Surface Adhesion Research Articles

Mimicking Mytilus edulis foot protein: A versatile strategy for robust biomedical coatings

Universal coatings with versatile surface adhesion, good mechanochemical robustness, and the capacity for secondary modification are of great scientific interest. However, incorporating these advantages into a system is still a great challenge. Here, we report a series of catechol-decorated polyallylamines (CPAs), denoted as pseudo-Mytilus edulis foot protein 5 (pseudo-Mefp-5), that mimic not only the catechol and amine groups but also the backbone of Mefp-5. CPAs can fabricate highly adhesive, robust, multifunctional polyCPA (PCPA) coatings based on synergetic catechol-polyamine chemistry as universal building blocks. Due to the interpenetrating entangled network architectures, these coatings exhibit high chemical robustness against harsh conditions (HCl, pH 1; NaOH, pH 14; H2O2, 30%), good mechanical robustness, and wear resistance. In addition, PCPA coatings provide abundant grafting sites, enabling the fabrication of various functional surfaces through secondary modification. Furthermore, the versatility, multifaceted robustness, and scalability of PCPA coatings indicate their great potential for surface engineering, especially for withstanding harsh conditions in multipurpose biomedical applications.

Pore confinement enhances but surface adhesion reduces bacterial cell-to-cell conjugation

Pore confinement enhances but surface adhesion reduces bacterial cell-to-cell conjugation

The Role of Platelet Activation in the Development and Metastasis of Solid Tumors

The blood coagulation system is actively involved in the development of cancer. It is known that many solid tumors express tissue factor, a “trigger” of the cascade of plasma coagulation reactions, which leads to an increased risk of cancer-associated thrombosis and venous thrombosis in cancer patients. It has also long been known that platelets - small cellular fragments that are the basis of blood clots - play a critical role in metastasis by binding to the tumor cell after it enters the blood vessel, “shielding” it from the immune system and promoting the adhesion and extravasation of the tumor cell into tissues and the formation metastasis. In addition, platelets, being mobile “storehouses” of growth factors, are actively attracted and, in some cases, consumed by the tumor, which contributes to its development and vascularization. Platelet attraction occurs both through activation of the blood coagulation system in the tumor area and through exposure of the adhesive surface by the tumor. Activated in the tumor vicinity, platelets attract and induce neutrophil activation and the formation of neutrophil extracellular traps (NETs), thereby modulating the tumor microenvironment. When activated, platelets are known to secrete a variety of growth factors that promote both tumor development and vascularization. In addition to direct interaction, platelets and tumor cells exchange mRNA, micro-RNA and other regulatory molecules through microvesicles, while platelets are containers for the spread of tumor genetic material (circulating nucleic acids) throughout the body. In this review, we consider the molecular mechanisms of platelet participation in the development and metastasis of solid tumors, and also discuss possible options for pharmacological interruption of this interaction.

Investigation of adhesion status of Candida species to the surface of resin materials produced at different angles with additive manufacturing

BackgroundThe aim of this study was to evaluate the adhesion of Candida glabrata, Candida albicans, Candida krusei, Candida parapsilosis and Candida tropicalis yeasts to disk-shaped resin materials produced from resin which used in the production of surgical guide with 0, 45 and 90-degrees printing orientations by Liquid Crystal Display additive manufacturing technology.MethodsDisk-shaped specimens were printed with surgical guide resin using the Liquid Crystal Display production technique in 3 printing orientations (0, 45 and 90-degrees). Surface roughness and contact angle values were evaluated. Real-Time PCR analysis was performed to evaluate Candida adhesion (C. glabrata, C. albicans, C. krusei, C. parapsilosis and C. tropicalis) Field emission scanning electron microscope (FESEM) images of the materials were obtained.ResultsSpecimens oriented at 45-degrees demonstrated higher surface roughness (P < .05) and lower contact angle values than other groups. No significant difference was found in the adhesion of C. glabrata, C. albicans, and C. parapsilosis among specimens printed at 0, 45, and 90-degrees orientations (P > .05). A higher proportion of C. krusei and C. tropicalis was found in the specimens printed at orientation degrees of 45 = 90 < 0 with statistical significance. Analyzing the adhesion of all Candida species reveals no statistical disparity among the printing orientations.ConclusionsThe surface roughness, contact angle, and adhesion of certain Candida species are affected by printing orientations. Hence, careful consideration of the printing orientation is crucial for fabricating products with desirable properties. In 45-degree production, roughness increases due to the layered production forming steps, whereas in 0-degree production, certain Candida species exhibit high adhesion due to the formation of porous structures. Consequently, considering these factors, it is advisable to opt for production at 90-degrees, while also considering other anticipated characteristics.

