Methods of providing a nanostructure formation coating layer on a medical device, coated medical devices, and kits for such coatings

Abstract

A method of forming a coated medical device, includes: providing a medical device having a polymer surface; contacting the polymer surface with an aqueous solution including water and one or more of retinin, retinin-like protein, and cuticular protein, and either not including a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20:1, to form a coating on the medical device; optionally, rinsing the coating with water or a saline solution; and optionally, drying the coating. The coating includes a nanostructure formation on the polymer surface, the nanostructure formation including the one or more of retinin, retinin-like protein, and cuticular protein attached to the polymer surface. Also provided are coated articles, and kits and methods for contact lens care.

Description

METHODS OF PROVIDING A NANOSTRUCTURE FORMATION COATING LAYER ON A MEDICAL DEVICE, COATED MEDICAL DEVICES, AND KITS FOR SUCH COATINGSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of U.S. Provisional Patent Application No. 63 / 748,608, filed January 23, 2025, and U.S. Provisional Patent Application No. 63 / 780,883, filed March 31, 2025, the contents of each of which are herein incorporated by reference in their entireties.INCORPORATION BY REFERENCE

[0002] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference in their entirety, as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.BACKGROUND

[0003] This disclosure relates generally to the field of medical devices and coatings, and more specifically to the field of treating or refreshing coated medical devices to form a nanostructure formation coating layer thereon. Described herein are systems, methods, and kits for such treatments.

[0004] Contact lenses are often used for vision correction, as an alternative to glasses or vision correction surgery. They may also be used for ocular pathologies and nonmedical or aesthetic purposes. Currently, contact lenses are typically classified as either “soft” or “hard” lenses. “Soft” lenses typically include hydrogel materials with greater than 30% water content. Soft lens hydrogels may be formed from traditional hydrophilic polymers or from silicone-containing polymers to enhance oxygen permeability. Soft lenses can also be composed of silicone elastomers, which have a low water content. “Hard” lenses are comprised of a rigid, gas-permeable material. “Hybrid” lenses also exist, which combine a rigid center with a soft material skirting, circumferentially disposed around the rigid center.

[0005] Contact lenses may often be uncomfortable to wear as they become dry and an irritant in the eye, either over the course of daily wear, or over the lifetime of the lens. Silicone-based lenses have a hydrophobic core, which can lead to abrasion of ocular tissue and infection. Rigid lenses can sometime be too hard to be comfortable. Finally, “hybrid” lenses, like soft and hard lenses, are prone to protein build-up.

[0006] Recent changes in contact lenses have attempted to address these issues. Substances and materials can be deposited onto a contact lens surface to improve the biocompatibility of the lens and therefore improve the interaction of the lens with the ocular region. For example, hydrophilic agents or wetting agents may be added to the lens solution to temporarily coat the lens such that upon wearing, the wetting angle of the lens is reduced. However, these temporary hydrophilic “coatings” are short-lived and quickly rinsed from the surface either with a cleaning solution or over the course of a day of wear.

[0007] Additionally, hydrophilic polymer solutions have been bound to the lens surface to create an outer coating on the lens surface that reduces the wetting angle and remains on the lens surface during rinsing with cleaning solution and through several or a plurality of wearing cycles. However, over time, these hydrophilic coatings are subject to wear, such that a thickness of the coating diminishes over time and limits the useful life of the contact lens.

[0008] Further, with such treated lenses, there is a possibility for enhanced treatment of several pathologies, that include, but are not limited to, dry-eye disease, glaucoma, corneal ulcers, scleritis, keratitis, iritis, and corneal neovascularization. In particular, such lenses may be useful for preventing dry-eye like symptoms in contact lens wearers.

[0009] Dry eye disease is considered to be a consequence of a disruption of the tear film that covers the surface of the eye or a particular vulnerability to such disruption. This tear film is an aqueous layer disposed between an underlying mucous layer that is secreted by corneal cells, and an overlying lipid layer that is secreted by Meibomian glands on the conjunctival surface of the eyelids. The tear film includes an aqueous pool that transits across the eye surface, having a flow path that, to some degree, may be independent of the lipid layers that it is disposed between at any point in time. The improved lenses diminish or substantially eliminate contact lens disruption of the tear film.

[0010] Such lenses may lose their effectiveness over time as the coating is either rinsed from the surface or reduced over time (reduced thickness) as a result of continued wear or use. Some systems recommend placing drops in the eye to coat the lens with additional hydrophilic moieties; however, such systems are ineffective and incapable of truly regenerating the coating on the lens.

[0011] Myopia is one of the fastest-growing vision disorders worldwide, with its prevalence projected to reach nearly 50% of the global population by 2050. The increasing burden of myopia necessitates innovative solutions for vision correction, particularly inchildren and young adults. Orthokeratology (Ortho-K) lenses offer a non-surgical approach to myopia management by reshaping the cornea, thereby reducing refractive error. Despite their efficacy, the adoption of lenses is significantly limited by challenges related to microbial contamination, biofilm formation, oxidative stress, and overall lens discomfort. The development of advanced biomaterials capable of enhancing lens biocompatibility and longterm user comfort is therefore of critical importance.

[0012] Ortho-K contact lens materials are inherently hydrophobic, which poses a challenge for both comfort and cleanliness. Hydrophilicity plays a crucial role in enhancing wearability by improving tear film stability and reducing friction between the lens and the ocular surface. Studies have shown that proper in vitro wettability or an anti-dryness function obtained through surface coatings, rather than lubricants, significantly improves subjective comfort in contact lens wearers. Beyond comfort, hydrophilicity is also essential for minimizing biofouling, as it reduces the adhesion of tear proteins, lipids, and microbial contaminants. Therefore, modifying lens surfaces to achieve long-lasting hydrophilic properties is a key to enhancing both safety and comfort of user experience.

[0013] The ocular surface presents a highly dynamic and sensitive environment, where tear film stability, protein-lipid interactions, and microbial exposure influence lens performance. One of the major limitations of Ortho-K lenses is their susceptibility to biofouling, as tear proteins such as lysozyme denature upon contact with the lens material, reducing their natural antimicrobial properties and contributing to inflammatory responses. Furthermore, lipid deposits on the lens surface create hydrophobic patches that exacerbate bacterial adhesion and biofilm formation. These factors not only compromise lens clarity and comfort but also increase the risk of complications such as microbial keratitis and conjunctival inflammation.

[0014] Traditional cleaning regimens rely on mechanical rubbing combined with chemical disinfection. However, even with the use of strong detergents and protein removers, a significant portion of deposits remains adhered to the lens surface. Moreover, frequent mechanical cleaning can lead to surface scratches, which further promote microbial adhesion and long-term biofouling. Therefore there is a need for an alternative and sustainable solution that enhances lens wettability, prevents protein and lipid deposition, and provides antimicrobial protection.

[0015] Beyond bacterial contamination, Ortho-K lenses can also serve as vectors for viral transmission, particularly in the case of highly infectious agents such as SARS-CoV-2. Viral particles can adhere to lens surfaces and persist for extended periods, increasing therisk of ocular transmission. The tear film, which serves as a protective barrier, is particularly vulnerable to disruption in contact lens wearers, potentially facilitating viral entry. Studies have shown that certain surface modifications can reduce viral adhesion and inactivate pathogens upon contact, highlighting the need for antiviral functionalities in next-generation lens coatings.

[0016] It is thus apparent that additional systems, devices, and methods may be contemplated to increase contact lens comfortability.SUMMARY

[0017] In view of the above problem and technical development, the present invention provides a method capable of effectively forming and maintaining a nanostructure formation coating layer on a contact lens.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Generally, the nomenclature and the laboratory procedure used herein are known in the art and usually used. Conventional methods are used in the processes, for example, methods mentioned in the art and various common references. When the terms are provided in singular forms, it is understood that the plural forms of the terms also apply. As used in the entire disclosure, unless otherwise specified, the following terms shall be understood as having the following meaning.

[0019] A dry lens refers to a lens that has been subjected to curing but has not yet been subjected to hydration during a contact lens production process.

[0020] A wet lens refers to a lens that has been subjected to hydration during a contact lens production process.

[0021] A contact lens packaging solution refers to an aqueous solution that undergoes a sterilization process along with a wet lens during a contact lens production process. The contact lens formed through the sterilization process is usually continued to be preserved in the aqueous solution, so the aqueous solution is also called a contact lens packaging solution. In addition, the aqueous solution usually has a buffering function, that is, has a pH value and an osmotic pressure within a certain range. Therefore, buffer is also used herein to refer to a contact lens packaging solution.

[0022] A core lens body refers to a portion of a lens that is formed from a contact lens formulation after subjected to curing.

[0023] A contact lens refers to a conventional contact lens suitable for patient use. The contact lens generally includes a core lens body coated with a polymer layer to provideimproved comfort wear. This polymer layer, or shell, is generally formed on the surface of the core lens body in other processes after curing.

[0024] In an embodiment, the present disclosure provides a lens conditioning kit comprising:a first component comprising a sterile lyophilized protein comprising one or more of retinin, retinin-like protein, and cuticular protein; anda second component comprises a sterile aqueous solution, wherein the kit optionally comprises a long-chain inhibitor of the sterile lyophilized protein such that a weight ratio of the sterile lyophilized protein to the long-chain inhibitor of the sterile lyophilized protein is greater than 20: 1.

[0025] In an embodiment, the present disclosure provides a coated contact lens comprising:a contact lens comprising a polymer surface; anda nanostructure formation formed on said polymer surface, said nanostructure formation comprising one or more of retinin, retinin-like protein, and cuticular protein and formed in an absence of a long-chain inhibitor or in presence of a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1.

[0026] In a further embodiment, the present disclosure provides a method of forming a coated medical device, comprising, in order:a) providing a medical device comprising a polymer surface;b) contacting the polymer surface with an aqueous solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a coating on the medical device;c) optionally, rinsing the coating with water or a saline solution; and d) optionally, drying the coating,wherein the coating comprises a nanostructure formation on said polymer surface, said nanostructure formation comprising the one or more of retinin, retinin-like protein, and cuticular protein attached to said polymer surface.

[0027] In a still further embodiment, the present disclosure provides a method for cleaning a contact lens, comprising in order:a) contacting a contact lens with a protein cleaning solution to remove protein deposits from the contact lens;b) optionally rinsing the contact lens with a rinsing solution;c) contacting the contact lens with an aqueous conditioning solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a nanostructure formation coating layer of the one or more of retinin, retinin-like protein, and cuticular protein attached to an outer surface of the contact lens; andd) rinsing the coating layer with a rinsing solution.BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The above-mentioned aspects, as well as other aspects, features, and advantages of the present technology are described below in connection with various embodiments, with reference made to the accompanying drawings.

[0029] Fig. 1 A shows a contact lens having a concave and convex surfaces.

[0030] Fig. IB is a cross-sectional view of an exemplary contact lens with a nanostructure formation coating.

[0031] Fig. 2 is a cross-sectional view of the contact lens shown in Fig. IB on the cornea.

[0032] Figs. 3 A and 3B are TEM images of cryo-cross-sections of coated RGP lenses with different magnifications.

[0033] Figs. 4A, 4B, and 4C are AFM time-courses of nanostructure formation. Each scan area is 2 * 2 pm.

[0034] Figs. 5A, 5B, and 5C are adhesion force mapping of uncoated (control) versus nanostructure formation-coated lenses, (quantified in the central plot Fig. 5B).

[0035] Figs. 6A and 6B are results of a wettability test and lipid adhesion test comparing 5- and 10-minute coating protocols to uncoated controls.

[0036] Fig. 6C shows quantitative analysis demonstrating a significant decrease in hydrophobicity and lipid accumulation with increasing coating duration. Statistical significance (two-tailed t-test) is reported relative to the non-coated control, n = 8 for wettability test, and 4 for lipid adhesion test. All data are presented as mean ± SD .“****” represents P value <0.0001.

[0037] Fig. 7 is protein deposition test showing that the nanostructure formation coating (labeled as AEGIS) protects the lens surface from nonspecific binding of BSA. Data represent contact angles of treated surfaces (solid bars) and after Progent™ cleaning (striped bars). P-values are shown relative to the nanostructure formation-coated lens, n = 6-10.

[0038] Fig. 8 is representative AFM scans of nanostructure formation-coated and BSA-treated RGP lens surfaces. Each scan area: 3 x 3 pm.

[0039] Fig. 9 shows wettability of nanostructure formation-coated RGP lenses during a 7-day simulated wear-and-wash cycle. The dashed line marks the 60° threshold, above which wettability is considered suboptimal. P-values are shown relative to day 0. n = 20.

[0040] Fig. 10 shows AFM scans of nanostructure formation-coated RGP lenses at different time points during a 7-day simulation, revealing gradual degradation of the coating. Each scan area: 3 x 3 pm.

[0041] Fig. 11 A to 1 ID show testing results for nanostructure formation-coated lenses (shown as AEGIS or RetL). Fig. 11 A show a slight but significant bactericidal effect against E. coli after 1 hour of incubation, n = 2-3; N = 2 (replicates). Fig. 1 IB shows antiviral activity showing that the nanostructure formation nanocoating reduces viral particle viability by 30% after 6 hours of incubation, n = 5. Fig. 11C shows cytotoxicity assessment showing that nanostructure formation treatment does not affect cell viability, n = 12. Fig. 1 ID shows antioxidant activity of nanostructure formation coating, measured using the DPPH assay, n = 3. All data are presented as mean ± SD. “ns” stands for non-significant, “*” indicates the p value < 0.05, “**” - p value < 0.01,“***” - p value < 0.001,“****” - p value < 0.0001.

[0042] Figs. 12A and 12B show TEM images showing polymer brush structures on uncoated RGP lenses, visible as a thin surface layer (Fig. 12A) or thread-like extensions (Fig.12B). Scale bars: 50 nm.

[0043] Fig. 13 shows a schematic illustration of polymer brush relaxation: collapsed chains in PBS, swelled and extended in isopropanol (IP A).

[0044] Figs. 14A and 14B shows AFM scans performed at varying tip forces in 20% IPA (Fig. 14A) and 80% IPA (Fig. 14B). In 20% IP A, the rigid surface of the lens is consistently detected, indicating that the polymer brushes are collapsed and do not obscure the substrate. In 80% IPA, increasing the applied force from 10 to 100 pN improves resolution of the underlying surface, consistent with splaying and penetration of swollen polymer brush layers.

