WO2008091386A2

WO2008091386A2 - Biomimetic modular adhesive complex: material, methods and applications therefore

Info

Publication number: WO2008091386A2
Application number: PCT/US2007/075298
Authority: WO
Inventors: Phillip B. Messersmith; Haeshin Lee; Bruce P. Lee
Original assignee: Northwestern University
Priority date: 2006-08-04
Filing date: 2007-08-06
Publication date: 2008-07-31
Also published as: WO2008091386A3; US8563117B2; US20080169059A1

Abstract

Nano-scale or micro-scale adhesive structures comprising an array of nano-fabricated, pillars, the pillars having coated upon, or having disposed on a working surface thereof, a protein- mimetic, marine-adhesive coating. Methods of fabricating the nano-scale pillars, synthesis of the protein-mimetic coating or wet adhesive and application of the adhesive to the pillars are described.

Description

Biomimetic Modular Adhesive Complex: Materials, Methods and Applications Therefore

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application claims the priority date of U.S. Provisional Application 60/835,615 filed 08-04-06, entitled "Geckel-Mimetic Nanostructures, Materials, Methods and Applications

Therefore." The entirety of the aforementioned 60/835,615 Provisional Patent Application, including all references and attachments incorporated by reference therein, are incorporated by reference into this application.

BACKGROUND OF THE INVENTION The ability of the gecko to climb vertical surfaces, seemingly in defiance of gravity, has been a source of amazement and scientific inquiry for many years. Since about 1965, it has been known that the adhesive strategy of the gecko relies on foot pads composed of specialized keratinous foot-hairs called setae, which are subdivided into terminal spatulae of dimensions approximately 200 nm.¹ Contact between the gecko foot and an opposing surface generates adhesive forces that are sufficient to allow the gecko to cling onto vertical and even inverted surfaces. Although strong, the adhesion is temporary, permitting rapid detachment and reattachment of the gecko foot during locomotion. Researchers have attempted to capture the unique properties of gecko adhesive in synthetic mimics with nanoscale surface features reminiscent of setae,^2"7 however maintenance of adhesive performance over many cycles has been elusive,²' ⁸ and gecko adhesion is dramatically diminished upon full immersion in water.⁹' ¹⁰

Many organisms found in water use chemical bioglues for various purposes such as adhesion, defense mechanism, and symbiosis. Those glues are found in barnacles, mussels, and microbes colonized on inorganic surfaces at an early stage of biofouling. Among these, the adhesion mechanism of mussels has been well characterized; 3,4-dihydroxy-L-phenylalanine, DOPA, is found in adhesive pads and has been considered as a key component responsible for the strong holdfast under water. The present invention provides a unique "mimetic" functional combination of the two unique natural adhesion mechanisms inspired by geckos and mussels. BRIEF SUMMARY OF THE INVENTION

Briefly, in one aspect, the present invention is a multi-component, modular adhesive complex or apparatus comprising 1) an adhesive coating, 2) a go-between nanostructural or microstructural array, and 3) a bottom-most supporting material, or means, or substrate. This modular adhesive complex is capable of achieving renewable adhesive contacts or retaining adhesion characteristics in a high humidity or a wet, as well as an ambient, dry or arid environment. Single adhesive or permanent use, i.e., a single mating or application of adhesive working surfaces, is also contemplated.

The adhesive coating is to be broadly interpreted as a coating or film of sufficient thickness and area so as to provide a complete or partial coverage of the nanostructural array, especially a working surface or exposed surface, for purposes of creating an adhesive or adhesion relationship. The coating provides interfacial binding ability to the modular adhesive complex through various adhesive mechanisms i.e. chemical (ionic, hydrogen, or colvalent) bond formation, van der Waals forces, capillary forces, electrostatic interactions, diffusion, mechanical interlocking, etc. The coating or film is applied to, and binds strongly to, the underlying nanostructural array, transferring interfacial stress to bulk material consisting of nanostructural array and the supporting material. In a preferred embodiment, the coating is comprised of DHPD (dihydroxyphenyl derivative) or DHPP i.e. a polymer comprising a variable number, distribution, or concentration of DHPD. DHPD and DHPP are further defined below.

In a further aspect the present invention is a microstructural array or microarray having a coating disposed or applied upon an exposed or working surface thereof. "Working surface" for purposes of this invention is any surface, or any portion thereof directly or indirectly involved in creating adhesion or an adhesive structure, array, or relationship. The layer or coating is preferably DHPD (dihydroxyphenyl derivative) or DHPP i.e., DHPD-containing polymer. The DHPD, which is to be broadly interpreted, is of sufficient thickness and area so as to provide the array with an optionally renewable adhesive or adhesion character in a high humidity or wet environment. Single adhesive or permanent use, i.e., a single mating or application of array working surfaces, is also contemplated. In one aspect, the microstructures (or nanopillars as they may be called) of the array are substantially columnar, setae-like, spatulae-like or elongate and upstand, upend, protrude, or extend from a substrate and have an exposed area, working area or workpiece, end or surface. The exposed area, end, or surface of the columnar microstructure has a DHPD coating or layer disposed thereon, the DHPD layer having sufficient thickness and area in one embodiment, so that the microstructure is adhesive in a high humidity or wet environment. In a preferred practice the adhesive character of the DHPD layer is renewable as discussed below. In one embodiment of this aspect of the invention the columnar microstructures have a working surface, the coating DHPD, e.g., being disposed thereon. The columnar, setae-like structure comprises a structural polymeric material.

