Systems, methods, and material compositions for use in correcting eye conditions.

Abstract

Methods, kits, and material compositions for use in correcting eye conditions are disclosed. This involves a selected three-dimensional patterning of the user's corneal surface. The pattern is selected to provide an optical effect according to a given visual impairment of the user. A dispersant in liquid (aqueous solution) form containing nanoparticles is applied onto the selected three-dimensional pattern, thereby dispersing the nanoparticles in the incision areas of the pattern. The nanoparticles include biocompatible protein-based nanoparticles. The dispersant may also be used as eye drops, thereby allowing the nanoparticles to occupy etching areas on the cornea, thereby functionalizing and maintaining the visual correction effect of the pattern on the eye condition. The disclosure further provides methods and kits for ablating corneal tissue in a selected pattern by using visible-spectrum light.

Description

[Technical Field]

[0001] The present invention relates to the field of vision correction technology, and more particularly to the correction of visual impairments, such as myopia, hyperopia, presbyopia, astigmatism, myopia control, and other visual impairments. [Background technology]

[0002] Various eye conditions are known to limit vision. These conditions commonly include nearsightedness (myopia), farsightedness (hyperopia), and age-related changes in the eye (typically associated with presbyopia). Various techniques are known to provide correction for these eye conditions, including eyeglasses, contact lenses, and intraocular lenses. The lenses used feature a selected one or more optical powers and / or patterns designed to increase the depth of focus of the lens depending on the user's eye condition. One of the approaches commonly used to overcome various eye conditions, such as presbyopia, is based on intraocular lens technology, which has evolved through either accommodation IOLs, multifocal IOLs, or, more recently, IOLs with extended depth of focus. Optical power can generally be provided using refraction of the optical path. However, diffraction techniques for manipulating the optical path can also provide suitable optical power. Such techniques include Fresnel zone plates or Fresnel zone rings, which provide light diffraction in a lens-like manner. Additional optical manipulation techniques may include the use of optical interference to extend the depth of focus. For example, U.S. Patent No. 7,859,769 provides an image-forming arrangement and method for extending depth of field. The image-forming arrangement includes an image-forming lens having a specific effective aperture and an optical element associated with the image-forming lens. The optical element is configured as a phase-influencing, non-diffractive optical element that defines a spatially low-frequency phase transition. The optical element and the image-forming lens define a predetermined pattern of substantially optically transparent features having different optical properties, which are spaced apart. The location of at least one phase transition region of the optical element within the image-forming lens plane is determined by at least the dimensions of the effective aperture. [Overview of the Initiative]

[0003] This invention provides a novel technique for correcting eye conditions that cause visual difficulties in users. The technique involves applying a pattern selected according to the user's eye condition to the user's cornea, and introducing a solution containing nanoparticles (NPs) of a selected material composition into the pattern. This optical pattern is configured to provide a transient, targeted correction of refractive errors without altering the shape of the cornea. Generally, patterns formed on the cornea affect local changes in refractive index. While the nanoparticles themselves have a different refractive index than the environment to which they are applied, the arrangement density of nanoparticles in the pattern is relatively low, and the average local refractive index is approximately the same as that of the solution to which they are placed, and therefore does not substantially affect the optical effect induced by the pattern. It should be noted that the effects induced by the arrangement of nanoparticles dispersed in the pattern include scattering of incident light in the visible spectral range due to the filling of the pattern's cutting / etching regions by the nanoparticles, as well as the functionalization and stabilization of the pattern's optical effects (which are selected depending on the visual correction applied). However, the density of the nanoparticles should be such that the arrangement of nanoparticles does not change the overall refractive index and therefore does not affect the refractive effect induced by the pattern structure. The selected material composition may generally be provided in the form of eye drops containing nanoparticles, which may contain biocompatible protein chains, such as albumin protein chains, e.g., human serum albumin (HSA) or recombinant human serum albumin. Human serum albumin or recombinant human serum albumin (rHSA) is the most abundant protein in human blood and offers advantages such as its non-toxicity, non-immunogenicity, water solubility, and in vivo conversion into harmless materials. As will be discussed later, the nanoparticles may also be metal particles with a biocompatible polymer shell (e.g., an HAS-based shell).

[0004] As mentioned above, for example, rHSA can be used as nanoparticles. Generally, the material composition of the nanoparticles is selected as a biocompatible, non-toxic material composition that has high stability both during storage and on the user's cornea. The nanoparticles may be human serum albumin (HSA)-based nanoparticles, such as rHSA-based nanoparticles; or metal nanoparticles, such as gold particles supported by a biocompatible material, such as HAS. However, preferably, the nanoparticles are HAS-based or rHSA-based particles. Suitable ophthalmic formulations may include, for example, recombinant human serum albumin nanoparticles, or any suitable nanoparticles containing gold supported by metal-based nanoparticles, such as biocompatible materials, such as a biocompatible polymer shell (e.g., an albumin shell). The biocompatible polymer shell (e.g., an albumin shell) typically exhibits a protein folded around the particle. A pattern is a surface relief in the form of a two-dimensional arrangement of spaced-out incisions / grooves (etched regions) at a specific depth. Thus, a pattern is actually three-dimensional in the sense that the critical dimensions of the pattern are defined by the arrangement of etched regions along the x and y axes and the depth of the etched regions.

[0005] Therefore, according to a broader embodiment, the present invention is a method for use in correcting the condition of the eye in question, To form the selected three-dimensional pattern on the target corneal surface, Applying a dispersant containing nanoparticles onto the selected pattern, wherein the nanoparticles are selected and applied such that (i) they generate conditions that are invariant with respect to one or more of the following: incident light within a predetermined spectral range on the cornea, the precise position of the nanoparticles in the pattern, and the liquid type of the dispersant; and (ii) they exhibit a dominant scattering effect with respect to incident light within the predetermined spectral range. This provides a method that includes this. The formation of the selected three-dimensional pattern is performed using a patterning system that includes a patterning utility (e.g., an etching utility) that operates in a controllable manner according to input data, which includes pattern data corresponding to the correction of the user's visual impairment. The patterning utility may be configured to transmit selected ultrasound to etch the pattern onto the user's cornea, or it may be configured to mechanically etch the cornea. In some other embodiments, the patterning utility includes at least one laser unit and a beam steering module, configured to provide optical energy to etch a selected pattern on the cornea of ​​the user's eye according to operating commands. The laser unit may be configured to provide etching to a depth of a single cell layer of the cornea.

[0006] According to some embodiments, the three-dimensional pattern includes a diffraction pattern. For example, the pattern may include a Fresnel ring pattern. According to some embodiments, the three-dimensional pattern includes an interference pattern that affects the phase. The pattern may include, for example, a depth-of-focus extension pattern. According to some embodiments, the selected pattern enables optical correction with an optical power resolution between 0.1 and 0.3 diopters. Generally, this allows for correction below 0.25 diopters, which is the limit of typical conventional optical correction. In some embodiments of the method, the particles are selected to exhibit the maximum scattering effect within the visible range, i.e., in the range of 380 nm to 700 nm. In some embodiments of the method, the particles are selected to exhibit the maximum scattering effect within a spectral range defined between 450 nm and 600 nm. In some embodiments of the method, the nanoparticles are selected to exhibit maximum absorption effect within a predetermined spectral range.

[0007] In some embodiments of the method, the particles are selected based on one or more properties of material, size, shape, charge, concentration, or any combination thereof. The nanoparticles may be spherical or rod-shaped, and with respect to any given shape, their size is selected to obtain a dominant scattering effect, particularly at a desired wavelength or wavelength range, so that the desired results are obtained. The nanoparticles may be selected so that their shape distribution between spherical and rod-shaped is known. For example, the nanoparticles may be selected such that at least X% are spherical and at least Y% are rod-shaped, and their average size is selected so as to exhibit a desired effect, e.g., a maximum scattering effect at a wavelength in the green spectrum, where X and Y represent any given number between 0 and 100. For example, the nanoparticles may be spherical rHSA particles with a diameter of 80 nm. In some embodiments of the method, by forming the aforementioned pattern and applying a dispersant, an invariant condition with respect to the point spreading function obtained on the target retina is generated. In some embodiments of the method, the application of the dispersant provides a distribution of at least two or more nanoparticles on a λ×λ area within the selected pattern, where λ is a wavelength in the visible spectral range, for example, about 530 nm. In this context, the term "about" refers to a range of ±30 nm.

[0008] In some embodiments, the application of the dispersant results in nanoparticles of about 80 nm size and λ of about 530 nm within the incision region of the selected pattern. 3 A distribution of up to 200 nanoparticles is provided within the volume space. In some embodiments of the method, the three-dimensional pattern includes a diffraction pattern. In some embodiments of the method, the pattern includes a Fresnel ring pattern. In some embodiments of the method, the three-dimensional pattern includes an interference pattern that affects the phase. In some embodiments of the method, the pattern includes a depth of field extension pattern. In some embodiments of the method, the selected pattern enables optical correction between 0.25 diopters and 10 diopters with a resolution of + / -0.25D. In some embodiments, the method further includes preparing the dispersant before application, the preparation including mixing the liquid with the nanoparticles. In some embodiments of the method, the nanoparticles include a biocompatible protein chain. In some embodiments of the method, the protein chain includes human serum albumin protein and / or recombinant human serum albumin protein.