[0045] Fig. 15 show force-distance curves acquired in 80% (upper line) and 20% (lower line) IPA. The long-range exponential repulsion observed in 80% IPA confirms brush-like chain behavior, while the weak short-range repulsion in 20% IPA indicates collapsed, non-brush-like conformations, n = 100; data shown as mean ± SD. The inset shows fitting with the Alexander-de Gennes model (sloping dashed line) gives values of length, L = 158 ± 4 nm and s = 15.5 ± 2 nm.

[0046] Figs. 16A and 18B show schematic visualizations of nanostructure formation nanocoating (labeled as RetL) self-assembly on polymer brushes. Initially, the uncoated contact lens surface consists of collapsed hydrophobic polymer chains. Upon binding to the amphiphilic RetL protein, hydrophilicity increases. This interaction activates the polymer brushes as reaction-diffusion morphogens, triggering the self-assembly of the nanocoating. In the final state, the polymer chains are fully coated and no longer exposed at the interface. The RetL protein structure shown in the scheme is based on an AlphaFold prediction. In Fig. 16A, protein hydrophobicity is illustrated as decreasing from bright (high) to dark (low).

[0047] Fig. 17 shows contact angle results for contact lenses according to Example 2 before and after coating, with and without washing. Data is shown as average +- SD, n=2.

[0048] Fig. 18A shows SEM images of non-coated contact lens taken with xl2000 and x25000 magnifications.

[0049] Fig. 18B shows SEM images of freshly coated contact lens taken with xl2000 and x25000 magnifications.

[0050] Fig. 18C shows SEM of the coated lens after seven days of the usage simulation with x25000 magnification.DETAILED DESCRIPTION OF EMBODIMENTS

[0051] The foregoing is a summary, and thus, necessarily limited in detail. The above mentioned aspects, as well as other aspects, features, and advantages of the present disclosure will now be described in connection with various embodiments. The inclusion of the following embodiments is not intended to limit the disclosure to these embodiments, but rather to enable any person skilled in the art to make and use the contemplated invention(s). Other embodiments may be utilized and modifications may be made without departing from the spirit or scope of the subject matter presented herein. Aspects of the disclosure, as described and illustrated herein, can be arranged, combined, modified, and designed in a variety of different formulations, all of which are explicitly contemplated and form part of this disclosure.

[0052] There is a need for a new and useful system and method for treating, retreating, or refreshing a specially coated medical devices such as contact lens. In particular, there is a need for one or more systems, devices, and methods that create and / or regenerate a nanostructure formation coating layer on a medical device such as a contact lens.

[0053] As used herein “nanostructure formation” or “nanostructure formation coating layer” refer to a structure formed from the assembly or organization of the retinin, retinin-like protein, and cuticular protein at the nanoscale level. The terms “nanostructure formation” or “nanostructure formation coating layer” may also be used to refer to such a structure formed as a coating (“nanocoating”) on an underlying substrate.

[0054] One aspect of the present disclosure is directed to a method of generating in the first instance a nanostructure formation coating layer on a newly-produced medical device such as a contact lens. Another aspect of the present disclosure is directed to a method of regenerating the nanostructure formation coating layer on a medical device such as a contact lens as the original nanostructure formation coating layer has begun to degrade and / or is fully or partially removed during a routine cleaning process of the medical device. Other aspects of the present disclosure are directed to materials, solutions, kits, and the like for carrying out these methods.

[0055] As used herein, treating comprises initially creating a coating on a medical device such as a contact lens and / or subsequently retreating or refreshing or rejuvenating a previously coated medical device to regenerate the existing coating or add to an existing coating on the medical device. As used herein, treating, refreshing, rejuvenating, and regenerating may generally be used interchangeably unless otherwise apparent from context.

[0056] As used herein, an existing coating or layer refers to a coating present on a surface, particularly an exposed surface, of a medical device such as a contact lens before rejuvenation or refreshing or retreating. The existing coating may have been deposited during the manufacturing phase, before the last use or wear cycle, after purchase, before a first use or wear, etc. The existing coating or layer may have an initial thickness before use or wear and a second or reduced thickness after use or wear. The reduced thickness may occur after one use or wear cycle or after n number of use or wear cycles (e.g., n equals 1 day, 1-3 days, 3-6 days, 1-5 days, 1-10 days, 7-14 days, 2-4 weeks, 1 month, etc.). After treatment with the solutions or kits to regenerate or rejuvenate the existing coating or layer, the second or reduced thickness may increase such that is approaches the initial thickness, exceeds the initial thickness, or substantially equals the initial thickness.

[0057] In some embodiments, the medical device is a contact lens adapted for placement in the eye. Exemplary, non-limiting lenses include contact lens, smart lenses, disposable soft lenses, daily lenses, extended wear lenses, spherical lenses, toric lenses, multifocal or bifocal lenses, diagnostic lenses, monovision lenses, conventional lenses, rigid gas permeable lenses, hard lenses, color variation lenses, opaque lenses, enhancers lenses, and visibility tinted lenses. For ease of the description herein, lenses will be commonly referred to herein as contact lenses.

[0058] As shown in Fig. 1 A, a contact lens 2 may be generally understood as having a body with a concave surface 4 and a convex surface 6. The lens body may include a periphery or a perimeter 8 between the surfaces. The periphery may also include a circumferential edge between the surfaces.

[0059] The concave surface 4 may also be referred to as a posterior surface and the convex surface 6 may also be referred to as an anterior surface, terms that refer to respective position when worn by a user. In practice, the concave surface of the lens is adapted to be worn against or adjacent to an ophthalmic surface. When worn, the concave surface may lie against a user's corneal surface 48 (see Fig. 2). The convex surface is outward-facing, exposed to the environment when the eye 40 is open. When the eye 40 is closed, the convex surface is positioned adjacent or against the inner conjunctival surface 44 of theeyelids 42 (see Fig. 2).

[0060] Because the convex and concave surfaces of a lens may be placed against or adjacent ophthalmic tissue such as the corneal surface, the properties of the surfaces can greatly affect a user's comfort and wearability of the lens as described above. For example, the lens may disrupt the tear film 16 of the eye 40 causing symptoms associated with dry eye. As such, embodiments described herein provide for a rejuvenated coated contact lens having a nanostructure formation coating layer rejuvenated on at least one of the lens's surfaces to improve the lens's wettability and wearability with minimal tear film disruption.

[0061] In one embodiment, the contemplated coated contact lens includes a contact lens with at least one surface having a nanostructure formation coating layer of the present disclosure. In some cases, the nanostructure formation coating layer is adapted for placement against an ophthalmic surface and / or is adapted for placement on a surface of a contact lens opposite the ophthalmic surface. The nanostructure formation coating layer may cover a portion of the contact lens. Alternatively, the nanostructure formation coating layer may completely or substantially completely cover the contact lens surface.

[0062] In other variations, more than one core surface of the contact lens has a nanostructure formation coating layer. For example, a nanostructure formation coating layer on both the concave and the convex surfaces of the contact lens may be provided. Each nanostructure formation coating layer on either concave or convex surfaces may independently completely or partially cover respective surfaces. In some cases, the nanostructure formation coating layer on each side of the contact lens forms a contiguous layer across both surfaces.

[0063] Referring to Fig. IB, a cross-section of an exemplary embodiment of a coated contact lens 10 is shown. Coated contact lens 10 includes a contact core 18 and a nanostructure formation coating layer 20 attached to the contact lens 18. As shown, in this embodiment a nanostructure formation coating layer 20 surrounds the contact lens 18. Both the concave and convex surfaces 12, 14 are coated by the same nanostructure formation coating layer 20 on both sides of the contact lens 18 with the nanostructure formation coating layer 20 extending to the peripheral edge 8 of the coated contact lens 10. As shown, the outer nanostructure formation coating layer 20 is substantially contiguous through or across a circumferential edge portion 8. A variety of different materials can be used as the contact lens core and polymer layer of the contact lens as described in detail below. In some embodiments, the contact lens can be a rigid gas permeable lens. In some embodiments, the lens core can be a hydrophobic material, such as silicone, coated with a polymer layer. As used herein, silicone includes polysiloxanes. In some embodiments, the contact lens can include a lens core including a hydrogel.

[0064] Returning to Fig. 2, the coated contact lens 10 of Fig. IB is positioned in a user's eye 40. The eye 40 is shown with eye lens 46 and iris 50. The concave surface 12 of the coated lens 10 is disposed and centered on the cornea. The convex surface 14 of the lens 10 is directed outwardly, facing the environment when the eye 40 is open. When the eyelid 42 closes, the convex surface 14 is adjacent to the inner or conjunctival surface 44 of the eyelid 42. As the eyelids 42 open and close, the conjunctival surface 44 slides across the convex surface 14 of the lens 10.

[0065] When placed on the cornea, the nanostructure formation coating layer 20 of the coated contact lens 10 interacts with the natural tear film 16 of the eye 40. The coated contact lens 10 may be positioned within the tear film 16 and / or substantially reside within the aqueous layer of the tear film 16 that covers the eye 40. In some cases, the coated contact lens 10 is immersed in the tear film 16. The nanostructure formation coating layer 20 may be adapted to minimize disruption of the tear film by the coated contact lens 10.

[0066] In some embodiments, the medical device is one or more coated contact lenses. The disclosed solutions, kits, and methods may be used to treat or retreat coated hard, soft, and / or hybrid contact lenses, although any suitable medical device or contact lens may be envisioned.

[0067] The contact lens envisioned may be any suitable contact lens, and in the present aspect, the surface of the contact lens comprises a polymer layer. A coated contact lens includes a lens core and a nanostructure formation coating layer attached to the polymer layer of the contact lens. A nanostructure formation coating layer thus surrounds the contact lens. Both the concave and convex surfaces are coated by the same nanostructure formation coating layer on both sides of the contact lens with the nanostructure formation coating layer extending to the peripheral edge of the contact lens. As shown, the outer nanostructure formation coating layer is substantially contiguous through or across a circumferential edge portion. A variety of different materials can be used as the lens core of the contact lens. In some embodiments, the lens core is a rigid gas permeable material. In some embodiments, the lens core is a hydrophobic material, such as silicone, which may include polysiloxanes. In some embodiments, the lens core comprises a soft lens material. In some embodiments, the lens core can include a hydrogel.

[0068] Any suitable contact lens with any lens core may be used, for example any lens core having an existing polymer coating or layer. For example, the lens core itself may be hydrophobic or hydrophilic. A hydrophilic core may include a hydrogel containing core such as a pure hydrogel lens. For example, the core may contain Polyhexyethyl methacrylate lenses (pHEMA).

[0069] In some embodiments, the lens core is a rigid gas permeable (RGP) material. In some embodiments the rigid gas permeable material is non-hydrophilic. In some embodiments the rigid gas permeable material is hydrophobic. Examples of rigid gas permeable materials include cellulose acetate butyrate, polyacrylate-silicone, non-hydrophilic silicone elastomers, polysiloxane, fluoro-silicon polymers, etc. As used herein silicone includes polysiloxanes. Examples of commercial RGP lenses that can be treated with the processes disclosed herein include: Bausch & Lomb Boston Lens, Paragon CRTlens, Menicon Rose K, Menicon Lagado Flosi, Menicon Lagado Tyro, Menicon Lagado Onsi, Contamac Optimum Classic, Contamac Optimum Comfort, Contamac Optimum Extra, Contamac Optimum Extreme, Contamac Optimum Infinite, HEXA100, ENFLU 18, Fluorperm 92, Fluoroperm 60 / Paragon HDS, Fluoroperm 30 / Paragon Thin, Fluoroperm 151 / HDS 100, Boston XO, Boston XO2, Boston ES, and Boston EO.

[0070] In some embodiments, the lens core includes a rigid gas permeable material in a center region with a soft outer skirt circumferentially disposed about the center region, the soft outer skirt comprising silicone or other soft material. RGP lenses having a soft outer skirt are known as hybrid lenses. The original nanostructure formation coating layer and regenerated or rejuvenated nanostructure formation coating layer described herein can be formed on one or both of the convex and concave surfaces of the hybrid RGP / soft coating contact lens, as described herein. Examples of commercial hybrid RGP lenses include those made by Synergeyes, such as the Synergeyes Duette Lens and the Synergeyes Ultra Health, and Laboratoire LCS, such as EyeBrid.

[0071] The RGP and hybrid RGP lenses are typically used by the patient for several months or more. In some cases, the RGP and hybrid RGP lenses can be used for a year or more. In contrast to the soft lenses, which are disposable and used for shorter amounts of time, the RGP and hybrid RGP lenses can be exposed to harsher cleaning processes than the disposable soft lenses. In order to meet the design requirements for RGP lenses and hybrid RGP lenses, it is desirable for any coatings on the contact lens, including the nanostructure formation coating layer of the present disclosure, to have a sufficiently long shelflife as well as the capability to withstand the more rigorous cleaning associated with those types of lenses. Alternatively, the coating, and particularly the nanostructure formation coating layer of the present disclosure, may be regenerated multiple times throughout the wearing cycle by systems, methods, and materials described elsewhere herein.

[0072] In some embodiments, a lens core comprising silicone and coated with a polymer layer can be used with any of the nanostructure formation coating layers described herein. The silicone lens core can comprise one or more polysiloxane compounds. In some embodiments, the polysiloxanes are cross-linked.

[0073] In some embodiments, the lens core can be primarily made of cross-linked polysiloxanes with trace impurities or trace additives. The lens core may consist substantially entirely of pure silicone (e.g. polysiloxane compounds), i.e. the core comprises about 100% silicone by weight. In other embodiments, the lens core can be made out of only polysiloxanes (e.g. 100% silicone by weight). In some embodiments, the lens core consists of polysiloxane. In other cases, the lens core, base, or substrate comprises about 10% to about 50% of silicone by weight. In some cases, the substrate or core comprises about 25% silicone by weight.

[0074] The silicone lens cores are resistant to water and do not absorb water. The lack of absorption of water can be described as the water equilibrium constant. In contrast to hydrogels, which by definition absorb water, silicone does not appreciably absorb water.

[0075] In some embodiments, the lens core is a soft contact lens. For example, a soft contact lens can include an elastic modulus of less than about 2.0 MPa. In some embodiments, the lens core has an elastic modulus of less than about 1.8 MPa. In some embodiments, an elastic modulus of a hydrogel coated contact lens before use and / or wear is substantially equal to an elastic modulus of a regenerated or rejuvenated hydrogel coating on a hydrogel coated contact lens.

