The effect of surface microtopography of poly(dimethylsiloxane) on protein adsorption, platelet and cell adhesion

Chemical homogeneous poly(dimethylsiloxane) (PDMS) surface with dot-like protrusion pattern was used to investigate the individual effect of surface microtopography on protein adsorption and subsequent biological responses. Fibrinogen (Fg) and fibronectin (Fn) were chosen as model proteins due to their effect on platelet and cell adhesion, respectively. Fg labeled with ¹²⁵I and fluorescein isothiocyanate (FITC) was used to study its adsorption on flat and patterned surfaces. Patterned surface has a 46% increase in the adsorption of Fg when compared with flat surface. However, the surface area of the patterned surface was only 8% larger than that of the flat surface. Therefore, the increase in the surface area was not the only factor responsible for the increase in protein adsorption. Clear fluorescent pattern was visualized on patterned surface, indicating that adsorbed Fg regularly distributed and adsorbed most on the flanks and valleys of the protrusions. Such distribution and local enrichment of Fg presumably caused the specific location of platelets adhered from platelet-rich plasma (PRP) and flowing whole blood (FWB) on patterned surface. Furthermore, the different combination of surface topography and pre-adsorbed Fn could influence the adhesion of L929 cells. The flat surface with pre-adsorbed Fn was the optimum substrate while the virgin patterned surface was the poor substrate in terms of L929 cells spread.     

Nanoscale organization of adsorbed collagen: influence of substrate hydrophobicity and adsorption time

The adsorption of collagen on polystyrene (PS) and polystyrene oxidized by oxygen plasma discharge (PSox) was studied as a function of time using radiolabeling, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). Radiolabeling and XPS indicated that the initial step of adsorption was faster on PS than on PSox. AFM imaging under water revealed very different supramolecular organization of the adsorbed films depending on time and on the nature of the substrate: PS showed patterns of collagen aggregates at all adsorption times (from 1 min to 24 h); PSox was covered with a smooth layer except at long adsorption times (24 h), for which a mesh of collagen structures was observed. After fast drying, the collagen layer remained continuous and showed a morphology which recalled that observed under water. The mechanical stability of the adsorbed films was assessed under water by scraping with the AFM probe at different loading forces: no perturbations were created on PSox; in contrast, the layer adsorbed on PS was sensitive to scraping, the minimum force required to alter the collagen layer morphology increasing with time. These differences in the film properties were correlated with force measurements upon retraction: multiple adhesion forces were observed with collagen adsorbed on PS samples, whereas such an effect was never observed on PSox. The results show that the amount adsorbed and the organization of the adsorbed film respond differently to the adsorption time and that this is influenced by surface hydrophobicity. The quick initial adsorption on PS, compared to PSox, is thought to leave dangling collagen segments that are responsible for the observed morphology, for adhesion forces, and for lower mechanical resistance of the adsorbed layer.     

Fibronectin conformation switch induced by coadsorption with human serum albumin

The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn ? HSA), and Fn was not adsorbed onto predeposited HSA (HSA ? Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)·(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fn, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.     

AFM analysis of organic silane thin films for bioMEMS applications

Designing surfaces that elicit the desirable response is essential for bioMEMS (biological microelectromechanical systems) applications. To this end, we have developed two different types of silane film—hydrophobic and hydrophilic—using vinyltrichlorosilane and poly(ethylene glycol) silane, respectively. As the surface topography plays a very important role in governing protein or cell interactions, these films were characterized extensively using atomic force microscopy. All the films developed were found to have a very low root‐mean‐square roughness value (<1.3 nm). Furthermore, the topography of protein‐adsorbed silane‐modified surfaces was investigated because cell adhesion is mediated primarily by proteins. Three‐dimensional and section plots were able to differentiate the way in which protein interacts with hydrophobic and hydrophilic surfaces. Copyright © 2003 John Wiley & Sons, Ltd.     