[0009] In some embodiments of the method, the formation is carried out according to one of the embodiments of the method for ablating corneal tissue described below. In another broad aspect of this disclosure, a formulation applied to a target eye is provided. The formulation comprises nanoparticles dispersed in a liquid and together forming a dispersant, the majority of which, when applied to a three-dimensional pattern formed on the target cornea, (i) generate conditions invariant on the cornea with respect to one or more of the incident irradiation light, nanoparticle concentration, and precise positions of the nanoparticles and liquid in a given (e.g., visible) spectrum, and (ii) exhibit a dominant scattering effect over absorption or an absorption effect over scattering within the given spectral range. It should be noted that the conditions invariant are generated by the combined effect of the intrinsic properties of the cornea and the nanoparticles applied to the three-dimensional pattern formed on the corneal surface. Nanoparticles containing a substance (macrostructure) may be provided in a regenerative solid powder form or as a read-for-use liquid ophthalmic formulation in an aqueous carrier. The aqueous carrier may be any one of the following: water, bacteriostatic water, sodium chloride solution, glucose solution, liquid surfactant, pH buffer solution, etc.

[0010] If the macrostructure is provided in an aqueous medium, it may be treated to obtain a reversible powder form. The powder form may be obtained by freeze-drying or spray-drying techniques. Preferably, the formulation is in the form of eye drops that can be applied to the ocular surface, i.e., the cornea. Preferably, the nanoparticles are selected such that, when applied to a three-dimensional pattern formed on the target cornea, they exhibit a dominant scattering effect within the visible spectral range, typically the maximum scattering effect within the visible spectral range. For example, this is the spectral range of 450 nm to 600 nm. Nanoparticles are selected based on one or more properties of material, size, shape, or any combination thereof. The nanoparticles may be human serum albumin (HSA)-based nanoparticles, such as rHSA-based nanoparticles. In yet another broader embodiment, the present invention provides a kit for use in correcting visual impairment, comprising the pattern-forming system described above and eye drops containing the formulation described above.

[0011] In some embodiments of the kit, the patterning system is comprised of one of the following embodiments of the kit for ablating corneal tissue. The kit may further include instructions for operating the patterning utility to form a selected pattern on the user's cornea and for applying eye drops to the eye. Generally, the technology of the present disclosure enables optical correction for users who are not suitable for modern laser vision correction surgery because the pattern etching is shallow on the cornea and can be performed on thin or weak corneas. Therefore, the kit may be suitable for use at home or in an eye care specialty clinic. Furthermore, the kit and technology enable correction for users who have already undergone cataract surgery, and moreover, additional correction on the intraocular lens that has already been used by the user is possible. Nanoparticles (e.g., those made from biocompatible protein chains such as HSA or rHSA) are configured, arranged / dispersed within the pattern to enhance the stability of the optical effect of the pattern, thereby enabling the persistence of the optical correction effect, for example, lasting from several days to several months.

[0012] The technology of the present disclosure provides correction for any one of myopia, myopia control, hyperopia, presbyopia, and astigmatism. The present disclosure further provides a method and a kit for ablating corneal tissue by using irradiation light within the range of the visible spectrum. To do so, an energy absorption material that can interact with the irradiation light of a selected wavelength to cause, for example, on-spot ablation of the cornea by thermally absorbing the irradiation light is used. The energy absorption material is mixed in a liquid for application to the corneal tissue as an eye drop. By ablating the cornea through this specific photothermal chemical reaction, very low energy can be used, and the wavelength that causes ablation of the cornea is not limited to the wavelength that needs to directly interact with the corneal tissue. In this process, the wavelength used may be determined by the energy absorption material or by its reaction with the cornea. In some non-limiting embodiments of the present disclosure, the energy absorbing material is a fluorescein-based material, such as sodium fluorescein dye, and the selected wavelength is in the visible spectrum. However, it should be noted that the energy absorbing material may be synthesized or produced to have an absorption profile at any selected wavelength, whether it is in the visible spectrum or in the IR spectrum.

[0013] Thus, aspects of the present disclosure provide a method for ablating corneal tissue or an energy absorbing material for use in a method for ablating corneal tissue. The method includes applying a liquid containing the energy absorbing material onto the corneal tissue. The energy absorbing material absorbs irradiation light at an absorption wavelength or within an absorption wavelength range. The absorption wavelength is a wavelength suitable for the energy absorbing material to absorb and cause a thermal reaction. It should be noted that the energy absorbing material may absorb irradiation light of more than one wavelength. Thus, the energy absorbing material absorbs irradiation light at the absorption wavelength, causing a local photothermal chemical reaction, thereby obtaining local ablation of the cornea. After the liquid is applied onto the corneal tissue, the method further includes irradiating the corneal tissue with irradiation light including the absorption wavelength to provide a selected ablation profile for the corneal tissue. Thus, this irradiation light causes a local photothermal chemical reaction, thereby obtaining local ablation of the cornea. It should be noted that the energy absorbing material can be synthesized to have any desired optical characteristics, i.e., optical absorption at any desired wavelength. It should be noted that any combination of the embodiments described with respect to any aspect of this present disclosure is applicable. In other words, any aspect of the present disclosure can be defined by any combination of the described embodiments.

[0014] It should be further noted that all of the following method embodiments may also be applicable to aspects of the energy absorbing material for use in a method for ablating corneal tissue. In some embodiments of the method, the energy-absorbing material is a biocompatible organic compound suitable for ophthalmic applications. In some embodiments of the method, the liquid comprises fluorescein. In some embodiments of the method, the liquid is sodium fluorescein dye. In some embodiments of the method, the irradiation includes focusing the irradiation light along a selected pattern on the corneal tissue. In some embodiments of the method, the selected pattern includes one or more annular rings or Fresnel rings. In some embodiments of the method, the energy-absorbing material absorbs the energy of the irradiated light in the visible spectrum. In some embodiments of the method, the energy-absorbing material absorbs irradiated light at multiple wavelengths or wavelength ranges.

[0015] In some embodiments of the method, the application is performed approximately 1 to 10 minutes before the irradiation. Throughout this application, the term "approximately" should be understood as a deviation of ±20% from the nominal value. Therefore, for example, if the value is approximately 10, it should be understood to lie within the range of 8 to 12. In some embodiments of the method, the application is performed approximately one minute before the irradiation. In some embodiments of the method, the application results in a surface made of the liquid being obtained on the corneal tissue, for example, a liquid reservoir being formed on the corneal surface. In some embodiments of the method, the irradiation is used to obtain the absorption of photons of the absorption wavelength. In some embodiments of the method, the absorption leads to a localized photothermochemical reaction, which in turn leads to localized ablation. In some embodiments of the method, the absorption wavelength is in the visible spectrum and is typically between 380 nm and 750 nm.

[0016] In some embodiments of the method, the irradiating light has a wavelength between 400 nm and 650 nm. In some embodiments of the method, the irradiated light has a wavelength of approximately 390 nm. In some embodiments of the method, the irradiated light has a wavelength of approximately 435 nm. In some embodiments of the method, the irradiated light has a wavelength of approximately 530 nm. In some embodiments of the method, the irradiation is performed by using a laser device. In some embodiments of the method, the laser device is a fiber laser. In some embodiments of the method, the irradiation light is continuous wave irradiation light. In some embodiments, the method further includes removing the fluid from the corneal tissue after the irradiation by washing the eye, for example, with a suitable cleaning agent. In some embodiments, the method further includes applying a dispersant containing nanoparticles onto the selected ablation profile, wherein the nanoparticles are selected to produce conditions that are invariant with respect to (i) incident light within a predetermined spectral range on the cornea, the precise position of the nanoparticles in the pattern, and the liquid type of the dispersant, and (ii) exhibit a dominant scattering effect with respect to incident light within the predetermined spectral range.

[0017] In some embodiments, the method further includes any of the above-described embodiments of the method for use in correcting the eye condition of the subject. A further aspect of this disclosure provides a kit for ablating corneal tissue. The kit includes a liquid containing an energy-absorbing material for application to corneal tissue, wherein the energy-absorbing material absorbs irradiation light at an absorption wavelength; and an irradiation unit configured to irradiate the corneal tissue with irradiation light including the absorption wavelength when the liquid is applied to the corneal tissue, thereby producing a selected ablation profile for the corneal tissue. The absorption wavelength is a wavelength suitable for the energy-absorbing material to absorb and produce a thermal reaction. It should be noted that the energy-absorbing material may absorb irradiation light of more than one wavelength. The energy-absorbing material absorbs irradiation light at the absorption wavelength, producing a localized photothermochemical reaction, thereby resulting in localized ablation of the cornea. In some embodiments of the kit, the energy-absorbing material is a biocompatible organic compound suitable for ophthalmic applications. In some embodiments of the kit, the liquid contains fluorescein. In some embodiments of the kit, the liquid is sodium fluorescein dye.

