Cell Adhesion Strength Research Articles

Nonthermal Plasma Technology as a Versatile Strategy for Polymeric Biomaterials Surface Modification: A Review

In modern technology, there is a constant need to solve very complex problems and to fine-tune existing solutions. This is definitely the case in modern medicine with emerging fields such as regenerative medicine and tissue engineering. The problems, which are studied in these fields, set very high demands on the applied materials. In most cases, it is impossible to find a single material that meets all demands such as biocompatibility, mechanical strength, biodegradability (if required), and promotion of cell-adhesion, proliferation, and differentiation. A common strategy to circumvent this problem is the application of composite materials, which combine the properties of the different constituents. Another possible strategy is to selectively modify the surface of a material using different modification techniques. In the past decade, the use of nonthermal plasmas for selective surface modification has been a rapidly growing research field. This will be the highlight of this review. In a first part of this paper, a general introduction in the field of surface engineering will be given. Thereafter, we will focus on plasma-based strategies for surface modification. The purpose of the present review is twofold. First, we wish to provide a tutorial-type review that allows a fast introduction for researchers into the field. Second, we aim to give a comprehensive overview of recent work on surface modification of polymeric biomaterials, with a focus on plasma-based strategies. Some recent trends will be exemplified. On the basis of this literature study, we will conclude with some future trends for research.

High-content profiling of cell responsiveness to graded substrates based on combinyatorially variant polymers.

We have developed a novel approach combining high information and high throughput analysis to characterize cell adhesive responses to biomaterial substrates possessing gradients in surface topography. These gradients were fabricated by subjecting thin film blends of tyrosine-derived polycarbonates, i.e. poly(DTE carbonate) and poly(DTO carbonate) to a gradient temperature annealing protocol. Saos-2 cells engineered with a green fluorescent protein (GFP) reporter for farnesylation (GFP-f) were cultured on the gradient substrates to assess the effects of nanoscale surface topology and roughness that arise during the phase separation process on cell attachment and adhesion strength. The high throughput imaging approach allowed us to rapidly identify the "global" and "high content" structure-property relationships between cell adhesion and biomaterial properties such as polymer chemistry and topography. This study found that cell attachment and spreading increased monotonically with DTE content and were significantly elevated at the position with intermediate regions corresponding to the highest "gradient" of surface roughness, while GFP-f farnesylation intensity descriptors were sensitively altered by surface roughness, even in cells with comparable levels of spreading.

Cell adhesive affinity does not dictate primitive endoderm segregation and positioning during murine embryoid body formation

The classical cell sorting experiments undertaken by Townes and Holtfreter described the intrinsic propensity of dissociated embryonic cells to self-organize and reconcile into their original embryonic germ layers with characteristic histotypic positioning. Steinberg presented the differential adhesion hypothesis to explain these patterning phenomena. Here, we have reappraised these issues by implementing embryoid bodies to model the patterning of epiblast and primitive endoderm layers. We have used combinations of embryonic stem (ES) cells and their derivatives differentiated by retinoic acid treatment to model epiblast and endoderm cells, and wild-type or E-cadherin null cells to represent strongly or weakly adherent cells, respectively. One cell type was fluorescently labeled and reconstituted with another heterotypically to generate chimeric embryoid bodies, and cell sorting was tracked by time-lapse video microscopy and confirmed by immunostaining. When undifferentiated wild-type and E-cadherin null ES cells were mixed, the resulting cell aggregates consisted of a core of wild-type cells surrounded by loosely associated E-cadherin null cells, consistent with the differential adhesion hypothesis. However, when mixed with undifferentiated ES cells, the differentiated primitive endoderm-like cells sorted to the surface to form a primitive endoderm layer irrespective of cell-adhesive strength, contradicting the differential adhesion hypothesis. We propose that the primitive endoderm cells reach the surface by random movement, and subsequently the cells generate an apical/basal polarity that prevents reentry. Thus, the ability to generate epithelial polarity, rather than adhesive affinity, determines the surface positioning of the primitive endoderm cells.

