In vitro binding of Campylobacter jejuni/coli outer membrane preparations to INT 407 cell membranes

Outer membrane fractions (OMs) of nine Campylobacter (C.) jejuni and two C. coli strains belonging to different serovars, from human and various animal origins, were extracted by treatment with sodium N-lauryl sarcosinate. Using n-octyl--d-glucopyranoside a 42-kDa protein and a flagella-enriched fraction were obtained. The capacity of the crude bacterial OM preparations, the purified 42-kDa protein and the flagella to bind to membranes of the human embryonic intestinal cell line INT 407 was tested by an enzyme-linked immunosorbent assay. The crude OM and the 42-kDa-enriched fraction were found to bind very well to the cell membranes, whereas the flagella preparation showed only a weak binding. Using monoclonal antibodies (mAbs) with HS 2-lipopolysaccharide (LPS) specificity, binding of crude HS 2 strain OM preparations to cell membranes was detected in a significant range, whereas with flagellin-specific mAbs binding of OMs and flagella to cell membranes was only detected to a very low extent. Binding of OMs to cell membranes was inhibited by preincubation of OMs with serovar-specific mouse hyperimmune serum, whereas on preincubation with mAbs directed against LPS or flagella binding was practically not inhibited. OMs extracted after pretreatment of the bacteria with proteinase K showed an altered SDS-PAGE pattern especially for the 42-kDa protein subunit and and their capacity to bind to cell membranes was significantly reduced. The binding was also reduced by preincubation of the OMs with l-fucose or d-mannose.     

Mast cell adhesion to fibronectin.

The MCP-5 murine mast cell line, as well as primary bone marrow-derived cultured mast cells (BMCMC), are demonstrated to bind to fibronectin, a ubiquitous adhesion protein of the extracellular matrix. BMCMC required activation by phorbol myristate acetate (PMA) to adhere to fibronectin, whereas MCP-5 displayed spontaneous adherence. The binding of both MCP-5 and BMCMC was dose dependent, with maximal adhesion at a fibronectin concentration of 20 micrograms/ml. The 120,000 molecular weight (MW) proteolytic fragment of fibronectin containing the RGDS cell attachment site was able to substitute for the native fibronectin molecule in promoting mast cell attachment. Mast cell adhesion to fibronectin, in addition, could be inhibited by the RGDS peptide alone. These data suggest that, in addition to the previously described mast cell-laminin interactions, mast cells also adhere to fibronectin, thus providing further insight into their tissue localization and possible roles in processes such as wound healing and fibrosis.     

Distinct mechanism of human neuroblastoma cell adhesion to fibronectin

We investigated the adhesion of three morphologically distinct human neuroblastoma cell lines (NCG, GOTO and SK-N-DZ) to intact fibronectin, central cell binding domain fragment (CBF) and CS peptide-IgG conjugates in the fibronectin molecule. Each cell line was found to express different integrin fibronectin receptors ( _{3 1}, _{4 1} and ₅₁), although similarly attached on intact fibronectin. To CBF, NCG attached well, while GOTO moderately and SK-N-DZ poorly attached. Only GOTO adhered to CS1-IgG. RGDS inhibited the spreading of NCG and SK-N-DZ on intact fibronectin, but it barely inhibited that of GOTO. The analysis by fluorescence-activated cell sorting (FACS) revealed that NCG expressed abundant ₃₁ and ₅₁, but little ₄₁, while GOTO expressed a large amount of ₄₁ as well as ₅₁. SK-N-DZ was undetectable in any of these molecules, but expressed _v1, which was identified by immunoprecipitation and immunoblotting. Polyclonal antibody to _v3 inhibited the adhesion of SK-N-DZ but not that of NCG or GOTO on intact fibronectin. These results suggest the existence of a distinct mechanism of cell adhesion to fibronectin among human neuroblastoma cell lines. It remains to be determined if such heterogeneous adhesion properties are related to the unique metastatic character of human neuroblastoma.     