[0076] Conventional silicone contact lens cores are known in the art to stick to the surface of the eye and are unsuitable for use in adults without additional processing and treatments. An uncoated conventional silicone lens can stick to the eye and damage the surface of the eye if the lens is moved or removed. Various hydrogel or polymer coatings known in the art can thus be applied to and coated on and used to coat both sides of silicone lenses to improve the lens properties and biocompatibility with the eye to provide a wearable contact lens. The hydrogel or polymer coatings allow the coated silicone lens to be adapted for on-eye movement without damage to the eye or ophthalmic surface. The contact lenses thereby are adapted to provide adequate on eye movement while maintaining the health of the ophthalmic surface and wearer comfort.

[0077] Another advantage of a silicone lens core is the high refractive index of silicone. Conventional hydrogel lenses have a much higher water content. The water content decreases the overall refractive index of the lens. A thin silicone core with a high refractive index can be used with a thin hydrogel or polymer coating to produce a contact lens with smaller thickness than conventional lenses and a higher refractive index. In some embodiments, the contact lens can have a refractive index of greater than about 1.420. The contact lens can have a thickness of less than 50 microns. In some embodiments, the contact lens has a thickness of less than 25 microns.

[0078] The silicone cores can be formed from and include a variety of different monomers. Examples of preferred silicone-containing vinylic monomers include, without limitation, N-[tris(trimethylsiloxy)silylpropyl]-(meth)acrylamide, N-[tris(dimethylpropylsiloxy)-silylpropyl]-(meth)acrylamide, N-[tris(dimethylphenylsiloxy)silylpropyl](meth)acrylamide, N-[tris(dimethylethylsiloxy)silylpropyl](meth)acrylamide, N-(2 -hydroxy-3 -(3-(bis(trimethylsilyloxy)methylsilyl)propyloxy)propyl)-2-methyl acrylamide; N-(2 -hydroxy-3-(3-(bis(trimethylsilyloxy)methylsilyl)propyloxy)propyl)acrylamide; N,N-bis[2-hydroxy-3-(3-(bis(trimethylsilyloxy)methylsilyl)propyloxy)propyl]-2-methyl acrylamide; N,N-bis[2-hydroxy-3-(3-(bis(trimethylsilyloxy)methylsilyl)propyloxy)propyl]acrylamide; N-(2-hydroxy-3-(3-(tris(trimethylsilyloxy)silyl)propyloxy)propyl)-2-methyl acrylamide; N-(2-hydroxy-3-(3-(tris(trimethylsilyloxy)silyl)propyloxy)propyl)acrylamide; N,N-bis[2-hydroxy-3-(3-(tris(trimethylsilyloxy)silyl)propyloxy)propyl]-2-methyl acrylamide; N,N-bis[2-hydroxy-3-(3-(tris(trimethylsilyloxy)silyl)propyloxy)propyl]acrylamide; N-[2-hydroxy-3-(3-(t-butyldimethylsilyl)propyloxy)propyl]-2-methyl acrylamide; N-[2-hydroxy-3-(3-(t-butyldimethylsilyl)propyloxy)propyl]acrylamide; N,N-bis[2-hydroxy-3-(3-(t-butyldimethylsilyl)propyloxy)propyl]-2-methyl acrylamide; N,N-bis[2-hydroxy-3-(3-(t-butyldimethylsilyl)propyloxy)propyl]acrylamide; 3 -methacryloxy propylpentamethyldisiloxane, tris(trimethylsilyloxy)silylpropyl methacrylate (TRIS), (3-methacryloxy-2-hydroxypropyloxy)propylbis(trimethylsiloxy)methylsilane), (3-methacryloxy-2-hydroxypropyloxy)propyltris(trimethylsiloxy)silane, 3 -methacryloxy -2-(2-hydroxyethoxy)-propyloxy)propylbis(trimethylsiloxy)methylsilane, N-2-methacryloxyethyl-O-(methyl-bis-trimethylsiloxy-3-propyl)silylcarbamate, 3 -(trimethyl silyl)propylvinyl carbonate, 3-(vinyloxycarbonylthio)propyl-tris(trimethyl-siloxy)silane, 3-[tris(trimethylsiloxy)silyl]propylvinyl carbamate, 3-[tris(trimethylsiloxy)silyl]propyl allyl carbamate, 3-[tris(trimethylsiloxy)silyl]propyl vinyl carbonate, t-butyldimethyl-siloxyethyl vinyl carbonate; trimethylsilylethyl vinyl carbonate, and trimethyl silylmethyl vinyl carbonate). Most preferred siloxane-containing (meth)acrylamide monomers of formula (1) are N-[tris(trimethylsiloxy)silylpropyl]acrylamide, TRIS, N-[2-hydroxy-3-(3-(t-butyldimethylsilyl)propyloxy)propyl]acrylamide, or combinations thereof.

[0079] A class of preferred silicone-containing vinylic monomers or macromers is polysiloxane-containing vinylic monomers or macromers. Examples of such polysiloxane-containing vinylic monomers or macromers are monomethacrylated or monoacrylated poly dimethyl siloxanes of various molecular weight (e.g., mono-3 -methacryloxypropyl terminated, mono-butyl terminated polydimethylsiloxane or mono-(3 -methacryloxy -2-hydroxypropyloxy)propyl terminated, mono-butyl terminated polydimethylsiloxane); dimethacrylated or diacrylated polydimethylsiloxanes of various molecular weight; vinyl carbonate-terminated polydimethylsiloxanes; vinyl carbamate-terminated polydimethylsiloxane; vinyl terminated polydimethylsiloxanes of various molecular weight; methacrylamide-terminated polydimethylsiloxanes; acrylamide-terminated polydimethylsiloxanes; acrylate-terminated polydimethylsiloxanes; methacrylate-terminatedpolydimethylsiloxanes; bis-3-methacryloxy-2-hydroxypropyloxypropyl polydimethylsiloxane; N,N,N',N'-tetrakis(3-methacryloxy-2-hydroxypropyl)-alpha,omega-bis-3-aminopropyl-polydimethylsiloxane; polysiloxanylalkyl (meth)acrylic monomers; siloxane-containing macromer selected from the group consisting of Macromer A, Macromer B, Macromer C, and Macromer D described in U.S. Patent No. 5,760,100; the reaction products of glycidyl methacrylate with amino-functional polydimethylsiloxanes; hydroxylfunctionalized siloxane-containing vinylic monomers or macromers; polysiloxane-containing macromers disclosed in U.S. Patents Nos. 4,136,250, 4,153,641, 4,182,822, 4,189,546, 4,343,927, 4,254,248, 4,355,147, 4,276,402, 4,327,203, 4,341,889, 4,486,577, 4,543,398, 4,605,712, 4,661,575, 4,684,538, 4,703,097, 4,833,218, 4,837,289, 4,954,586, 4,954,587, 5,010,141, 5,034,461, 5,070,170, 5,079,319, 5,039,761, 5,346,946, 5,358,995, 5,387,632, 5,416,132, 5,451,617, 5,486,579, 5,962,548, 5,981,675, 6,039,913, and 6,762,264; polysiloxane-containing macromers disclosed in U.S. Patents Nos. 4,259,467, 4,260,725, and 4,261,875. Di and triblock macromers consisting of polydimethylsiloxane and polyalkyleneoxides could also be of utility. For example, one might use methacrylate end capped polyethyleneoxide-block-polydimethylsiloxane-block-polyethyleneoxide to enhance oxygen permeability. Suitable monofunctional hydroxyl-functionalized siloxane-containing vinylic monomers / macromers and suitable multifunctional hydroxyl-functionalized siloxane-containing vinylic monomers / macromers are commercially available from Gelest, Inc, Morrisville, Pa.

[0080] In some embodiments, the lens core may comprise a silicone-hydrogel (SiHy). The silicone hydrogel lens core can have a higher water content than the silicone lens core embodiments since hydrogels absorb water. For example, the silicone hydrogel lens core can have an equilibrium water content greater than 2% and less than 60%. In such cases, the SiHy lens core can be coated by the described hydrophilic polymer layers to improve wettability and wearability of the lens core. In other variations, the core comprises about 10% to about 50% of silicone by weight. In some embodiments, the hydrophilic layer can have a thickness of less than 100 nm.

[0081] In an exemplary embodiment, the silicone-containing layer or core of the coated contact lens is lotrafilcon, balafilcon, galyfilcon, senofilcon, narafilcon, omafilcon, comfilcon, enfilcon, or asmofilcon. In some cases, the silicone-containing core is NuSil Med 6755.

[0082] Alternatively, a non-silicone based core may be used as the substrate for coating. For example, an oxygen permeable lens made from a non-silicone material with apolymer surface layer may also be coated with the described nanostructure formation coating layer.

[0083] In an exemplary embodiment, the thickness of the core or core layer is from about 25 microns to about 200 microns, or from about 50 microns to about 150 microns, or from about 75 microns to about 100 microns, or from about 20 microns to about 80 microns, or from about 25 microns to about 75 microns, or from about 40 microns to about 60 microns.

[0084] According to the present disclosure, a nanostructure formation coating layer is applied over the polymer coating of a medical device such as a contact lens. The nanostructure formation coating layer is made from one or more of retinin, retinin-like protein, and cuticular protein, and is formed in an absence of a long-chain inhibitor such as a lipid, a wax, and a surfactant. The nanostructure formation coating layer provides a wide range of benefits, including increased antimicrobial properties, increased resistance to lipid adhesion, and antioxidant surface activity.

[0085] As used herein, a “long-chain inhibitor” is a compound or moiety, generally of an elongated chain-like structure such as the lipids, waxes, and surfactants described herein, that provides an inhibitory effect upon the otherwise uncontrolled assembly of the retinin, retinin-like protein, and / or cuticular protein, allowing those proteins to assembly into an organized nanostructure.

[0086] Details of the nanostructure formation coating layer made from one or more of retinin, retinin-like protein, and cuticular protein are provided in U.S. Patent No.12,104,078, the entire disclosure of which is incorporated herein by reference. These and further details are provided herein.

[0087] In the present context, the term “retinin or retinin-like protein” refers to a protein expressed in the eye or cornea of a variety of insects and which preferably comprises a conserved region known as the retinin C-domain. However, “retinin-like proteins” also includes proteins that does not comprise the retinin C-domain, but wherein the remaining part of the amino acid sequence is similar to the middle and C-terminus parts of the Retinin sequence (disregarding the signal peptide sequence). The retinin C-domain is designated PF04527 in the PF AM database of protein families and is also known as Retinin-like domain designated IPR007614 in the InterPro database of proteins.

[0088] The term “retinin or retinin-like protein” as used herein allow also for a certain degree of homology in their common retinin C-domain region or other regions of theprotein. Therefore, the amino acid sequence of the protein may deviate from the sequence identified above and thus vary between retinin or retinin-like proteins described herein.

[0089] In the present context cuticular proteins are proteins found in the exoskeleton of arthropod organisms, hereunder insects. The cuticula or cuticle is a multilayered structure outside the epidermis of many invertebrates, notably arthropods, in which it forms an exoskeleton. Thus, cuticle proteins form the major part of the integument of arthropods. The integument is the natural covering of an organism or an organ, such as its skin, husk, shell, or rind. Thus, it includes most of the material of the exoskeleton of the insects.

[0090] In the present context, the term “lipid” refers to hydrophobic or amphiphilic molecules that are soluble in non-polar solvents and do not easily dissolve in water. Lipids include, but are not limited to, waxes, fats, oils, sterols, phospholipids, and mono-, di- and triglycerides. Generally, oils are lipids that are liquid at room temperature, whereas fats are lipids that are solid at room temperature.

[0091] In the present context, the term “wax” refers to compounds that are lipophilic, malleable solids near room temperature. Thus, at room temperature, waxes display an elevated ability to deform under compressive stress. Typically, waxes have melting points above 40°C. Waxes appear as unctuous, meltable, viscous liquids to solid substances, having a characteristic waxy luster, and may be of natural or synthetic origin or derived from natural or synthetic materials. Waxes include hydrophobic or amphiphilic higher alkanes that are soluble in non-polar solvents, but insoluble in water. Waxes may encompass various functional groups such as, but not limited to, fatty acids, alcohols, unsaturated bonds, aromatics, amides, ketones and aldehydes. Specifically, many waxes comprise fatty acid esters. These waxes are made from a fatty acid bonded through an ester linkage to an alcohol. In contrast, paraffin wax is an example of a wax comprised of a mixture of long-chain hydrocarbons without functional groups.

[0092] In the present context, the term “insect” refers to animals of the subclass Pterygota, which is a subclass to the class Insecta. The subclass Pterygota comprises all the winged insects.

[0093] More specifically, the retinin or retinin-like protein or cuticular protein may be obtained from any animal within the subclass Pterygota suitable for providing the retinin, retinin-like protein or cuticular protein. Similarly, the retinin or retinin-like protein or cuticular protein may be provided as a recombinant protein based on the amino acid (orinitially nucleic acid) of any animal within the subclass Pterygota, which express retinin, retinin-like or cuticular protein.

[0094] The present invention aims at providing means for in vitro coating of any type of surface, including soft matter materials sensitive to harsh treatment. Thus, the present invention relates to a simple and cheap method for producing functional nanostructure formation coating layers on natural or artificial surfaces of medical devices such as contact lenses using biologic reagents under mild conditions.

[0095] The nanostructure formation is derived from the observation that terrestrial arthropods carry elaborated nanocoatings on top of their corneal surfaces. Serving an antireflective function (and potentially anti-wetting, bactericidal, and dirt-removing purposes), these nanocoatings in different arthropods can be built by nanoscale paraboloid protrusions (nipples), ridges, mazes, dimples, or various transitions among these forms.

[0096] It was previously found that the corneal protein retinin in combination with lipids were identified as the minimal components for formation of natural nanocoatings. Cuticular proteins from insects were also found to behave similarly to retinin in this respect. It was found the nanocoatings could be recreated on artificial surfaces through the admixing of e.g. retinin and commercial lipids. By varying the admixing procedure, the method could be used to produce a diverse set of nanocoatings yielding different functionalities.Importantly, the bioinspired nanocoating was produced under mild conditions and used natural reagents that are easily accessible.

