Adhesiveness of opportunistic Staphylococcus aureus to materials used in dental office: In vitro study.

Staphylococcus aureus (S. aureus) is one of several opportunistic microbial pathogens associated with many healthcare problems. In the present study, S. aureus was assessed for its biofilm-forming ability on materials routinely used in dental offices, including stainless steel (SS), polyethylene (PE), and polyvinyl chloride (PVC). Materials that were tested were characterized for roughness (Ra) and surface free energy (SFE). The adhesion forces exerted by S. aureus to each substratum were investigated using atomic force microscopy (AFM), and biofilm formation was quantitatively assessed by crystal violet staining assay. AFM measurements demonstrated that the strongest adhesion forces (20 nN) were exerted on the PE surfaces (P < 0.05) and depended more on Ra. In addition, the results of biofilm formation capability indicated that S. aureus exhibited more affinity to SS materials when compared to the other materials (P < 0.05). This ability of biofilm formation seems to be more correlated to SFE (R = 0.65). Hence, control of the surface properties of materials used in dental practices is of crucial importance for preventing biofilm formation on dental materials to be used for patients' dental care.

Adhesive forces and surface properties of cold gas plasma treated UHMWPE.

Cold atmospheric plasma (CAP) treatment was used on ultra-high molecular weight polyethylene (UHMWPE), a common articulating counter material employed in hip and knee replacements. UHMWPE is a biocompatible polymer with low friction coefficient, yet does not have robust wear characteristics. CAP effectively cross-links the polymer chains of the UHMWPE improving wear performance (Perni et al., Acta Biomater. 8(3) (2012) 1357). In this work, interactions between CAP treated UHMWPE and spherical borosilicate sphere (representing model material for bone) were considered employing AFM technique. Adhesive forces increased, in the presence of PBS, after treatment with helium and helium/oxygen cold gas plasmas. Furthermore, a more hydrophilic surface of UHMWPE was observed after both treatments, determined through a reduction of up to a third in the contact angles of water. On the other hand, the asperity density also decreased by half, yet the asperity height had a three-fold decrease. This work shows that CAP treatment can be a very effective technique at enhancing the adhesion between bone and UHMWPE implant material as aided by the increased adhesion forces. Moreover, the hydrophilicity of the CAP treated UHMWPE can lead to proteins and cells adhesion to the surface of the implant stimulating osseointegration process.

Adhesion strength and anti-tumor agents regulate vinculin of breast cancer cells.

The onset and progression of cancer are strongly associated with the dissipation of adhesion forces between cancer cells, thus facilitating their incessant attachment and detachment from the extracellular matrix (ECM) to move toward metastasis. During this process, cancer cells undergo mechanical stresses and respond to these stresses with membrane deformation while inducing protrusions to invade the surrounding tissues. Cellular response to mechanical forces is inherently related to the reorganization of the cytoskeleton, the dissipation of cell-cell junctions, and the adhesion to the surrounding ECM. Moreover, the role of focal adhesion proteins, and particularly the role of vinculin in cell attachment and detachment during migration, is critical, indicating the tight cell-ECM junctions, which favor or inhibit the metastatic cascade. The biomechanical analysis of these sequences of events may elucidate the tumor progression and the potential of cancer cells for migration and metastasis. In this work, we focused on the evaluation of the spreading rate and the estimation of the adhesion strength between breast cancer cells and ECM prior to and post-treatment with anti-tumor agents. Specifically, different tamoxifen concentrations were used for ER+ breast cancer cells, while even concentrations of trastuzumab and pertuzumab were used for HER2+ cells. Analysis of cell stiffness indicated an increased elastic Young's modulus post-treatment in both MCF-7 and SKBR-3 cells. The results showed that the post-treatment spreading rate was significantly decreased in both types of breast cancer, suggesting a lower metastatic potential. Additionally, treated cells required greater adhesion forces to detach from the ECM, thus preventing detachment events of cancer cells from the ECM, and therefore, the probability of cell motility, migration, and metastasis was confined. Furthermore, post-detachment and post-treatment vinculin levels were increased, indicating tighter cell-ECM junctions, hence limiting the probability of cell detachment and, therefore, cell motility and migration.