Temporal Effect of Functional Blocking of β1 Integrin on Cell Adhesion Strength under Serum Depletion

Cell adhesion is generally concomitant to the formation of focal adhesion. Although it is well-known that focal adhesion plays an important role in the functional regulations of anchorage dependent cells, previous experimental studies have not provided quantitative description of the relation between focal adhesion and biophysical responses of cells. Furthermore, there is lack of knowledge on the importance of the beta1 integrin subunit to the dynamic responses of cells during initial cell seeding. In this study, we attempt to bridge the quantitative connection between focal adhesion density and cell-substrate interactions and evaluate the influence on functional blocking of beta1 integrin on adhesion strength. Total internal reflection fluorescence microscopy (TIRFM), fluorescence microscopy, and phase contrast microscopy was employed to study the time-dependent evolvement of vinculin pattern, distribution of actin filament, and morphological change, respectively, during 4 h of culture for porcine esophageal fibroblasts (non-blocked and beta1-blocked) on a fibronectin-coated surface. Micropipet aspiration technique was used to study the change of mechanotransduction through the determination of adhesion force and strength. It is shown in our experimental results that spread area, adhesion force, and adhesion strength increases over time on the two types of cells. Throughout the culture period, the two key mechanotransduction parameters of non-blocked cells is higher than those of beta1-blocked cells. Interestingly, adhesion strength initially ascends, then begins to diminish at a critical time point, and finally resumes increasing linearly against the increase of focal adhesion density. This variation as mentioned above can be explained by peeling and fracture models based on the dissimilar vinculin pattern of cells after being cultured for different time periods. Moreover, the averaged focal adhesion strength and non-focal adhesion strength of beta1-blocked cells are significantly less than those of non-blocked of cells. The weaker adhesion strength on beta(1)-blocked cells is directly caused by lower focal and non-focal adhesion strength, as well as by smaller focal adhesion density.

Interactions of Zoospores of Ulva linza with Arginine-Rich Oligopeptide Monolayers

We recently reported on the strong interactions of zoospores of the green alga, Ulva linza with an arginine-rich oligopeptide self-assembled monolayer (SAM) [Biofouling 2008, 24, 303-312], where the arginine-rich peptide induced not only high spore settlement, but also a form of abnormal settlement, or "pseudo-settlement", whereby a proportion of spores do not go through the normal process of surface exploration, adhesive exocytosis, and loss of flagella. Further, it was demonstrated that both the total number of settled spores and the fraction of pseudosettled spores were related to the surface density of the arginine-rich peptide. Here we present a further investigation of the interactions of zoospores of Ulva with a set of oligomeric, de novo designed, arginine-rich peptides, specifically aimed to test the effect of peptide primary structure on the interaction. Via variations in the peptide length and by permutations in the amino acid sequences, we gain further insight into the spore-surface interactions. The interpretation of the biological assays is supported by physicochemical characterization of the SAMs using infrared spectroscopy, ellipsometry, and contact angle measurements. Results confirm the importance of arginine residues for the anomalous pseudosettlement, and we found that settlement is modulated by variations in both the total length and peptide primary structure. To elucidate the causes of the anomalous settlement and the possible relation to peptide-membrane interactions, we also compared the settlement of the "naked" zoospores of Ulva (which present a lipoprotein membrane to the exterior without a discrete polysaccharide cell wall), with the settlement of diatoms (unicellular algae that are surrounded by a silica cell wall), onto the peptide SAMs. Cationic SAMs do not notably affect settlement (attachment), adhesion strength, or viability of diatom cells, suggesting that the effect of the peptides on zoospores of Ulva is mediated via specific peptide-membrane interactions.

Antifouling potential of Subtilisin A immobilized onto maleic anhydride copolymer thin films

The proteinaceous nature of the adhesives used by most fouling organisms to attach to surfaces suggests that coatings incorporating proteolytic enzymes may provide a technology for the control of biofouling. In the present article, the antifouling (AF) and fouling release potential of model coatings incorporating the surface-immobilized protease, Subtilisin A, have been investigated. The enzyme was covalently attached to maleic anhydride copolymer thin films; the characteristics of the bioactive coatings obtained were adjusted through variation of the type of copolymer and the concentration of the enzyme solution used for immobilization. The bioactive coatings were tested for their effect on the settlement and adhesion strength of two major fouling species: the green alga Ulva linza and the diatom Navicula perminuta. The results show that the immobilized enzyme effectively reduced the settlement and adhesion strength of zoospores of Ulva and the adhesion strength of Navicula cells. The AF efficacy of the bioactive coatings increased with increasing enzyme surface concentration and activity, and was found to be superior to the equivalent amount of enzyme in solution. The results provide a rigorous analysis of one approach to the use of immobilized proteases to reduce the adhesion of marine fouling organisms and are of interest to those investigating enzyme-containing coating technologies for practical biofouling control.