Campylobacter jejuni-Stimulated Secretion of Interleukin-8 by INT407 Cells

Incubation of INT407 cells with various clinical isolates of Campylobacter jejuni resulted in secretion of interleukin-8 (IL-8) at levels ranging from 96 to 554 pg/ml at 24 h. The strains which produced the highest levels of IL-8 secretion were 81-176 and BT44. Induction of IL-8 secretion required live cells of 81-176 and was dependent on de novo protein synthesis. Site-specific mutants of 81-176, which were previously shown to be defective in adherence and invasion, resulted in reduced levels of secretion of IL-8, and cheY mutants of strains 81-176 and 749, which are hyperadherent and hyperinvasive, resulted in higher levels of IL-8 secretion. Another mutant of 81-176, which adheres at about 43% of the wild-type levels but is noninvasive, also showed marked reduction in IL-8 levels, suggesting that invasion is necessary for high levels of IL-8 secretion. When gentamicin was added to INT407 cells at 2 h after infection with 81-176, IL-8 secretion 22 h later was equivalent to that of controls without gentamicin, suggesting that the events which trigger induction and release of IL-8 occur early in the interactions of bacteria and eukaryotic cells.     

Electrospray deposited fibronectin retains the ability to promote cell adhesion

Scaffolds for tissue engineering require the correct biochemical cues if the seeded cells are to migrate into the scaffold and proliferate. For complex tissues this would require precise patterning of the scaffold structure with the particular biochemical cue required at each location on the scaffold. Electrospray enables the deposition of a wide number of biomolecules onto surfaces and can be used for precise patterning. We assessed the functionality of a key cell-adhesion molecule, fibronectin, after depositing it onto a surface using the electrospray technique. The addition of polypropylene glycol allowed a stable spray to be obtained from solutions with a range of fibronectin concentrations. Immunoassay tests showed that the amount of fibronectin retained on the surface was proportional to that sprayed from the solution. Increasing the surface density of fibronectin deposited onto silicon surfaces enhanced fibroblast attachment. The fibronectin thus appears to have retained its cell attachment functionality after undergoing the electrospray process. Since recent advances allow electrospray to pattern material from solution with micrometre accuracy this may allow materials to be biologically functionalized on a similar scale.     

Adsorption characteristics of human plasma fibronectin in relationship to cell adhesion

Adsorption of human plasma fibronectin (FN) on nonsulfonated and sulfonated polymer surfaces was studied, by using a polyclonal antiserum to FN and the ELISA method. ELISA signal was recorded as a function of FN concentration in solutions. The concentration dependence of FN binding shows the saturation effect in the range 5-10 microg/mL. ELISA data are discussed in the terms of a self-assembled monolayer and different conformations of the FN molecule.The early adhesion of L1210 cells to polymer surfaces after prior adsorption of FN on these surfaces was studied under static conditions. In the case of FN adsorbed on sulfonated surfaces, the relative number of adhering cells increased with the increase of the interfacial surface tension (i.e., the cell adhesion depends on the surface density of sulfonic groups). However, in the case of FN adsorbed on nonsulfonated surfaces, the relative number of adhering cells was low and independent on the interfacial surface tension.The alpha(5)beta(1)-integrin blocking by a monoclonal antibody resulted in a strong inhibition of the cell adhesion to FN adsorbed on sulfonated polymer surfaces. This indicates that cell adhesion to FN adsorbed on these surfaces is mostly mediated by the alpha(5)beta(1)-integrin. In contrast, in the case of FN adsorbed on nonsulfonated surfaces the cell adhesion was not inhibited by the alpha(5)beta(1)-integrin blocking.     

Binding of outer membrane preparations of Campylobacter jejuni to INT 457 cell membranes and extracellular matrix proteins

Binding of outer membrane (OM) preparations of the thermophilic Campylobacter species C. jejuni to epithelial cell membranes and extracellular matrix proteins were studied in an in vitro model system using enzyme-linked immunosorbent assay. The OM preparations exhibited significant binding to INT 407 intestinal cell membranes. The process of adhesion was modulated by enzymatic, chemical or immunological pretreatment of the bacteria. Following oxidation of the lipopolysaccharide (LPS) with sodium meta-periodate, the OM preparations essentially retained their binding properties. After pretreatment with proteinase K, the OM preparations lost their binding capacity and the apparent molecular mass of the major OM protein shifted from 42 to 24 kDa. Preincubation of C. jejuni bacteria with C. jejuni-specific antiserum reduced adhesion significantly; preincubation with LPS-specific monoclonal antibodies only to a minimal extent. The OM preparations also bound significantly to the extracellular matrix proteins collagen and fibronectin; however, they bound virtually no bovine serum albumin or horse serum.     