[0018] In some embodiments of the kit, the irradiation unit includes a controller configured to control the irradiation unit, including when to irradiate and which pattern to follow, so as to focus the irradiation light along a selected pattern on the corneal tissue. That is, the controller is configured to autonomously create an irradiation light pattern on the cornea. In some embodiments of the kit, the selected pattern includes one or more annular rings or Fresnel rings. In some embodiments of the kit, the energy-absorbing material absorbs the energy of the irradiated light in the visible spectrum. In some embodiments of the kit, the energy-absorbing material absorbs irradiated light at multiple wavelengths or wavelength ranges. In some embodiments of the kit, the irradiation results in the absorption of photons of the absorption wavelength. In some embodiments of the kit, the absorption leads to a localized photothermochemical reaction, which in turn leads to localized ablation.

[0019] In some embodiments of the kit, the absorbance wavelength is in the visible spectrum and is typically between 380 nm and 750 nm. In some embodiments of the kit, the irradiation light has a wavelength between 400 nm and 650 nm. In some embodiments of the kit, the irradiated light has a wavelength of approximately 390 nm. In some embodiments of the kit, the irradiated light has a wavelength of approximately 435 nm. In some embodiments of the kit, the irradiated light has a wavelength of approximately 530 nm. In some embodiments of the kit, the irradiation unit includes a laser device for performing the irradiation. In some embodiments of the kit, the laser device is a fiber laser. In some embodiments of the kit, the irradiation light is continuous wave irradiation light.

[0020] In some embodiments, the kit further comprises eye drops comprising a dispersant for application to the selected ablation profile to functionalize and stabilize the optical effect of the pattern, the dispersant comprising (i) a condition that is invariant with respect to one or more of the following: incident irradiation light within a predetermined spectral range on the cornea, the precise position of nanoparticles in the pattern, and the liquid type of the dispersant, and (ii) nanoparticles selected to exhibit a dominant scattering effect with respect to incident irradiation light within the predetermined spectral range. In some embodiments, the kit further includes any of the above embodiments of a formulation applied to the target eye. To further understand the subject matter disclosed herein and to illustrate how it can be carried out in practice, embodiments are described only by non-limiting examples with reference to the accompanying drawings. [Brief explanation of the drawing]

[0021] [Figure 1A] A diagrammatic illustration shows a kit for use in the ocular vision correction technology of this disclosure. [Figure 1B] The principle of preparing a dispersant containing nanoparticles using the technology of this disclosure is illustrated. [Figure 1C] A schematic diagram illustrates a system for use in correcting eye conditions according to some embodiments of this disclosure. [Figure 2] This example illustrates a Fresnel zone plate pattern suitable for correcting eye conditions by increasing or decreasing optical power. [Figure 3-4] This illustrates the ocular correction pattern associated with the expansion of the lens's depth of focus. [Figure 5] The following are examples of albumin protein desolvation processes according to some embodiments of the present invention. [Figure 6A-6B] This shows the measurement of the size and electrical (zeta) potential of nanoparticles generated by some embodiments of the present invention. [Figure 7A-7C] TEM and cryoTEM micrographs, as well as a size distribution histogram, are shown for HA score NP. [Figure 8A-8B] Figure 8A shows the FTIR spectroscopy of an HSA core NP supported by maghemite nanoparticles according to some embodiments of the present invention. Figure 8B shows the FTIR results, and Figure 8B focuses on the peaks in the absorption spectrum of the maghemite supported by the HSA nanoparticles; [Figure 9A-9B] The X-ray photoelectron (XPS) spectra of crude HSA (Figure 9A) and HSA NP (Figure 9B) described herein are shown. [Figure 10A-10D] This report demonstrates that the morphological and size distribution of hybrid HSA / CAN-γ-Fe2O3 nanoparticles were characterized using TEM, cryoTEM, and HR-SEM. [Figure 11A-11D] Figure 11A shows HR-SEM images of HSA NPs according to some embodiments of the present invention, as well as measurements of the amounts of selected elements using line scan analysis for carbon (Figure 11B), oxygen (Figure 11C), and iron (Figure 11D). [Figure 12]This shows reflectance measurements from superreflective iron particles embedded in HAS NPs, indicating NPs within the etched region of the cornea. [Figures 13A-13B] The selected pattern is applied to the eye, and the measured changes in the optical power of the pig's eye are shown, provided by the provision of an eye drop solution according to some embodiments of the present invention. [Figures 14A-14B] This is a schematic block diagram illustrating non-limiting embodiments of a kit for ablating corneal tissue according to aspects of the present disclosure. [Figures 15a-15b] Figure 15a shows an ex-vivo pig eye image of an eye placed in a custom-made container, and Figure 15b shows the container connected to a stepping motor and fixed on an optical post. [Figures 16a-16c] The experimental setup is shown; Figure 16a shows the eye placed in a custom-made container with the fiber tip positioned close by; Figure 16b shows a top view of Figure 16a; and Figure 16c shows the optical system scheme. [Figures 17a-17k] The images show eye images. Figures 17a-17h show bright-field images of various sections of the eye irradiated with a laser (images were taken after surgical removal of the corneal layer), Figure 17i shows a photograph of the eye after laser treatment, Figure 17j shows a photograph of the eye before laser treatment, and Figure 17k shows a photograph of the eye after laser treatment. The ablation trajectory is not shown. [Figures 18a-18b] The images of eye 5 are shown; Figure 18a shows a bright-field microscopy image of the corneal surface of eye number 5 after ablation; and Figure 18b shows a fluorescence confocal image of the same eye using a Lecia confocal microscope with a 20x objective lens. [Figures 19a-19o] This image shows a Z-stacked confocal microscope image of eye 5 from depth Z=0 to Z=184 μm. Fluorescence signals are only observed up to a depth of 160 μm. [Figures 20a-20d]The images of eyes 6 and 7 are shown. Figure 20a shows a bright-field microscopy image of eye number 6 at 1.25x magnification of the corneal surface; Figure 20b shows a confocal image of the entire eye of eye number 6; Figure 20c shows a confocal image of eye number 7 with an ablation width of 300 μm and an ablation depth of 247 μm; and Figure 20d shows a reflection mode image of eye number 7 in the same field of view. [Figures 21a-21b] The images of eye 8 are shown. Figure 21a shows a bright-field microscopy image of eye number 8 at 1.25x magnification of the corneal surface; Figure 21b shows a confocal image of the whole eye of eye number 8 before sectioning with an ablation width of 570 μm and an ablation depth of 465 μm. [Modes for carrying out the invention]

[0022] The technology of this disclosure provides correction for visual impairments, such as myopia, myopia control, hyperopia, and presbyopia. Figure 1A schematically shows a kit 10 for carrying out the ocular vision correction technology according to this disclosure. Kit 10 includes a patterning system 20 and eye drops 50. The patterning system 20 includes a controllably operable patterning utility 30 configured to generate a selected three-dimensional pattern on the user's cornea. To achieve this, the patterning utility 30 is operable by a controller 40, which provides patterning utility input data indicating pre-stored pattern data corresponding to a particular user's visual impairment.

[0023] The patterning utility may be any known and suitable type for generating a two-dimensional arrangement of incisions / grooves placed at specific depth intervals, and thus for forming a three-dimensional pattern. The patterning utility may be configured to utilize operational commands to generate a pattern on the cornea of ​​the user's eye by applying ultrasound, via mechanical etching, or via laser-based etching. In the following description, such patterning utilities are sometimes referred to as “etching utilities,” but it should be understood that the principles of the technology described herein are not limited to this particular example. The eye drops 50 are formulated in the form of a dispersant containing nanoparticles dispersed in a liquid. The majority of the nanoparticles are selected so that, when the eye drops are applied to a three-dimensional pattern formed on the target cornea, the nanoparticles produce conditions that are invariant with respect to the incident irradiation light in a given (e.g., visible) spectrum on the cornea, and / or the concentration of the nanoparticles, and / or the precise position of the nanoparticles and the liquid. Furthermore, the majority of the nanoparticles are selected so that, when the eye drops are applied to a three-dimensional pattern, the nanoparticles exhibit a dominant scattering effect over nanoparticle absorption or an absorption effect over scattering within a given spectral range.

[0024] It should be noted that the parameters of the nanoparticles, such as shape and size, are selected to provide desired scattering-absorption properties, ensuring that the nanoparticles respond as scatterers rather than absorbers to incident light in a particular spectrum (the visible spectrum for the purposes of this disclosure). In this regard, the scattering and absorption properties of nanoparticles are defined by scattering and absorption resonance peaks, as well as by the size and shape of the nanoparticles. It should be understood that generally, spherical nanoparticles have one scattering resonance peak and one absorption resonance peak, while rod-shaped nanoparticles have two scattering peaks and two absorption resonance peaks. Therefore, for the purposes of this disclosure, the size and shape of nanoparticles of a given material composition are selected such that (1) the center of the scattering resonance peak is located at a desired wavelength (e.g., the wavelength of the green spectrum, e.g., about 530 nm) and the peak width is relatively broad, and (2) the amplitude of scattering at the spectral peak is much higher than the amplitude of absorption.