The nanostructural array is to be broadly interpreted as having an arrangement, collection, parade, of ordered or disordered, random, sequential, or hierarchical structural or geometric features, textures, projections or surfaces with dimensions in the nanoscale (e.g., 0.1 - 100 nm) and microscale (e.g., 100 nm - 100 μm) range or the combination thereof. Possible geometric features include but are not limited to fibers, columns, pillars, loops, tubes, cones, blocks, cubes, hemispheres, spheres, walls, grids, plains, holes, or indentations of regular, irregular, or uneven shape of the combination thereof. The nanostructurual arrays are anchored, attached, or glued to, or protrude or extend from the bottommost supporting material (e.g., substrate) and provide an exposed area, working area or workpiece, end or surface with the adhesive coating being applied or disposed thereon. The nanostructural array promotes interfacial contact through substantially increased surface area or the ability to make contact with defects of nanoscale and macroscale sizes on the adherent surface. The possibility of mechanical interlocking between the nanostructural array and a second array or other surface, especially the microstructure thereof, is also included within the present invention. The nanostructural array can be comprised of an organic or inorganic polymeric material, carbon in the form of rods or tubes or spheres, or inorganic solids such as metals, metal oxides and ceramics.

The supporting material or substrate is to be broadly interpreted as a macroscaled base, bulk, or backing material having sufficient cohesive properties, miscibility with the nanostructural array, and elasticity. It is preferable that the supporting material exhibits sufficient cohesive or bulk mechanical properties for holding the whole adhesive complex intact in the presence of applied stresses i.e. peeling or detaching forces. Sufficient miscibility with or allowing attachment or anchoring of nanostructural array is necessary for transferring interfacial stresses. Elasticity is needed for the deformation of the adhesive complex. Examples of material support include but are not limited to elastomers (i.e. silicone-, acrylate-, urethane-, polyester-, polyether-based polymers or natural and synthetic rubber), but could be semi-crystalline or noncrystalline polymer with sufficient flexibility as described. It is preferable that the supporting material is constructed from the same or dissimilar material as the nanostructural array. Another aspect of this invention comprises a method of adhering surfaces to one another, preferably in a high humidity, wet, ambient or dry environment. The method comprises the following steps:

(1) providing a modular adhesive complex comprising a bulk material constructed or disposed thereupon an array of nanoscaled features having an exposed area, end, or surface on which there is a further disposed superficial layer of adhesive coating preferably of DHPP;

(2) adhering the adhesive complex to a second surface with the DHPD layer being applied there between;

(3) detaching or removing the complex from the second surface, the DHPD layer adhering to nanostructural array which is attached the bottommost bulk material; and

(4) adhering the adhesive complex to the second surface (or to a third surface).

It is in the above sense that the present invention provides a "renewable" or "repositionable" adhesive or adhesion quality. A modular adhesive complex of this invention can be adhered to a second surface, removed from the surface, and re-adhered or reattached to the same or a different second surface, during which, each component remains intact. The renewable adhesion hereby permitted may be renewed or re-created two or more times, preferably multiple times, and most preferably hundreds to thousands of times with either the same or a new second surface. In the parlance of conventional adhesion or adhesive products, arrays of this invention could be described as "repositionable." A further aspect of this invention comprises a method of adhering surfaces to one another, preferably in a high humidity or wet environment. The method is comprised of the following steps:

(1) providing an array of columnar microstructures pending upstanding or projecting from a substrate or support, the microstructures having an exposed area, end, or surface or working surface on which there is disposed a layer or coating e.g., of DHPD;

(2) adhering the array to a second surface with the DHPD layer being disposed therebetween;

(3) detaching or removing the array from the second surface, the DHPD layer adhering to the columnar microstructure and to itself; and optionally (4) adhering the array to the second surface (or to a third surface in second or subsequent repositionings). In one version, the second surface is an array of this invention.

It is in the above sense that the present invention provides a "renewable" adhesive or adhesion quality. An array of this invention can be adhered to a second surface, removed from the surface, and re-adhered or reattached to the same or a different second surface. The renewable adhesion hereby permitted may be renewed or re-created two or more times, preferably multiple times, and most preferably hundreds to thousands of times with either the same or a new second surface. In the parlance of conventional adhesion or adhesive products, arrays of this invention could be described as "repositionable." Adhesive coating (or plaque) containing no DHPD is also contemplated, see e.g., Fig. 8 and associated discussion. Thus the term coating as used herein is to be broadly constructed to include adhesive coating, resistive coating (e.g., resistive to cellular adhesion), as well as protective coating.

It is noted that the present invention provides adhesion in high humidity, "highly humid" or "wet" environments. "Highly humid" or "high humidity" environments herein means environments having ambient relative humidity of at least 50%, preferably 70%, and most preferably 80% up to what is considered "wet," "saturated" or even "super-saturated" environments. Adhesion under dry environment is also contemplated herein. "DHPD" and its chemistry are discussed in greater detail below. "DHPP" as used herein is to be understood to mean polymeric, crosslinked, or network structures containing multiple i.e., two or more DHPD structures, units or moieties. DHPP has the following preferred characteristics:

• Weight average molecular weight between 10,000 and 5,000,000 Da but preferably between 100,000 and 1,000,000 Da. • The abundance of DHPD moieties in DHPP can vary between 0 to 100 weight percent, but is preferably between 10 and 50 weight percent.

• DHPP contains between one and 10 substituents but is preferably between one and 4.

• DHPP can consist of single, di-, tri-, and multi-block of singular, random, sequential, or ordered substituents.

• DHPP is preferably water insoluble, but can be water swellable.

• DHPP can have linear, brush, branched, hyper-branched, crosslinked, network, gel, or hydrogel architecture.

• DHPP is preferably acrylate-based, but can consist of ether, urethane, urea, amide, carbonate, or ester linkages, or any combination thereof.

• DHPP is preferably hydrophilic or amphiphilic but can be hydrophobic. • DHPP can include chemically linked, crosslinked, or polymerized forms of multiple DHPP and/or DHPD.

The terms "nanostructure" and "microstructure" are used herein. Nanostructures are features or textures having dimensions of nanoscale (e.g., 0.1 - lOOnm) in size. Microstructures are features or textures of dimensions of microscale (e.g., lOOnm - 100mm) in size. Unless otherwise indicated by the context, no criticality should be accorded to the use of one term versus the other.

