Nanoengineered Surfaces for Focal Adhesion Guidance Trigger Mesenchymal Stem Cell Self-Organization and Tenogenesis
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The initial conditions for morphogenesis trigger a cascade of events that ultimately dictate structure and functions of tissues and organs. Here we report that surface nanopatterning can control the initial assembly of focal adhesions, hence guiding human mesenchymal stem cells (hMSCs) through the process of self-organization and differentiation. This process self-sustains, leading to the development of macroscopic tissues with molecular profiles and microarchitecture reminiscent of embryonic tendons. Therefore, material surfaces can be in principle engineered to set off the hMSC program toward tissuegenesis in a deterministic manner by providing adequate sets of initial environmental conditions.Tissue morphogenesis during development is dependent on activities of the cadherin family of cell–cell adhesion proteins that includes classical cadherins, protocadherins, and atypical cadherins (Fat, Dachsous, and Flamingo). The extracellular domain of cadherins contains characteristic repeats that regulate homophilic and heterophilic interactions during adhesion and cell sorting. Although cadherins may have originated to facilitate mechanical cell–cell adhesion, they have evolved to function in many other aspects of morphogenesis. These additional roles rely on cadherin interactions with a wide range of binding partners that modify their expression and adhesion activity by local regulation of the actin cytoskeleton and diverse signaling pathways. Here we examine how different members of the cadherin family act in different developmental contexts, and discuss the mechanisms involved.
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Objective To study the effection of Echinacea polysaccharide(EP) on Escherichia coli adhesion to cell.Method Adhesion test and inhibiting adhesion test were carried out on PK-15 cell.Result EP had the best inhibition for E.coli adhesion to PK-15 cell at 1.6 mg/ml concentration,the number of bacteria adhesion to cells were reduced from 50 per cell to 6.8 per cell.Conclusion The cell adhesion of E.coli can be inhibited by EP,which may have the potential application value in regulating of gut microflora.
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Integrin-mediated adhesion to extracellular matrix proteins is dynamically regulated during morphological changes and cell migration. Upon cell adhesion, protein-protein interactions among molecules at focal adhesions (FAs) play major roles in the regulation of cell morphogenesis and migration. Although tyrosine phosphorylation of paxillin is critically involved in adhesion-mediated signaling, the significance of paxillin phosphorylation at Ser-85 and the mechanism by which it regulates cell migration remain unclear. In this study, we examined how Ser-85 phosphorylation of paxillin affects FA formation and cell migration. We found that paxillin phosphorylation at Ser-85 occurred during HeLa cell adhesion to collagen I and was concomitant with tyrosine phosphorylation of both focal adhesion kinase and talin. However, the non-phosphorylatable S85A mutant of paxillin impaired cell spreading, FA turnover, and migration toward collagen I but not toward serum. Furthermore, whereas the (presumably indirect) interaction between paxillin and the C-terminal tail of talin led to dynamic FAs at the cell boundary, S85A paxillin did not bind talin and caused stabilized FAs in the central region of cells. Together, these observations suggest that cell adhesion-dependent Ser-85 phosphorylation of paxillin is important for its interaction with talin and regulation of dynamic FAs and cell migration.
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Protein Adsorption
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Biomaterial
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In order to study the biophysical forces involved in cell-substrate (or cell-cell) adhesion, it is necessary to measure the strength of adhesion. Two questions can be addressed using the centrifugal cell adhesion assay provided in this unit: what is the ligand-receptor affinity for cells adhering at 4 degrees C and what is the strength of the ATP-dependent processes that strengthen adhesion at 37 degrees C. In both cases, the strength of adhesion is measured as the resistance to disruption in the presence of a measured centrifugal force.
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ABSTRACT Adhesion of BHK cells to a variety of polymer surfaces carrying measured densities of hydroxyl and carboxyl groups was studied. The effects on cell adhesion of blocking hydroxyl groups by acetylation and carboxyl groups with diazomethane were measured. Hydroxyl groups were required for cell adhesion, though the very high surface densities of these groups diminished cell adhesion. The optimal surface density of OH groups for BHK adhesion was 2000 per 1× 10−11 cm2. Carboxyl groups slightly inhibit cell adhesion, since blocking of these groups by methylation increased adhesion. The role of oxidizing systems of cellular origin in conditioning of the substrate, in serum-free conditions, was demonstrated for leucocytes and BHK cells, in particular by the result that oxidizable substrates such as phytane and poly(l,2-butadiene) could be made suitable for cell adhesion by contact with cells.
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Objective:The adhesion of cells to cells and cells to stroma play very important role in the cell-cell information exchanges.Methods:The cell adhesion assay based on the 3 H-TdR incorporation assay was introduced. Firstly, coated cells were cultured to confluence in 96 well plate. After incubated with 3 H-TdR for 6h, the isotope labeled cells were added into plate wells and incubated for another 4 h. Then the un-adhered cells were removed by gently washing. The cpm was counted after cell harvest.Results:The cell adhesion assay based on the 3 H-TdR incorporation assay was very sensitive in the detection of cell adhesion. It could reflect the relative of cell to cell adhesion.Conclusion:The cell adhesion assay based on the 3 H-TdR incorporation assay is an useful method in the detection of cell to cell adhesion.
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The use of force spectroscopy to study the adhesion of living fibroblasts to their culture substrate was investigated. Both primary fibroblasts (PEMF) and a continuous cell line (3T3) were studied on quartz surfaces. Using a fibronectin-coated AFM cantilever, it was possible to detach a large proportion of the 3T3 cells from the quartz surfaces. Their adhesion to the quartz surface and the effects of topography on this adhesion could be quantified. Three parameters characteristic of the adhesion were measured: the maximum force of detachment, the work of adhesion, and the distance of detachment. Few PEMF cells were detached under the same experimental conditions. The potential and limitations of this method in measuring cell/surface interactions for adherent cells are discussed.
Force Spectroscopy
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