New approach for processing chitosan as low cost protective hybrid coating for C-steel in acid media

The novelty of this study is that it the first time blending and formulation of chitosan as a new hybrid (organometallic) protective coatings for achieving synergistic protection for carbon steel alloy during acid pickling. The role of coated silica (by 0.1 wt % stearic acid lubricant) in the improvement of coating performance was highlighted. Variable weight percentage of chitosan and silica in addition to a fixed weight percentage (35 %) of guar gum natural plant resin, 5 × 10−6 mmol (2-Hydrazinyl-6-methyl (or phenyl) −4, 5-di-H pyrimidinone) as organic corrosion inhibitors were compounding as hot melt in the presence of a low cost surfactant as an emulsifying agent improved compatibility between coating constituents. Guar gum increased coating flow during application and grafted chitosan into high molecular copolymer resin insoluble in acid media. Phosphorous acid improved coating flexibility during application by hot dipping. Hybrid coating decreased corrosion potential of carbon steel and retarded both redox reactions of corrosion acting as adsorbed mixed-type inhibitor. Percentages protection (%P) approached hundred percentage as confirmed from the agreement between impedance and polarization parameters. Guar gum plant resin and slice powder increased gloss of coating. The coated silica filled the pores and increased stiffness of coating. Super hydrophobicity of coating was confirmed by the measured contact angle above 150oC indicating good spreading of coating sample as insulating adherent surface film.

Titanium surface treatments and their effects on the roughness factor

The surface pretreatment of a Ti substrate to produce metal oxide electrodes with electrocatalytic properties is of fundamental importance. The preparation of the substrate prior to the application of the coating critically affects the properties of the coating, including its surface roughness, adhesion, and corrosion resistance. In this work, different treatments were applied to the surfaces of Ti discs and their effects on surface roughness were studied. The treatments applied to Ti consisted of three stages: (1) mechanical treatment, (2) degreasing, and (3) chemical treatment. The first stage was carried out with 120 grit SiC abrasive sandpaper, the second stage was carried out with acetone (group A) or sodium hydroxide (NaOH) solution (group B), and the third stage was conducted using hydrochloric acid (HCl), oxalic acid, a mixture of hydrofluoric acid (HF)/phosphoric acid (H3PO4), or a sequence of all three chemical treatments (i.e., HCl, oxalic acid, and HF/H3PO4). The morphological properties, surface composition, and electrochemical behavior of the treated Ti discs were characterized by scanning electron microscopy (SEM), X-ray diffractometry (XRD) and cyclic voltammetry (CV) in a 0.5 M H2SO4 solution, respectively. The electroactive surface area (Aes) and roughness factor (frs) of each Ti disc were estimated from capacitance measurements of the electrochemical double layer. The morphology and voltammetric behavior of Ti varied markedly depending on the surface treatment applied. In addition, the XRD results showed that Ti surfaces subjected to different treatments exhibited different crystallographic planes. The trend in surface roughness with respect to the chemical reagent used was as follows: Group A: oxalic acid > HF/H3PO4 > global treatment > acetone > HCl; Group B: oxalic acid > HF/H3PO4 > NaOH > HCl > global treatment. The most severe chemical attack on the substrate was induced by oxalic acid, which led to the largest electroactive area and roughness factor among the treated Ti samples investigated in this study.