The terms "array", "nanoarray", and "microarray" are used herein. These terms are to be broadly interpreted to mean geometric features, textures, or surfaces having likeness of fibers, columns, pillars, loops, tubes, cones, blocks, cubes, hemispheres, spheres, walls, grids, plains, holes, or indentations of regular, irregular, or uneven shape, support, substrate, or the combination thereof, projecting from or attached, anchored, or glue to a support, member or backing member.

The preferred coating layer of this invention comprises DHPD of formula (I) wherein

R₁ and R₂ may be the same or different and are independently selected from the group consisting of hydrogen, saturated and unsaturated, branched and unbranched, substituted and unsubstituted C_1-4 hydrocarbon; x is a value between zero and four.

P is separately and independently selected from the group consisting of -NH₂, -COOH, -OH, -SH,

wherein R₁, R₂, and x are defined above, a single bond, halogen, A₁. .CH₂

N

H wherein A₁ is selected from the group consisting of H, C, a single bond, a protecting group, substantially alkyl, substantially poly(alkyleneoxide),

O

H₂C ^:

R₃ wherein R₃ is H or C_1-6 lower alkyl; CH₂, C C-poly(alkylene oxide)

wherein R₃ is defined above;

wherein A₂ is selected from the group of -OH, C, -NH-, in addition to the definition of Ai;

wherein Ai and A₂ are defined above;

wherein n ranges between 1 and about 3 and Ai and A₂ are defined above; spect the poly(alkylene oxide) has the structure

wherein R₄ and R₅ are separately and independently H, or CH₃ and m has a value in the range between 1 and about 250, A₃ is -NH₂ -COOH, —OH, and -SH, — H or a protecting group.

In a very preferred form, DHPD is

R₁, R₂, and P being defined as above.

In a further preferred form DHPD is of the structure:

wherein Ai is of the structure

H₂C:

CH₃

These dihydroxyphenyl derivative (DHPD) adhesives function well in an aqueous or a high humidity environment. To form the polymeric composition, a DHPD moiety which generally provides adhesive functionality is coupled to a polymer which provides the desired adhesive or surface effect. These components are described in detail in U.S. Application S.N. 11/068,298 at paragraphs [0054] and [0057] - [0070] as numbered in the application as filed. That disclosure is specifically incorporated by reference herein in its entirety.

BRIEF DESCRIPTION OF THE FIGURES Figure 1

Rational design and fabrication of wet/dry hybrid nanoadhesive. E-beam lithography (eBL) was used to create an array of holes in a polymethyl methacrylate (PMMA) thin film supported on Silicon (Si) (PMMA/Si master). Casting of poly(dimethylsiloxane) (PDMS) onto the master followed by curing and lift-off resulted in gecko-mimetic nanopillar arrays. Finally, a mussel adhesive protein mimetic polymer is coated onto the fabricated nanopillars. The topmost organic layer contains catechols, a key component of wet adhesive proteins found in mussel holdfasts.

Figure 2

Fabricated gecko and geckel adhesives. A. Scanning electron microscopy (SEM) image of eBL fabricated gecko nanopillar microarray, nanoarray, or simply or array (scale = 10 μm). B. Atomic force microscopy (AFM) line scan of the gecko nanopillars. The height and diameter of the pillars (sometimes referred to as projections, nanopillars, microscale projections, supports, or microstructures) used in this study were 600nm and 400nm, respectively. The apparent widening of the pillars near the base is believed to be an artifact arising from the pyramidal shape of the AFM tip used for imaging. C. Chemical structure of the mussel adhesive protein mimetic polymer, p(DMA-co-MEA), which is applied to the surface of the gecko nanopillars. D. SEM image of geckel adhesive after coating the nanopillar array with p(DMA-co-MEA). The coating has little effect on the pillar geometry (scale = 10 μm).

Figure 3 AFM (atomic force microscopy) method for adhesion measurement and imaging of contact area at the single pillar level. A. Experimental set-up for measuring adhesion. A tipless AFM cantilever is brought into contact with the nanopillar array (3A) and then retracted while the contact area is imaged from an objective located the underneath adhesive film. B. The number of pillars contacting the cantilever was controlled through the distance, d, between pillars (d = 1, 2, and 3 μm) and the angle, θ, between the cantilever and the axis of the pillar array (θ = either 0° or 45°). The inset shows an SEM image of a cantilever contacting a geckel pillar array to yield a five pillar contact condition (d = 1 μm and θ = 45°). C and D. Optical microscope images showing contact between AFM tip and pillar array. One pillar contact was achieved when d = 3 μm and θ = 45° (C), and six pillars were in contact when d = 1 μm and θ = 0°.

Figure 4

Force-distance curves and adhesion strength of geckel adhesive. All data are for contact with a Si₃N₄ cantilever. A, B, Retraction force-distance curves for uncoated (A) and p(DMA- co-MEA) coated (B) pillars in water. Force-distance curves were obtained for contact with one (red), two (blue), three (green), four (pink), and five (black) pillars. C, Retraction force- distance curve for contact between cantilever and flat p(DMA-co-MEA)-coated PDMS (contact area = 55.3 μm²). D, Mean separation force values versus number of pillars for gecko (triangle) and geckel (circle) in water (red) and air (black) (n > 60, for each data point). E, Adhesion force per pillar, obtained from the slopes of the regression lines shown in D. Wet adhesion was increased 15 -fold in water. Error bars represent standard deviation.

Figure 5. Long-term performance of geckel adhesive. Multiple cycles of attachment and detachment of geckel adhesive were performed in water (red) and air (black). Adhesion strength decreased by only 15% in water (red) and 2% in air (black) after 1100 successive cycles of contact and separation (two-pillar contact).

Figure 6.

X-ray photoelectron spectroscopy (XPS) of gecko and geckel adhesives. A. XPS was used to probe p(DMA)-co-MEA) coating thickness in a semi-quantitative way. Spincoating p(DMA-co-MEA) onto PDMS resulted in no silicon signals (2s, 153 eV and 2p 103 eV) indicating that the coating thickness is more than the x-ray penetration depth, typically around 20 nm. Dipcoating resulted in both silicon and nitrogen signals, thus indicating that the coated polymer thickness is <20nm. 6(B) Surface atomic composition of unmodified and modified PDMS substrates from the XPS data shown in 6(A). Dipcoated samples showed both silicon and nitrogen composition.