Adhesion force evolution of protein on the surfaces with varied hydration extent: Quantitative determination via atomic force microscopy.

The adhesion force evolution of protein on surfaces with continuously varied hydrophobicity/hydration layer has not been completely clarified yet, limiting the further development of environmental applications such as membrane anti-biofouling and selective adsorption of the functional surfaces. Herein, chemical force spectroscopy using atomic force microscopy (AFM) was utilized to quantify the evolution of the adhesion forces of protein on hydration surfaces in water, where bovine serum albumin (BSA) was immobilized on an AFM tip as the representative protein. The stiffness, roughness and charge properties of the substrate surfaces were kept constant and the hydrophobicity was the only variant to monitor the role of hydrated water layers in protein adhesion. The adhesion force increased non-monotonically as a function of hydrophobicity of substrate surfaces, which was related to the concentration of humic acid, and independent of pH values and ionic strength. The non-monotonic variation occurred in the range of contact angle at 60-80° due to the mutual restriction between solid-liquid interface energy and solid-solid interface energy. Hydrophobic attraction was the dominant force that drove adhesion of BSA to these model substrate surfaces, but the passivation of hydration layers at the interface could weaken the hydrophobic attraction. In contrast to the measurements in water, the adhesion forces decreased as a function of surface hydrophobicity when measured in air, because capillary forces from condensation water dominated adhesion forces. The passivation of hydration layers of protein was revealed by quantitatively determining the evolution of adhesion forces on the hydration surfaces of varying hydrophobicity, which was ignored by traditional adhesion theory.

The effect of granules characters on mechanical properties of press-coated tablets: A comparative study.

The aim of this study was to investigate the correlation between critical granules characters (including particle size, surface roughness, and apparent porosity) and mechanical properties of press-coated tablets. Granules of a model formulation were prepared through Roll Compaction Granulation (RCG), High Shear Granulation (HSG), and Fluidized Bed Granulation (FBG) to prepare granules with different surface roughness and apparent porosity. The surface roughness and porosity of granules had a significantly greater effect on mechanical properties than the particle size of granules. Whether for brittle or plastic materials, FBG granules with the roughest surface and the greatest apparent porosity exhibited the best compression properties. The elastic recovery test, the interlayer adhesion forces study, the break pattern test, and the X-ray microcomputed tomography investigation suggested that granules with great apparent porosity and rough surfaces could contribute to the production of stable press-coated structures. Moreover, for press-coated tablets prepared using granules, the proper granules in the coat layer could eliminate the side effect of the rigid core on the mechanical strength. The above understandings will be conducive to the selection of compatible and appropriate granules characters, which can enhance mechanical properties and extend the application of press-coated tablets.

Adhesion of LHRH/EphA2 to human Triple Negative Breast Cancer tissues.

The adhesive interactions between molecular recognition units (such as specific peptides and antibodies) and antigens or other receptors on the surfaces of tumors are of great value in the design of targeted nanoparticles and drugs for the detection and treatment of specific cancers. In this paper, we present the results of a combined experimental and theoretical study of the adhesion between Luteinizing Hormone Releasing Hormone (LHRH)/Epherin type A2 (EphA2)-AFM coated tips and LHRH/EphA2 receptors that are overexpressed on the surfaces of human Triple Negative Breast Cancer (TNBC) tissues of different histological grades. Following a histochemical and immuno-histological study of human tissue extracts, the receptor overexpression, and their distributions are characterized using Immunohistochemistry (IHC), Immunofluorescence (IF), and a combination of fluorescence microscopy and confocal microscopy. The adhesion forces between LHRH or EphA2 and human TNBC breast tissues are measured using force microscopy techniques that account for the potential effects of capillary forces due to the presence of water vapor. The corresponding adhesion energies are also determined using adhesion theory. The pull off forces and adhesion energies associated with higher grades of TNBC are shown to be greater than those associated with normal/non-tumorigenic human breast tissues, which were studied as controls. The observed increase in adhesion forces and adhesion energies are also correlated with the increasing incidence of LHRH/EphA2 receptors at higher grades of TNBC. The implications of the results are discussed for the development of targeted nanostructures for the detection and treatment of TNBC.