[0025] For example, according to the Mie model, a nanosphere with diameter d is λ res It has a resonance peak at the wavelength, which can be determined according to the following relationship:

number

number

[0026] It should be noted that the invariant conditions are generated by the combined effect of the inherent properties of the cornea and the nanoparticles applied to the three-dimensional pattern formed on the corneal surface. Therefore, the kit can be used as follows: operate the patterning utility as described above to form the selected three-dimensional pattern on the corneal surface; and apply the aforementioned dispersant to the pattern using eye drops. The formation of the selected three-dimensional pattern is performed using a patterning utility (e.g., an etching utility) that operates in a controllable manner according to input data indicating the user's visual impairment. The patterning utility may be configured to transmit selected ultrasound to etch the pattern onto the user's cornea, or it may be configured to mechanically etch the cornea. In some other embodiments, the patterning utility includes at least one laser unit and a beam steering module, configured to provide optical energy to etch a selected pattern on the cornea of ​​the user's eye according to operating commands. The laser unit may be configured to provide etching to a depth of a single cell layer of the cornea.

[0027] As schematically shown in Figure 1B in an obvious manner, pattern data, as well as data indicating the operational spectral range (typically the visible range in a particular application of the eye drop), are used to select the optimal formulation (dispersant) containing nanoparticles. This includes the material composition of the nanoparticles, the size and shape of the nanoparticles, and the amount / concentration of nanoparticles in the liquid. The selection of nanoparticles aims to provide the desired results of the combined effect of the pattern and nanoparticles, particularly obtaining a dominant scattering effect at a desired wavelength or wavelength range, and thus functionalizing and stabilizing the optical effect of the pattern itself. It should be understood that the selection of formulation data can be performed automatically using a computer system that utilizes data from a specific database. Such data may include the scattering vs. absorption properties of nanoparticles of any given shape and size, as well as the material composition. Figure 1C is a schematic diagram of system 100 that defines the operation of the pattern forming system 20 for further use in the kit 10 for correcting the eye conditions described above. System 100 may generally be configured as a computer system and, in conjunction with the pattern forming system 20, provide output pattern data stored in the storage utility 400 of the pattern forming system 20 (for example, in the controller 40 of system 20). Thus, system 100 includes a processing utility 200 and typically also includes input / output utilities and a user interface, which are not specifically shown in the figure.

[0028] In this non-limiting example, the patterning utility 30 is illustrated as including an etching utility 300 configured and operable to form a pattern by appropriately applying laser radiation. The etching utility 300 includes a laser unit 310 and a beam steering unit 320. The operation of the etching utility is described below. The processing utility 200 may generally include one or more processors and further include one or more hardware or software modules, such as a corrective pattern module 210. The corrective pattern module 210 is configured to receive and process input data indicating, for example, the thickness of the epithelium, the optical correction required for the user's eye (each eye may require a different optical correction), and to determine, for example, an optical wavefront or an automated refractometer, a selected three-dimensional pattern (such as those described above) suitable for optical correction, or to retrieve it from a storage utility (e.g., an external database). The processing utility 200 is further configured to generate pattern data that forms operational commands for a patterning utility to etch the selected pattern onto the user's cornea to provide the selected optical correction. The processing utility 200 generates and transmits the pattern data (commands) to the etching utility 300, instructing it to provide optical illumination and generate the selected pattern on the cornea of ​​the user's eye.

[0029] Figures 2, 3, and 4 schematically illustrate three possible patterns selected according to the user's eye condition. Figure 2 shows a Fresnel zone plate pattern containing multiple concentric rings of varying widths; Figures 3 and 4 illustrate patterns selected for extending the depth of focus, suitable, for example, for correcting presbyopia. Generally, the optical properties of such patterns and the precise three-dimensional arrangement of each pattern may be determined according to the selected ocular correction, e.g., the optical power required for correction, or the required increase in depth of focus. Generally, the etching utility 300 is configured to apply a selected pattern to the corneal epithelial layer, Bowman's layer, or the anterior corneal stroma as an incision / ablation. Etching may generally be provided using optical etching, ultrasonic etching, and / or mechanical incision of the cornea with a selected wavelength range. For simplicity, the etching utility is exemplified herein as an optical etching utility 300 comprising a laser light source and a beam steering unit 320. The laser light source 310 is configured to provide optical radiation at a wavelength and power suitable for etching within epithelial cells, Bowman's layer, or the anterior corneal stroma, and the steering unit 320 is configured to change the path of the output light beam so that the selected pattern is projected onto the user's cornea. The operating speed of the steering unit 320 and the power of the light source unit 310 are typically selected to provide corneal photoetching within the epithelial surface, for example, to the depth of a single-cell layer, Bowman's layer, or the anterior corneal stroma.

[0030] More specifically, the pattern should generally be etched within or directly beneath the epithelial layer. It should also be noted that the selected optical pattern can be effectively imprinted within the corneal epithelial layer. As described above, the pattern is selected to provide the desired optical effect to enable the correction of visual impairment. When applied to the pattern, the nanoparticle-containing dispersant actually functions / activates and stabilizes (gives persistence) this optical effect. Also, as described above, the eye drop solution (formulation / dispersant) contains selected nanoparticles having a different refractive index than the cornea and / or periocular tears, and the dispersant is applied to the pattern so that the nanoparticles are arranged / dispersed within the pattern (the incision / ablation area of ​​the pattern) at substantially low density (i.e., so that the arrangement of nanoparticles does not affect the refractive effect induced by the structure of the pattern). The effects induced by the arrangement of nanoparticles dispersed in the pattern include the scattering (or scattering and absorption) of light of a desired spectral range (e.g., the visible spectral range) incident on the nanoparticles located within the cutting / etching regions of the pattern, which functionalizes and stabilizes the optical effects of the pattern (which are selected depending on the visual correction applied) resulting from the scattering nanoparticles filling the cutting / etching regions of the pattern.

[0031] More specifically, the material composition of the nanoparticles is selected such that, with respect to a given size and shape of the nanoparticles, it exhibits dominant scattering with respect to incident and irradiated light in a given spectral range (e.g., visible light) (by performing excessive absorption, thereby preventing unwanted heating effects in the eye); and the arrangement of the nanoparticles is selected to be substantially low density. For example, several nanoparticles (generally at least two nanoparticles in the visible range, e.g., four nanoparticles) are selected for a λ×λ area region within the pattern. This means that only a small number of nanoparticles are needed to disperse within the pattern (i.e., the concentration of nanoparticles in the eye drops is reduced). Such selection of nanoparticles for a given pattern provides an invariant condition for the pattern with nanoparticles on the cornea with respect to incident light in a given spectrum, and thus generates an invariant condition for the point spreading function obtained on the target retina through the combined effect of the intrinsic properties of the cornea, the pattern, and the dispersion of nanoparticles within the pattern. Generally, the amount of nanoparticles required to be dispersed in a pattern and their arrangement in the pattern depend, in particular, on the critical dimensions of the pattern, such as the ablation depth, diameter, and width. For example, considering a pattern containing a ring, the width relates to the width of the ring itself (e.g., 375 microns in the case of presbyopia treatment), and the diameter relates to the outer diameter of the pattern, such as 3 mm.

[0032] The inventors have shown that, with relatively low density selected nanoparticles (i.e., the nanoparticles scatter most of the light in the selected visible range), the nanoparticles function as scatterers that do not affect / modify the refractive optical effect induced by the pattern structure itself, but rather function and stabilize the effect of this pattern due to the optical effect of scattering induced by the nanoparticles. For example, considering substantially spherical nanoparticles with a diameter of about 80 nm and an operational wavelength in the visible spectral range (e.g., λ = 530 nm), the arrangement of nanoparticles in the pattern is such that λ in the pattern 3 This is a volume space that contains up to 200 nanoparticles. As described above, suitable ophthalmic formulations may include, for example, HAS-based nanoparticles (e.g., recombinant human serum albumin nanoparticles), or metal-based nanoparticles, such as biocompatible materials, such as biocompatible shells (e.g., albumin shells), i.e., any suitable nanoparticles containing gold supported by folded proteins around the particle. It should be noted that the particles in the ophthalmic solution are configured to ensure that the optical effect of the engraved pattern stabilizes over time. More specifically, the nanoparticles are fixed within the pattern (within the incised / etched area) by etching to a specific depth, functionalizing / activating the pattern, preserving the pattern shape in "optical activity," and thus maintaining the optical effect of the pattern, providing the optical effect for several days to several months.

[0033] Generally, protein-based NPs are often used as drug carrier particles and may be of interest for eye drop solutions due to their high stability during storage, as well as their biocompatibility, such as non-toxicity and non-antigenicity. Albumin proteins provide polymer carriers that have been shown to be biodegradable, non-immunogenic, non-toxic, and in vivo metabolizable. In some embodiments of the present invention, the nanoparticles may contain human serum albumin (HSA, 66.5 kDa). HAS is the most abundant plasma protein (35-50 g / L human serum) and has a mean half-life in the blood of 19 days, which may make it preferable, and therefore it is used below. However, it should be noted that the nanoparticles of the present invention may typically be associated with any albumin type protein, including various animal serums and / or synthetic or recombinant albumins. Generally, albumin contains 35 cysteinyl residues forming one sulfhydryl group and 17 disulfide bridges. HSA is robust to pH and stable in the pH range of 4-9. Furthermore, HSA can be heated at 60°C for up to 10 hours.