Figure 7.

Substrate-dependent wet adhesion of geckel. AFM force measurements revealed changes in wet adhesion of the geckel adhesive on different substrates (Si₃N₄, TiO_x, and Au.) 86.3 ± 5 nN for Si₃N₄ (the data from Figure 4D, Geckel- Water), 130.7 ± 14.3 nN for TiO_x (n = 50), and 74.3 ± 4.1 nN forAu (n = 65). Figure 8.

Wet adhesion of p(MEA)-coated gecko. (A) Adhesion of a p(MEA)-coated pillar array to Si₃N₄ in water (5 pillar contact, 400nm pillar diameter, d=l μm and θ=45°). A significant decay in adhesion was observed with successive contacts. The force traces shown in the figure represent every 10^th cycle: 1(127.9 nN, black); 11(93.8 nN, blue), 21(86.6 nN, green), 31(82.7 nN, pink), 41(73.1 nN, orange), and 51(59.3 nN, red). (B) Carbon Is high resolution XPS spectra of bare PDMS (panel a), p(MEA)-coated PDMS (panel b), and p(MEA)-PDMS after incubation in water at room temperature for 18 hrs (panel c).

DETAILED DESCRIPTION OF THE INVENTION

Described herein is a new class of hybrid biologically-inspired adhesives comprising an array of nanofabricated polymer columnar pillars coated with a thin layer of a synthetic polymer that mimics the wet adhesive proteins found in mussel holdfasts. Wet adhesion of the nanostructured polymer pillar arrays of this invention increased nearly 15-fold when coated with mussel adhesive protein mimetic polymer. The adhesive can function like a sticky note (a repositionable partially adherent note structure), maintaining its adhesive performance for over

1000 contact cycles in both dry and wet environments. This hybrid adhesive, which uniquely combines the salient design elements of both gecko and mussel adhesives, provides a useful reversible attachment means for a variety of surfaces in many environments. "Single use" or single attachments also are contemplated.

The adhesive forces of the gecko have been observed to be on the order of 40 μN or more per seta¹¹' ¹² and 10 nN per spatula.¹³ Gecko adhesion has been explained as arising from weak secondary bond forces such as van der Waals.¹¹ However, adhesion of a single spatulae varies as a function of humidity and is dramatically reduced under water,⁹' ¹⁰ suggesting some contribution from capillary forces. Contact mechanics arguments have been invoked to explain the subdivision of the setal contact surface into multiple independent nanosized spatulae, giving rise to enhancement of the mechanical behavior.¹⁴ For the idealized case of a circular contact area, theory suggests that the adhesion strength scales as Vn , where n is the number of independent contacts into which the area is subdivided. The contact splitting theory qualitatively explains the scaling of dry adhesive systems employed by some amphibians and insects, and provides guidance for development and optimization of synthetic gecko mimics.⁶' ^{15 16} Synthetic gecko adhesives that exhibit dry adhesion have been fabricated from polymers^2" ⁴ as well as multiwalled carbon nanotubes.⁵ However, maintenance of adhesion during repetitive contacts has only been demonstrated for a few contact cycles,²' ⁸ and none have been shown to function under water or in high humidity environments.

A celebrated biological model for wet adhesion is the mussel, which is well known for its ability to cling to wet surfaces.¹⁷' ¹⁸ Mussels secrete specialized adhesive proteins containing a high content of the catecholic amino acid 3,4-dihydroxy-L-phenylalanine (DOPA).^19"21 Both natural and synthetic adhesives containing DOPA and its derivatives have demonstrated strong interfacial adhesion strength.^22"25 Using single molecule measurements in aqueous media, we recently demonstrated that DOPA formed extraordinarily strong yet reversible bonds with surfaces.²⁶ In fact, the force necessary to dissociate DOPA from an oxide surface (-800 pN) was the highest ever observed for a reversible interaction between a small molecule and a surface.²⁶ It was theorized that the incorporation of mussel adhesive protein mimetic polymer onto a gecko-mimetic nanoadhesive structure would yield strong yet reversible wet/dry adhesion - a property that existing materials do not exhibit. Arrays of gecko foot-mimetic nanoscale pillars coated with a thin MAP-mimetic polymer film are shown in Figure 1. Designs of both the pillar array and the coating polymer were undertaken in view of current knowledge of the respective biological systems. For the pillar array, primary design criteria include the dimensions of the pillars and their spacing, as well as the stiffness of the pillar material.¹⁵' ¹⁶ For flexibility in adapting to rough surfaces, both the supporting substrate and the pillar material were fabricated from poly(dimethylsiloxane) (PDMS) elastomer, which is a well-known organic material with a long history of use in microfabrication.²⁷ Arrays of PDMS pillars 200, 400, and 600 nm in diameter, 1-3 μm center-to-center distance, and 600-700 nm in height were successfully fabricated using e-beam lithography (eBL) (see Figure 1). The pillar arrays are supported on a continuous film of PDMS 2-3 mm in thickness, with each PDMS pillar representing a single spatula found at the surface of a gecko foot (Figure 2A, B). Pillar arrays of 400nm diameter and 600nm height were tested for adhesion.