Characterization of biomaterials polar interactions in physiological conditions using liquid–liquid contact angle measurements: Relation to fibronectin adsorption

Wettability of biomaterials surfaces and protein-coated substrates is generally characterized with the sessile drop technique using polar and apolar liquids. This procedure is often performed in air, which does not reflect the physiological conditions. In this study, liquid/liquid contact angle measurements were carried out to be closer to cell culture conditions. This technique allowed us to evaluate the polar contribution to the work of adhesion between an aqueous medium and four selected biomaterials widely used in tissue culture applications: bacteriological grade polystyrene (PS), tissue culture polystyrene (tPS), poly(2-hydroxyethyl methacrylate) film (PolyHEMA), and hydroxypropylmethylcellulose-carboxymethylcellulose bi-layered Petri dish (CEL). The contributions of polar interactions were also estimated on the same biomaterials after fibronectin (Fn) adsorption. The quantity of Fn adsorbed on PS, tPS, PolyHEMA and CEL surfaces was evaluated by using the fluorescein-labeled protein. PolyHEMA and CEL were found to be hydrophilic, tPS was moderately hydrophilic and PS was highly hydrophobic. After Fn adsorption on PS and tPS, a significant increase of the surface polar interaction was observed. On PolyHEMA and CEL, no significant adsorption of Fn was detected and the polar interactions remained unchanged. Finally, an inverse correlation between the polarity of the surfaces and the quantity of adsorbed Fn was established.     

Influence of surface topography on osteoblast response to fibronectin coated calcium phosphate thin films

The ability to engineer biomaterial surfaces that are capable of a dynamic interaction with cells and tissues is central to the development of medical implants with improved functionality. An important consideration in this regard is the role played by the extracellular proteins that bind to an implant surface in vivo. Deliberate use of an ad-layer of such proteins on an implant surface has been observed to guide and direct cell response. However, the role that changes in surface topography might play in determining the nature of this cell–protein–surface interaction has not been investigated in detail. In this study, calcium phosphate (CaP) thin films have been deposited onto substrates with varying topography such that this is reflected in the (conformal) CaP surface features. A fibronectin (FN) ad-layer was then deposited from solution onto each surface and the response of MG63 osteoblast-like cells investigated. The results revealed that in all cases, the presence of the adsorbed FN layer on the CaP thin films improved MG63 cell adhesion, proliferation and promoted early onset differentiation. Moreover, the nature and scale of the response were shown to be influenced by the underlying CaP surface topography. Specifically, MG63 cell on FN-coated CaP thin films with regular topographical features in the nanometer range showed statistically significant differences in focal adhesion assembly, osteocalcin expression and alkaline phosphase activity compared to CaP thin films that lacked these topographical features. As such, these data indicate that surface topography can be used to further influence cell adhesion and downstream differentiation by enhancing the effects of a surface adsorbed FN layer.     

Does the nanometre scale topography of titanium influence protein adsorption and cell proliferation?

To investigate the influence of titanium films with nanometre scale topography on protein adsorption and cell growth, three different model titanium films were utilized in the present study. The chemical compositions, surface topographies and wettability were investigated by using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and water contact angle measurement, respectively. The films share the same surface chemistry but exhibit different topographies on a nanometre scale. Thus, they act as model systems for biological studies regarding surface topography effects. The films were obtained by varying the deposition rate and the film thickness, respectively. These films displayed nanometre scale surface roughness (root mean square roughness, R_rms) from 2 to 21 nm over areas of 50 μm × 50 μm, with different grain sizes at their surfaces. Albumin and fibrinogen adsorption on these model titanium films were performed in this study. Bicinchoninic acid assay was employed to determine the amount of adsorbed protein on titanium film surfaces. No statistically significant differences, however, were observed for either albumin or fibrinogen adsorption between the different groups of titanium films. No statistically significant influence of surface roughness on osteoblast proliferation and cell viability was detected in the present study.     