Linkage to the actin cytoskeleton is critical for α4β1‐mediated leukocyte rolling and chemokine‐induced arrest, but not prolonged adhesion

The avidity or strength of integrin‐mediated cell adhesion is attributed to affinity (strength of individual bonds) and valency (number of bonds formed). We investigated the role of the actin cytoskeleton in α4β1 integrin mediated adhesion to VCAM‐1 in assays that model distinct steps of leukocyte emigration.In U937 cells stably transfected with the formyl peptide receptor, α4β1 integrin affinity up‐regulation induced by GPCR signaling remained intact following disruption of the actin cytoskeleton by cytochalasin D (cytoD) or latrunculin B (latB) (2 µM, 30 min incubation). Parallel plate flow chamber adhesion assays revealed that U937 cells pretreated with cytoD or latB failed to roll on surfaces coated with VCAM‐1 and arrest on surfaces co‐coated with VCAM‐1 and SDF‐1α. High affinity α4β1 induced by Mn2+ independent of GPCR signaling mediated spontaneous U937 arrest, which was blocked by cytoD or latB pretreatment. In contrast, U937 cell adhesion in 15 min assays was not inhibited by cytoD or latB. Fluorescence recovery after photobleaching demonstrated increased diffusion of α4β1 integrins in cells pretreated with cytoD, consistent with a disruption of integrin linkage to the actin cytoskeleton.These studies suggest that constitutive linkage of α4β1 integrins to the actin cytoskeleton is critical for mediating α4β1‐dependent rolling and arrest, but not prolonged leukocyte adhesion.Funded by CIHR.

Design and synthesis of biomimetic hydrogel scaffolds with controlled organization of cyclic RGD peptides.

We report on the rational design and synthesis of a new type of bioactive poly(ethylene glycol) diacrylate (PEGDA) macromers, cyclic Arg-Gly-Asp (cRGD)-PEGDA, to mimic the cell-adhesive properties of extracellular matrix (ECM), aiming to create biomimetic scaffolds with controlled spatial organization of ligands and enhanced cell binding affinity for tissue engineering. To attach the cRGD peptide in the middle of PEGDA chain, a tailed cRGD peptide, c[RGDfE(SSSKK-NH2)] (1), was synthesized with c(RGDfE) linked to a tail of SSSKK. The tail consists of a spacer with three serine residues and a linker with two lysine residues for conjugating with acryloyl-PEG-NHS (5) to create cRGD-PEGDA (6). cRGD-PEGDA possesses good photopolymerization ability to fabricate hydrogel scaffolds under UV radiation. Surface morphology and composition analysis demonstrates that cRGD-PEGDA hydrogels were well-constructed with porous three-dimensional (3D) structures and uniform distribution of cRGD ligands. Our results show that cRGD-PEGDA hydrogels facilitate endothelial cell (EC) adhesion and spreading on the hydrogel surfaces and exhibit significantly higher EC population in comparison with linear RGD-modified hydrogels at low peptide incorporation. Since ligand presentation in biomimetic scaffolds plays an important role in controlling cell behavior, cRGD-PEGDA has great advantages of controlling hydrogel properties and ligand spatial organization in the resulting scaffolds. Furthermore, cRGD-PEGDA is an attractive candidate for the future development of tissue engineering scaffolds with optimum cell adhesive strength and ligand density.