Physicochemical and Electrochemical Characterization of Electropolymerized Polydopamine Films: Influence of the Deposition Process.

Polydopamine (PDA) is a synthetic eumelanin polymer which is, to date, mostly obtained by dip coating processes. In this contribution, we evaluate the physical and electrochemical properties of electrochemically deposited PDA films obtained by cyclic voltammetry or pulsed deposition. The obtained PDA thin films are investigated with respect to their electrochemical properties, i.e., electron transfer (ET) kinetics and charge transfer resistance using scanning electrochemical microscopy and electrochemical impedance spectroscopy, and their nanomechanical properties, i.e., Young's modulus and adhesion forces at varying experimental conditions, such as applied potential or pH value of the medium using atomic force microscopy. In particular, the ET behavior at different pH values has not to date been investigated in detail for electrodeposited PDA thin films, which is of particular interest for a multitude of applications. Adhesion forces strongly depend on applied potential and surrounding pH value. Moreover, force spectroscopic measurements reveal a significantly higher percentage of polymeric character compared to films obtained by dip coating. Additionally, distinct differences between the two depositions methods are observed, which indicate that the pulse deposition process leads to denser, more cross-linked films.

Quantifying the relationship between surfaces' nano-contact point density and adhesion force of Candida albicans.

It has been recently recognized that controlled surface structuring on the nanometer scale is a successful strategy to endow different materials with antimicrobial properties. Despite many studies on bacterial interactions with nanostructured surfaces, a quantitative link between surface topography and bacterial adhesion is still missing. To quantitatively link cell adhesion data with topographical surface parameters, we performed single-cell spectroscopy on chemically identical surfaces with controlled nano-contact point density achieved by immobilization of gold nanoparticles (AuNP) on gold thin films. Such materials surfaces have previously shown antimicrobial (anti-adhesive) efficacy towards Gram-negative Escherichia coli cells. In the current study, the influence of nano-structured surfaces on the surface coverage and adhesion forces of clinically relevant Candida albicans (C. albicans), the fungus primarily associated with implant infections, was investigated to validate their antimicrobial potency against different microbial cells. The adhesion forces of C. albicans cells to nanostructured surfaces showed a decreasing trend with decreasing contact-point density and correlated well with the results of the respective C. albicans cell counts. The surfaces with the lowest contact-point density, 25 AuNP/µm², resulted in an average adhesion force of 5 nN, which was up to 5 times lower compared to control and 61 AuNP/µm² surfaces. Further, detailed analyses of force-distance curves revealed that the work of adhesion, and thus the energy required to remove the C. albicans cell from the surface is up to 10 times lower on 25 AuNP/µm² surfaces compared to unstructured surfaces. These findings show that a controlled tuning of nanostructured surfaces in terms of accessible nano-contact points is crucial to generate surface structures with enhanced antimicrobial properties. The gained knowledge can be further exploited for the design of biomaterials surfaces to prevent adhesion of some most commonly encountered pathogens.

Effect of cholesterol on monolayer structure of different acyl chained phospholipids.

In this work we have investigated the effect of cholesterol (CHOL) in phospholipid monolayers on a series of phosphatidylcholines differing in acyl chain composition. We have used the CHOL proportion that abolishes the gel (Lß)-to-liquid-crystalline (Lß) transition in bilayers in order to investigate the mixing properties and laterally-segregated domains formed by specific phospholipid-CHOL ratios at the air-water interface. The binary monolayers where formed by mixing CHOL with 1,2-palmitoyl-sn-glycero-3-phos-phatidylcholine (DPPC);1,2-distearoyl-sn-glycero-3-phosphatidylcholine (DSPC); 1-pal-mitoyl-2-stearoyl-sn-glycero-3-phosphatidylcholine (PSPC); 1-palmitoyl-2-oleoyl-sn-gly-cero-3-phosphatidylcholine (POPC) and 1-palmitoyl-2-linoleyl-sn-glycero-3-phosphatidyl-choline (PLPC), respectively. From surface pressure-area (π-A) isotherms the isothermal compression modulus were calculated, and the mixing properties of the monolayers obtained by performing a basic surface thermodynamic analysis. From the excess Gibbs energy, the interaction parameter and the activity coefficients were also calculated. The study of the monolayers was complemented by determining the molecular dipole moment normal to the plane of the monolayer. The existence of laterally segregated domains was assessed by atomic force microscopy (AFM) of Langmuir-Blodgett films (LBs) extracted at 30 mNm-1. To get insight into the nature and composition of the observed domains force spectroscopy (FS) based on AFM was applied to the LBs.