Fibronectin adsorption onto polyelectrolyte multilayer films

The Layer-by-layer deposition of positively and negatively charged macromolecular species is an ideal method for constructing thin films incorporating biological molecules. We investigate the adsorption of fibronectin onto polyelectrolyte multilayer (PEM) films using optical waveguide lightmode spectroscopy (OWLS) and atomic force microscopy (AFM). PEM films are formed by adsorption onto Si(Ti)O2 from alternately introduced flowing solutions of anionic poly(sodium 4-styrenesulfonate) (PSS) and cationic poly(allylamine hydrochloride) (PAH). Using OWLS, we find the initial rate and overall extent offibronectin adsorption to be greatest on PEM films terminated with a PAH layer. The polarizability density of the adsorbed protein layer, as measured by its refractive index, is virtually identical on both PAH- and PSS-terminated films; the higher adsorbed density on the PAH-terminated film is due to an adsorbed layer of roughly twice the thickness. The binding of monoclonal antibodies specific to the protein's cell binding site is considerably enhanced to fibronectin adsorbed to the PSS layer, indicating a more accessible adsorbed layer. With increased salt concentration, we find thicker PEM films but considerably thinner adsorbed fibronectin layers, owing to increased electrostatic screening. Using AFM, we find adsorbed fibronectin layers to contain clusters; these are more numerous and symmetric on the PSS-terminated film. By considering the electrostatic binding of a segmental model fibronectin molecule, we propose a picture of fibronectin adsorbed primarily in an end-on-oriented monolayer on a PAH-terminated film and as clusters plus side-on-oriented isolated molecules onto a PSS-terminated film.     

Probing fibronectin-surface interactions: a multitechnique approach

The development of adhesive as well as antiadhesive surfaces is essential in various biomaterial applications. In this study, we have used a multidisciplinary approach that combines biological and physicochemical methods to progress in our understanding of cell-surface interactions. Four model surfaces have been used to investigate fibronectin (Fn) adsorption and the subsequent morphology and adhesion of preosteoblasts. Such experimental conditions lead us to distinguish between anti- and proadhesive substrata. Our results indicate that Fn is not able to induce cell adhesion on antiadhesive materials. On adhesive substrata, Fn did not increase the number of adherent cells but favored their spreading. This work also examined Fn-surface interactions using ELISA immunoassays, fluorescent labeling of Fn, and force spectroscopy with Fn-modified tips. The results provided clear evidence of the advantages and limitations of each technique. All of the techniques confirmed the important adsorption of Fn on proadhesive surfaces for cells. By contrast, antiadhesive substrata for cells avoided Fn adsorption. Furthermore, ELISA experiments enabled us to verify the accessibility of cell binding sites to adsorbed Fn molecules.     

Water in extremely narrow planar pores with crystalline walls. 1. Structure

Computer simulation has been employed to study the structure of water condensate filling planar pores 1.25 and 0.62 nm wide located parallel to the basal face in a silver iodide crystal at 260 K. All stages of adsorption of single molecules up to complete pore filling have been described. At an initial stage, strong clustering of molecules is observed on the walls; then, the walls are covered with a monomolecular film; and, at the final stage, molecules are adhered to the surface of the film, thus filling the internal space of the pore. First, adsorption occurs at the wall containing positive ions on the surface and, then, on the opposite wall with negative ions. On both walls, adsorbed molecules are adhered to the surface via the interaction with ions of the second crystallographic layer; given this, two types, α and β, of molecule plane orientation are realized on opposite walls. The adhesion of an adsorbed molecular film to molecules filling the interior of the pore requires the partial transition of film molecules from the α- to the β-type orientation on one wall and the inverse transition on the other wall. The deficiency of α-oriented molecules on one wall and β-oriented ones on the other is the main reason for poor wettability of the surface of the monomolecular films adsorbed on the walls. In an extremely narrow pore, molecules are simultaneously captured in the field of both walls. The forces acting from the sides of both walls result in the separation of a film into spots having structures matched to the crystalline structure of each wall, with the film being on the verge of breakage.     

Regulation of cellular responses to macroporous inorganic films prepared by the inverse-opal method

Regenerative medicine for repairing damaged body tissues has recently become critically important. Cell culture scaffolds are required for the control of cell attachment, proliferation, and differentiation in in vitro cell cultures. A new strategy to control cell adhesion, morphology, and proliferation was developed by culturing mouse osteoblast-like MC3T3-E1 cells on novel cell culture scaffolds fabricated using ordered nanometer-sized pores (100, 300, 500, and 1000 nm). Results of this study indicate that after 72 h of incubation, the number of cells cultured on a silica film with a pore size of 1000 nm was similar to or slightly lower than that cultured on a non-porous control silica film. Films with 100-500 nm pore sizes, however, resulted in the cell growth inhibition. Morphology of the cultured cells revealed increased elongation and the formation of actin stress fibers was virtually absent on macroporous silica films with 100-500 nm pore size. Vinculin molecules expressed in cells cultured on the non-porous silica films showed many clear focal adhesions, whereas focal contacts were insufficiently formed in cells cultured on macroporous films. The influence of hydroxyapatite (HAp) and alumina scaffolds on the behavior of MC3T3-E1 cells was also evaluated. The proliferation rate of MC3T3-E1 cells cultured on HAp films with 1000 nm pore size was increased to approximately 20% above than that obtained of cells cultured on non-porous HAp films. These results demonstrate that the pore size and constituents of films play a role in controlling the morphology and proliferation rate of MC3T3-E1 cells.     

