations has been used for the first time to deposit single and multi-layer biological scaffolds of fibrinogen. Fibrinogen is a very well established tissue engineering scaffold material, as it improves cellular interactions and allows scaffold remodeling compared to synthetic polymers. Current state-of-the-art fiber deposition techniques lack the ability to fabricate scaffolds of desired fiber dimensions and orientations and in this study we present fabrication and aligned deposition of fibrinogen fiber arrays with diameters ranging from sub-200 nm to sub-microns and several millimeters in length. The fabricated scaffolds are then cultured with pluripotent mouse C2C12 cells for seven days and cells on the scaffolds are observed to elongate resembling myotube morphology along the fiber axis, spread along intersecting layers and fuse into bundles at the macroscale. Additionally, we demonstrate the ability to deposit poly (lactic-co-glycolic acid) (PLGA), Polystyrene (PS) biomaterial scaffolds of different diameters to investigate the effects of topological variations on cellular adhesion, proliferation and migration. Previous studies have indicated cells making right angle transitions upon encountering perpendicular double layer fibers and cellular motion is thwarted in the vicinity of diverging fibers. Current ongoing studies are aimed at determining the effects of fiber diameter and fiber spacing on mouse C2C12 cellular adhesion and migration, which are envisioned to aid in the design of future scaffolds for tissue engineering possessing appropriate material and geometrical properties.

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Small heat shock proteins in cellular adhesion and migration

Cell Adhesion & Migration ◽

10.4161/cam.20101 ◽

2012 ◽

Vol 6 (2) ◽

pp. 78-84 ◽

Cited By ~ 6

Author(s):

Georgina N. Montagna ◽

Kai Matuschewski ◽

Carlos A. Buscaglia

Keyword(s):

Heat Shock ◽

Heat Shock Proteins ◽

Cellular Adhesion ◽

Small Heat Shock Proteins ◽

And Migration ◽

Shock Proteins

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The regulation of the cell surface proteome by amp-activated protein kinase

10.32920/ryerson.14662851.v1 ◽

2021 ◽

Author(s):

Eden Ross

Keyword(s):

Protein Kinase ◽

Cell Surface ◽

Metabolic Regulation ◽

Cultured Cells ◽

Cellular Adhesion ◽

Ampk Activation ◽

And Migration ◽

Surface Proteome ◽

Amp Activated Protein Kinase ◽

Cell Surface Proteome

The cell-surface proteome controls numerous cellular functions and is dynamically controlled by endocytosis and recycling under different cellular conditions. Energy stress is a state in which a cell must engage adaptive responses to ensure survival, including remodelling of the cell-surface proteome. AMP-activated protein kinase (AMPK) is an important metabolic regulator in the cell. Recent studies suggest AMPK activation may alter the endocytosis of a few specific proteins. How increased AMPK activity globally regulates the cell surface proteome is not known. I have developed a method to isolate the cell surface proteome from cultured cells. Coupling this method to quantitative mass spectrometry has allowed systematic identification of changes in the cell-surface proteome upon metabolic regulation. I found that activation of AMPK results in robust changes in the cell surface proteome, including cell adhesion and migration proteins. I confirmed that AMPK activation elicits a decrease in the cell surface abundance of the adhesion and migration protein β1-integrin, and that this is correlated with altered function of the endocytosis protein Dab2. Thus, my research furthers our understanding of how AMPK regulates the cell surface proteome and the specific mechanism by which AMPK regulates cellular adhesion and migration.

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Corrigendum to “TGF-β1 targets a microRNA network that regulates cellular adhesion and migration in renal cancer” [Canc. Lett. 412 (2018) 155–169]

Cancer Letters ◽

10.1016/j.canlet.2020.01.006 ◽

2020 ◽

Vol 480 ◽

pp. 52-55

Author(s):

Joanna Bogusławska ◽

Katarzyna Rodzik ◽

Piotr Popławski ◽

Hanna Kędzierska ◽

Beata Rybicka ◽

...

Keyword(s):

Renal Cancer ◽

Cellular Adhesion ◽

And Migration ◽

Tgf Β1

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The Suppressing Effects of Dkk3 Expression on Aggressiveness and Tumorigenesis of Colorectal Cancer

Frontiers in Oncology ◽

10.3389/fonc.2020.600322 ◽

2020 ◽

Vol 10 ◽

Author(s):

Shuang Zhao ◽

Chang-lai Hao ◽

En-hong Zhao ◽

Hua-mao Jiang ◽

Hua-chuan Zheng

Keyword(s):