Analysis of mussel adhesive protein compositions gave insight into a rational design for a mussel-mimetic polymer. First, the synthetic polymer should have a high catechol content since DOPA accounts for as much as 27% of amino acids in the adhesive proteins found at the interface between mussel byssal pads and their substrate.²¹ Second, long-lasting waterproof adhesion requires polymers with low water solubility to prevent their loss into the aqueous medium. Poly(dopaminemethacrylamide-co-methoxyethylacrylate) (p(DMA-co-MEA),

(Figure 2C) was synthesized using free-radical polymerization where the adhesive monomer, DMA, accounts for 17% of this copolymer by weight (¹H NMR). p(DMA-co-MEA) has a high molecular weight and is insoluble in water. p(DMA-co-MEA) was applied to the PDMS pillar array by dip coating in an ethanol solution of p(DMA-co-ME A). X-ray photoelectron spectroscopy (XPS) analysis of the coated substrate indicated a thin coating (<20 nm) as demonstrated by the presence of both silicon (103 eV, Si 2p) from the PDMS and nitrogen (399 eV, N Is) from the p(DMA-co-MEA) (Figure 6). Spin-coating p(DMA-co-MEA) onto PDMS resulted in no silicon signals (2s, 153 eV and 2p 103 eV) indicating that the coating thickness is more than the x-ray penetration depth, typically around 20 nm (Figure 6A). Dip-coating resulted in both silicon and nitrogen signals, thus indicating that the coated polymer thickness is <20 nm. The surface atomic compositions of unmodified and dip-coated PDMS substrates are shown in Figure 6B. Dip- coated sample showed both silicon and nitrogen compositions. A thin coating was desired for minimizing the change in pillar dimensions during coating, which was confirmed by scanning electron microscopy after coating with p(DMA-co-MEA) (Figure 2D). We refer to the resulting flexible organic nanoadhesive as 'geckeP, reflecting inspiration from both the gecko and the mussel.

The performance of geckel adhesive was evaluated using an atomic force microscopy (AFM) system fully integrated with optical microscopy, which permitted simultaneous measurement of the adhesive contact force along with clear visualization of the nanoscale contact area down to the single pillar level. In a typical adhesion experiment (Figure 3), the AFM piezo was used to bring a tipless cantilever (Si₃N₄) into contact with the geckel pillar array, and upon retraction the force necessary to separate the cantilever from the pillar array was measured. Furthermore, independently changing the spacing (d) between pillars (d = 1, 2, and 3 μm; Figure 3A) and the angle of orientation (θ) between the pillar array and the cantilever axis (Figure 3B) allowed us to control the number of pillars contacting the cantilever precisely from one to six. For example, a geckel adhesive with d = 3 μm and θ = 45° resulted in a single pillar contact (Figure 3C), whereas d = 1 μm and θ = 0° resulted in six pillars interacting with the cantilever simultaneously (Figure 3D, movie 1).

Adhesion experiments were performed both in air and under water for uncoated (hereafter 'gecko') and p(DMA-co-MEA) coated ('geckel') pillar arrays (Figure 4). Pillar- resolved (i.e. area-defined) force measurements showed strong adhesive forces when the cantilever was pulled away from the pillar surface. Figures 4A and 4B show typical force- distance (F-D) curves, with each curve representing a specific number (1-6) of 400nm diameter pillars interacting with the Si₃N₄ cantilever surface. The pull-off force was determined from each F-D curve, and mean values from multiple experiments were plotted in Figure 4D as a function of the number of contacting pillars. The observed linear increase in force with pillar number indicates constructive force accumulation, i.e. simultaneous detachment of individual pillars from the cantilever. The adhesive force per pillar (nN/pillar) was calculated from the individual slopes (Figure 4E): 39.8 ± 2 (gecko in air), 5.9 ± 0.2 (gecko in water), 120 ± 6 (geckel in air), and 86.3 ± 5 (geckel in water).

Although the addition of p(DMA-co-MEA) coating on the pillars significantly increased dry adhesion, the enhancement of wet adhesion was particularly dramatic, as the wet adhesive force per pillar increased nearly 15 times (5.9 -> 86.3 nN/pillar, Si₃N₄) when coated with p(DMA-co-MEA). The geckel wet-adhesion strength was also high when tested against other surfaces: titanium oxide (130.7 ± 14.3 nN/pillar) and gold (74.3 ± 4.1 nN/pillar) (Figure 7). The versatility of geckel is not surprising given recent single molecule force experiments showing the ability of DOPA to interact strongly with both organic and inorganic surfaces.²⁶ These interactions can take many forms, including metal coordination bonds, pi electron interactions, and covalent bonds. The lower adhesion strength of geckel on gold is in qualitative agreement with our earlier single molecule pull-off and polymer adsorption studies that indicated DOPA interacts less strongly with gold than with titanium oxide.²⁶' ²⁹

The ability of the bond between DOPA and a metal oxide surface to rupture upon pulling, and then re-form when brought back into contact with the surface,²⁶ is an important aspect of this invention. Repetitive AFM measurements showed that geckel adhesive 's wet- and dry-adhesion power was only slightly diminished during many cycles of adhesion, maintaining 85% in wet (red) and 98% in dry (black) conditions after 1100 contact cycles (Figure 5). To our knowledge no other gecko-mimetic adhesive has demonstrated efficacy for more than a few contact cycles,²' ⁸ and none have been shown to work under water. This surprising and unexpected advantage of the present invention suggests many possible applications. Control experiments involving pillar arrays coated with catechol-free polymer, p(MEA), showed lower adhesion strength (26 nN/pillar for the first contact cycle) as well as rapid decay in the adhesion performance under cyclic testing occurred over 5 adhesive contacts (Figure 8A). From XPS spectra shown in Figure 8B, carbonyl peak for the p(MEA)-coated surface disappeared over 18 hours of incubation suggesting the detachment of the polymer. Although repeatable adhesion can be achieved underwater using a DHPD-free polymer, the adhesive performance is significantly reduced emphasizing the importance of the mussel- mimetic catechol groups in enhancing wet adhesion as well as anchoring the p(DMA-co-MEA) polymer to the pillar array. At the same time, it appears that the nanostructured surface is essential to the observed geckel adhesive behavior. Force measurements on flat substrates coated with p(DMA-co-MEA) indicated a complex peeling behavior initiating at low adhesive strength (Figure 4C), which is in distinct contrast to the linear force accumulation behavior exhibited by the geckel adhesive (Figure 4D).