Comparison of the influence of titanium and chromium adhesion layers on the properties of sol–gel derived NKN thin films

Lead-free (Na_0.5K_0.5)NbO₃ (NKN) thin films were prepared on Pt/X/SiO₂/Si substrates (with the adhesion promoters X = Ti, Cr) by a sol–gel process with and without post-annealing treatment. The effect of the diffusion of the adhesion layer elements Ti and Cr into the NKN film was analysed by secondary ion mass spectrometry, scanning electron microscopy pictures, X-ray diffraction (XRD), and leakage current measurements. It turned out that Cr diffuses into the films to a higher extent than Ti. The high amount of Cr diffusion led to the formation of a secondary phase, as seen in the XRD pattern, and to pore formation on the surface of the NKN films. In contrast, the films with Ti adhesion layer were single phase NKN without pore formation. Also, the leakage current measurements showed a strong influence of the Cr diffusion. The leakage current of the films with Cr adhesion layer was about four orders of magnitude higher than that of the films with Ti adhesion layer. The study shows the strong influence of the adhesion layer of the substrate on the properties of NKN films.     

Hydrodynamic thickness of petroleum oil adsorbed layers in the pores of reservoir rocks

The hydrodynamic thickness delta of adsorbed petroleum (crude) oil layers into the pores of sandstone rocks, through which the liquid flows, has been studied by Poiseuille's flow law and the evolution of (electrical) streaming current. The adsorption of petroleum oil is accompanied by a numerical reduction in the (negative) surface potential of the pore walls, eventually stabilizing at a small positive potential, attributed to the oil macromolecules themselves. After increasing to around 30% of the pore radius, the adsorbed layer thickness delta stopped growing either with time or with concentrations of asphaltene in the flowing liquid. The adsorption thickness is confirmed with the blockage value of the rock pores' area determined by the combination of streaming current and streaming potential measurements. This behavior is attributed to the effect on the disjoining pressure across the adsorbed layer, as described by Derjaguin and Churaev, of which the polymolecular adsorption films lose their stability long before their thickness has approached the radius of the rock pore.     

A careful examination of the adsorption step in the alternate layer-by-layer assembly of linear polyanion and polycation

In order to elaborate alternate layer-by-layer assembly as a means to prepare ultrathin films, details of conventional polyion assemblies have been quantitatively analyzed by quartz crystal microbalance (QCM) technique with the aid of scanning electron microscopy (SEM) and atomic force microscopy (AFM). The alternate adsorption of poly(styrenesulfonate) (PSS) and poly(allylamine) (PAM) onto oppositely-charged surfaces displayed the pseudo first-order kinetics and was saturated within 10–20 min at pH 3 and 22°C. It was revealed that drying at every step increased the thickness of adsorbed films due to enhanced surface roughness of the films. Therefore, frequent drying is not profitable for preparing films in a good quality. Non-contact AFM observation revealed that drying of the film with nitrogen stream, forced polymer chains to align to one direction with increasing surface roughness. In contrast, water washing between the consecutive adsorptions was effective for successful alternate adsorption. About 10% of an adsorbed polyion layer was removed by 5-min water washing probably due to removal of the loosely-attached materials.     

