WO2021236579A1

WO2021236579A1 - Delivery of amniotic membrane components to ocular epithelia for wound healing

Info

Publication number: WO2021236579A1
Application number: PCT/US2021/032878
Authority: WO
Inventors: Christopher P. Adams; Neal R. COOK
Original assignee: Arlington Vision, Llc
Priority date: 2020-05-18
Filing date: 2021-05-18
Publication date: 2021-11-25

Abstract

Ocular devices comprising a conformer having a concave surface and a convex surface with a layer of amniotic particles, characterized by an average diameter less than or equal to 1 mm, and/or stem cells disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface are disclosed, along with methods for making and using such devices.

Description

DELIVERY OF AMNIOTIC MEMBRANE COMPONENTS TO OCULAR EPITHELIA

FOR WOUND HEALING

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Appl. No. 63/026,249, filed May 18, 2020, which is hereby incorporated by reference in its entirety.

GOVERNMENT RIGHTS

[0002] None.

BACKGROUND

[0003] Chemical injury, a potentially blinding condition constitutes 11.5-22.1% of ocular traumas. Inflammation followed by scarring is the major sequel of alkali burn, leading to vision impairment.

[0004] The antifibrotic and anti-inflammatory properties of pirfenidone, an antifibrotic agent used to treat idiopathic pulmonary fibrosis, have brought the drug to the interest of ocular surgeons as a post-operation anti-scarring agent. Also, amniotic membranes are used for treatment of cornea injuries, including chemical burns.

[0005] The amniotic membrane (AM) is the innermost avascular layer of the placenta about 0.02-0.05 mm thick located next to the fetus. The AM consists of 3 different layers: the epithelium, basement membrane and stroma which further consists of three layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has anti-inflammatory, anti-fibrotic, anti-angiogenic and anti-microbial properties. Additionally, it is transparent and lacks immunogenicity.

[0006] The mechanism through which AM causes healing is complex and involves a combination of factors including providing a substrate for growth of the regenerating ocular epithelia and providing anti-fibrotic, anti-inflammatory, anti- angiogenetic and anti-microbial properties. For example, fetal hyaluronic acid present in the AM stroma suppresses TGF b signaling with reduced expression of TGF b-1, b-2, and b~3 isoforms in addition to reduced expression of TGF-Receptor P, which inhibits proliferation of corneal, limbal and conjunctiva! fibroblasts, which reduces scarring. The presence of AM also inhibits expression of pro-inflammatory cytokines from the damaged ocular surface, e.g., interleukin (IL) 1a, IL-2, IL-8, interferon-g, tumor necrosis factor-b, basic fibroblast growth factor and platelet derived growth factor. The substrate of the AM could also promote regeneration of the epithelium by facilitating migration and differentiation, and preventing apoptosis.

[0007] AM is typically transplanted onto the cornea in the first week following an ocular surface burn. Using the AM can relieve pain, accelerate healing and reduce scarring. The surgery involves applying a patch of AM over the entire ocular surface up to the eyelid margins, which is a tedious and problematic procedure. The AM is typically placed on the damaged epithelia and sutured. Based on the typical thickness of 0.02-0.05 mm for AM and approximate area of cornea, the implanted AM weights about 2-5 mg if it covers the entire cornea. The epithelia side of the membrane faces away from the cornea with the idea that the AM will act as a substrate for the progenitor epithelial cells. In this approach, the surrounding 1-2 mm of the patient’s epithelium is debrided to facilitate growth of the regenerating epithelia over the amniotic membrane, effectively incorporating the AM inside the patient’s eye. In some cases where the defect size is not too large, an AM larger in size than the defect is sutured far away from the edge of the damage. In this case, the AM serves as a cover and the regenerating epithelia grows under the membrane.