Fabrication of a durable omniphobic thin film and its application in precision nozzles

The ultrafine nozzles used in micropatterning processes easily encounter pollution on the nozzle surfaces, such as clogging, liquid clumping, and bond tailing. In this study, an omniphobic functional film was developed by an initiated chemical vapor deposition (iCVD) method and used to fabricate an antifouling nozzle. In the iCVD process, polymeric fluorine films were grown under optimized conditions, and their durability was analyzed through abrasion and nanoscale cutting tests to evaluate the surface wear resistance and adhesion strength, respectively. Two omniphobic films, prepared using homopolymer p(C6FMA) and copolymer p(C6FMA-co-DVB), were compared; the p(C6FMA-co-DVB) film was determined to have a superior durability than the p(C6FMA) film. Consequently, the p(C6FMA-co-DVB) film was deposited on both the inside and outside surface of a nozzle of diameter 0.3 mm. The oleophobicity was examined by comparing the discharge rates of a high-viscosity epoxy paste containing metal powders in coated and bare nozzles. The bare nozzle exhibited significant variations in discharge volume and frequently experienced clogging, while coated nozzles demonstrated consistent discharge rates and remained unclogged. These findings suggest that the proposed thin film coating has the potential to increase the lifespan and efficiency of ultra-fine nozzles in the automated micro-patterning industry.

Strategy to prevent calcification by restricting surface adhesion of Ca2+: Reduced affinity of extracellular polymeric substances for Ca2+ by mild acidic conditions

Controlling CaCO3 precipitation within anaerobic granular sludge (AnGS) is crucial for the anaerobic treatment of paper recycling wastewater. A viable strategy was proposed to control calcification by adjusting a mild acidic condition in an anaerobic reactor without hindering organic degradation. The results indicated that lowering the bulk pH (6.5 to 6.8) reduced calcium precipitation by 60.1 % in calcium-rich influent (Ca2+ 1200 mg/L) and eradicated CaCO3 deposition on AnGS. Extracellular polymeric substances (EPS) have proven to be crucial participants in Ca2+ migration. The acidic solution weakens the interactions between EPS and Ca2+ and then diminishes the EPS adsorption capacity and affinity for Ca2+. The mild acidic environment goes beyond reducing CaCO3 formation in wastewater. EPS protonation reduced the probability of Ca2+ adhering to the AnGS surface, which halted calcium transportation from bulk liquid to granule. This work offers a feasible strategy to prevent AnGS calcification in high-calcium wastewater.

Sustainable adhesives: Exploring boronic ester vitrimers containing lignin microparticles

The market of epoxy resin-based adhesives is constantly growing in the automotive, electronics, and healthcare industries thanks to their unique features such as high bonding strength, durability, and corrosion resistance. In this work, alternative sustainable vitrimer adhesives, containing from 20 to 50 wt% of lignin microparticles, were prepared from epoxidized linseed oil (ELO) and a boronic ester dithiol crosslinker (DBEDT), in the absence of solvents and catalysts. The addition of unmodified commercial Kraft lignin to the composites directly influences their thermal and mechanical properties and determines their bonding capacity between several adherent substrates, also affecting the rewelding potential. The lignin-vitrimer composites and the corresponding neat vitrimer matrix exhibited values of lap shear strengths in the range of 9 to 17 MPa, when tested as adhesives with aluminum, stainless steel, and wood specimens, and good to excellent rewelding capability. The amount of lignin microparticles influences the balance between cohesive and adhesive forces during the separation of the adhered aluminum surfaces and the eventual joint failure. In the case of the composites with 20 wt% of lignin, lap shear strength remains constant even after four cycles of rebonding via compression molding, indicating that the best rewelding performance is associated with previous cohesive failure when the adhesive remains on both aluminum sheets' surfaces. In addition, the adhesion was preserved for 83 % of the initial value after 24 h immersed in water. Importantly, biodegradability in both soil and seawater was enhanced by the presence of the lignin filler. In summary, the simple preparation strategy of bio-based vitrimer composites coming from two natural sources could pave the way to green alternatives for industrial applications, such as epoxy-based adhesives.

Wetting transitions in adhesive surfaces of polystyrene: The petal effect

HypothesisThe petal effect is a well-known natural phenomenon in surface science and has served as inspiration for the creation of several materials with superhydrophobic qualities and high adhesion. As surface roughness has a crucial role in these properties, being able to modulate it could help us design materials at will. Capillary penetration frustrates diffusion and promotes large contact angles as well as high adhesion. ExperimentsPolystyrene surfaces were created using the spin-coating technique. By varying the polymer concentration, the surface roughness was modified. To determine the roughness parameters, atomic force microscopy was used. We recorded advancing and receding contact angles using water and glycerol as test liquids to study contact angle hysteresis, the work of adhesion and the apparent surface energy, which was determined with the Chibowski and Perea–Carpio method. For the purpose of elucidating the wetting states, captive bubble experiments were conducted. FindingsUsing an easy method, we create polystyrene surfaces with both superhydrophobicity and strong adhesion, where the roughness area factor regulates wetting transitions from Cassie–Baxter to Wenzel. The receding contact angle suggests capillary penetration, which we demonstrate by captive bubble experiments. In addition, a link was found between the surface roughness and apparent surface energy.