[0097] The present inventors have now surprising found that a nanostructure formation coating layer can be formed expressly in an absence of a long-chain inhibitor such as a lipid, a wax, and a surfactant on a polymer surface. In some embodiments, some lipid, wax, and / or surfactant that otherwise acts as an added long-chain inhibitor may be present with the retinin, retinin-like protein, and / or cuticular protein during formation of the nanostructure formation coating layer, although in amounts less than previously deemed required in U.S. Patent No. 12, 104,078. The polymer surface according to the present disclosure behaves very differently from other surfaces such as metal or glass. On such metal and glass surfaces, an added long-chain inhibitor is required in order to form the nanostructure formation coating layer, but that long-chain inhibitor is not required for polymer surfaces, or its amount may be significantly reduced. This discovery is thus particularly applicable to medical devices such as contact lenses.

[0098] As used herein, “polymer surface” generally refers to an exposed out outer surface of an object such as a contact lens or other medical device, which is made from or comprises a polymer material as described herein.

[0099] Thus, an aspect of the present invention relates to a method for preparation of a coated surface, said method comprising: providing a protein solution comprising water and retinin, retinin-like protein, or cuticular protein; and contacting a polymer surface with the protein solution, thereby providing a coated surface.

[0100] In embodiments, the protein solution can comprise one of retinin, retinin-like protein, or cuticular protein. In other embodiments, the protein solution can comprise two or all three of retinin, retinin-like protein, and cuticular protein. Preferably, the retinin, retinin-like protein, or cuticular protein is retinin or retinin-like protein.

[0101] Cuticular proteins may be found throughout the fauna of nature and include, but are not limited to, cuticular proteins from arthropod organisms, hereunder insects. These proteins are found in the surface of the exoskeleton of arthropods including insects, and may thus preferably be insect cuticular protein.

[0102] Another embodiment relates to the methods as described herein, wherein the retinin, retinin-like protein, or cuticular protein originates from an arthropod or insect. A further embodiment of the present invention relates to the methods as described herein, wherein the retinin, retinin-like protein, or cuticular protein comprises a secretory signal peptide and / or have a size in the range of 10 kDa to 35 kDa.

[0103] Retinin is a protein that is restricted in its expression to the cornea of insects. It belongs to the group of small insect proteins possessing the functionally uncharacterized retinin C-domain. Although insects constitute a diverse group of animals, the retinin protein is predicted to be present in several distinct orders of insect whose genomes have been sequenced to-date (see e.g. PF AM database of EMBL-EBI).

[0104] Thus, an embodiment of the present invention relates to the methods as described herein, wherein the retinin, retinin-like protein, or cuticular protein originates from an insect belonging to the order Diptera. In another embodiment, the insect belongs to a family selected from the group consisting of Drosophilidae, Culicidae, Calliphoridae, Phoridae, Tephritidae, Psychodidae, Muscidae, Chironomidae, Corethrellidae, and Glossinidae. In a further embodiment, the insect belongs to a genus selected from the group consisting of Drosophila, Anopheles, Culex, Aedes, Wyeomiya, Lucilia, Megaselia, Bactrocera, Lutzomiya, Phlebotomus, Musca, Stomoxys, Clunio, Corethrella, Ceratitis, and Glossina.

[0105] In yet another embodiment, the insect is a species selected from the group consisting of D. melanogaster, D. virilis, D. busckii, D. erecta, D. simulans, D. yakuba, D. sechellia, D. pseudoobscura, D. ananassae, D. persimilis, D. suzukii, D. grimshawi, D. willistoni, D. mojavensis, D. ficusphila, Anopheles gambiae, Anopheles darlingi, Anopheles sinensis, Anopheles aquasalis, Anopheles albimanus, Anopheles arabiensis, Anopheles atroparvus, Anopheles christyi, Anopheles coluzzii, Anopheles culicifacies, Anopheles dims, Anopheles epiroticus, Anopheles farauti, Anopheles funestus, Anopheles maculatus, Anopheles melas, Anopheles merus, Anopheles minimus, Anopheles quadriannulatus, Anopheles stephensi, Culex quinquefasciatus, Aedes aegipty, Aedes albopictus, Wyeomiya smithii, Lucilia cuprina, Megaselia scalaris, Bactrocera cucurbitae, Bactroteca latifrons, Bactrocera dorsalis, Lutzomiya longipalsis, Phlebotomus papatasi, Musca domestica, Stomoxys calcitrans, Clunio marinus, Corethrella appendiculata, Ceratitis capitata, Glossina brevipalpis, Glossina fuscipes, Glossina pallidipes, Glossina palpalis, Glossina morsitans, and Glossina austeni.

[0106] In gambiae, the protein CPR10 found in the eye was identified as a retinin-like protein involved in nanostructure formation. An embodiment of the present invention relates to the method as described herein, wherein the insect is Anopheles gambiae.

[0107] An extensively studied genus of insects is Drosophila, an in particular D. melanogaster, which has served as model for many basic research studies. Proteomic characterization of a large group of species within the Drosophila genus revealed a protein composition similar to that of D. melanogaster, including the presence of the retinin protein. These findings support the notion that retinin is an important structural protein found in a large variety of insects. Therefore, in an embodiment, the insect belongs to thegenus Drosophila.

[0108] In yet other embodiments, the insect is a species selected from the group consisting of D. melanogaster, D. virilis, D. busckii, D. erecta, D. simulans, D. yakuba, D. sechellia, D. pseudoobscura, D. ananassae, D. persimilis, D. suzukii, D. grimshawi, D. willistoni, D. mojavensis, and D. ficusphila. In a preferred embodiment, the insect is D. melanogaster.

[0109] In the Drosophila species, numerous retinin-like genes are present on the same locus of the chromosome arm 3L, suggestive of recent duplication events. The data available from the Drosophila gene expression atlas (Fly Atlas) shows that the retinin-like genes from this cluster are expressed in other tissues than the eye, where retinin is strongly expressed. Many of these encode the retinin C-domain, including, but not limited to,CG13044, CG13062, CG13040, CG4962, CG13041, CG13042, CG13043, CG13060, CG4982 and CG13063. Thus, in an embodiment, the retinin or retinin-like protein is selected from the group consisting of CG13044, CG13062, CG13040, CG4962, CG13041, CG13042, CG13043, CG13060, CG4982 and CG13063.

[0110] Furthermore, inD. melanogaster , the protein CGI 3059 was identified as a retinin-like protein involved in nanostructure formation.[oni] In an additional embodiment, the retinin, retinin-like protein, or cuticular protein originates from an insect belonging to the subclass Pterygota.

[0112] By recombinant expression of retinin, retinin-like protein, or cuticular protein in a suitable expression system followed by purification of the recombinant protein, relatively large yields of recombinant retinin, retinin-like protein, or cuticular protein can be obtained. This protocol is based on well-known techniques, such as E. coli recombinant protein expression and standard Ni-NTA purification, and therefore should be readily accessible without need for advanced and expensive equipment. Thus, in embodiments, the retinin, retinin-like protein, or cuticular protein is recombinant retinin, retinin-like protein, or cuticular protein.

[0113] Without being bound by theory, it is hypothesized that the importance of retinin in the formation of corneal nanocoatings is linked to the common retinin C-domain that is shared among retinin and retinin-like proteins of a variety of insects. Therefore, in an embodiment of the present disclosure, the retinin or retinin-like protein comprises the common Retinin C-domain designated PF04527 as described in the PF AM database of protein families, also known as Retinin-like domain designated IPR007614 as described in the InterPro database of proteins.

[0114] The nanostructure formation coating layer described herein provides the coated surface with properties directly derivable from the formed nanopattems on the surface, e.g. anti-reflectivity, anti-wetting, etc. However, it is possible to endow the coated surface with additional properties through modification of the recombinant retinin, retinin-like protein, or cuticular protein. Thus, the recombinant production of retinin, retinin-like protein, or cuticular protein may be preceded by a design phase in which modification to the protein is schemed to obtain a desirable functionality. Modification of the protein may be carried out using standard molecular biology techniques known to the person skilled in the art.

[0115] Thus, in an embodiment of the present disclosure, the retinin, retinin-like protein, or cuticular protein comprises one or more modifications. The one or moremodifications can be selected from the group consisting of a fluorescent label, a radioactive label, a metal capturing moiety, an antibody -binding site, and an enzyme-binding site.

[0116] The proteins of the invention can be readily obtained, e.g. by recombinant expression in bacteria, as described herein. For efficient formation of nanocoatings, only relatively low concentrations of protein is required. The low consumption of material ensure that costs of the method are kept at a minimal. Thus, for example, in embodiments, the concentration of retinin, retinin-like protein, or cuticular protein in the aqueous protein solution is from 0.1 mg / mL to 2.0 mg / mL, such as from 0.2 mg / mL to 1.5 mg / mL, such as 0.3 mg / mL to 1.0 mg / mL, such as 0.4 mg / mL to 0.8 mg / mL, such as 0.5 mg / mL to 0.7 mg / mL, preferably 0.6 mg / mL.

[0117] A solvent of the protein solution may be adjusted to induce proper nanocoating formation. The solvent may comprise, for example, salts to regulate stability and interaction of the main constituents of the nanocoating. In embodiments, the protein solution is provided in aqueous solutions.

[0118] As desired, the aqueous solutions can be a solution comprising at least one salt, such as saline solutions.

[0119] Also as desired, the aqueous solution can comprise a buffering system. Suitable buffering systems can be selected from the group consisting of TRIS, phosphate, HEPES, MOPS, and PIPES, although others may also be suitable.

[0120] The formation and final morphology of the nanocoating may be affected by a range of protocol parameters that can be adjusted to induce formation of a homogeneous nanocoating.

[0121] The contacting of the protein solution to the substrate or surface to be coated can be conducted at ordinary temperatures, humidities, and pressure. For example, the contacting can be performed at a temperature in the range of 0-50°C, such as 10-40°C, such as 15-35°C, such as 20-30°C, such as at or around 25°C. The contacting can also be performed at a humidity in the range of 10-90%, such as 35-75%, such as 40-70%, such as 45-65%, such as at or around 50%. The contacting can also be performed at a pressure of about 1 atmosphere. In general, the contacting is conducted at ordinary room temperature conditions such as standard temperature and pressure.

[0122] According to embodiments, the contacting of the protein solution to the substrate or surface to be coated can be conducted for any suitable time that permits adequate and desired formation of the nanostructure formation coating layer. For example, the contacting can be conducted for as little as 1 minute, 2 minutes, 3 minutes, 4 minutes, 5minutes, or more to as much as or 10 minutes, 15 minutes, 20 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 180 minutes, or more. The contact time can be adjusted, for example, depending upon the structure to be coated, the concentration of the protein in the protein solution, and the like. Contacting can be conducted, for example, by immersing the substrate or surface into the protein solution, or the like.

[0123] Optionally, the coated surface can be subjected to drying once the contacting is completed. For example, the coated surface can be dried for at least 5 minutes, such as for at least 10 minutes, such as for at least 15 minutes, or for approximately 20 minutes.

[0124] In other embodiments, the coated surface can be rinsed following the contacting. Rinsing had the effect of removing any unreacted protein and other materials. The coated surface can then be suitably stored in a dry or wet environment, as desired, or may be subsequently employed for the products intended purpose. For example, in the case of the coated surface being a coated contact lens, the coated contact lens can be rinsed with a conventional contact rinsing solution. The coated contact lens can then be stored in a suitable case, or can be inserted in a person’s eye.

[0125] The methods described herein are suitable for use with any type of surface to which the liquid mixture comprising the retinin, retinin-like protein, or cuticular protein can adhere. In the case of the express absence of a lipid, wax, or surfactant, or in the case where such a lipid, wax, or surfactant is not present in amounts otherwise necessary for formation of the nanostructure formation coating layer on non-polymeric surfaces, the surface is generally a polymer surface. Thus, products made from a wide range of materials may benefit from the nanostructure formation coating layer described herein. Contact lenses, sunglasses, displays and solar panels are all examples of products in which optical coatings are extensively used to adjust e.g. anti-reflectivity, absorbance etc. Another group of products suited for nanocoating is biological implants on which nanocoatings may regulate in vivo cell and / or tissue attachment, growth and differentiation. Regulation of cell and / or tissue attachment, growth and differentiation may also be achieved in vitro by nanocoating of surfaces and proper laboratory protocols. Especially regulation of attachment, growth and differentiation of cells and / or tissues, such as bone and cartilage cells and / or tissues, as well as stem cells of various types are applications that would benefit from the methods described herein. A broad palette of materials is used for the products as exemplified above.

[0126] For some applications, depending on the constituents of the nanostructure formation coating layer and the targeted surface, it may be favorable to extend the protocol toencompass additional process steps, such as washing, drying and material deposition on the surface.

[0127] In one exemplary embodiment, the present disclosure can be integrated into a routine care regimen for contact lenses, such as by an end-user consumer at home. Typical lens care routines involve daily rinsing and storage, such as rinsing the contact lenses when the user removes them from their eyes, followed by storage (such as overnight) of the lenses in a storage solution. That daily rinsing and storage can, for example, involve the following steps:1) Clean the lens with a cleaning solution (e.g. Menicare Plus®) after removing it from the eye.2) Rinse the lens with a rinsing solution (e.g. Menicare Plus®).3) Store the lens in a suitable case or container immersed in a soaking solution (e.g. Menicare Plus®).4) The next day, or at some later point in time, remove the lens from the lens case / soaking solution and wear it as is.

[0128] The typical lens care routines also generally include a recommended periodic more thorough cleaning of the lenses, such as semi-weekly, weekly, biweekly, or the like. That more thorough cleaning can, for example, involve the following steps:1) Soak the lens in an intensive cleaner (e.g. Progent®) to remove protein and other deposits. This step can remove protein and other deposits that conventionally accumulate during wearing of contact lenses, and can also remove any remaining portions of a previously-applied nanostructure formation coating layer so as to provide a clean contact lens surface.2) Rinse the lens with a rinsing solution (e.g. Menicare Plus®). This step can be conducted to remove any remaining intensive cleaner, which might interfere with subsequent formation of a new nanostructure formation coating layer.3) Soak the lens in the nanostructure formation coating solution. Drying of the lens before and / or after the soaking step is not needed.4) Rinse the lens with a rinsing solution (e.g. Menicare Plus®). This step removes any residual unreacted protein solution from the contact lens surface, without affecting the newly-formed nanostructure formation coating layer.5) Store the lens in a suitable case or container immersed in a soaking solution (e.g. Menicare Plus®).6) The next day, or at some later point in time, remove the lens from the lens case / soaking solution and wear it as is.