[0008] The need for suturing the AM can be eliminated by using a commercial device, sold under the tradename PROKERA^®, which contains the AM mounted in a device that allows placing the lens on the eye without any sutures if is a cryopreserved AM mounted in a polycarbonate ring or an elastomeric band. Over time the membrane dissolves releasing components beneficial for healing. The device provides the benefits of AM-facilitated healing while allowing for ease of application to the eye. However, as the AM in the device dissolves, a fraction is lost to tear drainage. [0009] Another existing device uses dehydrated AM placed on the eye and covered with a soft contact lens to minimize the loss of AM to tear drainage. Based on the device’s 15 mm diameter, about 6 g or 18 mg of amnion is provided from a 35 micron or 100 micron thick device, respectively.

[0010] Versions of these devices with multiple layers of AM are available for serious cases, and placement of either existing device involves application of topical proparcaine (anesthetic) on the ocular surface.

SUMMARY

[0011] It is a purpose of the present invention to provide a device to facilitate contact between amniotic material and an injured cornea or other ocular anatomy to overcome the difficulties associated with applying an amniotic patch on the eye. It is a further goal to enhance the healing potential of AM by keeping it and any material that leaches out of the AM on the surface of the damaged tissue for at least a few hours to maximize benefits. Yet another goal is to design a device that has greater flexibility in the amount of amnion that is incorporated into the device compared to current devices. To achieve this, the disclosed devices and methods allow for depositing powdered amnion in a conforming device (conformer) that can be overlaid on the cornea. The conformer is designed such that placing it on the cornea results in formation of a pool of liquid between the underside of the conformer and the cornea with limited leakage of the contents from inside the pool to outside the device. The outer surface of the conformer is smooth to provide a suitable surface for a tear film to form thereon and an eyelid to glide over. The amnion powder is typically deposited on the inner concavity of the conformer in a manner to be in close proximity to the cornea but without pushing the amniotic material onto the cornea. The amnion powder is also deposited such that the deposited material can break apart and detach from the conformer after placement of the device on the eye. It is also a goal of the present invention to incorporate pharmaceuticals and/or stem cells into the device to enhance healing and provide other benefits, such as antibiotic activity.

[0012] In an aspect, an ocular device comprises a conformer having a concave surface and a convex surface and a layer of amniotic particles characterized by an average diameter less than or equal to 1 mm disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface.

[0013] In an embodiment, the conformer is transparent and/or colorless.

[0014] In an embodiment, the concave surface comprises a pocket circumscribed by a skirt, the pocket having a steeper curvature gradient than the skirt. In an embodiment, the amniotic particles are disposed within the pocket.

[0015] In an embodiment, the amniotic particles are characterized by an average diameter between 1 micron and 1 mm, or between 10 microns and 0.5 mm, or between 20 mm and 0.5 mm, or between 25 microns and 250 microns, or between 50 microns and 100 microns. In an embodiment, the amniotic particles are nanoparticles characterized by an average diameter of 100 nm or less. In an embodiment, the amniotic particles are characterized by an average diameter between 100 nm and 5 nm, or between 75 nm and 10 nm, or 50 nm and 20 nm.

[0016] In an embodiment, a mass of the amniotic particles is between 100 micrograms and 10 grams, or between 1 milligram and 5 grams, or between 10 milligrams and 1 gram, or between 50 milligrams and 0.5 gram, or between 0.1 grams and 0.3 grams.

[0017] In an embodiment, the layer of amniotic particles consists of the amniotic particles.

[0018] In an embodiment, the amniotic particles are dispersed within a hydrophilic polymer film. For example, the hydrophilic polymer may dissolve in biological fluid to release the amniotic particles.

[0019] In an embodiment, a concentration of the amniotic particles in the hydrophilic polymer film is between 10% and 90% by weight, or between 20% and 80% by weight, or between 25% and 70% by weight, or between 30% and 50% by weight.

[0020] In an embodiment, a pharmaceutical agent may be dispersed within the hydrophilic polymer film. In an embodiment, the pharmaceutical agent may be at least partially encapsulated within a microparticle, such as a microparticle formed from a biologically compatible polymer like PLA, PLGA, PEG or combinations thereof.

[0021] In an embodiment, one or more pharmaceutical agents and/or amniotic particles are at least partially encapsulated within the conformer.

[0022] In an embodiment, a conformer is a scleral contact lens, a rigid gas permeable contact lens, or a soft polymer contact lens.