High-dose pemphigus antibodies against linear epitopes of desmoglein 3 (Dsg3) can induce acantholysis and depletion of Dsg3 from keratinocytes

We have previously demonstrated that serum autoantibodies of patients with pemphigus vulgaris (PV) may affect desmoglein 3 (Dsg3)-mediated adhesion by decreasing its half-life and inducing Dsg3 cleavage. Here we sought to gain more insights into the role of Dsg3-targetting IgG in acantholysis. To do so, alterations of keratinocyte morphology and cell–cell adhesion strength were investigated in the presence of PV serum, PV IgG, and IgG purified from PV patients’ sera against linear epitopes of Dsg3 (anti-Dsg3-L IgG). Changes in Dsg3 protein levels were assessed by Western blotting. Results showed that both PV serum and PV IgG were able to induce acantholysis and decrease the total amount of Dsg3 in cell lysates. Polyclonal anti-Dsg3-L IgG displayed Dsg3-depleting activity solely when used at 1 μg/ml, i.e. under non-physiologic conditions. Furthermore, cell–cell detachment induced by PV IgG and anti-Dsg3-L IgG seemed to precede the loss of Dsg3 from keratinocytes, suggesting that depletion/degradation of Dsg3 represents a late event in acantholysis. Collectively, the data presented here demonstrate that PV IgG recognizing non-conformational epitopes of Dsg3 are pathogenic when administered on doses largely exceeding those found in PV sera.

Tumor suppressor KAI1 affects integrin αvβ3-mediated ovarian cancer cell adhesion, motility, and proliferation

The tetraspanin KAI1 had been described as a metastasis suppressor in many different cancer types, a function for which associations of KAI1 with adhesion and signaling receptors of the integrin superfamily likely play a role. In ovarian cancer, integrin αvβ3 correlates with tumor progression and its elevation in vitro provoked enhanced cell adhesion accompanied by significant increases in cell motility and proliferation in the presence of its major ligand vitronectin. In the present study, we characterized integrin αvβ3-mediated tumor biological effects as a function of cellular KAI1 restoration and proved for the first time that KAI1, besides its already known physical crosstalk with β1-integrins, also colocalizes with integrin αvβ3. Functionally, elevated KAI1 levels drastically increased integrin αvβ3/vitronectin-dependent ovarian cancer cell adhesion. Since an intermediate level of cell adhesive strength is required for optimal cell migration, we next studied ovarian cancer cell motility as a function of KAI1 restoration. By time lapse video microscopy, we found impaired integrin αvβ3/vitronectin-mediated cell migration most probably due to strongly enhanced cellular immobilization onto the adhesion-supporting matrix. Moreover, KAI1 reexpression significantly diminished cell proliferation. These data strongly indicate that KAI1 may suppress ovarian cancer progression by inhibiting integrin αvβ3/vitronectin-provoked tumor cell motility and proliferation as important hallmarks of the oncogenic process.

Regionally-selective cell colonization of micropatterned surfaces prepared by plasma polymerization of acrylic acid and 1,7-octadiene

Micropatterned surfaces have been used as a tool for controlling the extent and strength of cell adhesion, the direction of cell growth and the spatial distribution of cells. In this study, chemically micropatterned surfaces were prepared by successive plasma polymerization of acrylic acid (AA) and 1,7-octadiene (OD) through a mask. Rat vascular smooth muscle cells (VSMC), bovine endothelial cells (EC), porcine mesenchymal stem cells (MSC) or human skeletal muscle cells (HSKMC) were seeded on these surfaces in densities from 9,320 cells/cm(2) to 31,060 cells/cm(2). All cell types adhered and grew preferentially on the strip-like AA domains. Between day 1 and 7 after seeding, the percentage of cells on AA domains ranged from 84.5 to 63.3 % for VSMC, 85.3 to 73.5 % for EC, 98.0 to 90.0 % for MSC, and 93.6 to 55.0 % for HSKMC. The enzyme-linked immunosorbent assay (ELISA) revealed that the concentration of alpha-actin per mg of protein was significantly higher in VSMC on AA. Similarly, immunofluorescence staining of von Willebrand factor showed more apparent Weibel-Palade bodies in EC on AA domains. MSC growing on AA had better developed beta-actin cytoskeleton, although they were less stained for hyaluronan receptor (CD44). In accordance with this, MSC on AA contained a higher concentration of beta-actin, although the concentration of CD44 was lower. HSKMC growing on AA had a better developed alpha-actin cytoskeleton. These results based on four cell types suggest that plasma polymerization is a suitable method for producing spatially defined patterned surfaces for controlled cell adhesion, proliferation and maturation.