Adhesion forces of biofilms developed in vitro from clinical strains of skin wounds.

A biofilm is a very complex consortium formed by a mix of different microorganisms, which have become an important health problem, because its formation is a resistance mechanism used by bacteria against antibiotics or the immune system. In this work, we show differences between some physicochemical properties of biofilms in mono- and multi-species, formed by bacteria from clinical samples of infected chronic wounds. Of the most prevalent bacteria in wounds, two mono- and one multi-species biofilms were developed in vitro by Drip Flow Reactor: one biofilm was developed by S. aureus, other by P. aeruginosa, and a third one by the mix of both strains. With these biofilms, we determined microbial growth by plate counting, and their physicochemical characterization by Atomic Force Microscopy, Raman Micro-Spectroscopy and Scanning Electron Microscopy. We found that the viability of S. aureus was less than P. aeruginosa in multi-species biofilm. However, the adhesion force of S. aureus is much higher than that of P. aeruginosa, but it decreased while that of P. aeruginosa increased in the multi-species biofilm. In addition, we found free pyrimidines functional groups in the P. aeruginosa biofilm and its mix with S. aureus. Surprisingly, each bacterium alone formed single layer biofilms, while the mix bacteria formed a multilayer biofilm at the same observation time. Our results show the necessity to evaluate biofilms from clinically isolated strains and have a better understanding of the adhesion forces of bacteria in biofilm multispecies, which could be of prime importance in developing more effective treatments against biofilm formation.

Adhesive forces between two cleaved calcite surfaces in NaCl solutions: The importance of ionic strength and normal loading.

The mechanical strength of calcite bearing rocks is influenced by pore fluid chemistry due to the variation in nano-scale surface forces acting at the grain contacts or close to the fracture tips. The adhesion of two contacting surfaces, which affects the macroscopic strength of the material, is not only influenced by the fluid chemistry but also by the surface topography. In this paper, we use Atomic Force Microscope (AFM) to measure the interfacial forces between two freshly cleaved calcite surfaces in CaCO3-saturated solutions with varying NaCl concentration. We show that calcite contacts become stronger with increasing NaCl concentration (>100 mM), as a result of progressively weaker secondary hydration and increasing attraction due to instantaneous ion-ion correlation. Moreover, we discuss the effect of normal applied force (Fn) and surface roughness on the measured adhesion forces (Fad). We show that the measured pull-off force (adhesion) is linearly correlated with the magnitude of Fn, where an increase in applied force results in increased adhesion. This is attributed to a larger number of contacting surface asperities and thus increase in real contact area and the contact-bond strength. We discuss that the possible variation in local topography at contacts, together with strong dependence on ionic strength of the solution, can explain the inconsistent behavior of calcite rocks in NaCl solutions.

Aqueous-Based Fabrication of Low-VOC Nanostructured Block Copolymer Films as Potential Marine Antifouling Coatings.