Experimental investigation of adhesion force, folding factor and surface roughness of chronic lymphocytic leukemia cell using AFM

In this paper, the Morphological properties, Contact Mechanics, Adhesion, Roughness and Folding of Chronic Lymphocytic Leukemia (CLL) cells have been examined utilizing Atomic Force Microscopy (AFM). The results show that the morphology of the CLL cell is spherical, with an average surface roughness of 23.06 nm and 5.34% folds. Adhesion force and work of adhesion of CLL cells is in the range of 23.2 ± 15.6% nN and 1484.6 ± 13.1% nJ, respectively. DMT and BCP contact theories have been more accurate than Hertz theory in characterizing the mechanical contact behavior of CLL cells due to the consideration of adhesion properties. Also, applying the folding factor in the Hertz, DMT and BCP contact theories, for the same contact force predicts a larger contact radius and a smaller indentation depth, and the accuracy of its results has increased compared to the smooth state.

Feasibility study of adhesively bonded drills with a small diameter for cutting of CFRP

Adhesively bonded cutting tools possess many advantages over classic (insert) tools due to different manufacturing processes: With reduced manufacturing temperatures, the damage to the cutting edge decreases, and the damping effects of the adhesive enhance the tool’s life. In contrast to standard joining technologies like soldering, adhesive bonding enables the joining of various cutting materials and tool base bodies, like ceramic and steel. Furthermore, the usage of expensive, abrasive-resisted cutting materials, like carbide, has to be reduced in future because of the classification of the components (cobalt, wolfram carbide) as a critical resource. Therefore, this article conducts a feasibility study of adhesively bonded drills with a ceramic tip for cutting carbon-fiber-reinforced plastics. The main contents are investigations of different surface preparation methods (laser surface texturing and SACO-blasting), examining four different adhesives for cutting tool usage, and manufacturing and validating prototype drills for cutting operations. The experimental results show a high scattering of the toughness of the joints between 2.8 MPa and 27.8 MPa, depending on the combination of surface preparation and adhesives. Also, the choice of adhesive influences the performance of the manufactured drills. The maximum number of drilled holes scatters between 15 and 67 holes before breakage. A linear relation within the analysis of axial forces and factors of the covered area is observed. For further studies, the surface preparation needs to be improved, the glass transition temperature must be maximized, and the drill geometry must be optimized. Furthermore, a cooling concept for the cutting tools needs to be investigated.

Electric-field activating on-surface tailored (OST) coarse polyester fibers for efficient airborne particle removal: Interfacial morphologies and electrical response

Fibrous filtration is the most-used method for indoor particulate matter (PM) removal. The filtration performances are governed by filters’ intrinsic geometries, displaying the tradeoff among efficiency, resistance, and lifespan. In this contribution, we provided a chemically-synthesized strategy to improve the performance of coarse filters while preserving their ultralow initial resistances. After elucidating PM-fiber interactions, the fibrous interfacial nano-morphologies were tailored by different dielectric coatings based on commercially-available substrates. Activated by fiber polarizing, the tuned morphologies with ∼100 nm surface roughness and enhanced electrostatic potential are considered the drivers of boosted filtration efficiency. In practice, a 10-mm-thick polydopamine (PDA)-MnOx coated polyester filter possessed an improved efficiency of 97.7 % for 300–500 nm particles, and the ultralow resistance of 11.2 Pa at 0.5 m/s filtration velocity. The strong surface adhesion facilitated long-term efficiency of 95.6 % in a continuous 25-day period without any decay. The nanoscale coatings, which were just one-thousandth in thickness of the fiber gaps, enabled more than 50-fold improvement of the filters’ quality factor. We further established dimensionless factors to evaluate the cost-performance effect. We expect the strategy and techniques can pave the way to better understand electrostatic air filtration, being competitive candidates in the air filtration community.