Characterizing components of the Saw Palmetto Berry Extract (SPBE) on prostate cancer cell growth and traction

Saw Palmetto Berry Extract (SPBE) is applied for prostate health and treatment of urinary tract infections, nonbacterial prostitis and Benign Prostatic Hyperplasia (BPH) in man. An assumption is that SPBE affects tumor cell progression and migration in breast and prostate tissue. In this work, DU-145 cells were used to demonstrate that SPBE and its sterol components, β-sitosterol and stigmasterol, inhibit prostate cancer growth by increasing p53 protein expression and also inhibit carcinoma development by decreasing p21 and p27 protein expression. In the presence of cholesterol, these features are not only reversed but increased significantly. The results show for the first time the potential of SPBE, β-sitosterol and stigmasterol as potential anti-tumor agents. Since the protein p53 is also regarded as nuclear matrix protein facilitating actin cytoskeletal binding, 2D tractions were measured. The cell adhesion strength in the presence of SPBE, β-sitosterol and cholesterol and the observation was that the increase in p53 expression triggered an increase in the intracellular force generation. The results suggest a dual function of p53 in cells.

Investigation of effects of Argenine–Glycine–Aspartate (RGD) and nano-scale titanium coatings on cell spreading and adhesion

This paper examines the effects of physical and chemical surface modifications on the biocompatibility of silicon surfaces that are relevant to implantable silicon Bio-micro-electro-mechanical systems (BioMEMS). Two types of surface modifications were explored. The first involved the deposition of nano-scale biocompatible layers of pure titanium on silicon, while the second explored the covalent attachment of the binding peptide Argenine–Glycine–Aspartic acid (RGD) for improved cell adhesion. Improvements in biocompatibility were assessed through examination of cell areas after culture, as well as the measurements of adhesion strengths, as determined by shear assay techniques. The titanium nanolayers and the RGD coating resulted in improvements in biocompatibility. Increased cell spreading areas and improved adhesion strength were obtained from short and long-term studies of Human Osteosarcoma (HOS) cells cultured on the coated surfaces. RGD functionalization resulted in the greatest improvement in cell spreading area and adhesion strength for short culture times. The effects of the titanium, while less than those of RGD for short culture times, appeared to be greater after 48 h of culture.

Mixed extracellular matrix ligands synergistically modulate integrin adhesion and signaling

Cell adhesion to extracellular matrix (ECM) components through cell-surface integrin receptors is essential to the formation, maintenance and repair of numerous tissues, and therefore represents a central theme in the design of bioactive materials that successfully interface with the body. While the adhesive responses associated with a single ligand have been extensively analyzed, the effects of multiple integrin subtypes binding to multivalent ECM signals remain poorly understood. In the present study, we generated a high throughput platform of non-adhesive surfaces presenting well-defined, independent densities of two integrin-specific engineered ligands for the type I collagen (COL-I) receptor alpha(2)beta(1) and the fibronectin (FN) receptor alpha(5)beta(1) to evaluate the effects of integrin cross-talk on adhesive responses. Engineered surfaces displayed ligand density-dependent adhesive effects, and mixed ligand surfaces significantly enhanced cell adhesion strength and focal adhesion assembly compared to single FN and COL-I ligand surfaces. Moreover, surfaces presenting mixed COL-I/FN ligands synergistically enhanced FAK activation compared to the single ligand substrates. The enhanced adhesive activities of the mixed ligand surfaces also promoted elevated proliferation rates. Our results demonstrate interplay between multivalent ECM ligands in adhesive responses and downstream cellular signaling.