Colorectal Cancer ◽

Cellular Adhesion ◽

Migration And Invasion ◽

Signal Pathways ◽

Protein Levels ◽

Cancer Aggressiveness ◽

Pattern Differentiation ◽

And Migration ◽

Patient Prognosis ◽

Epithelial Lesions

Dkk3 has been discovered during comparison of immortalized and parental cells. Its expression has been shown to reduce colony formation and induce apoptosis of cancer cells, acting as a tumor suppressor. Herein, we demonstrate that Dkk3 overexpression or protein treatment may inhibit colorectal cancer cell proliferation, migration, and invasion and that they may promote apoptosis and G2 phase arrest with hypoexpression of Bcl-2, cdc25B, cdc25c, N-cadherin, slug, and twist and hyperexpression of Bax and E-cadherin. This effect is consistent with that of recombinant Dkk3 exposure and blocked with anti-Dkk3 antibody. Dkk3 deletion in intestinal cells was not associated with the emergence of epithelial lesions; however, adenoma emerged after sodium desoxycholate treatment. At both mRNA and protein levels, Dkk3 expression was higher in normal than in cancer tissues (p<0.05). Dkk3 mRNA expression was negatively associated with its promoter methylation, growth pattern, differentiation, and favorable prognosis in the patients with colorectal cancer (p<0.05). Dkk3-related signal pathways in colorectal cancer included those of cellular adhesion and migration, melanogenesis, chemokine, Hedgehog, JAK-STAT, TOLL-like receptor, TGF-β, MAPK, and calcium signaling (p<0.05). These findings indicate that Dkk3 expression levels can help assess cancer aggressiveness and patient prognosis. It might also suppress aggressive phenotypes and tumorigenesis as a molecular target in gene therapy.

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Vitronectin acts as a key regulator of adhesion and migration in umbilical cord-derived MSCs under different stress conditions

10.1101/2021.11.18.468035 ◽

2021 ◽

Author(s):

Malancha Ta ◽

Ankita Sen

Keyword(s):

Cellular Therapy ◽

Cellular Adhesion ◽

Stress Conditions ◽

Cellular Migration ◽

Serum Starvation ◽

Erk Pathway ◽

Stress Fibre ◽

Spread Area ◽

And Migration ◽

Cell Spread

Mesenchymal stem cell (MSC)-based cellular therapy gets compromised as adverse microenvironmental conditions like nutrient deprivation, ischemia, hypoxia affect migration and engraftment, in addition to viability, of MSCs at the target site post transplantation. To improve the treatment efficacy, it is critical to identify factors involved in regulating migration and adhesion of MSCs under such microenvironmental stress conditions. In our study, we observed that Wharton's jelly-MSCs (WJ-MSCs) exhibited increase in cell spread area and adhesion with reduction in cellular migration under serum starvation. The changes in adhesion and migration characteristics were accompanied by extensive stress fibre formation and altered ECM gene expression with notable induction in vitronectin (VTN) expression and reduction in MMP-1 expression. Molecular and phenotypic correlative studies advocated the possible role of VTN and not MMP-1, in regulating adhesion and migration of WJ-MSCs. NF-kb was found to be the positive regulator of VTN expression while ERK pathway regulated it negatively. Further investigation with inhibition of these signalling pathways or VTN knockdown studies under serum starvation established the correlation between increase in VTN expression and increased cellular adhesion with corresponding reduction in migration. VTN knockdown under serum starvation also led to reduction in actin stress fibre along with reversal in expression of several ECM genes. Additionally, VTN induction being absent in hypoxia-treated WJ-MSCs, the hypoxic cells showed no significant change in the adhesion and migration properties. However, when VTN expression was induced under hypoxia by ERK pathway inhibition, similar increase in cell spread area and adhesion was observed. Our study thus highlights VTN as a factor which is induced under serum starvation stress and possibly affects the adhesion and migration properties of WJ-MSCs.

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Stimulation of β1-Integrin Function by Epidermal Growth Factor and Heregulin-β Has Distinct Requirements for erbB2 but a Similar Dependence on Phosphoinositide 3-OH Kinase

Molecular Biology of the Cell ◽

10.1091/mbc.10.9.2861 ◽

1999 ◽

Vol 10 (9) ◽

pp. 2861-2878 ◽

Cited By ~ 58

Author(s):

Margaret A. Adelsman ◽

James B. McCarthy ◽

Yoji Shimizu

Keyword(s):

Cell Adhesion ◽

Growth Factor ◽

Breast Carcinoma ◽

Carcinoma Cell ◽

Critical Role ◽

Metastatic Breast ◽

Cellular Adhesion ◽

Breast Carcinoma Cell Line ◽

Β1 Integrin ◽

And Migration

Integrins and growth factor receptors are important participants in cellular adhesion and migration. The EGF receptor (EGFR) family of tyrosine kinases and the β1-integrin adhesion receptors are of particular interest, given the implication for their involvement in the initiation and progression of tumorigenesis. We used adhesion and chemotaxis assays to further elucidate the relationship between these two families of transmembrane signaling molecules. Specifically, we examined integrin-mediated adhesive and migratory characteristics of the metastatic breast carcinoma cell line MDA-MB-435 in response to stimulation with growth factors that bind to and activate the EGFR or erbB3 in these cells. Although ligand engagement of the EGFR stimulated modest β1-dependent increases in cell adhesion and motility, heregulin-β (HRGβ) binding to the erbB3 receptor initiated rapid and potent induction of breast carcinoma cell adhesion and migration and required dimerization of erbB3 with erbB2. Pharmacologic inhibitors of phosphoinositide 3-OH kinase (PI 3-K) or transient expression of dominant negative forms of PI 3-K inhibited both EGF- and HRGβ-mediated adhesion and potently blocked HRGβ- and EGF-induced cell motility. Our results illustrate the critical role of PI 3-K activity in signaling pathways initiated by the EGFR or erbB3 to up-regulate β1-integrin function.

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