The geckel nanoadhesive was shown to be highly effective at adhering reversibly to surfaces under water, and with functional performance resembling that of a sticky note. Although we must be cautious in extrapolating our results to larger areas because of the challenges associated with maintaining equal load sharing among a large number of pillars, in its current form (400 nm pillar diameter and 1 μm spacing) a 1 cm² surface area of geckel adhesive would transmit 9 N of force under water (90 kPa). It is interesting to note that this value is similar to estimates for the strength of gecko dry adhesion,⁹' ^{u> I2} suggesting that under wet conditions our hybrid geckel adhesive may perform as well as gecko adhesives do under dry conditions. Further refinement of the pillar geometry and spacing, the pillar material, and mussel mimetic polymer may lead to even greater improvements in performance of this nanostructured adhesive. We believe geckel type adhesives will prove useful in a great variety of medical, industrial, consumer and military settings.

Example 1 - Preparation of PDMS nanoscaled arrays coated with p(DMA-co-MEA)

For the fabrication of gecko-mimetic adhesive arrays, e-beam lithography was used to create a pattern of holes in a PMMA film supported on a silicon wafer (negative mold). Solid phase PDMS was then cast onto the negative mold, thermally solidified, and then lifted off from the substrate to yield a positive array of PDMS pillars (~400 nm in diameter and 600 nm in height) supported on by a continuous PDMS film. Mussel-mimetic polymer, p(DMA-co- MEA), was synthesized by radical copolymerization of dopamine methacrylamide (DMA) and methoxyethylacrylate (MEA) monomers. Finally, the geckel adhesive was prepared by dip- coating PDMS pillar arrays into an ethanol solution of p(DMA-co-MEA) for 3hrs. Surface chemical compositions were analyzed by X-ray photoelectron spectroscopy (XPS) and time-of- flight secondary ion mass spectrometry (ToF-SIMS). Pillar arrays were imaged by AFM and scanning electron microscopy (SEM). Adhesive forces under dry and wet conditions were determined by AFM equipped with tipless cantilevers. The contact area between tip and the pillar array was precisely controlled by the distance between pillars (d = 1, 2, and 3 μm) and the angle between cantilever and pillar axis (θ), and was determined by optical imaging using a 4Ox objective and fiber-optic illumination.

Example 2 - Synthesis of dopamine methacrylamide (DMA)

2Og of sodium borate and 8g of NaHCCb were dissolved in 20OmL of deionized water and bubbled with Ar for 20 min. 1Og of dopamine-HCl (52.8 mmol) was then added followed by the dropwise addition of 9.4 mL of methacrylate anhydride (58.1 mmol) in 50 mL of THF, during which the pH of solution was kept above 8 with addition of IN NaOH as necessary. The reaction mixture was stirred overnight at room temperature with Ar bubbling. The aqueous mixture was washed twice with 100 mL of ethyl acetate two times and then the pH of the aqueous solution was reduced to less than 2 and the solution extracted with 100 mL of ethyl acetate 3 times. The final three washes were combined and dried over MgSO₄ to reduce the volume to around 5OmL. 450 mL of Hexane was added with vigorous stirring and the suspension was held at 4⁰C overnight. The product was recrystalized from hexane and dried to yield 9.1 g of grey solid. ¹H NMR (400 MHz, DMSO-c//TMS): δ 6.64-6.57 (m, 2H, C₆HH₂(OH)₂-), 6.42 (d, IH, C₆H₂H(OH)₂-), 5.61 (s, IH, -C(=O)-C(-CH₃)=CHH), 5.30 (s, IH, -C(=O)-C(-CH₃)=CHH), 3.21 (m, 2H, C₆H₃(OH)₂-CH₂-CH₂(NH)-C(=O)-), 2.55 (t, 2H, C₆H₃(OH)₂-CH₂-CH₂(NH)-C(=O)-), 1.84 (s, 3H, -C(=O)-C(-CH₃)=CH₂). ¹³C NMR (400 MHz, DMSO-dOTVJS): δ 167.3 (s, 1C, -NH-C(O)-C(CH₃)OH₂), 145.0 (s, 1C, -NH-C(O)- C(CH₃)=CH₂), 143.5-115.5 (6C, C₆H₃(O-C(O)-CHs)₂), 130.3 (s, 1C, -NH-C(O)- C(CHs)=CH₂), 41.0 (s, 1C, C₆H₃(OH)₂-CH₂-CH₂(NH)-C(O)-), 34.6 (s, 1C, C₆H₃(OH)₂-CH₂- CH₂(NH)-C(O)-), 18.7 (s, 1C, -C(O)-C(-CH₃)OH₂).

Example 3 - Synthesis of p(DMA-co-MEA)

12.5mL of MEA was passed through a column packed with 3Og Of Al₂O₃ to remove inhibitors. 7.5g of purified MEA (57.9 mmol), 1.7g of DMA (7.4 mmol), and 106 mg of AIBN (0.64 mmol) were added to 20 mL of DMF in an AirFree® flask. The solution mixture was degassed through pump-freeze-thaw cycles 3 times. While sealed under vacuum, the solution was heated to 6O⁰C and stirred overnight. The reaction mixture was diluted with 5OmL of methanol and added to 40OmL of Et₂O to precipitate the polymer. After precipitating in DCM/ethyl ether two more times and drying in a vacuum desicator, 5.7 g of white, sticky solid was obtained. ¹H NMR (400 MHz, CDC1₃/TMS): δ 6.81-6.70 (d, br, 2H, C₆HH₂(OH)₂-), 6.58 (s, br, IH, C₆H₂H(OH)₂-), 4.20 (s, br, 2H, CH₃-O-CH₂-CH₂-O-C(O)-), 3.57 (s, br, 2Η, CH₃- 0-CH₂-CH₂-O-C(O)-), 3.36 (s, br, 3H, CH₃-O-CH₂-CH₂-O-C(O)-), 2.69 (s, br, 2H, C₆H₃(OH)₂-CH₂-CH₂(NH)-C(O)-), 2.39 (s, br, IH, -0-C(O)-CH(CH₂-)-CH₂-), 2.14 (s, br, 2H, C₆H₃(OH)₂-CH₂-CH₂(NH)-C(O)-), 1.93 (s, 3H, -NH-C(O)-C(CH₃)(CH₂-)-CH₂-), 1.68 (m, br, -0-C(O)-CH(CH₂-)-CH₂-), 0.98 (m, br, -NΗ-C(O)-C(CΗ₃)(CΗ₂-)-CH₂-). GPC- MALLS (Wyatt Technology, Santa Barbara, CA with mobile phase of 2OmM LiBr in DMF and Shodex-OΗ Pak columns): M_n = 252 kDa, PD = 1.73. For control experiments, a catechol- free p(MEA) homopolymer (M_w = 100 kDa, Scientific Polymer Products, Ontario, NY) was used.