[0129] Benefits of an additional washing and / or drying step or steps include more homogeneous nanostructure formation coatings and the removal of residual unbound constituents of the liquid protein solution. The solvent used for washing of the surface is preferably an aqueous solvent, such as water or alcohols. In embodiments, the solvent for washing is selected from the group consisting of aqueous solutions, ethanol, and polar solvents, or water.

[0130] If desired, the step of contacting the surface to be coated with the protein solution can be repeated one or more additional times.

[0131] In embodiments, the protein (retinin, retinin-like protein, or cuticular protein) can be provided in an already -prepared solution, such as an already prepared sterilized solution of the protein, or can be provided in powder form, such as a sterilized lyophilized protein powder form. If in solution form, the solution may be in any suitable form, such as aqueous solution optionally including additives such as buffers, diluents, pH / osmotic adjustment agents, and the like. The protein solution can also be provided in any suitable concentration, such as in a concentration ready for use to form a desired nanostructure formation coating, or in a more concentrated form that is to be further diluted prior to use.

[0132] However, in embodiments it is preferred that the protein be provided in powder form and then reconstituted prior to use, as this provides improved stability and shelflife. Thus, in this embodiment a kit can be provided that includes a first component being the protein (retinin, retinin-like protein, or cuticular protein) in powder to concentrated solution form, and a second component being a diluent such as water or an aqueous solution of water and one or more additives such as buffers, diluents, salts, pH / osmotic adjustment agents, and the like. Preferably, such as where the kit is intended for use with contact lenses worn by a person, the components of the kit are sterilized.

[0133] In such embodiments, the protein component and / or the diluent component can be provided in single-use or multi-use form. For example, in consideration of maximum stability, shelf-life, and sterility, the two components can be provided in single-use form, where the two components are mixed, used, and then the residual is discarded. Multiple sets of kits can also be included in a larger kit, such as in the case of multiple single-use kits packaged together. In other forms, a kit may include, for example, multiple single-use forms of the protein component and a larger multi-use form of the diluent.

[0134] If desired, the kit may also include one or more containers suitable for use for mixing the protein component and the diluent component. Alternatively, a containercontaining the protein component or a container containing the diluent component can be provided in such a size as to permit mixing of the two components in that container.

[0135] The kit may also include suitable instructions for a user to reconstitute or further dilute the protein component and how to use a final aqueous solution of protein to form the desired nanostructure formation coating.

[0136] The material of the surface to be coated depends largely on the nature of the product. Some products require the material to be rigid, curvy, elastic or transparent, thereby indirectly predisposing the use of some materials. The coated surfaces as described herein is not limited to a specific group of materials and the nanostructured patterns making up the nanostructure formation coating layer may be formed on any material and on all geometries as opposed to some conventional techniques for formation of coatings that may be restricted to, for example, flat geometry or hard materials.

[0137] The nanocoating can be further functionalized by inclusion of one or more modifications. The modifications of the coated surface may be either positioned and / or attached directly of the surface or as part of the nanocoating. Thus, the modification may be part of one of the constituents of the nanocoating, e.g. a part of the recombinant retinin, retinin-like protein, or cuticular protein. For example, and depending upon the desired use of the coated product, suitable modifications can include but are not limited to a fluorescent label, a radioactive label, a surface-bound metal, an antibody, and an enzyme.

[0138] A preferred embodiment of the present invention relates to the use as described herein, wherein the coated product is contact lenses.

[0139] Also provided herein are various methods for forming, retreating, refreshing, and / or rejuvenating a nanostructure formation coating layer on a surface.

[0140] In one embodiment is provided a method of forming a coated medical device such as a lens. The method comprises providing a medical device such as a lens comprising a polymer surface; coating the polymer surface with an aqueous solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a coating on the lens. Following the coating step, the coated medical device can be optionally rinsed with an appropriate rinsing solution such as water, a saline solution, or the like, and optionally dried. The result is a coating comprising a nanostructure formation on the polymer surface, the nanostructure formation comprising the one or more of retinin, retinin-like protein, and cuticular protein attached to said polymer surface.

[0141] The disclosure is not limited to methods for initially forming a nanostructure formation coating layer on a surface. In addition, the disclosure encompasses methods where an existing nanostructure formation coating layer is retreated, refreshed, or rejuvenated to restore the nanostructure formation coating layer and counter any degradation that may have occurred.

[0142] Thus, in another embodiment is provided a method for cleaning a medical device such as an ophthalmic lens. Such a method initially comprises contacting the medical device such as an ophthalmic lens with a protein cleaning solution to remove protein deposits from the ophthalmic lens. This step removes from the medical device any protein contaminants that may be present on the surface and / or may remove part or all of any preexisting nanostructure formation coating layer from the surface. Next, the medical device such as an ophthalmic lens is contacted with an aqueous conditioning solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a nanostructure formation coating layer of the one or more of retinin, retinin-like protein, and cuticular protein attached to an outer surface of the ophthalmic lens. The coated medical device may thereafter optionally be rinsed with a rinsing solution. Such rinsing solutions are known in the art for various types of medical devices including for contact lenses.

[0143] In the above method, it is also sometimes desirable to rinse the medical device with a rinsing solution between the steps of using a protein cleaning solution and using the aqueous conditioning solution. This removes any leftover protein cleaning solution prior to the use of the protein solution that forms the nanostructure formation coating layer.

[0144] In each of the above methods, and particularly in the case of the medical device being a contact lens, the coated medical device may subsequently be stored in an appropriate storage solution until the medical device is to be used. Such contact lens storage solutions are known in the art.

[0145] A feature of embodiments is that the nanostructure formation is formed on an underlying polymer surface from a protein solution comprising the one or more of retinin, retinin-like protein, and cuticular protein and specifically in the absence of a long-chain inhibitor such as a lipid, wax, or surfactant. It has been discovered that the polymer surface has polymer brushes on the lens surface, which interact with the retinin, retinin-like protein, and cuticular protein to enable formation of the nanostructure formation coating layer. Thisis a behavior that occurs on the polymer surface, but does not occur on other surfaces such as metal, glass, or other materials.

[0146] Thus, in embodiments, the nanostructure formation is formed on an underlying polymer surface from a protein solution comprising the one or more of retinin, retinin-like protein, and cuticular protein and specifically in the absence of a long-chain inhibitor such as a lipid, wax, or surfactant. That is, no lipid, wax, or surfactant that would have an inhibiting effect on the retinin, retinin-like protein, and cuticular protein is present, either in a protein solution of the retinin, retinin-like protein, and cuticular protein, in an aqueous solution that might be mixed with such a protein solution, or otherwise added to or present in the system of nanostructure formation.

[0147] In other embodiments, it is understood that a minor amount of such a lipid, wax, or surfactant may be present, albeit in an amount that would not be expected to have any significant effect on the nanostructure formation system. For example, U.S. Patent No. 12,104,078 discloses the required presence of a lipid emulsion or suspension in a substantial amount, such as a ratio of protein solution to lipid emulsion or suspension in a range of 1 : 1 to 1:10 vol / vol, where the concentration of the protein solution is rather low, such as from 0.1 mg / mL to 2.0 mg / mL, and the concentration of the lipid emulsion or suspension is 0.1 g / mL.

[0148] Polymer brushes generally consist of polymer chains that are anchored at one end to a surface, either through physical adsorption or covalent bonding. The other end of the polymer chains is free to extend into and interact with materials in the surrounding environment, forming a brush-like structure. This configuration allows the polymer chains to stretch out when exposed to a good solvent, reducing the overall free energy of the system.

[0149] The properties of polymer brushes are influenced by several factors, including chain length, surface attachment, and responsive capabilities. The length of the polymer chains affects the thickness and density of the brush, impacting its flexibility and load-bearing capacity. The method of tethering the polymer chains (for example, grafting-to or grafting-from) influences the stability and uniformity of the brush. Incorporating stimuli-responsive monomers can enable the brushes to change their characteristics in response to external stimuli, such as pH or temperature.

[0150] In embodiments, any suitable polymer materials conventionally used for forming contact lenses or other medical devices may be used as the polymer surface on which the nanostructure formation coating layer is to be formed. In embodiments, these polymers have the polymer brush feature described herein. Generally, the present disclosure is applicable to polymeric contact-lens materials formed from monomers or comonomerscomprising aliphatic hydrocarbon segments, including those containing -(CH2)n- moieties, where n represents a number of such moieties, which give rise to polymer chains capable of forming surface-associated polymer brushes. These polymer brushes may arise from grafted, surface-enriched, or chain-extended polymer segments and are not limited to a specific polymer chemistry.

[0151] Examples of such suitable polymers include styrene-(meth)acrylate copolymers, styrene-silyl / siloxanyl copolymers, silicone-containing copolymers such as silicone-based (meth) acrylate or silicone-based styrene copolymers, and the like. More particularly, examples include but are not limited to: crosslinked copolymers including a (meth)acrylic monomer, a styrene monomer, and other monomers, such as disclosed in U.S. Patent No. 5,041,511; a polymer formed using a styrene monomer containing a silyl or siloxanyl group, such as disclosed in U.S. Patent No. 4,594,401; a copolymer obtained by polymerizing a copolymerization component including specific silicone-containing monomers, such as disclosed in Japanese Patent Publication No. JP 2005-181729 A; and the like. The entire disclosures of these patents and patent publications are incorporated herein by reference. Of course, the present disclosure is not restricted to these specific polymers, but may be implemented using a wide range of polymeric contact-lens materials having surface polymer chains capable of exhibiting brush-like behavior.

[0152] Investigation focusing on how nanostructures formed led to the conclusion that the nanostructures are formed by the Turing Model, which relates the action of a slowly diffusing activator (u) and a fast diffusing inhibitor (v). The Turing Model is:

[0153] According to the present disclosure, it was found that the activator is the retinin, retinin-like protein, and / or cuticular protein, and the inhibitor is the polymer brushes on the underlying polymer substrate surface. According to the present disclosure it is thus possible to form the nanostructure formation coating directly on the polymer substrate surfacein the absence of a long-chain inhibitor such as a lipid, wax, or surfactant that is required for other surfaces such as glass, metal, and the like. That is, the polymer substrate surface itself provides the inhibitor to the retinin, retinin-like protein, and / or cuticular protein activator. This was surprising, because according to the Turing Model, the inhibitor must diffuse across space. Therefore, the polymer surface being the inhibitor should its surface formed as tightly packed and cross-linked polymers should not be mobile to provide an inhibitory action.

[0154] Investigation discovered that under conditions such as salt, pH, and the like, the end of the tightly packed and cross-linked polymers unfold in space to form the polymer brushes. The free ends of these polymer brushes are thus able to move in space, and interact with and provide an inhibitory effect upon the retinin, retinin-like protein, and / or cuticular protein, allowing formation of the nanostructure formation coating as the polymer brushes collapse into their original tightly packed configuration. This is shown generally in Figs. 16A and 16B.

[0155] Existence of the polymer brushes was confirmed by Atomic Force Microscopy of a contact lens. In an air environment, Atomic Force Microscopy showed mushroom-like structures on the lens surface, indicative of the existence of collapsed polymer brushes. The structures were further investigated under hydrophobic and hydrophilic environments. Atomic Force Microscopy showed that in a hydrophobic solution, the polymer brushes were compacted and collapsed on the lens surface; however, in a hydrophilic solution, the polymer brushes were relaxed and extended into space from the lens surface.

[0156] Under contact mode measurement, application of force to the contact lens surface led to collapse of the outer layer. This phenomenon was also clearly evidenced by changes in the lens topography, conclusively indicating presence of a soft layer on the contact lens surface, indicative of polymer brushes on the lens surface.

[0157] This phenomenon was further investigated using electron microscopy, which indicated a less dense layer on the lens surface, likewise indicative of polymer brushes on the lens surface. Following application of the nanostructure formation coating of the present disclosure, electron microscopy showed that the outer layer of the lens was less transparent to electrons and an absence of polymer brushes on the lens surface, indicating that the polymer brushes had collapsed during formation of the nanostructure formation coating.

[0158] The above embodiments are described as using an aqueous solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and not comprising a long-chain inhibitor to form a coating on the lens. However, it will beunderstood that in some embodiments, some amount of a long-chain inhibitor such as lipid, wax, and / or surfactant may be present in the coating solution and / or on the surface to be coated. However, the amount of such long-chain inhibitor present is such that it does not itself fully cause formation of the nanostructure formation coating layer. That is, the polymer brushes present on the underlying polymer surface positively contribute to the formation of the nanostructure formation coating layer, thereby helping to anchor the nanostructure formation coating layer to the polymer surface.

[0159] For example, in an embodiment an amount of such long-chain inhibitor present may be limited such that at least 25% of formation of the nanostructure formation coating layer is attributable to presence of polymer brushes on the underlying polymer surface. In embodiments, an amount of such long-chain inhibitor present is limited such that at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of formation of the nanostructure formation coating layer is attributable to presence of polymer brushes on the underlying polymer surface. For example, U.S. Patent No. 12,104,078 describes that nanostructure formation coating layers can be provided on surfaces such as metal and glass using a ratio of protein solution to lipid emulsion or suspension in a range of 1 : 1 to 1 : 10 vol / vol. The concentration of the protein solution is described as being such as from 0.1 mg / mL to 2.0 mg / mL. A concentration range for the lipid emulsion or suspension is not described, although the Examples use concentrations of 0.1 g / mL. In such concentrations, embodiments of the present disclosure may encompass an amount of such long-chain inhibitor being present in a ratio of protein solution to lipid emulsion or suspension in a range of greater than 1:1, such as from 1,000,000: 1 or from 500,000: 1 or from 100,000:1 or from 50,000:1 or from 10,000:1 or from 1,000:1 or from 500:1 or from 100:1 to 2:1 or to 3:1 or to 4:1 or to 5:1 or to 10:1 or to 20:1 or to 25:1 or to 50:1 or to 100:1 vol / vol. Higher ratios and ratios within these ranges are encompassed.