In-situ monitoring of breast cancer cell (MCF-7) growth and quantification of the cytotoxicity of anticancer drugs fluorouracil and cisplatin

A wireless sensing device was developed for the in-situ monitoring of the growth of human breast cancer cells (MCF-7) and evaluation of the cytotoxicity of the anticancer drugs fluorouracil and cisplatin. The sensor is fabricated by coating a magnetoelastic ribbon-like sensor with a layer of polyurethane that protects the iron-rich sensor from oxidation and provides a cell-compatible surface. In response to a time-varying magnetic field, the magnetoelastic sensor longitudinally vibrates, emitting magnetic flux that can be remotely detected by a pick-up coil. No physical connections between the sensor and the detection system are required. The wireless property facilitates aseptic biological operation, especially in cell culture as illustrated in this work. The adhesion of cells on the sensor surface results in a decrease in the resonance amplitude, which is proportional to the cell concentration. A linear response was obtained in cell concentrations of 5×104 to 1×106cellsml−1, with a detection limit of 1.2×104cellsml−1. The adhesion strength of cells on the sensor is qualitatively evaluated by increasing the amplitude of the magnetic excitation field. And the cytotoxicity of the anticancer drugs fluorouracil and cisplatin is evaluated by the magnetoelastic biosensor. The cytostatic curve is related with the quantity of cytostatic drug. The lethal concentration (LC50) for cells incubated in the presence of drugs for 20h is calculated to be 19.9μM for fluorouracil and 13.1μM for cisplatin.

Formation of E-Cadherin/β-Catenin–based Adherens Junctions in Hepatocytes Requires Serine-10 in p27(Kip1)

The adhesion between epithelial cells at adherens junctions is regulated by signaling pathways that mediate the intracellular trafficking and assembly of its core components. Insight into the molecular mechanisms of this is necessary to understand how adherens junctions contribute to the functional organization of epithelial tissues. Here, we demonstrate that in human hepatic HepG2 cells, oncostatin M-p42/44 mitogen-activated protein kinase signaling stimulates the phosphorylation of p27(Kip1) on Ser-10 and promotes cell-cell adhesion. The overexpression of wild-type p27 or a phospho-mimetic p27S10D mutant in HepG2 cells induces a hyper-adhesive phenotype. In contrast, the overexpression of a nonphosphorylatable p27S10A mutant prevents the mobilization of E-cadherin and beta-catenin at the cell surface, reduces basal cell-cell adhesion strength, and prevents the stimulatory effect of oncostatin M on cell-cell adhesion. As part of the underlying molecular mechanism, it is shown that in p27S10A-expressing cells beta-catenin interacts with p27 and is prevented from interacting with E-cadherin. The intracellular retention of E-cadherin and beta-catenin is also observed in hepatocytes from p27S10A knockin mice that express the p27S10A mutant instead of wild-type p27. Together, these data suggest that the formation of adherens junctions in hepatocytes requires Ser-10 in p27.

Simple forces, complex shapes

Form follows physics in the fly eye, say Sascha Hilgenfeldt, Sinem Erisken, and Richard Carthew (Northwestern University, Evanston, IL). Figure 1 Shapes predicted by energy minimization modeling (red lines) match the outlines of cells in the fly eye. The idea that a small set of physical forces governs the shape of tissues goes back at least a hundred years, but there has been little evidence to support it. Now, Hilgenfeldt et al. show that just two parameters—cell elasticity and adhesion strength—can account for the arrangement of cells in the ommatidium, the 20-cell subunit of the fly's compound eye. The geometry of the cone cells in the center of the ommatidium resembles groups of soap bubbles, says Hilgenfeldt, who studies the physics of bubbles, foams, and other “soft solids.” This arrangement suggested that, as in bubble aggregates, adhesion and elasticity might help determine shape. And like bubbles, the cells might be minimizing the surface energy at their interface. To test this theory, the authors created a computer-simulated model of ommatidium geometry. Adhesion in these structures is supplied by cadherins at the cells' apicolateral surfaces. The elasticity of the cell membrane arises from a number of complex interactions between the lipid bilayer and the cytoskeleton but can be characterized by its resistance to deformation. The team found that beginning with cells of random shapes, and then adjusting the relative strengths of cadherin binding and membrane elasticity to minimize the total surface energy of all the cells, they could precisely reproduce the observed geometries of the eye. The model also predicted the abnormal cellular arrangements that are found in various cadherin mutants. “Even a small variation in adhesion can alter cell packing dramatically,” Carthew says. Can other tissue geometries be explained as simply? “We think these results stand as an example for less-ordered epithelia,” Carthew says, though some element of random variation may be needed in the mix to account for these structures. They also hope to model developmental changes with the same principles. “You could imagine that tissues undergoing morphogenesis go through a series of minimal energetic states, dictated by changes in a few factors that regulate cell interactions,” Carthew says, such as placement and number of adhesion molecules or cytoskeletal alterations. Reference: Hilgenfeldt, S., et al. 2008. Proc. Natl. Acad. Sci. USA. 10.1073/pnas.0711077105 [PubMed] [Cross Ref]