Example 4 - e-beam lithography e-beam resist (950PMMA A3, MicroChem) was spin-coated (4000 rpm, 40 sec) on silicon wafer several times until the resist thickness, as measured by ellipsometry (Woolam Co. Lincoln, NE), reached 600-700 nm. The resist was patterned at 30 kV with an area dose between 650-800 μC/cm² using Quanta 600F (FEI Co. Hillsboro, OR). Resist development was performed for 1 min with a solution of methyl isobutyl ketone/isopropanol (1/3, v/v), followed by rinsing with water. The patterned substrates were treated with oxygen plasma (Harrick, Pleasantville, NY) for 30 sec and repeated 2-3 times to completely remove residual resist from the exposed Si regions. The patterned substrates were then exposed to a triethoxyoctylsilane vapor for 30 min. PDMS was prepared as follows: 4 μL of Pt-catalyst (platinum-divinyl tetramethyl-disiloxane in xylene) and 4 μL of modulator (2,4,6,8- tetramethyl -2,4,6, 8-tetravinylcyclotetrasioxane) were added to a 7-8% vinylmethylsiloxane solution (3.5 g). The solution was subsequently mixed with a 25-30% methylhydrosiloxane (Ig) solution. Finally the solution was cured (80 ⁰C) after spin-coating (1000 rpm for 1 min) onto the PMMA/Si master. The spin-coated substrate was covered either by thin cover glass for force measurements or sylgard-184 PDMS for other experiments such as optical imaging or x-ray photoelectron spectroscopy (XPS). Gecko adhesive was obtained by PDMS pattern liftoff and brief exposure to oxygen plasma (100 W, 30 sec) and used within 2-3 hrs after plasma treatment. Geckel adhesive was prepared by dip-coating gecko adhesive in a 1 mg/mL solution of p(DMA-co-MEA) in ethanol at 70⁰C. Unstructured controls were fabricated in the same manner using flat PDMS.

Example 5 - X-ray photoelectron spectroscopy The presence of p(DMA-co-MEA) and p(MEA) on PDMS surfaces was confirmed by x-ray photoelectron spectroscopy (XPS) (Qmicron, Taunusstein Germany) equipped with a monochromatic Al Ka (1486.8 eV) 300 W x-ray source and an electron gun to eliminate charge build-up.

Example 6 -Atomic force and optical microscopy

All force data were collected on an Asylum Mfp-1D AFM instrument (Asylum Research, Santa Barbara, CA) installed on a Nikon TE2000 microscope. Spring constants of individual cantilevers (Veecoprobes, NP-20 tipless SisN₄ tips, Santa Barbara, CA) were calibrated by applying the equipartition theorem to the thermal noise spectrum.³⁰ Due to the large forces exhibited by the adhesive, only tips exhibiting high spring constants (280 - 370 pN/nm) were used. Metal and metal oxide coated cantilevers were formed by sputter coating -10 nm of Au or Ti (a native oxide formed at the Ti surface, TiO_x) using a Denton Vacuum Desk III (Moorestown, NJ). The surface composition of each cantilever was confirmed by time-of-flight secondary ion mass spectrometry (ToF-SIMS), using a PHI-TRIFT III (Ga⁺, 15keV, Physical Electronics, Eden Prairie, MN). Cantilevers were treated by oxygen plasma (100 W, 150 mTorr) for 3 min before use. Force measurements were conducted either in deionized water or ambient (air) conditions at a cantilever pulling speed of 2 μm/sec. In wet experiments, optical microscopic examination of the contact region indicated the absence of air bubbles trapped between nanopillars and on the nanopillar surface (not shown). Tapping mode AFM images were obtained using a multimode Veeco Digital Instrument (San Diego, CA) with a Si cantilever (resonance frequency of 230-280 kHz). Contact area was imaged by an inverted optical microscope using a 4Ox objective illuminated by a fiber-optic white light source perpendicular to the objective.

The following list of references, including the references themselves, is incorporated by reference herein.

References

1. Ruibal, R. & Ernst, V. The structure of the digital setae of lizards. J Morphology 117, 271- 293 (1965).

2. Geim, A. K. et al. Microfabricated adhesive mimicking gecko foot-hair. Nat. Materials 2, 461-463 (2003).

3. Northen, M. T. & Turner, K. L. A batch fabricated biomimetic dry adhesive. Nanotechnology 16, 1159-1166 (2005).

4. Sitti, M. & Fearing, R. Synthetic gecko foot-hair micro/nano-structures as dry adhesives. J. Adhes. Sci. Technol. 17, 1055-1073 (2003).

5. Yurdumakan, B., Raravikar, N. R., Ajayan, P. M. & Dhinojwala, A. Synthetic gecko foot- hairs from multiwalled carbon nanotubes. Chem. Commun. 30, 3799-3801 (2005).

6. Peressadko, A. & Gorb, S. N. When less is more: Experimental evidence for tenacity enhancement by division of contact area. J. Adhesion 80, 1-5 (2004).

7. Crosby, A. J., Hageman, M. & Duncan, A. Controlling polymer adhesion with "Pancakes". Langmuir 21, 11738-11743 (2005).

8. Northen, M. T. & Turner, K. L. Meso-scale adhesion testing of integrated micro- and nano- scale structures. Sensors and Actuators A 130-131, 583-587 (2006).