[0034] The inventors of the present invention have identified a technique that enables the provision and characterization of HSA-based nanoparticles (NPs) having robust and controllable particle size. The technique utilizes a nanofabrication method by a desolvation process and a crosslinking process with divinyl sulfone (DVS), as illustrated in Figure 5. During the desolvation process, spherical protein NPs are formed and then cured / stabilized by crosslinking facilitated by DVS. In this context, a covalent Michael reaction occurs between the free nucleophilic functional groups of the HSA protein and the strongly electrophilic bifunctional DVS reagent, resulting in intra-chain and / or inter-chain bonds within the NPs. The preparation of such HSA NPs may make available various free functional groups on their surface, which can be used for a further second step of functional group modification.

[0035] In some examples, the nanoparticles may be based on a hybrid organic / inorganic nanosystem consisting of HSA NPs encapsulated with metal nanoparticles (gold nanoparticles or maghemite (γ-Fe2O3) nanoparticles). Hybrid organic / inorganic nanosystems containing HSA NPs encapsulated with hydrophilic (NH4)2Ce(lV)(NO3)6, modified with cerium ammonium nitrate (CAN) and called CAN-maghemite or CAN-γ-Fe2O3NP, have also been fabricated and characterized. Several suitable nanoparticles have been generated and characterized, these being the nanoparticle variants / species and / or nanoparticles associated with the intermediate generation steps in this technique, as exemplified below.

[0036] (Example 1) Manufacturing of HSA core nanoparticles HSA core nanoparticles were prepared by dissolving 50.0 mg of HSA in 1.0 mL of purified water (ddH2O). Ethanol was added during the desolvation process to a total volume of 5.0 mL, resulting in a final HSA concentration of 10 mg / ml. Protein chain crosslinking was induced by adding 140.0 μL of DVS (5% w / w in EtOH). The crosslinking process was carried out by stirring the suspension at 55°C for 1 hour, followed by 20 minutes in an ultrasonic bath. The resulting HSA nanoparticles were then purified by fractional centrifugation (at 13,500 rpm, 60 minutes, at 4°C) for 3 cycles, and then redispersed in the original volume of ddH2O. Each redispersion step was performed in an ultrasonic bath for 10-15 minutes. To enable long-term storage and avoid aggregation, the nanoparticles were stored in a refrigerator at 4°C.

[0037] (Example 2) Characterization of HSA core nanoparticles The obtained HSA core NPs were further characterized by various combinations of analytical, spectroscopic, and microscopic methods. Figures 6A and 6B show the analysis of the nanoparticle size distribution (Figure 6A) and charge distribution (zeta potential, Figure 6B). The obtained fabricated HSA NPs exhibit a NP hydrodynamic size (DLS) of 149.56 ± 1.8 nm and a strong negative zeta (electrical) potential of -35.4 ± 2.4 mV. Furthermore, the nanoparticles exhibit low dispersibility, which is associated with a polydispersity index (PDI) value of 0.17. Figures 7A–7C show TEM micrographs, cryo-TEM micrographs, and size distribution histograms of HSA core NPs. These measurements demonstrate the formation of spherical, substantially homogeneous HSA core NPs with an average size of 23.05 ± 5.3 nm. Additional measurements of nanoparticles were performed, and the crosslinking of HSA polymer chains was determined using DVS reagent. Figures 8A and 8B show that FTIR spectral measurements revealed structural differences between standard HAS G2 and maghemite G1 supported with HSA nanoparticles as described herein. Figure 8A is at 4050 cm⁻¹. -1 ~550cm -1 Figure 8B shows the relative intensity measured along the spectrum between 800 and 1100 cm⁻¹, focusing on the region indicated by the rectangle in Figure 8A, and in particular showing the peak in the absorption spectrum G1. Structural differences between particles are illustrated by FT-IR, and the changes in the absorption peak are generally attributed to the presence of DVS in the core HSA NPs. The spectral difference between the crude HSA and HSA NP spectra is shown in Figure 8B from 800 to 1100 cm⁻¹. -1 This can be easily observed within this range. The figure shows several peaks in the HSA NP spectrum G1 that do not appear in the standard HSA FTIR spectrum G2. 880 cm -1 The first peak in the region corresponds to the stretching vibration of the CS coupling at 1047 and 1083 cm. -1 The two other peaks that appeared in this sample may be due to the stretching vibration of the sulfoxide group (RS=O).

[0038] Furthermore, X-ray photoelectron (XPS) spectroscopy was used to confirm the involvement of this electrophilic DVS reagent during HSA polymer chain crosslinking. Figures 9A and 9B show the XPS spectra of crude HAS (Figure 9A) and HAS NPs described herein (Figure 9B). XPS analysis of the HSA NPs showed that the S element exists in two oxidation states, i.e., the first state corresponds to the R-S species (binding energy of BE = 164.0 eV), and the second state is in a high binding energy state, typically at 168.5 eV, which has been shown to correspond to the SO2 sulfoxide group derived from the DVS sulfone group. This peak at 168.5 eV does not appear in the XPS analysis of the starting crude HSA protein.

[0039] The main functional groups generally containing primary amines (NH2) and carboxyl groups (COOH) were successfully quantified using the UV-sensitive Kaiser test (1,3-diaminopropane was used to amidate / derivatize the EDC-activated COOH groups). The resulting differential EDC- and non-EDC-based Kaiser results showed that for 1.0 g of HSA NPs, the NH2 and COOH groups were present at 0.632 and 1.127 millimoles, respectively (these quantified surface functionalities enable the future use of HSA NPs as biocompatible biodegradable drug carriers to allow for additional therapeutic modalities and / or further binding of drugs targeting tumors and diseased cells).

[0040] (Example 3) Preparation of CAN maghemite nanoparticles The preparation of CAN maghemite NPs ((CeL n ) 3 / 4+ -γ-Fe2O3) was carried out in a two-step procedure involving basic coprecipitation of two types of Fe 2+ / 3+ salts. This procedure yielded magnetite (Fe3O4) NPs as the starting NP material with a low degree of oxidation, which was then oxidized with the monoelectronic oxidant cerium ammonium nitrate (CAN), (CeL n ) 3 / 4+By modifying the surface of cation / composite NPs through surface doping, we obtained CAN-γ-Fe2O3-NPs with a size of 6.61±2.04 nm and a positive charge (+45.7 mV).

[0041] (Example 4) Preparation of Hybrid CAN Maghemite-Containing HSA Nanoparticles To provide a desired nanoparticle structure with a refractive index suitable for acting on vision according to a selected pattern on the cornea, HSA and CAN-γ-Fe2O3NP, both nanoscale components, were assembled, and CAN-γ-Fe2O3NP was captured within HSA nanocomposite particles during the same DVS-mediated NP fabrication / component crosslinking. For this purpose, hybrid CAN maghemite containing HSA nanoparticles was prepared by the same procedure as described above for HSA NP. Briefly, CAN-γ-Fe2O3NP was added to HSA (50.0 mg in 1.0 mL of ddH2O) in a mass ratio of 25:1, incubated at room temperature for 1 hour, followed by desolvation and crosslinking processes. The resulting composite nanoparticles were then purified by fractional centrifugation (13,500 rpm, 60 min, 4°C) for 3 cycles, magnetically decanted (using a strong external magnet), and redispersed in the original volume of ddH2O. In each redispersant step, the reaction vessel was placed in a low-power ultrasonic bath for 15 minutes before processing. To ensure long-term storage and to avoid aggregation, the nanoparticles were stored in a refrigerator (4°C).

[0042] It should be noted that the optimal mass ratio between the HSA phase and the CAN-γ-Fe2O3NP phase for optimal encapsulation of the maghemite composition within the HSA nanoshell was found to be 25:1. Generally, the mass ratio may be in the range of 15:1 to 40:1. The hydrodynamic diameter, size distribution, and electrical potential of the DLS of the corresponding composite particles were determined. A DLS hydrodynamic diameter of 130 nm was measured, and the polydispersity index was less than 0.3. Electrical potential analysis was also performed to confirm the colloidal stability of the obtained hybrid NPs for aggregation control (capture method), which indicated that the composite particles were consistently negatively charged, with an average electrical potential of -25 mV. Referring to Figures 10A-10D, the morphological and size distribution measurements of hybrid HSA / CAN-γ-Fe2O3 nanoparticles were characterized using TEM, cryo-TEM, and HR-SEM. Figures 10A and 10B show TEM and cryo-TEM images, respectively, of each HSA NP phase (somewhat low-contrast gray areas) containing captured CAN maghemite NPs (dark spots) that are easily visible and identifiable within the HSA phase. Thus, it was observed that the electron-high density metal NPs were successfully incorporated into the surrounding HSA matrix, and at the same time, the distribution of CAN maghemite NPs between and within each particle was very homogeneous, which is thought to result from the promotion of interaction between the positively charged CAN-γ-Fe2O3 NP phase and the negatively charged HSA phase. The NP crystallinity was further confirmed by TEM / selective-region electron diffraction (SA ED) as shown in Figure 10D, and furthermore, the nanoparticle size distribution was also measured by TEM in Figure 10C, revealing an average size of 44.6 ± 4.18 nm.