Negative azeotropy in mixed adsorption of hexanol and dodecylammonium chloride at water/air interface

Miscibility of hexanol and dodecylammonium chloride (DAC) in adsorbed films and micelles was investigated by evaluating the compositions of the adsorbed films and micelles from surface tension measurements. Judging from the phase diagram of adsorption, negative azeotropy of adsorption was observed for the mixed adsorption of hexanol and DAC at water/air interfaces. The nonideal mixing in the adsorbed film was clarified using excess functions of adsorption. Interaction between hexanol and DAC in the adsorbed film was compared with that between other alkanols and surfactants. It was found that the range of azeotropes is narrower for the hexanol-DAC mixture than for the heptanol-octylsulfinylethanol mixture, because interaction between different species in an adsorbed film is weaker in the former than in the latter.     

Characterization of IgG Langmuir-Blodgett films immobilized on functionalized polymers

The bioactivity of anti-human IgG Langmuir-Blodgett (LB) films, the non-specific adsorption of protein and the topography of anti-IgG LB films have been studied for application in immunosensors. The antibody (AB) LB films were horizontally deposited on glass and functionalized polymers, such as carboxy-poly(vinyl chloride) (PVC-COOH), chloropropyl and aminopropyl sol-gel. The LB films were characterized by means of ellipsometry, atomic force microscopy (AFM) and bicinchoninic acid (BCA) protein test. The interpretation of ellipsometric data was performed using a one-layer model. Non-specifically adsorbed protein was desorbed by washing the IgG film in 0.5 M NaCl, 2 M NaCl and 1% N-cetyl-N,N,N-trimethylammoniumbromide detergent solution resulting in a 50% reduction of the film thickness. The mean thickness of an anti-IgG film on glass measured by ellipsometry, PVC-COOH and aminopropyl sol-gel was 9+/-2, 11+/-1 and 23+/-8 nm, respectively. According to the BCA test 6-8 mug antibody (AB) per slide was bound to the functionalized polymers, but only 3 mug AB per slide was adsorbed on glass. The average distance of anti-IgG granules as indicated by AFM measurements on PVC-COOH, chloropropyl and aminopropyl sol-gel was 42+/-20, 34+/-3 and 23+/-4 nm. The average distance of granular AB structures on glass, however, was 150+/-50 nm.     

Self-organization of a wedge-shaped surfactant in monolayers and multilayers

The self-organization behavior of a wedge-shaped surfactant, disodium-3,4,5-tris(dodecyloxy)phenylmethylphosphonate, was studied in Langmuir monolayers (at the air-water interface), Langmuir-Blodgett (LB) monolayers and multilayers, and films adsorbed spontaneously from isooctane solution onto a mica substrate (self-assembled films). This compound forms an inverted hexagonal lyotropic liquid crystal phase in the bulk and in thick adsorbed films. Surface pressure isotherm and Brewster angle microscope (BAM) studies of Langmuir monolayers revealed three phases: gas (G), liquid expanded (LE), and liquid condensed (LC). The surface pressure-temperature phase diagram was determined in detail; a triple point was found at approximately 10 degrees C. Atomic force microscope (AFM) images of LB monolayers transferred from various regions of the phase diagram were consistent with the BAM images and indicated that the LE regions are approximately 0.5 nm thinner than the LC regions. AFM images were also obtained of self-assembled films after various adsorption times. For short adsorption times, when monolayer self-assembly was incomplete, the film topography indicated the coexistence of two distinct monolayer phases. The height difference between these two phases was again 0.5 nm, suggesting a correspondence with the LE/LC coexistence observed in the Langmuir monolayers. For longer immersion times, adsorbed multilayers assembled into highly organized periodic arrays of inverse cylindrical micelles. Similar periodic structures, with the same repeat distance of 4.5 nm, were also observed in three-layer LB films. However, the regions of organized periodic structure were much smaller and more poorly correlated in the LB multilayers than in the films adsorbed from solution. Collectively, these observations indicate a high degree of similarity between the molecular organization in Langmuir layers/LB films and adsorbed self-assembled films. In both cases, monolayers progress through an LE phase, into LE/LC coexistence, and finally into LC phase as surface density increases. Following the deposition of an additional bilayer, the film reorganizes to form an array of inverted cylindrical micelles.     