Intestinal mucus gel and secretory antibody are barriers to Campylobacter jejuni adherence to INT 407 cells.

An in vitro mucus assay was developed to study the role of mucus gel and secretory immunoglobulin A (sIgA) in preventing attachment of Campylobacter jejuni to INT 407 cells. An overlay of rabbit small intestinal mucus was found to impede the attachment of C. jejuni to a monolayer of INT 407 cells. Mucus from rabbits previously colonized with C. jejuni was found to completely inhibit bacterial adherence to the underlying cells. Anti-Campylobacter sIgA was readily detected in mucus samples from previously exposed rabbits and was responsible for eliminating bacterial adherence to the INT 407 cells. This was shown by loss of inhibition after mucus absorption with Campylobacter cells. sIgA-containing mucus caused aggregation of the C. jejuni cells within the mucus layer of the assay system. Nonimmune mucus and sIgA alone were unable to cause bacterial aggregation, suggesting a cooperative role for mucus and sIgA. Antibodies responsible for adhesion inhibition were cross-reactive among several Campylobacter strains and were not directed solely against flagellar antigens.     

Inhibitory effect of wheat germ agglutinin on mouse mast cell adhesion to fibronectin

BACKGROUND: Mast cells play a critical role in allergic and inflammatory responses. The interactions between these cells and extracellular matrix components influence the distribution of mast cells in tissues and their biological responsiveness. It has been reported that the lectin wheat germ agglutinin (WGA) inhibits mast cell mediator release. We decided to investigate whether adhesion to fibronectin (FN), another mast cell function, which is upregulated following FcepsilonRI cross-linking, is also inhibited by WGA. METHODS: Mouse bone-marrow-derived mast cell line MCP5/L was used. For FcepsilonRI-dependent mast cell activation, MCP5/L cells were sensitized with mouse IgE antibodies. WGA was added to cell suspensions simultaneously with a challenging agent and, after an appropriate incubation period, beta-hexosaminidase release and adhesion to FN were determined. RESULTS: Both FcepsilonRI cross-linking-dependent mast cell adhesion to FN and mediator release were dose-dependently inhibited by WGA; however, the lectin concentrations required to induce maximum inhibition of adhesion were significantly lower. Furthermore, WGA inhibited phorbol-myristate-acetate- and A-23187-mediated mast cell adhesion to FN, i.e. processes that do not engage FcepsilonRI. The effect of WGA on FcepsilonRI-mediated secretion was reversed by GlcNAc. In contrast, combination of GlcNAc and NeuNAc or N, N'-diacetylchitobiose was required to reverse the inhibitory effect of WGA on mast cell adhesion. CONCLUSION: The characteristics of WGA-mediated inhibition of MCP5/L mast cell adhesion to FN suggest that mast cell integrins are targets of the inhibitory action of WGA and the sugar moieties on these receptors might be important for receptor function.     

Effects of fibronectin cleaved by neuropsin on cell adhesion and migration

Neuropsin is a serine protease cloned from the mouse hippocampus. Since neuropsin is a secreted protein which effectively cleaves fibronectin, it may affect cell adhesion or cell migration by modulating the content and/or chemical characteriscs of fibronectin in extracellular matrix (ECM). In adhesion assays, alpha5B2 cells expressing integrin alpha5beta1 bound less effectively to fibronectin teated with neuropsin than intact fibronectin. In Boyden chamber chemotaxis assays, the fibronectin-induced migration of alpha5B2 cells was not affected by neuropsin treatment. These findings suggest that neuropsin regulates the local microenvironment by modulating the interaction between cells and fibronectin in ECM.     

Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion

Integrin-mediated cell adhesion to proteins adsorbed onto synthetic surfaces anchors cells and triggers signals that direct cell function. In the case of fibronectin (Fn), adsorption onto substrates of varying properties alters its conformation/structure and its ability to support cell adhesion. In the present study, self-assembled monolayers (SAMs) of alkanethiols on gold were used as model surfaces to investigate the effects of surface chemistry on Fn adsorption, integrin binding, and cell adhesion. SAMs presenting terminal CH(3), OH, COOH, and NH(2) functionalities modulated adsorbed Fn conformation as determined through differences in the binding affinities of monoclonal antibodies raised against the central cell-binding domain (OH > COOH = NH(2) > CH(3)). Binding of alpha(5)beta(1) integrin to adsorbed Fn was controlled by SAM surface chemistry in a manner consistent with antibody binding (OH > COOH = NH(2) > CH(3)), whereas alpha(V) integrin binding followed the trend: COOH > OH = NH(2) = CH(3), demonstrating alpha(5)beta(1) integrin specificity for Fn adsorbed onto the NH(2) and OH SAMs. Cell adhesion strength to Fn-coated SAMs correlated with alpha(5)beta(1) integrin binding (OH > COOH = NH(2) > CH(3)), and experiments with function-perturbing antibodies demonstrated that this receptor provides the dominant adhesion mechanism in this cell model. This work establishes an experimental framework to analyze adhesive mechanisms controlling cell-surface interactions and provides a general strategy of surface-directed control of adsorbed protein activity to manipulate cell function in biomaterial and biotechnological applications.     

Involvement of fungal cell wall components in adhesion of Sporothrix schenckii to human fibronectin

Systemic sporotrichosis is an emerging infection potentially fatal for immunocompromised patients. Adhesion to extracellular matrix proteins is thought to play a crucial role in invasive fungal diseases. Here we report studies of the adhesion of Sporothrix schenckii to the extracellular protein fibronectin (Fn). Both yeast cells and conidia of S. schenckii were able to adhere to Fn as detected by enzyme-linked immunosorbent binding assays. Adhesion of yeast cells to Fn is dose dependent and saturable. S. schenckii adheres equally well to 40-kDa and 120-kDa Fn proteolytic fragments. While adhesion to Fn was increased by Ca(2+), inhibition assays demonstrated that it was not RGD dependent. A carbohydrate-containing cell wall neutral fraction blocked up to 30% of the observed adherence for the yeast cells. The biochemical nature of this fraction suggests the participation of cell surface glycoconjugates in binding by their carbohydrate or peptide moieties. These results provide new data concerning S. schenckii adhesion mechanisms, which could be important in host-fungus interactions and the establishment of sporotrichosis.     

Semiquantitative evaluation of fibronectin adsorption on unmodified and sulfonated polystyrene, as related to cell adhesion

The process of human fibronectin (FN) adsorption on nonsulfonated and sulfonated polystyrene surfaces was studied in relation to mechanisms of L1210 cell adhesion. Radioisotope assays directed towards FN, as well as ELISA measurements of adsorbed FN and bovine serum albumin (BSA) were carried out. (125)I radioisotope assays led to linear FN adsorption isotherms. When combined to ELISA measurements for FN, they revealed the multilayer adsorption. Results indicated a large difference in the saturating first-layer surface density of FN adsorbed on sulfonated and nonsulfonated polystyrene surfaces: significantly (ca. factor of 5) less FN molecules are necessary to complete a monolayer on sulfonated than on nonsulfonated polystyrene. This suggests an unfolded conformation of FN on sulfonated polystyrene, and a more compact one on the nonsulfonated polymer. Significant conformational changes of FN are also indicated by the following: (1) early phase of cell adhesion to FN adsorbed on sulfonated polystyrene surfaces is significantly (ca. factor of 6) higher than to FN on nonsulfonated surfaces, and in the former case adhesion proceeds mostly via alpha(5)beta(1) integrins; (2) RGD, the crucial fragment within central cell binding domain, seems to be partially hidden in the protein structure adopted on nonsulfonated surfaces; (3) patterns of F-actin organization differ in cells adhering to FN on sulfonated and nonsulfonated surfaces. The ELISA study directed against BSA (this protein always present on the surface after the adsorption of FN), showed the importance of "free area," uncovered by both proteins, which influence the cell adhesion processes.     