9. Huber, G. et al. Evidence for capillary contributions to gecko adhesion from single spatula nanomechanical measurements. Proc. Nat. Acad. Sci. USA 102, 16293-16296 (2005).

10. Sun, W., Neuzil, P., Kustandi, T. S., Oh, S. & Samper, V. D. The nature of the gecko lizard adhesive force. Biophys. J. 89, L14-16 (2005).

11. Autumn, K. et al. Evidence for van der Waals adhesion in gecko setae. Proc. Nat. Acad. Sci.

USA 99, 12252-12256 (2002).

12. Autumn, K. et al. Adhesive force of a single gecko foot-hair. Nature 405, 681-685 (2000).

13. Huber, G, Gorb, S. N., Spolenak, R. & Arzt, E. Resolving the nanoscale adhesion of individual gecko spatulae by atomic force microscopy. Biology Lett. 1, 2 (2005).

14. Arzt, E., Gorb, S. & Spolenak, R. From micro to nano contacts in biological attachment devices. Proc. Nat. Acad. Sci. USA 100, 10603-10606 (2003). 15. Arzt, E. Biological and artificial attachment devices: Lessons for materials scientists from flies and geckos. Materials Sci. and Eng. C: Biomimetic and Supramol. Systems 26, 1245- 1250 (2006).

16. Spolenak, R., Gorb, S. & Arzt, E. Adhesion design maps for bio-inspired attachment systems. Acta biomaterialia 1, 5-13 (2005).

17. Waite, J. H. Nature's underwater adhesive specialist. Chemtech 17, 692-697 (1987).

18. Waite, J. H. Adhesion a Ia moule. Meg. and Comp. Biol. 42, 1172-1180 (2002).

19. Waite, J. H. & Tanzer, M. L. Polyphenols substance of mytilus edulis: novel adhesive containing L-Dopa and hydroxyproline. Science 212, 1038-1040 (1981).

20. Papov, V. V., Diamond, T. V., Biemann, K. & Waite, J. H. Hydroxyarginine-containing polyphenols proteins in the adhesive plaques of the marine mussel Mytilus edulis. J. Biol. Chem. 270, 20183-92 (1995).

21. Waite, J. H. & Qin, X. X. Polyphenols phosphoprotein from the adhesive pads of the common mussel. Biochemistry 40, 2887-2893 (2001).

22. Yu, M. & Deming, T. J. Synthetic polypeptide mimics of marine adhesives. Macromolecules 31, 4739-4745 (1998).

23. Frank, B. P. & Belfort, G. Adhesion of Mytilus edulis Foot Protein 1 on Silica: Ionic Effects on Biofouling. Biotech. Prog. 18, 580-586 (2002).

24. Hwang, D. S., Yoo, H. J., Jun, J. H., Moon, W. K. & Cha, H. J. Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli. Appl. Environ. Microbiol. 70, 3352-9 (2004).

25. Lee, B. et al. Rapid Gel Formation and Adhesion in Photocurable and Biodegradable Block Copolymers with High DOPA Content. Macromolecules 39, 1740-1748 (2006).

26. Lee, H., Scherer, N. F. & Messersmith, P. B. Single molecule mechanics of mussel adhesion. Proc. Nat. Acad. Sci. USA 103, 12999-13003 (2006).

27. Whitesides, G. M. The origins and the future of microfluidics. Nature 442, 368-373 (2006).

28. Waite, J. H., Andersen, N. H., Jewhurst, S. & Sun, C. Mussel adhesion: finding the tricks worth mimicking. J. Adhesion 81, 1-21 (2005).

29. Dalsin, D. L., Hu, B.-H., Lee, B. P. & Messersmith, P. B. Mussel adhesive protein mimetic polymers for the preparation of nonfouling surfaces. J. Am. Chem. Soc. 125, 4253-4258 (2003). 0. Hutter, J. L. & Bechhoefer, J. Calibration of atomic-force microscope tips. Rev. ScL Instr. 64, 1868-1873 (1993).

Claims

CLAIMS What is claimed is:

1. An array of microstructures, the array comprising a substrate having upstanding or pending therefrom a plurality of microscale projections, the projections having a coating disposed on a working surface thereof.

2. An array of microstructures, the array comprising a substrate, the substrate having attached thereto a plurality of microscale projections, the projections having DHPP disposed on a working surface thereof.

3. An array of claim 1 wherein the projections are substantially columnar.

4. An array of claim 2 wherein the DHPP is a coating.

5. An array of claim 1 wherein the projections comprise a material other than DHPD.

6. A method comprising the steps of: providing an array of microstructures, the array comprising a support, the support having attached thereto a plurality of microscale elongate projections, the projections having DHPD disposed on a working surface thereof; adhering the array of microstructures to a surface with the DHPD layer being disposed between the surface and the support; detaching the array from the surface, the DHPD layer adhering to the array; and adhering the array to a second surface.

7. A method according to claim 6 wherein the array is attached to a first surface and the second surface is a cooperating second array having DHPD on a working surface thereof, the DHPD surface(s) being interposed between the first and second surfaces.

8. A method according to claim 6 wherein both the first and second surfaces have arrays of microstructures disposed therebetween.

9. A method according to claim 6 wherein the first and second surfaces are repositionable with respect to each other, their respective arrays being disposed therebetween with the DHPD (or DHPP) coatings being more cohesive than adhesive, and cooperating with each other.

10. A method according to claim 6 wherein the array has DHPP on a working surface thereof.

11. A microarray having a coating disposed on a working surface thereof.

12. A microarray of claim 11 wherein the coating is DHPP.

13. A microarray of claim 11 wherein the microarray comprises a substrate, the substrate supporting a plurality of microstructures.

14. A method of making a microarray comprising the steps of: providing a microarray having a plurality of working surfaces; and applying a coating to at least a portion of at least one of the working surfaces.

15. A method according to claim 14 wherein the applying step is accomplished by dip coating the microarray.

16. A method according to claim 15 wherein the applying step is accomplished by spin- coating the microarray.