Influence of NaCl on the behavior of PEO-PPO-PEO triblock copolymers in solution, at interfaces, and in asymmetric liquid films

The solution behavior of the polymeric surfactant Pluronic F127 (PEO(99)PPO(65)PEO(99)) and its adsorption behavior on aqueous-silica and aqueous-air interfaces, as well as the disjoining pressure isotherms of asymmetric films (silica/aqueous film/air) containing F127, are studied. The interfacial properties of adsorbed F127 layers (the adsorbed amount Gamma and the thickness h) as well as the aqueous wetting film properties [film thickness (h) and refractive indexes] were studied via ellipsometry. The solution properties of F127 were investigated using surface tensiometry and light scattering. The interactions between the air-water and silica-water interfaces were measured with a thin film pressure balance technique (TFB) and interpreted in terms of disjoining pressure as a function of the film thickness. The relations between the behaviors of the asymmetric films, adsorption at aqueous air, and aqueous silica interfaces and the solution behavior of the polymeric surfactant are discussed. Special attention is paid to the influence of the concentrations of F127 and NaCl. Addition of electrolyte lowers the critical micelle concentration, diminishes adsorption on silica, and increases the thickness of the asymmetric film.     

Polyelectrolyte blend multilayer films: surface morphology, wettability, and protein adsorption characteristics

We report the influence of polyelectrolyte (PE) multilayer films prepared from poly(styrene sulfonate)-poly(acrylic acid) (PSS-PAA) blends, deposited in alternation with poly(allylamine hydrochloride) (PAH), on film wettability and the adsorption behavior of the protein immunoglobulin G (IgG). Variations in the chemical composition of the PAH/(PSS-PAA) multilayer films, controlled by the PSS/PAA blend ratio in the dipping solutions, were used to systematically control film thickness, surface morphology, surface wettability, and IgG adsorption. Spectroscopic ellipsometry measurements indicate that increasing the PSS content in the blend solutions results in a systematic decrease in film thickness. Increasing the PSS content in the blend solutions also leads to a reduction in film surface roughness (as measured by atomic force microscopy), with a corresponding increase in surface hydrophobicity. Advancing contact angles (theta) range from 7 degrees for PAH/PAA films through to 53 degrees for PAH/PSS films. X-ray photoelectron spectroscopy measurements indicate that the increase in film hydrophobicity is due to an increase in PSS concentration at the film surface. In addition, the influence of added electrolyte in the PE solutions was investigated. Adsorption from PE solutions containing added salt favors PSS adsorption and results in more hydrophobic films. The amount of IgG adsorbed on the multilayer films systematically increased on films assembled from blends with increasing PSS content, suggesting strong interactions between PSS in the multilayer films and IgG. Hence, multilayer films prepared from blended PE solutions can be used to tune film thickness and composition, as well as wetting and protein adsorption characteristics.     

Effect of interfacial proteins on osteoblast-like cell adhesion to hydroxyapatite nanocrystals

A quartz crystal microbalance with dissipation (QCM-D) technique was employed to detecting the protein adsorption and subsequent osteoblast-like cell adhesion to hydroxyapatite (HAp) nanocrystals. The interfacial phenomena with the preadsorption of three proteins (albumin (BSA), fibronectin (Fn), and collagen (Col)), the subsequent adsorption of fetal bovine serum (FBS), and the adhesion of the cells were investigated. The QCM-D measured the frequency shift (Δf) and dissipation energy shift (ΔD), and the viscoelastic properties of the adlayers were evaluated using ΔD-Δf plot and Voigt-based viscoelastic model. The Col adsorption significantly showed higher Δf, ΔD, elasticity, and viscosity values as compared to the BSA and Fn adsorption, and the subsequent FBS adsorption depended on the preadsorbed proteins. The ΔD-Δf plot of the cell adhesion also showed a different behavior depending on the surfaces, and the Fn- and Col-modified surfaces showed the rapid mass and ΔD changes by forming the viscous interfacial layers with cell adhesion, indicating that the processes were affected by the cellular reaction through the extracellular matrix (ECM) proteins. The confocal laser scanning microscope images of adherent cells showed a different morphology and pseudopod on the surfaces. The cells adhered to the surfaces modified with the Fn and Col had significantly uniaxially expanded shapes and fibrous pseudopods, and those modified with the BSA had a round shape. Therefore, the different cell-protein interactions would cause the arrangement of the ECM and the cytoskeleton changes at the interfaces, and these phenomena were successfully detected by the QCM-D and Voigt-based model.