Adhesion pilus retraction powers twitching motility in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius

Type IV pili are filamentous appendages found in most bacteria and archaea, where they can support functions such as surface adhesion, DNA uptake, aggregation, and motility. In most bacteria, PilT-family ATPases disassemble adhesion pili, causing them to rapidly retract and produce twitching motility, important for surface colonization. As archaea do not possess PilT homologs, it was thought that archaeal pili cannot retract and that archaea do not exhibit twitching motility. Here, we use live-cell imaging, automated cell tracking, fluorescence imaging, and genetic manipulation to show that the hyperthermophilic archaeon Sulfolobus acidocaldarius exhibits twitching motility, driven by retractable adhesion (Aap) pili, under physiologically relevant conditions (75 °C, pH 2). Aap pili are thus capable of retraction in the absence of a PilT homolog, suggesting that the ancestral type IV pili in the last universal common ancestor (LUCA) were capable of retraction.

Cell adhesion molecule CD44 is dispensable for reactive astrocyte activation during prion disease

Prion diseases are fatal, infectious, neurodegenerative disorders resulting from accumulation of misfolded cellular prion protein in the brain. Early pathological changes during CNS prion disease also include reactive astrocyte activation with increased CD44 expression, microgliosis, as well as loss of dendritic spines and synapses. CD44 is a multifunctional cell surface adhesion and signalling molecule which is considered to play roles in astrocyte morphology and the maintenance of dendritic spine integrity and synaptic plasticity. However, the role of CD44 in prion disease was unknown. Here we used mice deficient in CD44 to determine the role of CD44 during prion disease. We show that CD44-deficient mice displayed no difference in their response to CNS prion infection when compared to wild type mice. Furthermore, the reactive astrocyte activation and microgliosis that accompanies CNS prion infection was unimpaired in the absence of CD44. Together, our data show that although CD44 expression is upregulated in reactive astrocytes during CNS prion disease, it is dispensable for astrocyte and microglial activation and the development of prion neuropathogenesis.

Comparative study of surface preparation for paint adhesion on CF-PEKK composites: Plasma, Chemical, and Flame treatment

Polyether ketone-ketone (PEKK) is a high-performance thermoplastic, and its fiber composites have various engineering applications. Fiber reinforced composites in long-term applications require a protective layer of coatings to avoid environmental degradation. In this work, the surface energy of carbon fiber-PEKK composites was modified by flame, chemical, and plasma treatments to improve paint adhesion. A detailed study was carried out to understand the physical, morphological, and chemical surface characteristics due to different surface treatments. We found through XPS that highly active surface functionalities (− C-Ȯ/−C = Ȯ) due to oxidation at the ketone/ether groups were for surface adhesion. In support of the enhanced surface functionality, we have also evaluated the wetting behavior of paint with respect to different treated surfaces using drop shape analyzer and found that the polar component of the surface free energy increased for all treatments compared to pristine. Surface crystallinity after surface treatment was assessed using XRD; flame and plasma showed reduced surface crystallinities (25.92 % and 38.18 %, respectively), compared to pristine (46.78 %), and yielded a good adherend for coatings. Surfaces with good wettability and functionalities show good adhesion between the paint and PEKK composites as found through adhesion tests following ASTM D3359. The findings demonstrate that plasma surface treatment outperformed flame and chemical treatments in terms of adhesion of the paint to the surface.

Anti-ageing and rheological performance of bitumen modified with polyaniline nanofibres

Asphalt pavements represent the great majority of roads worldwide. However, the bitumen physico-chemical properties change due to ageing processes during both the production and service life of the asphalt mixes. In recent years, a completely new trend has been observed in the road engineering industry oriented to extending the service life of roads, bringing direct economic, ecological and social benefits. This work presents the possibility of inhibiting ageing of road bitumen by using polyaniline nanofibers (PANI_NFs) as well as enhancing its rheological properties examined with Linear Amplitude Sweep (LAS) as well as Multiple Stress Creep and Recovery (MSCR) tests. These results show that the application of PANI_NFs maintains the primary penetration value of bitumen short-term and long-term aged road bitumen at 88 % and 52 %, respectively. Moreover, bitumen modification with PANI_NFs reduces the softening temperature increase by 33 % and 24 %, respectively, and the viscosity increase of road asphalt by 50 % and 25 %, respectively. PANI_NFs also stabilised the bitumen surface adhesion force, determined with AFM, consequently confirming the enhanced resistance to ageing processes. In addition, changes in the carbonyl index, sulphoxide index and aromaticity of bitumen demonstrated that PANI_NFs inhibit oxidation and aromatisation reactions of its components. The tests indicated a 4-fold decrease in the degree of bitumen component oxidation after short-term ageing and a 33 % decrease after long-term ageing. FTIR analysis also showed a reduction in the degree of bitumen component aromatisation process of 79 % and 76 % for short-term and long-term ageing, respectively. The LAS and MSCR test results also showed that PANI_NFs had a beneficial impact on the resistance of the bitumen to fatigue and rutting. Overall, the study results suggest that PANI_NFs consitute a promising modifier for bitumen in terms of ageing inhibition and performance characteristics.