[0043] Figures 11A–11D show HR-SEM images (Figure 11A) and line scan analyses of carbon (Figure 11B), oxygen (Figure 11C), and iron (Figure 11D) obtained in a scanning transmission electron microscope (STEM) style. Scan lines for elemental analysis are indicated as L1–L3 for carbon, oxygen, and iron, respectively. The figures show that CA maghemite NPs exist as fully encapsulated clusters within the HSA matrix, with the HSA phase appearing simultaneously as a surrounding cloud. The line scan elemental analysis, based on energy-dispersive X-ray spectroscopy (EDS), verifies that CAN-γ-Fe2O3 NP encapsulation primarily occurred within the resulting HSA nanocomposite particles. Indeed, the elements iron (upper right), carbon (lower left), and oxygen (lower right) are present simultaneously. The peak for iron is much fainter than the others, thus demonstrating that CAN-γ-Fe2O3 NPs were encapsulated within the HSA NPs. Furthermore, Figure 12 shows the reflection of iron particles embedded in the HSA NP. As shown by the reflection of the iron particles, the HSA NP is positioned within the corneal incision area and enhances the etched pattern.

[0044] ICP-AES elemental analysis was also performed to confirm that iron was encapsulated during hybrid NP formation. ICP measurements demonstrated a capture efficiency of 95% for elemental iron in the HSA phase. Additional concentration measurements of both the hybrid NP and CAN-maghemite NP showed that the mass percentage of the CA-maghemite NP phase was approximately 40% of the total mass of the corresponding hybrid NP. Therefore, this technology utilizes a solution formulation as eye drops containing CAN-γ-Fe2O3 nanoparticles with biocompatible protein chains, e.g., albumin-based NPs, e.g., rHSA-based NPs, and / or biocompatible protein shells, e.g., albumin shells (rHSA shells), so that the nanoparticles are dispersed in a selected etched pattern on the user's cornea and, when positioned in the incised area of ​​the pattern, produce a scattering effect with respect to incident light within a given spectral range, providing functionalization and stability of the refractive effect induced by the pattern structure. The nanoparticles generally remain on the cornea for several days to several months, allowing for correction of the user's eye condition according to the optical manipulation of the selected pattern. The inventors conducted tests in the eyes of pigs and rabbits to verify the optical effects of the etched pattern and selected nanoparticles for correcting optical visual impairment.

[0045] To achieve this objective, the refractive error of hundreds of fresh pig eyes was measured before and after the realization of corneal surface patterns, using both an automated refractometer and a wavefront abberometer, and then after the administration of eye drops filled with selected nanoparticles at specific known concentrations between 0.1 mg / ml and 15 mg / ml. The nanoparticle concentration used in this example was 10 mg / ml. Both myopia and presbyopia correction were tested in these ex-vivo experiments. Patterns were applied to pig corneas using a mechanical stamper, constructed using a 3D printer with a calibrated optical pattern, or with laser assistance. The laser-assisted pattern used in this embodiment provides surface ablation on the cornea, allowing nanoparticles to penetrate the cornea and thus enabling the desired change in optical power determined by the pattern. For myopia correction, the pattern itself was selected to be an optimized non-binary Fresnel plate zone diffraction pattern that provides 3 diopters of optical correction in this specific experiment. For presbyopia, a depth-of-focus extension pattern was used to achieve an increase in depth of focus equivalent to a 1.5D addition. The optical power of the pig eye system was measured and recorded five times at each time point in the experiment, as follows: baseline optical power, immediately after the pattern was etched into the cornea, after applying eye drops into the pattern, and then 5, 10, 20, and 30 minutes after dropping eye drops into the pattern. Various measurements were taken to avoid bias due to large standard deviations.

[0046] Therefore, after selecting the desired pattern according to the required optical correction, the 3D dimensional pattern was applied to the cornea. Once the pattern was etched onto the cornea, the eye was washed with an eye drop solution containing human serum albumin nanoparticles conjugated with Cy5 to allow for identification by confocal fluorescence microscopy and histopathology. Referring to Figures 13A and 13B, the results of changes in optical power for pig eyes corrected using the techniques described above are shown. Figure 13A shows the results of 3-diopter myopia correction for 64 pig eyes obtained using the techniques of this disclosure. As can be seen in Figure 13A, no change in induced refraction was observed immediately after pattern generation ("Pattern" in the graph), whereas when nanoparticles were added to the pattern, the refraction changed according to the planned correction ("NP" in the graph). The results were similar in in vivo and ex-vivo models, and a change in refraction occurred only when the pattern was filled with recombinant human serum albumin nanoparticles. Figure 13B shows similar results for depth-of-focus extension treatment suitable for presbyopia correction. The criterion for "focus shift" is based on the visual Strehr ratio by through focus and is measured to evaluate the effect generated by the depth-of-focus extension (EDOF) element (surface relief pattern) etched onto the corneal epithelium and optically activated with rHSA nanoparticles.

[0047] As control parameters, the inventors also analyzed changes in the overall spherical component of the eye ("spherical value") and the overall refraction of the optical system ("spherical equivalent value"). As shown in Figure 13A, the 3-diopter myopia pattern applied in combination with nanoparticle eye drops provided an average correction of -3.12 ± 0.58 diopters at the time of the final measurement (30 minutes after nanoparticle application), while the change induced by generating a myopia pattern in the eye alone (without nanoparticles) was not statistically significant (0.1 D ± 0.13). In Figure 13B, an increase of 1.22 ± 0.2 D in depth of focus was observed with respect to presbyopia correction, and the overall spherical value and spherical equivalent value of the eye were -0.08 D and -0.04 D, respectively, with no significant changes. The displacement of focus measured in the presbyopia test is based on the visual Strehr ratio due to through focus. Therefore, as shown, the use of a selected pattern applied / etched onto the cornea, with a suitable eye drop containing a selected sufficient concentration of nanoparticles as described above, provides a selected visual correction effect (refractive effect). As a result, when the nanoparticles are positioned / dispersed in the incised area of ​​the pattern, they induce a scattering effect with respect to light in a given spectral range (typically the visible range), functionalizing the optical effect of the pattern and allowing correction of various eye conditions by being sustained over a moderately long period. Thus, the technique of the present invention is minimally invasive. Generally, the pattern may be diffractive (e.g., Fresnel zone plate / ring) or configured to introduce optical interference and allow for an extension of the depth of focus of the eye.

[0048] Typically, the eye drops and corresponding nanoparticles described herein may be suitable for use in a variety of additional conditions. For example, the eye drop solutions described herein may be used to treat dry eye disease (DED), in which the nanoparticles provide lubrication to the eye's movement as synthetic tears. DED represents a heterogeneous group of conditions characterized by a lack of tear film and signs and / or symptoms of ocular surface irritation. These conditions may be associated with a variety of factors, including, for example, meibomian gland dysfunction (MGD). The use of nanoparticles, including the eye drops described herein, may be used to create biological bonds with lipid molecules that make up the lipid layer of the user's eye. As a result, the stability of the tear film is increased and the effect on the corneal surface is extended. Furthermore, it should be noted that the aforementioned nanoparticles are present in the eye drop solution and may be used as drug delivery carriers. Generally, the anatomical barriers and physiological clearance mechanisms of the ocular surface present challenges for the development of ocular drug delivery devices. More invasive methods, such as intravitreal injection, can improve the ocular bioavailability of therapeutic agents, but often result in serious side effects on vision.

[0049] The use of selected nanoparticles described herein may enhance the ocular bioavailability of one or more suitable therapeutic agents (e.g., drugs, proteins, peptides). The aforementioned nanoparticles may offer significant potential benefits, including, for example, improved penetration / bioavailability / delivery of any bioactive agent into deep tissues to treat more severe conditions, such as bacterial, fungal, or parasitic infections. Additional significant benefits may relate to improved bioavailability, which may lead to a significant reduction in the frequency of eye drop administration in chronic diseases, such as glaucoma or dry eye, as well as potentially externally controllable drug delivery via portable devices. Accordingly, the present invention provides an eye drop material composition comprising a desired (relatively low) concentration of nanoparticles of a selected size and shape, for example, biocompatible protein chains (preferably HSA-based), or metal-based nanoparticles accompanied by a biocompatible protein shell (e.g., an albumin shell), thereby enabling the arrangement of relatively low-density nanoparticles within the pattern, thereby functionalizing, maintaining / stabilizing the optical effect for which the pattern structure is designed. Thus, the eye drop composition is used in combination with a suitable patterning applied to the cornea of ​​the user's eye to provide activation, sustainment, and stabilization of selected optical correction for various eye conditions.

[0050] Referring to Figures 14A–14B, these are schematic block diagrams illustrating non-limiting embodiments of a kit for ablating corneal tissue according to aspects of the present disclosure. Figure 14A illustrates a kit 1400 comprising a liquid containing an energy-absorbing material 1402. The liquid 1402 may be contained in a container, e.g., a bottle or tube or disposable packaging. The energy-absorbing material is selected to interact with a particular irradiation wavelength, i.e., an absorption wavelength, to result in highly precise ablation of corneal tissue, for example, by causing explosive ablation. Thus, the energy-absorbing material absorbs the irradiation light at the absorption wavelength and reacts thermochemically with the cornea. The liquid 1402 is intended to be applied onto the corneal tissue of interest to form a layer of the energy-absorbing material-containing liquid on a portion of the cornea of ​​interest. The kit 1400 further comprises an irradiation unit 1404 for providing irradiation light including the absorption wavelength. The absorption wavelength may be in the IR or visible spectrum, as long as it interacts with the fluid 1402 on the corneal tissue and results in the desired ablation of the cornea. The irradiation unit 1404 can be controlled to produce a desired irradiation light pattern and profile, i.e., a pattern on the cornea along which the irradiation light is applied. This forms the desired ablation pattern on the target corneal tissue.