[0160] Put another way, in embodiments where a long-chain inhibitor such as lipid, wax, and / or surfactant is present, the long-chain inhibitor may be present such that a weight ratio of protein (retinin, retinin-like protein, and / or cuticular protein) to long-chain inhibitor in a range of greater than 1:1, such as from 1,000,000:1 or from 500,000:1 or from 100,000:1 or from 50,000:1 or from 10,000:1 or from 1,000:1 or from 500:1 or from 100:1 to 2:1 or to 3:1 or to 4:1 or to 5:1 or to 10:1 or to 20:1 or to 25:1 or to 50:1 or to 100:1. Higher ratios and ratios within these ranges are encompassed. In other embodiments, a long-chain inhibitor such as lipid, wax, and / or surfactant may be absent or substantially absent.

[0161] A particular advantageous use according to the present disclosure is in providing improved Orthokeratology (Ortho-K) lenses. Orthokeratology (Ortho-K) lenses are rigid gas-permeable (RGP) lenses worn overnight to reshape the cornea that offer a non-invasive approach for myopia control. However, their extended wear time imposes high demands related to surface comfort, hygiene, and resistance to biofouling. It has been found that a previously unrecognized feature of Ortho-K lenses is the presence of surface-bound brush-like polymer structures on the exterior polymer surface. These brush-like polymer structures have been found to be highly mobile and significantly contribute to hydrophobicity, protein adsorption, and microbial adhesion. These properties are detrimental to satisfactory usage of Ortho-K lenses.

[0162] A drawback of Ortho-K lenses is that their design includes a reverse curve in its surface structure. This reverse curve provides an ideal location for the build-up of deposits during use. These deposits can include protein, foreign matter such as dust, and the like. These deposits can in turn lead to onset of microbial keratitis and inflammatory reactions, which can hinder the widespread adoption of orthokeratology. To ensure the continued safe and comfortable use of Ortho-K lenses, it is crucial to clean, rinse, and disinfect the lenses after each removal.

[0163] One solution to this deposit build-up problem is to subject the Ortho-K lenses to rigorous periodic cleaning. For example, it has been strongly recommended to use potent protein removers on a weekly basis for thorough cleaning of the lenses. Even with these strong protein removers, intensive rubbing of the lens during the washing process is necessary to achieve optimal results, especially in the reverse zone of the Ortho-K lens. This often leads to unsuccessful deposit removal, such as where the cleaning process is not rigorous r is not performed as often as necessary, and / or appearance of scratches or other damage to the lens as a result of the rigorous cleaning.

[0164] With the materials and methods of the present disclosure, it is possible to further address the deposit build-up problem while providing additional benefits.

[0165] The nanostructure formation coating layer applied to the surface of Ortho-K lenses not only helps facilitate the cleaning process, but it also enhances comfort during lens insertion and removal, and improves safety, even in cases of poor care compliance by the user. These benefits are provided because the presence of the nanostructure formation coating layer means that less rigorous rubbing of the lens is required, which in turn means that there is less risk of accidental breakage caused by rubbing. By minimizing or potentially eliminating the need for a rigorous rubbing step, the cleaning process for Ortho-K and otherlenses can be made more routine and easier for the user. This may in turn lead to better compliance by users to routine cleaning procedures, reducing or eliminating the risk of microbial contamination. The present disclosure thus provides a process for easy and safe deposit removal, added comfort to the user, less damage to the lens, plus additional antibacterial and antiviral benefits.

[0166] The nanostructure formation coating layer also imparts a cushioning effect to the lenses, enhancing user comfort during lens insertion and especially during lens removal.

[0167] Another characteristic of the nanostructure formation coating layer of the present disclosure is that it provides a reduced wettability contact angle compared to the uncoated polymer substrate or surface. For example, a wettability contact angle of the nanostructure formation coating layer can be from about 20, about 25, about 30, or about 35 to about 45, about 50, about 55, about 60, about 65, about 70, or about 75 degrees (as measured using deionized water). Contact angle values within these ranges are encompassed.

[0168] These advantages and beneficial effects provided to Ortho-K lenses, and to other lenses and medical devices, provide improved hygiene and safety to users. This allows more widespread use of such lenses and medical devices, and provide improved quality of life to users.

[0169] In some embodiments, the medical device is not a contact lens. In such embodiments, the medical device may be configured to be implantable within a mammalian body. In a non-limiting example, the medical device is a stent configured to keep a cavity open. In another non-limiting example, the stent is configured to keep a blood vessel, bile duct, intestine, nasal passage or cavity, sinus cavity, or intraocular channel open.

[0170] In some embodiments, the medical device is a sensor, camera, vital sign monitor, drug depot device, neurostimulator, ultrasound, silicone implant, saline implant, hernia mesh, penile implant, intrauterine device, orthopedic rod or plate or pin or nails, pacemaker, cardiac valve, ear tube, aneurysm coil, or intraocular lens.

[0171] In some embodiments, the medical device is a test strip. Various nonlimiting examples includes a drug, salivary, urine, blood, interstitial fluid, genetic testing, or semen test strip.

[0172] In some embodiments, the medical device is a tool configured to be inserted within a mammalian body. Various non-limiting examples include a catheter, trocar, endoscope, probe (e.g., interstitial, microdialysis, etc.), or laparoscope.

[0173] In some embodiments, the medical device is configured to be used externally on a mammalian body, for example, for use as a bandage, wound dressing, external sensor, hearing aid, or artificial skin.

[0174] In addition to other benefits described herein, the nanostructure formation coating layer unexpectedly provides any of a range of benefits to the underlying medical device, such as a contact lens, and to users of such medical devices. For example, the nanostructure formation coating layer has been found to provide benefits including one or more of increased antimicrobial properties, increased resistance to lipid adhesion, and antioxidant surface activity. In the case of contact lenses in particular, the nanostructure formation coating layer provides higher levels of hygiene and enhanced user comfort during lens insertion and especially during lens removal.

[0175] Examples

[0176] Example 1 - Detailed Study

[0177] 1. Results

[0178] 1,1. Development and characterization of nanostructure formation coating

[0179] The above biomimetic protein-based nanocoatings (nanostructure formation coating layer) inspired by insect corneal nanostructures can be easily fabricated and adapted for various surfaces and applications. The present inventors developed a standardized protocol for the uniform coating of rigid gas-permeable (RGP) contact lenses with a nanostructure formation coating layer, mimicking natural nanostructures. This protocol involves immersing the lenses in a solution of one or more of retinin, retinin-like protein, and cuticular protein (hereafter referred to as a Retinin-like or RetL protein solution) for 10 minutes, resulting in a stable and homogeneous nanocoating. Since RGP lenses share the same material composition as Ortho-K lenses but are more cost-effective and easier to handle, they were used as a model system for the screening of coating conditions and protein variants.

[0180] 1,1.1. Structural analysis of the nanocoating

[0181] To evaluate the uniformity and structural integrity of the nanostructure formation coating, the inventors conducted transmission electron microscopy (TEM) on cryosectioned lenses. TEM images confirmed the presence of a continuous nanocoating on the lens surface. See Figs. 3 A through 6C. Cross-sectional profiles showed a reduction in electron permeability in the outermost layer following coating application, consistent with the deposition of a material chemically distinct from the lens substrate. See Figs 3 A and 3B. This distinction arises from the stronger interactions between the protein coating andosmium / uranium staining agents, compared to the lens polymers, which also enables precise measurement of the nanocoating thickness.

[0182] Atomic force microscopy (AFM) further characterized the formation of nanocoatings, revealing the nucleation of nanostructures at the 5-minute time point and their full development into a uniform layer within 10 minutes. The resulting surface exhibited characteristic nipple- and maze-like nanostructures, which significantly increased the adhesion force of the lens surface. See Figs. 4A to 5C. This adhesion force, measured directly by AFM, is largely governed by interactions with the surface-bound water layer, making it a sensitive indicator of surface hydrophilicity. The observed increase thus reflects the enhanced hydrophilic character of the nanocoated surface, contributing to improved lens wettability.

[0183] 1,1.2. Wettability and hydrophilic properties

[0184] RGP contact lenses are inherently hydrophobic, making the establishment of a stable hydrophilic surface critical for both wearer comfort and resistance to biofouling. To assess the effect of the AEGIS nanocoating on surface wettability, the inventors performed static water contact angle measurements before and after coating application. The 10-minute coating process led to a dramatic reduction in contact angle from 108.5° to 46.5°, indicating a substantial enhancement in surface hydrophilicity. See Figs. 6A and 6C. A contact angle below 60° is generally considered optimal for ensuring the biocompatibility of contact lenses, and the modified surfaces clearly met this criterion. Enhanced wettability not only improves comfort but also plays a key role in minimizing the accumulation of protein and lipid deposits, thereby supporting better ocular health during lens wear.

[0185] To further validate the hydrophilic properties of the nanostructure formation coating, the inventors performed a lipid adhesion assay using oleic acid, a representative tear film lipid. This test simulates real-world conditions in which contact lenses are exposed to lipid contaminants during wear. The improvement in surface wettability resulted in remarkable lipid-repellent behavior: while non-coated lenses readily accumulated oleic acid deposits, nanostructure formation-coated lenses showed virtually no lipid adhesion. See Figs.6B and 6C. The inability of lipids to adhere to the highly hydrophilic nanocoated surface underscores the dual benefit of the coating: enhancing wearer comfort and effectively preventing contamination by tear film components.

[0186] 1,1.3. Renewability and deposit-resistance

[0187] Given that Ortho-K lenses are typically worn for a year or even a longer time period, it is crucial for any applied coating to be both renewable and resistant to long-term biofouling. To assess the reapplication potential of a nanostructure formation coating layer, lenses were treated with Progent™ (Menicon) solution, an intensive enzymatic cleaner formulated to remove deposits from the contact lenses. As expected, Progent™ effectively stripped the nanostructure formation coating layer, restoring the lens to its original hydrophobic state. See Fig. 7. The nanocoating was then successfully reapplied, reestablishing both its hydrophilic and lipid-repellent properties.

[0188] The inventors also evaluated the resistance of the coating to protein deposition using a bovine serum albumin (BSA). See Fig. 8. Uncoated lenses exhibited persistent protein fouling even after thorough mechanical washing by using MeniCare™ (Menicon) solution, whereas nanostructure formation-coated lenses remained largely resistant to BSA accumulation. Remarkably, following BSA exposure and subsequent washing, nanostructure formation-coated lenses reverted to a hydrophobic state, similar to fresh, untreated lenses, indicating effective removal of the protein. In contrast, uncoated lenses retained a hydrophilic character after BSA treatment and cleaning, suggesting incomplete removal of protein deposits. These residual proteins may serve as nucleation sites for further fouling by additional proteins and microorganisms. AFM analysis confirmed the distinction between the two deposition patterns: the nanostructure formation coating layer formed a uniform, continuous nanolayer, whereas BSA appeared as randomly distributed nano- and microscale aggregates on the lens surface. See Fig. 8.

[0189] 1,1.4. Coating resilience

[0190] Previous studies have shown that protein-based nanocoatings derived from RetL proteins exhibit a Young’s modulus (a measure of material stiffness) in the range of 3-9 GPa whereas RGP contact lenses typically fall within the 1-3 GPa range. The relatively soft core material of RGP lenses allows them to undergo mechanical deformation during routine handling and care without breaking, which is beneficial for lens durability. However, this softness also makes the surface more susceptible to scratches. Given the higher stiffness of the nanostructure formation coating layer, its application is expected to further reinforce the lens surface, significantly enhancing resistance to abrasion and contributing to the overall mechanical robustness of the lens.

[0191] To evaluate the long-term stability of the nanostructure formation coating under mechanical and chemical stress, the inventors subjected coated lenses to a simulated wear protocol involving repeated washing cycles, intended to mimic daily lens care over a week of use. Hydrophilicity was monitored throughout this process by measuring changes of the contact angle after each wash. The measurements showed a gradual decline in wettabilityover the course of seven washing cycles. See Fig. 9. During the first five days, the contact angle exhibited only minor changes, suggesting that the nanocoating maintained its hydrophilic functionality. However, a statistically significant increase in contact angle, indicating reduced wettability, was observed starting from the fifth wash. By the seventh wash, although degradation had progressed, the contact angle still remained below 60°, suggesting that a residual functional layer persisted on the surface, continuing to offer partial hydrophilic and anti-fouling properties.

[0192] To further probe the physical integrity of the coating, microscopy imaging was employed. AFM topography scans revealed that the nanostructured coating remained largely uniform and intact after the initial washes. See Fig. 10. However, as the number of washing cycles increased, progressive deterioration of the nanostructures became apparent. By the sixth and seventh cycle, significant surface roughening and partial delamination were observed, consistent with the hypothesis of gradual mechanical erosion and chemical wear.

[0193] Taken together, these observations indicate that the nanostructure formation coating maintains its structure and function for several days of simulated wear, but begins to degrade noticeably after extended use. This degradation correlates with a measurable decline in hydrophilicity, highlighting the importance of periodic reapplication to preserve coating performance over longer durations.

[0194] 1,1.5. Antimicrobial, antiviral, and antioxidant properties

[0195] Beyond its influence on lens wettability, the nanostructure formation coating was also evaluated for its capacity to reduce microbial and viral contamination. The nanocoating exhibited antimicrobial activity, resulting in a modest reduction in bacterial presence on the lens surface. See Fig. 11 A. In viral adhesion assays, the nanostructure formation coating neutralized approximately 30% of viral particles within 6 hours of exposure. See Fig. 11B. Although this anti-infective effect is less potent than that of oxidative care solutions, it is comparable to the intrinsic antiviral and Gram-negative antibacterial activity of human tear fluid. Given that contact lens wear can disrupt the natural cleansing action of the eye and compromise tear film functionality, incorporating the nanostructure formation coating may help mitigate these negative impacts. Thus, these findings underscore the potential of the nanostructure formation coating layer to reduce the microbial load on contact lenses and lower the risk of infection during lens use.

[0196] To ensure the biocompatibility of the nanostructure formation coating, the inventors conducted cytotoxicity assessments using the MTT assay, a standard method for evaluating cell metabolic activity as a proxy for cell viability. As shown in Fig. 11C, therewas no statistically significant reduction in mitochondrial activity in cells exposed to the RetL protein, indicating that the nanostructure formation coating does not elicit cytotoxic effects under the tested conditions. The results demonstrated cell viabilities consistently above 95%, comparable to control levels, confirming that the nanocoating is well tolerated by ocular surface cells.