Endothelial cell adhesion to the fibronectin CS5 domain in artificial extracellular matrix proteins

This study examines the spreading and adhesion of human umbilical vein endothelial cells (HUVEC) on artificial extracellular matrix (aECM) proteins containing sequences derived from elastin and fibronectin. Three aECM variants were studied: aECM 1 contains lysine residues periodically spaced within the protein sequence and three repeats of the CS5 domain of fibronectin, aECM 2 contains periodically spaced lysines and three repeats of a scrambled CS5 sequence, and aECM 3 contains lysines at the protein termini and five CS5 repeats. Comparative cell binding and peptide inhibition assays confirm that the tetrapeptide sequence REDV is responsible for HUVEC adhesion to aECM proteins that contain the CS5 domain. Furthermore, more than 60% of adherent HUVEC were retained on aECM 1 after exposure to physiologically relevant shear stresses (相似文献   

Correlating fibronectin adsorption with endothelial cell adhesion and signaling on polymer substrates

Fibronectin (Fn) adsorption was studied on different commercial polymer surface chemistries, including tissue culture polystyrene (TCPS), bacteriologic polystyrene (BPS), fluoropolymer Teflon AF, and poly-L-lactide (PLLA). Antibody probes detected the availability of Fn's cell binding domain on adsorbed Fn in the competitive presence and absence of bovine serum albumin (BSA). Domain availability was highest for Fn adsorbed on TCPS, especially in the presence of either serum albumin or dilute serum. Attachment and growth efficiencies for human umbilical venous endothelial cells (HUVECs) cultured on surfaces preadsorbed with Fn in serum and serum-free media correlated with antibody cell-binding domain availability: TCPS > BPS, Teflon AF > PLLA. Intracellular signaling from the GTPase, RhoA, was highest (RhoA:RhoGDI inhibitor ratio) in cells cultured on the Teflon AF surfaces, indicating that despite lower attached cell numbers on Teflon AF compared to TCPS, cell signaling remained activated after 24 h of growth. Up-regulated cellular Fn mRNA messages, assessed using RT-PCR techniques, supported HUVECs' producing the endogenous extracellular matrix (ECM) protein Fn in order to attach and survive on the suboptimal Teflon AF culture surfaces.     

Surface properties of and cell adhesion onto allylamine-plasma-coated polyethylenterephtalat membranes

Allylamine was plasma polymerised onto a polyester (PET) membrane to obtain a surface with good cell adhesive properties. Samples were coated using a microwave plasma source operating at different process parameters. The effect of process parameters on the physical, and chemical properties of plasma-polymerised-allylamine (PPAa) was evaluated by studying elemental composition, amine concentration, wettability, and surface morphology. A relatively high amine concentration was measured (up to 50 nmol/cm2). In parallel, nitrogen enrichment was observed after exposure to high-energetic plasma. Irrespective of the treatment conditions, oxygen was incorporated into the polymer structure. PPAa surfaces were found to be more hydrophilic than PET. The wettability of the samples increased with increasing amine concentration. Pictures from scanning electron microscopy indicated that homogeneous pinhole-free PPAa layers were deposited on PET membranes, without a significant change of permeability. In vitro evaluation of biocompatibility was carried out by studying human skin fibroblast interaction with surfaces. Cell attachment and viability on PPAa layers were found to be more intensive than on the control PET, based on the higher metabolic activity of adhering cells, but also on morphological criteria including overall cell morphology.     

Homologous serum increases fibronectin expression and cell adhesion in airway smooth muscle cells

This study describes altered patterns of growth and upregulation of fibronectin expression of cultured canine airway smooth muscle cells grown in homologous serum, which provides a model of the vascular leakage occurring in asthma, compared to fetal bovine serum (FBS). Cells were incubated in increasing concentrations of serum (2.5–40%) for 72 hours. Both homologous serum and FBS caused cellular proliferation which reached a maximum increase at 2.5–5% serum concentration. Differences in the cellular responses to the two types of sera were noted at higher concentrations of sera. At a concentration of 40% FBS, airway smooth muscle cells increased in number by 307±16% (n=5) compared to serum-free control cells, whereas in canine serum the increase in growth was significantly smaller, 239±25% (n=7) (P<0.05). Airway fibrocytes similarly treated increased in number by 256±43% (n=3) in 40% FBS, but exhibited a reduction in cell number to 80 ±10% (n=3) of controls in 40% homologous serum (P<0.05). Smooth muscle cells demonstrated a dose-dependent increase in fibronectin expression when grown in homologous serum, but not in FBS, suggesting phenotypic change occurred in these cells when exposed to homologous serum. These data suggest that the leakage of plasma in the asthmatic airway may trigger phenotypic change in both airway smooth muscle cells and airway fibrocytes leading to cellular proliferation and expression of extracellular matrix molecules. These in vitro changes are consistent with the histological findings in clinical asthma.     