Endothelial Cell Migration on RGD-Peptide-Containing PEG Hydrogels in the Presence of Sphingosine 1-Phosphate

Sphingosine 1-phosphate (S1P) is a potent chemokinetic agent for endothelial cells that is released by activated platelets. We previously developed Arg-Gly-Asp (RGD)-containing polyethylene glycol biomaterials for the controlled delivery of S1P to promote endothelialization. Here, we studied the effects of cell adhesion strength on S1P-stimulated endothelial cell migration in the presence of arterial levels of fluid shear stress, since an upward shift in optimal cell adhesion strengths may be beneficial for promoting long-term cell adhesion to materials. Two RGD peptides with different integrin-binding specificities were added to the polyethylene glycol hydrogels. A linear RGD bound primarily to β 3 integrins, whereas a cyclic RGD bound through both β 1 and β 3 integrins. We observed increased focal adhesion formation and better long-term adhesion in flow with endothelial cells on linear RGD peptide, versus cyclic RGD, even though initial adhesion strengths were higher for cells on cyclic RGD. Addition of 100 nM S1P increased cell speed and random motility coefficients on both RGD peptides, with the largest increases found on cyclic RGD. For both peptides, much of the increase in cell migration speed was found for smaller cells (<1522 μm 2 projected area), although the large increases on cyclic RGD were also due to medium-sized cells (2288–3519 μm 2). Overall, a compromise between high cell migration rates and long-term adhesion will be important in the design of materials that endothelialize after implantation.

Hybrid Superporous Scaffolds: An Application for Cornea Tissue Engineering

Engineering a cell-based keratoprosthesis often requires a struggle between two essential parameters: natural 3-D biological adhesion and mechanical strength. A novel hybrid scaffold of natural and synthetic materials was engineered to achieve both cell adhesion and implantable strength. This scaffold was characterized in terms of cell adhesion, cell migration, swelling, and strength. While the study was focused on engineering a biointegrable prosthetic skirt, a clear central core with an appropriate refractive index and light transmission was also incorporated into the design for potential functionality. The hybrid scaffold was tested in rat corneas. This uniquely designed scaffold was well tolerated and encouraged host cell migration into the implant. The hybrid superporous design also enhanced cell adhesion and retention in a superporous scaffold without altering the bulk mechanical properties of the hydrogel.

Nitrogen starvation affects bacterial adhesion to soil.

One of the main factors limiting the bioremediation of subsoil environments based on bioaugmentation is the transport of selected microorganisms to the contaminated zones. The characterization of the physiological responses of the inoculated microorganisms to starvation, especially the evaluation of characteristics that affect the adhesion of the cells to soil particles, is fundamental to anticipate the success or failure of bioaugmentation. The objective of this study was to investigate the effect of nitrogen starvation on cell surface hydrophobicity and cell adhesion to soil particles by bacterial strains previously characterized as able to use benzene, toluene or xilenes as carbon and energy sources. The strains LBBMA 18-T (non-identified), Arthrobacter aurescens LBBMA 98, Arthrobacter oxydans LBBMA 201, and Klebsiella sp. LBBMA 204–1 were used in the experiments. Cultivation of the cells in nitrogen-deficient medium caused a significant reduction of the adhesion to soil particles by all the four strains. Nitrogen starvation also reduced significantly the strength of cell adhesion to the soil particles, except for Klebsiella sp. LBBMA 204–1. Two of the four strains showed significant reduction in cell surface hydrophobicity. It is inferred that the efficiency of bacterial transport through soils might be potentially increased by nitrogen starvation.