[0197] Additionally, the nanostructure formation coating was tested for its antioxidant capacity using the DPPH radical scavenging assay - a well-established method to quantify the ability of materials to neutralize free radicals. The eye is particularly susceptible to oxidative stress due to high metabolic activity, constant exposure to light, and elevated oxygen tension. As the cornea serves as the first barrier to environmental hazards, deficiencies in antioxidant defense are linked to the development of disease conditions such as cataract, glaucoma, keratoconus, and dry eye disease. The assay results showed that the nanostructure formation coating significantly reduced the presence of free radicals, demonstrating notable radical scavenging activity. See Fig. 1 ID. This antioxidative function could help counteract oxidative stress on the ocular surface, potentially reducing inflammation and enhancing long-term comfort for contact lens wearers.

[0198] 1,2, Mechanism of nanocoating formation and polymer brushes

[0199] The experimental data, combined with established principles of the Reaction-Diffusion mechanism, suggest that the formation of the nanostructure formation coating is driven by the interaction between RetL protein and a rapidly diffusing or moving inhibitor, specifically involving extended carbon-carbon chains. Based on this, it was proposed the presence of highly mobile polymer chains or polymer brushes on the surface of contact lenses, which enable the formation of the coatings.

[0200] To confirm the presence of polymer brushes on uncoated contact lenses and characterize their behavior, the inventors employed TEM of cryo-cross-sections and AFM in solution. Cryo-cross-sections of contact lenses were prepared and analyzed using TEM to visualize polymer brush layers. The outermost layer of uncoated lenses displayed increased electron permeability, indicative of a lower-density structure, consistent with the characteristics of polymer brushes. See Fig. 12A. Furthermore, this layer demonstrated a high degree of flexibility, as evidenced by the formation of column-like protrusions that appeared when the sample experienced slight displacement on the grid during the preparation process. See Fig. 12B. This observation suggests a less rigid, dynamic nature of the surface layer. Based on this preliminary visual analysis, the polymer chains in their relaxed,extended state are estimated to be at least as long as these columns, approximately 50 nm in length.

[0201] To assess the dynamic behavior of polymer brushes, AFM was performed in liquid environments with solvents of varying polarity, including phosphate-buffered saline (PBS) and isopropanol (IP A). See Fig. 13. In aqueous environments with low IP A concentrations (20%), considered as poor solvents for hydrophobic polymer chains, the brushes adopted a collapsed and compact conformation. In contrast, in an 80% IPA solution, which acts as a good solvent, the polymer brushes on the lens surface relaxed into an extended conformation. This solvent-dependent transition was clearly observed in AFM topography scans acquired under different applied forces, as well as in the force-distance relationship curves, consistent with classical polymer brush behavior. See Figs. 14A and 14B.

[0202] The force-distance curve recorded in 80% IPA exhibited characteristics typical for polymer brush compression, displaying a steric repulsion response described by the Alexander-de Gennes model. This model applies within the theoretical range of normalized tip-sample distances, specifically when the tip - sample distance D satisfies 0.2 < D / L < 0.9 (see Fig. 15, inset, vertical dashed line), where L is the effective brush height. The repulsive force F(D) as a function of distance D is given by: F(I ) «(100 n RtipD / s3)kBT exp (— 2nD / L), where As is Boltzmann’s constant, 7' is the absolute temperature, Rnpis the AFM tip radius, and .s is the lateral distance between polymer chains. Fitting the experimental force data (see Fig. 15, inset, sloped dashed line) to this equation yielded values ofZ =158 ± 4 nm and .s = 15.5 ± 2 nm, assuming an AFM tip radius of Rnp= 7 ± 3 nm. These values support the presence of a well -organized polymer brush layer with significant extension under good solvent conditions. The consistency between independently obtained experimental data confirms the brush-like nature of the polymer chains on the contact lens surface.

[0203] 2, Discussion

[0204] The nanostructure formation coating technology introduced in this study represents a novel, biomimetic strategy for modifying the surface properties of contact lenses, particularly those used in Ortho-K. Inspired by functional protein coatings found on insect surfaces, this self-assembling, protein-based nanolayer forms a multifunctional, nanopattemed interface that significantly enhances lens performance. Its key benefits include improved wettability, reduced biofouling, and broad-spectrum antimicrobial activity — properties that are essential for long-term lens comfort and safety.

[0205] At the molecular level, the inventors propose a self-assembly mechanism driven by interactions between RetL proteins and collapsed polymer molecules present on the lens surface in an aqueous environment. RetL amphiphilic nature of RetL, comprising both hydrophobic and hydrophilic regions, allows it to weakly bind to these polymer chains, gradually increasing their hydrophilicity. This interaction facilitates a transition from a collapsed state to an extended, brush-like conformation. Once mobilized, the polymer brushes further interact with RetL, leading to the formation of a self-organized, nanopatterned coating that defines the nanostructure formation coating architecture. See Fig. 16. This proposed mechanism aligns with the known behavior of natural and synthetic polymer brushes, which form dynamic interfacial layers characterized by high mobility, environmental responsiveness, and chemical reactivity.

[0206] To validate the presence of polymer brushes on the lens surface, the inventors employed TEM and AFM. TEM cross-sections revealed a peripheral zone of lower electron density compared to the bulk, consistent with a layer composed of less densely packed material. See Figs. 3A, 3B, 12A, and 12B. This outer layer exhibited notable mechanical flexibility, evidenced by the formation of column-like protrusions upon slight mechanical displacement during sample preparation. Such localized reorganization stands in contrast to the uniform deformation typically observed in homogeneous, gel-like networks. These observations strongly support the existence of a flexible, chain-like molecular architecture on the lens surface, composed of long polymer strands rather than a densely crosslinked network that can adapt its conformation while preserving overall structural integrity.

[0207] Based on force-distance curves obtained via AFM, calculations estimate the polymer brush length to be approximately 160 nm. Taking into account an effective monomer length (Kuhn length) of 0.25 nm, this corresponds to approximately 640 monomer units per freely extending polymer chain. This chain length falls well within the realistic range for synthetic polymers. For example, polymethyl methacrylate (PMMA) typically consists of up to 10000 monomer units per chain under standard polymerization conditions. In practical applications, PMMA and related materials are often lightly crosslinked using 0.5 - 1 wt % of ethylene glycol dimethacrylate (EGDMA). Assuming a crosslinking efficiency of 20 - 50%, which accounts for the fraction of EGDMA molecules that actually form effective crosslinks, the resulting ratio between linear (non-crosslinked) and crosslinked monomer subunits in the chain corresponds to roughly 200 - 1000 monomer units.Topological effects can be effectively ignored when considering the maximum availablecontour length in a randomly cut polymer network. This length includes contributions from segments that may contain short branches and loops, as long as they remain part of a continuous chain extending from a crosslinked anchor to a free end. This approximation supports the physical plausibility of the observed polymer brush length and suggests that the material achieves a functional balance: providing polymer chain mobility at the polished lens surface while maintaining structural stability within the bulk through sparse crosslinking.

[0208] Building on this, a key innovation of the nanostructure formation coating is its ability to harness this mobility of surface polymers through a reaction-diffusion-driven self-assembly mechanism. This process involves the interaction between RetL proteins and the highly mobile carbon chains that naturally act as polymer brushes on the lens surface. These brushes dynamically transition from collapsed to extended conformation in response to environmental stimuli. The inventors’ analyses, combining cryo-TEM cross-sectioning and AFM measurements in varying solvent conditions, confirmed responsive behavior of these brushes on uncoated lenses. Upon application of the nanostructure formation coating, the brush layer undergoes structural modification; the low-electron-density outer layer is eliminated (compare Figs. 12A and 12B with Figs. 3A and 3B), indicating a substantial alteration in the surface chemistry. This transformation is likely driven by specific interactions between RetL proteins and the polymer brushes, ultimately leading to the formation of a stable, functional nanolayer. Theoretical estimates of the diffusion coefficient (D) for these brush-like structures (106- 108cm2 / s) closely match experimentally reported values of approximately 7.5- 108cm2 / s, supporting their ability to act as morphogens within a reaction-diffusion system and reinforcing the validity of our proposed self-assembly mechanism.

[0209] Importantly, the nanostructure formation coating preserves essential physiological functions: Its protein-based, nanoscale architecture (<100 nm) maintains oxygen permeability, ensuring continuous corneal gas exchange critical for corneal health. Even after repeated simulated mechanical and chemical stresses, the coating retained its structural integrity and functional performance over at least seven wash cycles, consistently maintaining a contact angle below 60°- a hallmark of sustained hydrophilicity. AFM imaging further revealed partial self-healing behavior, with nanostructures reappearing along prior scratches and certain surface defects gradually diminishing. This remarkable durability, likely stemming from the cohesive RetL protein network, adds significant value for long-term lens performance. This is the first nanocoating developed for RGP lenses that not only reduces surface irritation but also forms a structurally resilient layer with a higher Young’smodulus than the lens substrate itself, enhancing resistance to abrasion and wear while preserving both optical clarity and physiological function.

[0210] 3, Conclusions

[0211] This study identifies hydrophobic polymer brushes on RGP lenses as a significant, previously underrecogized contributor to ocular irritation and inflammation, particularly during extended wear such as in Ortho-K use. These surface structures, prone to protein and lipid adsorption, can compromise both comfort and corneal health. The nanostructure formation coating directly addresses this issue by forming a thin, hydrophilic, and durable layer that neutralizes the chemical and mechanical aggressiveness of the lens surface. Its nanoscale protein architecture ensures oxygen permeability, mechanical resilience, and partial self-healing. Moreover, AEGIS provides broad-spectrum protective effects: antibacterial, antiviral, antioxidant, and anti-deposit properties, likely mediated by the biochemical activity of RetL proteins. Designed for easy post-cleaning application using biocompatible materials, the nanostructure formation coating integrates seamlessly into existing lens care routines. Its sustainable, non-toxic composition and demonstrated clinical benefits position it as a promising alternative to conventional lens coatings. Beyond ophthalmic applications, the mechanistic insights gained by this study into polymer brush dynamics and protein-polymer interactions provide a valuable foundation for the development of next-generation biointerfaces across diverse biomedical fields.

[0212] 4, Materials and methods

[0213] 4,1, Protein expression and purification

[0214] The RetL protein was expressed in E. coli BL21(DE3)pLysS cells transformed with the pET23b (RGSHisRetL) plasmid. Cultures were grown in Lauria Broth (LB) medium (Condalab) supplemented with ampicillin (100 pg / mL) at 37°C while shaking (GFL 3031 incubator shaker) until reaching mid-log phase (OD600 ~ 0.5). The incubation temperature was then reduced to 19°C for 1 hour, after which protein expression was induced with the final concentration of 1 mM isopropyl P-D-l -thiogalactopyranoside (IPTG) (Fluorochem). The culture was incubated for additional 12 hours post-induction at 19°C.

[0215] Cells were harvested by centrifugation at 4000 g for 15 minutes at 4°C using a Sorvall Lynx 6000 centrifuge (Thermo Scientific) equipped with an F9-6xl000 LEX rotor. The bacterial pellet was resuspended in TBS buffer (20 mM Tris, 150 mMNaCl) containing 1 mM PMSF (PanReac AppliChem) and lysed using a One-Shot Cell Disruptor (Constant Systems). The lysate was clarified by centrifugation at 18000 g for 30 minutes at 4°C.

[0216] The supernatant (~80 mL) was incubated with 400 pL of HisPur™ Ni-NTA Resin (Thermo Scientific) at 4°C for 12 hours with gentle agitation (IK A Loopster basic), in the presence of 20 mM imidazole to minimize nonspecific binding. The resin was washed three times with TBS containing 20 mM imidazole, and bound protein was eluted using TBS supplemented with 300 mM imidazole.

[0217] Eluted fractions were concentrated using Vivaspin 500 centrifugal filters (Sartorius) at 15 °C to prevent aggregation. The final purification step was performed using AKTA machine for size exclusion chromatography with Superdex™ 200 Increase 10 / 300 GL column (Cytiva) equilibrated with TBS. Active, oligomer-forming protein fractions were pooled and reconcentrated using Vivaspin 500 units (Sartorius) at 15 °C.

[0218] 4,2, Nanocoating protocol

[0219] Clean rigid gas-permeable contact lenses were dipped into a freshly prepared coating solution composed of 0.5 mg / mL RetL protein in TBS, supplemented with 1% MeniCare™ solution (Menicon). Lenses were incubated in the coating solution for 10 minutes at room temperature, then rinsed in sterile deionized water prior to characterization or testing.

[0220] 4,3, TEM

[0221] TEM analysis was made by the Electron Microscopy Facility (Pole Facultaire de Microscopic Ultrastructurale (PFMU)) at the Medical faculty of the University of Geneva, Switzerland. Lenses, poststained with 1% osmium tetroxide, embedd in gelatine, infiltrated with sucrose as cryoprotectant and freezed in liquid nitrgen were cut transversal (with 50 nm thickness) by cryo-sectioning. Resulting samples were analyzed by TEM using Tecnai 12 G2microscope (FEI, Eindhoven, Netherlands).

[0222] 4,4, AFM

[0223] Topographical imaging in both air and liquid environments was performed in tapping mode using an XE-100 AFM (Park Systems) with a CONTR-W cantilever (NanoWorld) (length: 450 pm; thickness: 2 pm; spring constant: 0.2 N / m; resonance frequency: 13 kHz; tip radius R<8 nm).

[0224] Adhesion force mapping and force-distance measurements were carried out using an NX7 AFM system (Park Systems) in PinPoint™ mode, equipped with an NCHR-W cantilever (NanoWorld) (length: 125 pm; thickness: 4 pm; spring constant: 42 N / m; resonance frequency: 320 kHz; tip radius R<8 nm).

[0225] All AFM experiments were conducted at the Micro-Nanotechnology Facility of Haute Ecole Specialisee de Suisse Occidentale (Geneva, Switzerland). Gwyddion software (version 2.55) was used for image processing and quantitative surface analysis.

[0226] 4,5, Wettability test

[0227] Lenses were gently blotted dry with a lint-free paper tissue and placed on a custom contact lens half-mold holder (Menicon). A 3 pL droplet of deionized water was deposited on the central surface of each lens. Static contact angles were measured using a Drop Shape Analyzer DSA-25 (Kriiss GmbH, Germany), and averaged across multiple replicates.

[0228] 4,6, Lipid adhesion test

[0229] A 0.1% (w / w) solution of Sudan Red dye in oleic acid was prepared at room temperature. Lenses were preconditioned in ISO saline solution at room temperature for 4 hours. For staining, each lens was briefly blotted to remove excess saline, then immersed for 1 minute in 4 mL of the oleic acid solution contained in a 12-well plate.