The ability to fabricate nanostructured films by exploiting the phenomenon of microphase separation has made block copolymers an invaluable tool for a wide array of coating applications. Standard approaches to engineering nanodomains commonly involve the application of organic solvents, either through dissolution or annealing protocols, resulting in the release of volatile organic compounds (VOCs). In this paper, an aqueous-based method of fabricating low-VOC nanostructured block copolymer films is presented. The reported procedure allows for the phase transfer of water insoluble triblock copolymer, poly(styrene-block-2 vinylpyridine-block-ethylene oxide) (PS-b-P2VP-b-PEO), from a water immiscible phase to an aqueous environment with the assistance of a diblock copolymeric phase transfer agent, poly(styrene-block-ethylene oxide) (PS-b-PEO). Phase transfer into the aqueous phase results in self-assembly of PS-b-P2VP-b-PEO into core-shell-corona micelles, which are characterized by dynamic light scattering techniques. The films that result from coating the micellar solution onto Si/SiO2 surfaces exhibit nanoscale features that disrupt the ability of a model foulant, a zoospore of Ulva linza, to settle. The multilayered architecture consists of a pH-responsive P2VP-"shell" which can be stimulated to control the size of these features. The ability of these nanostructured thin films to resist protein adsorption and serve as potential marine antifouling coatings is supported through atomic force microscopy (AFM) and analysis of the settlement of Ulva linza zoospore. Field trials of the surfaces in a natural environment show the inhibition of macrofoulants for 1 month.

Mapping molecular adhesion sites inside SMIL coated capillaries using atomic force microscopy recognition imaging.

Capillary zone electrophoresis (CZE) is a powerful analytical technique for fast and efficient separation of different analytes ranging from small inorganic ions to large proteins. However electrophoretic resolution significantly depends on the coating of the inner capillary surface. High technical efforts like Successive Multiple Ionic Polymer Layer (SMIL) generation have been taken to develop stable coatings with switchable surface charges fulfilling the requirements needed for optimal separation. Although the performance can be easily proven in normalized test runs, characterization of the coating itself remains challenging. Atomic force microscopy (AFM) allows for topographical investigation of biological and analytical relevant surfaces with nanometer resolution and yields information about the surface roughness and homogeneity. Upgrading the scanning tip to a molecular biosensor by adhesive molecules (like partly inverted charged molecules) allows for performing topography and recognition imaging (TREC). As a result, simultaneously acquired sample topography and adhesion maps can be recorded. We optimized this technique for electrophoresis capillaries and investigated the charge distribution of differently composed and treated SMIL coatings. By using the positively charged protein avidin as a single molecule sensor, we compared these SMIL coatings with respect to negative charges, resulting in adhesion maps with nanometer resolution. The capability of TREC as a functional investigation technique at the nanoscale was successfully demonstrated.

Sticky Matrix: Adhesion Mechanism of the Staphylococcal Polysaccharide Intercellular Adhesin.

The development of bacterial biofilms on surfaces leads to hospital-acquired infections that are difficult to fight. In Staphylococci, the cationic polysaccharide intercellular adhesin (PIA) forms an extracellular matrix that connects the cells together during biofilm formation, but the molecular forces involved are unknown. Here, we use advanced force nanoscopy techniques to unravel the mechanism of PIA-mediated adhesion in a clinically relevant methicillin-resistant Staphylococcus aureus (MRSA) strain. Nanoscale multiparametric imaging of the structure, adhesion, and elasticity of bacteria expressing PIA shows that the cells are surrounded by a soft and adhesive matrix of extracellular polymers. Cell surface softness and adhesion are dramatically reduced in mutant cells deficient for the synthesis of PIA or under unfavorable growth conditions. Single-cell force spectroscopy demonstrates that PIA promotes cell-cell adhesion via the multivalent electrostatic interaction with polyanionic teichoic acids on the S. aureus cell surface. This binding mechanism rationalizes, at the nanoscale, the well-known ability of PIA to strengthen intercellular adhesion in staphylococcal biofilms. Force nanoscopy offers promising prospects for understanding the fundamental forces in antibiotic-resistant biofilms and for designing anti-adhesion compounds targeting matrix polymers.

Harnessing tunable scanning probe techniques to measure shear enhanced adhesion of gecko-inspired fibrillar arrays.

The hierarchical arrays of mesoscale to nanoscale fibrillar structures on a gecko's foot enable the animal to climb surfaces of varying roughness. Adhesion force between the fibrillar structures and various surfaces is maximized after the gecko drags its foot in one direction, which has also been demonstrated to improve the adhesion forces of artificial fibrillar arrays. Essential conditions that influence the magnitude of these interactions include the lateral distance traveled and velocity between the contacting surfaces, as well as the velocity at which the two surfaces are subsequently separated. These parameters have, however, not been systematically investigated to assess the adhesion properties of artificial adhesives. We introduce a systematic study that investigates these conditions using a scanning probe microscope to measure the adhesion forces of artificial adhesives through a process that mimics the mechanism by which a gecko climbs. The measured adhesion response was different for arrays of shorter and longer fibrils. These results from 9000 independent measurements also provide further insight into the dynamics of the interactions between fibrillar arrays and contacting surfaces. These studies establish scanning probe microscopy techniques as a versatile approach for measuring a variety of adhesion properties of artificial fibrillar adhesives.