[0051] Figure 14B differs from the example in Figure 14A in that the laser 1406 and control unit 1408 are included in the irradiation unit 1404. The laser 1406 is an irradiation device that provides desired irradiation light onto the cornea. In this embodiment, the laser is controlled by the control unit 1408, which applies a desired irradiation light pattern to the target corneal tissue. The desired irradiation light pattern applied by the control unit 1408 can be individualized according to the specific properties of the target. That is, for each target, different irradiation light patterns and profiles are applied, which may be different patterns along the cornea, or the irradiation light may have different irradiation light wavelength profiles.

[0052] (Example 5) Enhanced ablation of fluorescein-doped corneas using visible wavelength lasers Materials and methods Preparation of pig eye specimens Fresh pig eyes were obtained from a local CRO (Lahav CRO, Israel). Pig eyes are anatomically similar to human eyes and are readily available in large quantities in excellent, reproducible biological condition. Extraction was performed in-house and the eyes were used within 4 hours of extraction. Each eye was placed in a rotating cylindrical container as shown in Figure 15a. The cylindrical container was connected to a rotary stepper motor operated using an Arduino board. The container was fixed to an optical post using the clamp structure shown in Figure 15b, maintaining the eye in a forward-facing position, similar to a human sitting and looking directly at an optical post. Experimental System The experimental setup is shown in Figures 16a-16c. It consists of a 532nm CW (maximum power 800mW) single-mode fiber laser. The fiber tip was positioned very close to the tip of the eye (less than 1mm away). A stepper motor was controlled by an Arduino board. The motor speed could be changed. The objective was to rotate the eye and simultaneously turn on the laser to ablate a circle of a predetermined radius on the cornea.

[0053] Fluorescein pigment Fluorescein was extracted from a clinically used strip (BIO FLUORO, 1 mg sodium fluorescein [C20H10O5.2Na.2H2O] USP strip (lot FL15G1605, CE 0434)) immersed in physiological saline to obtain a concentration of 4.68 mg / ml. This is the clinical concentration from a clinical strip and the amount of water typically used to moisten the strip during clinical application of the dye. The dye is a clinically used solution, approved by the FDA for topical application (NDA208582), and its safety has been established for all ages, including pediatric patients. A single application of the dye solution, similar to generally clinically accepted practice, is sufficient to produce a photoablation effect, and any excess dye is washed away by tears, eliminating the need for additional washing. The dye was either applied as a regular eye drop (i.e., one drop to create a tear film-like coating on the corneal surface) one minute before the laser procedure, or applied in large quantities until the eye was positioned forward and the laser procedure was immediately administered, i.e., the corneal surface was immersed for one minute in an excess amount of dye that formed a pool on the corneal surface.

[0054] Principle of Ablation The interaction between laser photons and corneal tissue can be classified into three main principles based on how the photon energy is deposited onto tissue molecules. When the photon flux is high enough to heat the substrate by converting electronic excitation into vibrational energy, it is called thermal ablation. Photochemical ablation occurs when damage is extremely low due to the localization of energy. The process occurs at a very fast rate because the energy conversion does not generate vibrational molecules that would cause a heating effect. The third principle is called explosive ablation, which is similar to thermal ablation but is a controlled process that does not rely on heated particles to ablate the surroundings. This type of ablation occurs on a short-time scale in which energy is converted into kinetic energy, without simultaneously damaging unirradiated tissue, and is based on the effect of enhancing laser energy absorption by introducing a dopant into the tissue. By adding fluorescein (an organic dye) as an energy-absorbing dopant into the tissue, effective ablation with 485 nm visible light can be obtained.

[0055] The chemical structure of fluorescein (F) can be written as shown in Scheme 1. [ka] Previous studies have shown that in a dark room, fluorescein undergoes a simple one-electron transfer to form a stable semifluorescein group (S * It has been shown that this forms (see Scheme 2). [ka]

[0056] When the dye is irradiated with 390 nm light, it undergoes photochemical reduction, resulting in the production of leucofluorescein (LH) as shown in Scheme 3. 26 It has been reported that it forms [a specific structure]. [ka] This is due to the flow of photocurrent during the reduction process. Ablation is enhanced by this resulting photochemical reaction. Here, using the same principle, the inventors treat the cornea with an eye-safe sodium fluorescein dye to enhance the tissue's absorption properties and achieve explosive ablation.

[0057] microscope Confocal imaging of whole-portion pig eyeballs was performed using a Lecia confocal STED microscope with 20x magnification. The entire cornea was surgically removed and examined using a bright-field microscope (Leica LMD) with 1.25x magnification. Results and Discussion In the first series of experiments, the corneal surface was irradiated using a 532 nm fiber-coupled laser, as shown in Figures 16a-16c. For each power setting, the stepper motor was rotated at either a speed of 2 mm / sec or 4 mm / sec. A total of 800 mW was applied, 50 mW at each setting. No corneal ablation was generated (Table 1, eyes numbered 1-4). Furthermore, when the cornea was irradiated at a single point with 800 mW without rotation, no ablation was observed.

[0058] Figures 17a-17k show examples of different regions of the cornea, demonstrating that no ablation was observed after laser irradiation (Figures 17a-17h). Figures 17j and 17k show high-resolution images of the eye before and after laser treatment, demonstrating that no ablation was observed. In the second series of experiments, the eyes were treated with a freshly prepared solution of fluorescein dye used clinically. One minute after the dye was applied as a standard eye drop (see Methods), the eyes were irradiated with a laser at maximum power at a speed of 2 ms / exposure step. Corneal ablations were generated as shown in Figures 18a-18b (eye number 5). The width and depth of the ablations were measured using whole-eye fluorescence confocal microscopy. In this example, the measured values ​​were 36 μm and 180 μm, respectively (see Figures 19a-19o for Z-stacked images at depths Z=0 to Z=184 μm). Fluorescence signals were only observed up to a depth of 160 μm.

[0059] The same procedure was repeated on the other eye of eye number 6, which had been immersed in the fluorescent dye solution (see Method). Ablation was generated with a width of 123 μm and an ablation depth of 100 μm. Figures 20a and 20B show 1.25x bright-field microscope images of the corneal surface and confocal images of the whole eye, respectively. In the third series of experiments, the exposure time was increased to 4 ms / exposure cycle by changing the motor speed. Microscopic images are shown in Figure 20c (showing a confocal image of eye number 7 with an ablation width of 300 μm and an ablation depth of 247 μm), Figure 20d (showing a reflection mode image of eye number 7 in the same field of view), Figure 21a (showing a 1.25x bright-field microscope image of the corneal surface of eye number 8), and Figure 20b (showing a confocal image of the whole eye of eye number 8 before sectioning with an ablation width of 570 μm and an ablation depth of 465 μm).

[0060] The complete experimental results are established in Table 1. [Table 1]

Claims

1. A method for ablating corneal tissue, The application of a liquid containing an energy-absorbing material onto corneal tissue, wherein the energy-absorbing material absorbs the irradiated light at the absorption wavelength, After the above application, the corneal tissue is irradiated with light containing the absorbance wavelength to produce a selected ablation profile for the corneal tissue. A method that includes this.

2. The method according to claim 1, wherein the energy absorbing material is a biocompatible organic compound.

3. The method according to claim 1 or 2, wherein the liquid contains fluorescein.

4. The method according to claim 3, wherein the liquid is sodium fluorescein dye.

5. The method according to any one of claims 1 to 4, wherein the irradiation includes focusing the irradiation light along a selected pattern on the corneal tissue.

6. The method according to claim 5, wherein the selected pattern includes one or more annular rings or Fresnel rings.

7. The method according to any one of claims 1 to 6, wherein the energy-absorbing material absorbs the energy of the irradiated light in the visible spectrum.

8. The method according to any one of claims 1 to 7, wherein the application is performed about 1 to 10 minutes before the irradiation.

9. The method according to any one of claims 1 to 8, wherein the application thereof results in a surface made of the liquid being obtained on corneal tissue.

10. The method according to any one of claims 1 to 9, wherein the irradiating light has a wavelength between 400 nm and 650 nm.

11. The method according to claim 10, wherein the irradiating light has a wavelength of approximately 390 nm (the term "approximately" in the claims should be understood as ±20 nm).

12. The method according to claim 10, wherein the irradiating light has a wavelength of approximately 435 nm (the term "approximately" in the claims should be understood as ±20 nm).

13. The method according to claim 10, wherein the irradiating light has a wavelength of approximately 530 nm (the term "approximately" in the claims should be understood as ±20 nm).