[0230] Following staining, lenses were gently dipped twice into each of three wells containing clean ISO saline to remove unbound dye. The imaging was conducted by transferring lenses into a Petri dish filled with ISO saline and capturing images using a Leica S9i optical microscope (Germany).

[0231] 4,7, Progent™ cleaning with protein deposits model

[0232] To evaluate the efficacy of the nanostructure formation coating in resisting protein deposition and its compatibility with commercial lens cleaning protocols, a bovine serum albumin (BSA; fraction V, Millipore) deposition model was used in combination with Progent™ (Menicon) treatment. RGP lenses (both coated and uncoated) were incubated in 1% (w / v) BSA solution in PBS at 37°C for 4 hours to simulate protein fouling. After incubation, lenses were rinsed briefly in PBS and subjected to cleaning with Progent™, following the manufacturer’s protocol. Briefly, lenses were immersed in the Progent™ dualsolution mixture (A + B) and incubated for 30 minutes at room temperature.

[0233] After treatment, lenses were rinsed thoroughly with deionized water and analyzed for residual protein: changes in surface wettability were assessed by static contact angle measurements, and surface morphology was visualized by using AFM. Comparisons were made between uncoated and nanostructure formation-coated lenses with and without BSA deposition and Progent™ treatment, in order to assess both the protective effect of the coating and its post-cleaning renewability.

[0234] 4,8, Coating wearing stability

[0235] To simulate extended wear of the contact lenses and assess the durability of the AEGIS nanocoating under repeated use conditions, coated RGP lenses were subjected to daily washing cycles. Each cycle consisted of lens immersion in MeniCare™ solution for 24 hours, followed by mechanical agitation and manual rinsing, mimicking typical contact lens cleaning routines.

[0236] The protocol was conducted over seven consecutive days. After each cycle, a subset of lenses was collected for analysis. Changes in hydrophilicity were evaluated via static contact angle measurements, while nanostructural integrity was assessed using AFM. Morphological degradation was further monitored by comparing surface topography across time points.

[0237] 4,9, Anti-Infective Activity Measurement

[0238] The antibacterial and antiviral properties of the nanostructure formation coating were evaluated in accordance with ISO 22196:2011 guidelines (iso.org / standard / 54431.html), with minor modifications as described below.

[0239] 4,9.1. Bactericidal activity assay

[0240] To assess bactericidal activity, 10 pL of a Topi OF E. coli suspension (Thermo Fisher Scientific) at a concentration of 104CFU / mL in LB medium was applied to a custom lens-shaped mold. The inoculum was covered with either an uncoated or nanostructure formation-coated RGP contact lens and incubated at 25°C for 1 hour.

[0241] Following incubation, the liquid was collected, and each lens surface was gently rinsed with 100 pL of sterile LB medium. The original inoculum and the wash solution were combined, serially diluted if necessary, and plated onto LB agar plates. After 16 hours of incubation at 37°C, bacterial colonies were counted manually, and the reduction in colony-forming units (CFU) was used as a quantitative measure of antibacterial efficacy.

[0242] 4,9.2. Antiviral activity assay

[0243] Antiviral activity was assessed using lentiviral particles produced in HEK-293T cells via triple transfection with pPAX2, pVSVG, and pSD28-GFP plasmids. A 10 pL aliquot of lentiviral suspension with GFP gene was applied to a lens-shaped mold, covered with either coated or uncoated RGP lenses, and incubated for 6 hours at 37°C.

[0244] Post incubation, the liquid was recovered, and each lens was washed with 100 pL of Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher). The collected viral suspensions were pooled and concentrated using polyethylene glycol (PEG-8000) in PBS, following standard precipitation protocols. The viral concentrate was then added to HEK-293 cells (10,000 cells / well) seeded in 96-well transparent plates (Thermo Fisher), in DMEMsupplemented with 10% fetal bovine serum (FBS) and 1% gentamicin with 10 pg / mL Polybrene (Sigma-Aldrich).

[0245] Lentiviral infection efficiency was quantified by measuring GFP fluorescence 48 hours post-infection using an Infinite M Plex multifunctional plate reader (Tecan, Switzerland). The degree of fluorescence reduction in samples treated with AEGIS-coated lenses was used to assess antiviral activity.

[0246] 4.10. Cytotoxicity assessment (MTT assay).

[0247] Cytotoxicity of the RetL-based nanostructure formation coating was evaluated using the MTT assay on human embryonic kidney HEK293T cells. Cells were maintained in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% gentamicin.

[0248] For the assay, 3000 cells per well were seeded into a 384-well, flat-bottom transparent plate (Greiner Bio-One, Monroe, NC, USA) in 30 pL of complete DMEM and incubated overnight at 37°C in a humidified 5% CO2 incubator. The next day, the culture medium was removed and replaced with 50 pL of fresh DMEM containing RetL, at a concentration of 0.05 pg / mL. Cells were exposed to the protein solutions for 72 hours under standard incubation conditions.

[0249] Following the incubation period, wells were emptied and 30 pL of MTT reagent (0.5 mg / mL in PBS; Sigma-Aldrich) was added to each well. After 3.5 hours of incubation at 37°C, the resulting formazan crystals were solubilized by adding 50 pL of dimethyl sulfoxide (DMSO) to each well. The optical density was measured at 590 nm using an Infinite M Plex microplate reader (Tecan, Switzerland) within 5 minutes of solubilization.

[0250] Data were statistically analyzed using a two-tailed t-test, and graphical representations were generated using Prism 10 software (Dotmatics, USA).

[0251] 4,11, Antioxidant activity measurement

[0252] A 0.2 mM solution of DPPH (2,2-diphenyl-l-picrylhydrazyl) (Sigma-Aldrich) was prepared in absolute ethanol and stored in the dark for 30 minutes to ensure full radical stabilization. RGP lenses, either uncoated or coated with a nanostructure formation, were gently blotted dry and placed in a 24-well plate. To each well, 200 pL of the DPPH solution was added, ensuring complete submersion of the lens.

[0253] Samples were incubated in the dark at 37°C for 1 hour. Following incubation, 100 pL of the DPPH solution from each well was transferred to a 96-well plate for absorbance measurement. The decrease in DPPH absorbance at 517 nm was recorded using an Infinite M Plex multifunctional plate reader (Tecan, Switzerland).

[0254] Radical scavenging activity was calculated using the following equation: DPPH Scavenging = ( 40— ls) / 40) • 100%, whereto is the absorbance of the control (DPPH solution without lens), and As is the absorbance of the sample.

[0255] Example 2 - Coating Resiliency Test

[0256] RGP contact lenses were gently washed with detergent and treated with Progent™ solution according to the manual. These lenses were then coated with nanostructure formation coating layer described herein (90 min, 4°C) and stored in the Menicare™ solution. Every 24 hours all lenses were washed by applying “massage.” Every time after washing, one lens was rinsed with water and dried for further investigation.

[0257] Wettability of the surface was measured by static contact angle. The contact lens was wiped with a paper tissue and put on the CL half mold. A drop of DI water: 5 pl was put on the top of the lens. Contact angle was measured with Drop Shape Analyzer DSA-25 (Kruss; Germany). The results are shown in Fig. 17. As can be seen, the contact angle increased straight after the first wash and reached a plateau of around 60 degrees.

[0258] Samples were also investigated using a scanning electron microscope.Samples were immobilized with double side glue tape and covered by gold using Q150T SZEZES Sample Preparation System (Quorum Technologies). 20 sec of gold plasma was applied under 20 mA current. SEM was done using Helios 660 Nanolab DualBeam FIBSEM (FEI, Eindhoven, Netherlands). The results are shown in Figs. 18A and 18B. The result of the SEM investigation of non-coated lenses can be seen in Fig. 18 A. Scratches can be seen; however, the rest of the lens surface is very smooth. As shown in Fig. 18B, the nanostructure formation coating layer not only fills the scratches but also produces visible nanostructures. Unfortunately, only the highest protrusions can be seen due to gold coating; however, the difference between uncoated and freshly coated surfaces is noticeable. Cracks appeared during the microscopy and must be ignored. After seven days of the usage simulation, the lens still shows the presence of nanoprotrussions. This is shown in Fig. 18C. The protrusions cannot be seen on the whole surface, meaning they were partially washed out; however, scratches cannot be seen on a lens, which shows both scratches filled by coating and anti-scratch protection due to higher Young's modulus.

[0259] These results show that the nanostructure formation coating layer can withstand wearing for seven days without losing functionality. For the first time, it was demonstrated that a coating can protect the surface of an RGP lens from scratches due to the formation of a thin layer with a higher Young's modulus. Even after seven days of the usagesimulation, the resulting lenses were found to be hydrophilic, with a contact angle of approximately 60°.

[0260] It will be apparent to those skilled in the art that variations of the methods and compositions described herein are possible and are intended to be encompassed within the scope of the present disclosure.

Claims

WHAT IS CLAIMED IS:

1. A lens conditioning kit comprising:a first component comprising a sterile lyophilized protein comprising one or more of retinin, retinin-like protein, and cuticular protein; anda second component comprises a sterile aqueous solution,wherein the kit optionally comprises a long-chain inhibitor of the sterile lyophilized protein such that a weight ratio of the sterile lyophilized protein to the long-chain inhibitor of the sterile lyophilized protein is greater than 20: 1.

2. The lens conditioning kit according to claim 1, wherein the sterile lyophilized protein is in powder form.

3. The lens conditioning kit according to claim 1, wherein an amount of the first component is configured for a single one-time use, and optionally an amount of the second component is configured for a single one-time use.

4. The lens conditioning kit according to claim 1, wherein the second component comprises one or more of a buffer, a diluent, a salt, a pH adjustment agent, and an osmotic adjustment agent.

5. The lens conditioning kit according to claim 1, wherein the first component and second component, when mixed together and contacted with a polymer surface comprising a polymer having a structure including polymer brushes, are configured to form a nanostructure formation on said polymer surface, said nanostructure formation comprising the one or more of retinin, retinin-like protein, and cuticular protein attached to said polymer surface.

6. The lens conditioning kit according to claim 1, wherein the long-chain inhibitor is selected from the group consisting of a lipid, a wax, and a surfactant.

7. A coated contact lens comprising:a contact lens comprising a polymer surface; anda nanostructure formation formed on said polymer surface, said nanostructure formation comprising one or more of retinin, retinin-like protein, and cuticular protein and formed in an absence of a long-chain inhibitor or in presence of a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1.

8. The coated contact lens of claim 7, wherein the polymer surface comprises a polymer having a structure including polymer brushes, a free end of which interact with theone or more of retinin, retinin-like protein, and cuticular protein to attach said nanostructure formation to said polymer surface.

9. The coated contact lens of claim 8, wherein the polymer surface comprises a polymer selected from the group consisting of styrene-(meth)acrylate copolymers, styrene-silyl / siloxanyl copolymers, silicone-containing copolymers, and polymers formed from monomers or comonomers comprising aliphatic hydrocarbon segments.

10. The coated contact lens of claim 7, wherein the lens is a rigid gas-permeable contact lens.

11. The coated contact lens of claim 7, wherein the coated contact lens exhibits a wettability contact angle of 75 degrees or less.

12. The coated contact lens of claim 7, wherein the long-chain inhibitor is selected from the group consisting of a lipid, a wax, and a surfactant.

13. A method of forming a coated medical device, comprising, in order:a) providing a medical device comprising a polymer surface;b) contacting the polymer surface with an aqueous solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a coating on the medical device;c) optionally, rinsing the coating with water or a saline solution; and d) optionally, drying the coating,wherein the coating comprises a nanostructure formation on said polymer surface, said nanostructure formation comprising the one or more of retinin, retinin-like protein, and cuticular protein attached to said polymer surface.

14. The method according to claim 13, wherein the coated medical device exhibits one or more of increased antimicrobial properties, antiviral properties, increased resistance to lipid adhesion, antioxidant surface activity, and increased comfort to a user as compared to a similar medical device without the coating.

15. The method according to claim 13, wherein the medical device is a rigid gas-permeable contact lens.

16. The method according to claim 13, further comprising:before step b), soaking the medical device in an intensive cleaner to remove protein and other deposits from the polymer surface, and optionally rinsing the medical device.

17. The method according to claim 13, wherein the polymer surface comprises a polymer having a structure including polymer brushes, a free end of which interact with the one or more of retinin, retinin-like protein, and cuticular protein to attach said nanostructure formation to said polymer surface.

18. The method according to claim 17, wherein the polymer surface comprises a polymer selected from the group consisting of styrene-(meth)acrylate copolymers, styrene-silyl / siloxanyl copolymers, silicone-containing copolymers, and polymers formed from monomers or comonomers comprising aliphatic hydrocarbon segments.

19. The method according to claim 13, wherein the contacting comprises immersing the polymer surface in the aqueous solution for a time of from about 1 to about 180 minutes.

20. The method according to claim 13, wherein the long-chain inhibitor is selected from the group consisting of a lipid, a wax, and a surfactant.

21. A method for cleaning a contact lens, comprising in order:a) contacting a contact lens with a protein cleaning solution to remove protein deposits from the contact lens;b) optionally rinsing the contact lens with a rinsing solution;c) contacting the contact lens with an aqueous conditioning solution comprising water and one or more of retinin, retinin-like protein, and cuticular protein, and either not comprising a long-chain inhibitor or containing a long-chain inhibitor whereby a weight ratio of the one or more of retinin, retinin-like protein, and cuticular protein to the long-chain inhibitor is greater than 20: 1, to form a nanostructure formation coating layer of the one or more of retinin, retinin-like protein, and cuticular protein attached to an outer surface of the contact lens; andd) rinsing the coating layer with a rinsing solution.

22. The method of claim 21, further comprising between steps a) and b) a step of rinsing the contact lens with a rinsing solution.

23. The method of claim 21, further comprising after step c) a step of storing the contact lens in a storage solution.

24. The method of claim 21, wherein the contact lens is a rigid gas-permeable contact lens.

25. The method of claim 21, wherein the contacting comprises immersing the contact lens in the aqueous conditioning solution for a time of from about 1 to about 180 minutes.

26. The method of claim 21, wherein the long-chain inhibitor is selected from the group consisting of a lipid, a wax, and a surfactant.

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