Multi-scale modelling of the dynamics of cell colonies: insights into cell-adhesion forces and cancer invasion from in silico simulations.

Studying the biophysical interactions between cells is crucial to understanding how normal tissue develops, how it is structured and also when malfunctions occur. Traditional experiments try to infer events at the tissue level after observing the behaviour of and interactions between individual cells. This approach assumes that cells behave in the same biophysical manner in isolated experiments as they do within colonies and tissues. In this paper, we develop a multi-scale multi-compartment mathematical model that accounts for the principal biophysical interactions and adhesion pathways not only at a cell-cell level but also at the level of cell colonies (in contrast to the traditional approach). Our results suggest that adhesion/separation forces between cells may be lower in cell colonies than traditional isolated single-cell experiments infer. As a consequence, isolated single-cell experiments may be insufficient to deduce important biological processes such as single-cell invasion after detachment from a solid tumour. The simulations further show that kinetic rates and cell biophysical characteristics such as pressure-related cell-cycle arrest have a major influence on cell colony patterns and can allow for the development of protrusive cellular structures as seen in invasive cancer cell lines independent of expression levels of pro-invasion molecules.

Improved Adhesion and Compliancy of Hierarchical Fibrillar Adhesives.

The gecko relies on van der Waals forces to cling onto surfaces with a variety of topography and composition. The hierarchical fibrillar structures on their climbing feet, ranging from mesoscale to nanoscale, are hypothesized to be key elements for the animal to conquer both smooth and rough surfaces. An epoxy-based artificial hierarchical fibrillar adhesive was prepared to study the influence of the hierarchical structures on the properties of a dry adhesive. The presented experiments highlight the advantages of a hierarchical structure despite a reduction of overall density and aspect ratio of nanofibrils. In contrast to an adhesive containing only nanometer-size fibrils, the hierarchical fibrillar adhesives exhibited a higher adhesion force and better compliancy when tested on an identical substrate.

Feasibility of measuring antigen-antibody interaction forces using a scanning force microscope.

The molecular affinity scanning force microscopy (MASFM) described in this study was developed in an effort to test the possibility of antigen-antibody binding measurement using force-separation distance profiles. The MASFM configuration was comprised of a spherical glass bead as an MASFM probe, to which the fluorescein antigen has been covalently attached, and a silicon dioxide-based substrate, to which the antifluorescyl IgG antibody was covalently bound. The bead was glued to the tip of a commercial SFM cantilever. Adhesion forces have been measured between two different specific antigen-antibody pairs and between nonspecific surfaces bearing only glycidoxypropylsilane immobilization chemistry. In force-separation (F-s) measurements, nonspecific forces displayed relatively few force discontinuities and mean adhesion forces lower than those found for specific antigen-antibody measurements. Force-separation profiles measured between specific antigen-antibody pairs showed many discontinuities and had higher mean forces. Positive controls revealed that the mean forces were slightly reduced by the addition of free ligand. The magnitude of mean forces did not correlate with the respective activation enthalpies of the proteins, as would be expected. At lower force values the force histograms for the specific pairs and for positive controls were indistinguishable. None of the force-separation data sets could fit a Poisson discrete-force model. This statistical analysis showed a large relative contribution from nonspecific interactions. It is concluded that the use of the large sphere as an SFM probe is counterproductive: while the large sphere does sample a larger number of specific interactions during each measurement, it also samples at the same time a large proportion of nonspecific forces. The presence of the nonspecific force contributions is likely due to the deformation of the polymerized GPS spacer layer which is thought to be delaminated from the surface upon the application of tension across the specific antigen-antibody bonds.