14. The method according to any one of claims 1 to 13, wherein the irradiation is performed by using a laser device.

15. The method according to claim 14, wherein the laser device is a fiber laser.

16. The method according to any one of claims 1 to 15, wherein the irradiation light is continuous wave irradiation light.

17. The method according to any one of claims 1 to 16, comprising removing liquid from corneal tissue after the irradiation.

18. The method according to any one of claims 1 to 17, wherein the irradiation provides absorption of photons of the absorption wavelength.

19. The method according to claim 18, wherein the absorbance brings about a localized photothermochemical reaction.

20. The method according to any one of claims 1 to 18, wherein the absorption wavelength is in the visible spectrum.

21. This is a kit for ablating corneal tissue. A liquid containing an energy-absorbing material for application to corneal tissue, wherein the energy-absorbing material absorbs irradiated light at an absorption wavelength; An irradiation unit configured to irradiate the corneal tissue with light containing the absorption wavelength when the liquid is applied to the corneal tissue, thereby producing a selected ablation profile for the corneal tissue. A kit that includes this.

22. The kit according to claim 21, wherein the energy absorbing material is a biocompatible organic compound.

23. The kit according to claim 21 or 22, wherein the liquid comprises fluorescein.

24. The kit according to claim 21, wherein the liquid is sodium fluorescein dye.

25. The kit according to any one of claims 21 to 24, comprising a controller configured to control the irradiation unit so that the irradiation unit focuses the irradiation light along a selected pattern on the corneal tissue.

26. The kit according to claim 25, wherein the selected pattern includes one or more annular rings or Fresnel rings.

27. The kit according to any one of claims 21 to 26, wherein the energy-absorbing material absorbs the energy of the irradiated light in the visible spectrum.

28. The kit according to any one of claims 21 to 27, wherein the irradiation light has a wavelength between 400 nm and 650 nm.

29. The kit according to claim 28, wherein the irradiating light has a wavelength of approximately 390 nm.

30. The kit according to claim 28, wherein the irradiating light has a wavelength of approximately 435 nm.

31. The kit according to claim 28, wherein the irradiating light has a wavelength of approximately 530 nm.

32. The kit according to any one of claims 21 to 31, wherein the irradiation unit includes a laser device for performing the irradiation.

33. The kit according to claim 32, wherein the laser device is a fiber laser.

34. The kit according to any one of claims 21 to 33, wherein the irradiation light is continuous wave irradiation light.

35. The kit according to any one of claims 24 to 34, wherein the absorption of photons of the absorbance wavelength is obtained by irradiation.

36. The kit according to claim 35, wherein the absorbance brings about a localized photothermochemical reaction.

37. The kit according to any one of claims 21 to 36, wherein the absorbance wavelength is in the visible spectrum.

38. A kit for use in correcting the condition of the target eye, A pattern forming system configured and operable to use predetermined pattern data to form a three-dimensional pattern corresponding to the corneal surface of a selected target, thereby applying an optical effect to correct the condition of the eye; An eye drop comprising a dispersant for applying to the aforementioned pattern to functionalize and stabilize the optical effect of the pattern, wherein (i) it generates conditions that are invariant with respect to one or more of the following: incident light in a predetermined spectral range on the cornea, the precise position of nanoparticles in the pattern, and the liquid type of the dispersant, and (ii) the dispersant comprises nanoparticles selected to exhibit a dominant scattering effect with respect to incident light in the predetermined spectral range. A kit that includes this.

39. The kit according to claim 38, wherein the nanoparticles are selected to exhibit the maximum scattering effect within the visible spectral range.

40. The kit according to claim 39, wherein the nanoparticles are selected to exhibit the maximum scattering effect within a spectral range defined between 450 nm and 600 nm.

41. The kit according to claim 38, wherein the nanoparticles are selected to exhibit the maximum absorption effect within the visible spectral range.

42. The kit according to claim 38, wherein the nanoparticles are selected based on one or more properties among material, size, and shape.

43. The kit according to claim 38, wherein the three-dimensional pattern is configured as a diffraction pattern.

44. The kit according to claim 38, wherein the pattern includes a Fresnel ring pattern.

45. The kit according to claim 38, wherein the three-dimensional pattern includes an interference pattern that affects the phase.

46. The kit according to claim 38, wherein the three-dimensional pattern includes a depth of field extension pattern.

47. The kit according to claim 38, wherein the three-dimensional pattern is selected to enable optical correction between 0.25 and 10 diopters with an optical power resolution of + / - 0.25 D.

48. The kit according to claim 38, wherein the nanoparticles include a biocompatible protein chain.

49. The kit according to claim 48, wherein the protein chain comprises albumin protein-based nanoparticles.

50. The kit according to claim 49, wherein the albumin protein-based nanoparticles comprise human serum albumin and / or recombinant human serum albumin.

51. The kit according to claim 38, wherein the nanoparticles include metal nanoparticles with a biocompatible polymer shell.

52. The kit according to claim 51, wherein the biocompatible polymer shell is a human serum albumin-based shell and / or a recombinant human serum albumin-based shell.

53. A method for use in correcting the condition of the eye, To form the selected three-dimensional pattern on the target corneal surface, Applying a dispersant containing nanoparticles onto the selected pattern, wherein the nanoparticles are selected and applied such that (i) they generate conditions that are invariant with respect to one or more of the following: incident light within a predetermined spectral range on the cornea, the precise position of the nanoparticles in the pattern, and the liquid type of the dispersant; and (ii) they exhibit a dominant scattering effect with respect to incident light within the predetermined spectral range. A method that includes this.

54. A method for use in correcting the condition of the eye, To provide the kit described in claim 38, To operate a pattern formation system to form a selected three-dimensional pattern on the target corneal surface. A dispersant is applied to the pattern, thereby functionalizing and stabilizing the optical effect of the pattern. A method that includes this.

55. The method according to claim 54, wherein the formation and application generate conditions that are invariant with respect to the point spreading function obtained on the retina of the subject.

56. The method according to claim 54, wherein the application of the dispersant provides a distribution of at least two nanoparticles over a λ × λ area within the incision region of the selected pattern, where λ is a wavelength in a predetermined spectral range.

57. With the application of the dispersant, the λ within the incision region of the selected pattern is as follows: for nanoparticles of about 80 nm size and λ of about 530 nm. 3 The method according to claim 54, wherein a distribution of up to 200 nanoparticles is provided in a volume space.

58. The method according to claim 54, further comprising preparing the dispersant before application, wherein the preparation comprises mixing the liquid with the nanoparticles.

59. A formulation used in eye drops applied to the three-dimensional pattern of the corneal surface of the target eye, A formulation in which nanoparticles are dispersed in a liquid and together form a dispersant, and when the nanoparticles are applied to a three-dimensional pattern formed on the cornea and positioned within the pattern incision area, the majority of the nanoparticles are selected to (i) generate conditions that are invariant with respect to one or more of the incident light in a predetermined spectral range on the cornea, the precise position of the nanoparticles, and the type of the liquid, and (ii) exhibit a dominant scattering or absorption effect with respect to the incident light in a predetermined spectral range, thereby providing the functionalization and stability of the optical effect of the three-dimensional pattern designed for visual acuity correction.

60. The formulation according to claim 59, wherein the nanoparticles are selected to exhibit a dominant scattering effect within the visible spectral range when applied to a three-dimensional pattern formed on the cornea of ​​a target.

61. The formulation according to claim 60, wherein the nanoparticles are selected to exhibit the maximum scattering effect within the visible spectral range.

62. The formulation according to claim 61, wherein the nanoparticles are selected to exhibit the maximum scattering effect within a spectral range defined between 450 nm and 600 nm.

63. The formulation according to claim 59, wherein the nanoparticles are selected based on one or more properties of material, size, shape, or any combination thereof.

64. The formulation according to claim 59, wherein the nanoparticles include human serum albumin (HAS)-based nanoparticles and / or recombinant human serum albumin (rHSA)-based nanoparticles.

65. The method according to any one of claims 53 to 58, wherein the formation is carried out according to the method according to any one of claims 1 to 17.

66. The method according to any one of claims 1 to 17, further comprising applying a dispersant containing nanoparticles onto the selected ablation profile, wherein the nanoparticles are selected to produce conditions that are invariant with respect to (i) incident light in a predetermined spectral range on the cornea, the precise position of the nanoparticles in the pattern, and the liquid type of the dispersant, and (ii) exhibit a dominant scattering effect with respect to the incident light in the predetermined spectral range.

67. The method of claim 66, further comprising the method of any one of claims 56 to 58.

68. A kit according to any one of claims 21 to 37, further comprising an eye drop containing a dispersant for applying to the selected ablation profile to functionalize and stabilize the optical effect of a pattern, the dispersant comprising (i) a dispersant that generates conditions invariant with respect to one or more of the following: incident irradiation light in a predetermined spectral range on the cornea, the precise position of nanoparticles in the pattern, and the liquid type of the dispersant, and (ii) a dispersant comprising nanoparticles selected to exhibit a dominant scattering effect with respect to incident irradiation light in the predetermined spectral range.

69. A kit according to any one of claims 21 to 37, further comprising the formulation according to any one of claims 59 to 64.

70. The kit according to any one of claims 38 to 52, wherein the pattern forming system is comprised of the kit according to any one of claims 21 to 37.

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