A new discrete element method for small adhesive non-spherical particles

Small adhesive particles with non-spherical shapes are common in natural and industrial processes. However, it is challenging to accurately simulate the dynamic behaviors of non-spherical particulate systems, not only due to the complex local geometry at the contact region but also to the non-linear coupling between the van der Waals adhesion and the elastic deformation. In this paper, we develop a discrete element method (DEM) framework for small non-spherical particles. Particularly, we present a simple algorithm for solving the implicit equations of the dimensionless generalized JKR model, to accurately resolve the local surface geometry at the contact region and the strong coupling between the elastic deformation and the surface adhesion. New explicit expressions are established for the pull-off force, overlap and minor-to-major ratio of the contact ellipse, to naturally capture the pull-off point upon collision. Then the actual contact parameters including the contact radius, curvatures and overlap are used to modify the interparticle interactions, including the normal elastic-adhesive-dissipative force and sliding-rolling-twisting resistances. Finally, the framework is quantitatively validated by four random packing problems, including adhesive spheres, ellipsoids, spherocylinders and dimers, and the representative non-adhesive ellipsoids. Both the microscopic and macroscopic properties of packings agree well with a large number of classic theories, experiments and simulations, showing the satisfactory accuracy and effectiveness. The newly developed adhesive DEM framework has the full capability to dynamically simulate the collision-related behaviors of complex shaped particles in the presence of van der Waals adhesion and frictional forces.

Identification of CD44 as a key mediator of cell traction force generation in hyaluronic acid-rich extracellular matrices.

Mechanical properties of the extracellular matrix (ECM) critically regulate a number of important cell functions including growth, differentiation and migration. Type I collagen and glycosaminoglycans (GAGs) are two primary components of ECMs that contribute to mammalian tissue mechanics, with the collagen fiber network sustaining tension, and GAGs withstanding compression. The architecture and stiffness of the collagen network are known to be important for cell-ECM mechanical interactions via integrin cell surface adhesion receptors. In contrast, studies of GAGs in modulating cell-ECM interactions are limited. Here, we present experimental studies on the roles of hyaluronic acid (HA, an unsulfated GAG) in single tumor cell traction force generation using a recently developed 3D cell traction force microscopy method. Our work reveals that CD44, a cell surface adhesion receptor to HA, is engaged in cell traction force generation in conjunction with β1-integrin. We find that HA significantly modifies the architecture and mechanics of the collagen fiber network, decreasing tumor cells' propensity to remodel the collagen network, attenuating traction force generation, transmission distance, and tumor invasion. Our findings point to a novel role for CD44 in traction force generation, which can be a potential therapeutic target for diseases involving HA rich ECMs such as breast cancer and glioblastoma.

Production of Cu/Diamond Composite Coatings and Their Selected Properties.

This article presents Cu/diamond composite coatings produced by electrochemical reduction on steel substrates and a comparison of these coatings with a copper coating without diamond nanoparticles (<10 nm). Deposition was carried out using multicomponent electrolyte solutions at a current density of 3 A/dm2 and magnetic stirring speed of 100 rpm. Composite coatings were deposited from baths with different diamond concentrations (4, 6, 8, 10 g/dm3). This study presents the surface morphology and structure of the produced coatings. The surface roughness, coating thickness (XRF), mechanical properties (DSI), and adhesion of coatings to substrates (scratch tests) were also characterized. The coatings were also tested to assess their solderability, including their spreadability, wettability of the solder, durability of solder-coating bonds, and a microstructure study.