The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications

Strain devices with expandable polydimethylsiloxane (PDMS) culture membranes are frequently used to stretch cells in vitro, mimicking mechanically dynamic tissue environments. To immobilize cell-adhesive molecules to the otherwise non-adhesive PDMS substrate, hydrophobic, electrostatic and covalent surface coating procedures have been developed. The efficacy of different coating strategies to transmit stretches to cells however is poorly documented and has not been compared. We describe a novel and simple procedure to covalently bind extracellular matrix proteins to the surface of stretchable PDMS membranes. The method comprises PDMS oxygenation, silanization, and covalent protein cross-linking to the silane. We demonstrate improved attachment (～2-fold), spreading (～2.5-fold) and proliferation (～1.2-fold) of fibroblasts to our new coating over established coating procedures. Further, we compared the efficiency of different PDMS coating techniques to transmit stretches. After 15% stretch, the number of maximally (15 ± 5%) stretched cells on our PDMS surface coating was ～7-fold higher compared with alternative coating protocols. Hence, covalent linkage of adhesive molecules is superior to non-covalent methods in providing a coating that resists large deformations and that fully transmit this stretch to cultured cells.     

Cholesterol-derivatized polyurethane: characterization and endothelial cell adhesion

Endothelialization of synthetic surfaces has been challenging with limited success thus far. We investigated the hypothesis that covalent attachment of cholesterol to polyurethane via the urethane nitrogen groups would create a high-affinity surface for attachment and adhesion of endothelial cells. Cholesterol was covalently bound to the polyether polyurethane, Tecothane, by first derivatizing the polyurethane nitrogen groups with bromoalkyl side chains, followed by reacting mercapto-cholesterol to the bromoalkyl sites. Cholesterol-modified polyurethane demonstrated a qualitatively smoother surface per atomic force microscopy than nonmodified and increased surface energy (contact angle measurements) compared with unmodified polyurethane. Cell attachment assays showed a significantly greater number of attached bovine arterial endothelial cells (p = 0.0003) after 45 min of seeding on cholesterol-modified polyurethane versus unmodified polyurethane. Bovine arterial endothelial cells cultivated on cholesterol-modified Tecothane showed significantly greater levels of cell retention compared with unmodified Tecothane when exposed to arterial level shear stress for 2 h (25 dynes/cm2) with 90.0 +/- 6.23% cells remaining adherent compared with unmodified polyurethane, 41.4 +/- 11.7%, p = 0.0070. Furthermore, ovine endothelial precursors, obtained as blood outgrowth endothelial cells, were seeded on cholesterol-modified polyurethane and exposed to 25 dynes/cm2 shear conditions for 2 h, with the retention of 90.30 +/- 3.25% of seeded cells versus unmodified polyurethane, which retained only 4.56 +/- 0.85% (p < 0.001). It is concluded that covalently linking cholesterol to polyurethane results in improved material properties that permit increased endothelial cell retention compared with unmodified polyurethane.     

PLGA nanometer surface features manipulate fibronectin interactions for improved vascular cell adhesion

The largest cause of mortality in the Western world is atherosclerotic vascular disease. Many of these diseases require synthetic vascular grafts; however, their patency rate is only 30% in small (<6 mm) diameter vascular grafts after 5 years of implantation. In an effort to increase small diameter vascular graft success, researchers have been designing random nanostructured surface features which enhance vascular cell functions. However, for the present study, highly-controllable, nanostructured features on poly(lactic-co-glycolic acid) (PLGA) surfaces were formulated. To create ordered nanostructured roughness on PLGA surfaces, either 500, 200, or 100 nm polystyrene nanospheres were separately placed onto mica. These were then used as a template for creating an inverse poly(dimethylsiloxane) mold which was utilized to cast PLGA. Compared to all other PLGA films formulated, AFM results demonstrated greater initial fibronectin spreading on PLGA which possessed spherical 200 nm features. Compared to smooth PLGA, PLGA with 500 or 100 nm surface features, results further showed that PLGA with 200 nm spherical features promoted vascular cell (specifically, endothelial, and smooth muscle cell) adhesion. In this manner, the present study demonstrated a specific nanometer surface feature size that promoted fibronectin spreading and subsequent vascular cell adhesion; criteria critical to vascular graft success.