[0023] In an embodiment, an ocular device disclosed herein further comprises stem cells disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface. In an embodiment, the stem cells are present in a quantity between 100 and 100,000, or between 250 and 10,000, or between 500 and 5,000.

[0024] In an embodiment, stem cells and/or amniotic particles are dispersed within a single layer formed by a single type of hydrophilic polymer film. In another embodiment, stem cells and/or amniotic particles are dispersed within separate layers, which may be formed by the same hydrophilic polymer or by different hydrophilic polymers. In an embodiment, an ocular device comprises a plurality of layers of amniotic particles, stem cells, or both amniotic particles and stem cells. For examples, the plurality of layers may alternate, regularly or irregularly, between amniotic particles and stem cells, or between layers having amniotic particles of different diameters, and/or between layers having different concentrations of active ingredients (amniotic particles, stem cells, and/or pharmaceutical agents).

[0025] In an embodiment, a layer of stem cells is disposed between the conformer and a layer of amniotic particles. In an embodiment, a layer of amniotic particles is disposed between the conformer and a layer of stem cells. In an embodiment, a conformer is disposed between a layer of stem cells and a layer of amniotic particles.

[0026] In an embodiment, stem cells and amniotic particles are disposed at physically distinct locations on the conformer relative to one another. For example, stem cells and amniotic particles may be disposed at different locations on the same surface of the conformer, or stem cells and amniotic particles may be disposed on different surfaces of the conformer relative to one another.

[0027] In an aspect, an ocular device comprises a conformer having a concave surface and a convex surface and a layer of stem cells disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface.

[0028] In an aspect, an ocular device kit comprises at least one conformer having a concave surface and a convex surface and a vessel comprising amniotic particles and/or stem cells, and optionally one or more pharmaceutical agents, dispersed within a hydrophilic polymer solution.

[0029] In an aspect, a method of making an ocular device comprises providing a conformer having a concave surface and a convex surface, mixing amniotic particles and/or stem cells in a solvent to form a dispersion, applying the dispersion to at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface, and removing the solvent from the dispersion to form a dry layer of amniotic particles on the conformer. In an embodiment, the step of mixing comprises sonicating. In an embodiment, the step of removing the solvent comprises air drying, reducing pressure and/or heating.

BRIEF DESCRIPTION OF DRAWINGS

[0030] FIG. 1 shows side-by-side photographs of a conformer containing amniotic powder (“AP”) and the same conformer containing sonicated powder dispersed in a hydroxy propyl methyl cellulose (HPMC) film.

[0031] FIG. 2 shows a convex surface of a conformer blank.

[0032] FIG. 3 shows a concave surface of the conformer blank of FIG. 2.

[0033] FIG. 4 shows a conformer containing amniotic powder (“AP”), according to an embodiment.

[0034] FIGS. 5, 6, and 7 show various angles of a conformer containing amnion powder dispersed in a saline film, according to an embodiment. DETAILED DESCRIPTION

[0035] Amniotic Membrane (AM) - AM is the innermost avascular layer of the placenta, about 0.02-0.05 mm thick located next to the fetus.

[0036] Amniotic Powder (AP) - AP is obtained by finely dividing dried AM into particles with an average diameter between 10 nm and 1 mm.

[0037] Conformer - A conformer is the device into or onto which the AP is deposited. A conformer has a concave surface and a convex surface, with the concave surface facing the anterior surface of a patient’s cornea when the device is placed on the eye of the patient. The AP is deposited on the concave surface (e.g., when corneal repair is desired), on the convex surface (e.g., when the backside of the eyelid is damaged), or on both surfaces (e.g., when multiple areas within the ocular anatomy are damaged).

[0038] Amniotic Conformer (AC) - An amniotic conformer is a conformer containing a film of AP on at least one surface of the conformer.

[0039] Embodiments of the present disclosure relate to methods for preparing and using a device that brings AP and/or stem cells in proximity to damaged ocular anatomy while minimizing loss of active ingredients due to leakage from the device.

[0040] Manufacturing of Amnion-Loaded Conformer

[0041] In general, in a method of preparing an AM-containing device, the AM is converted to AP by any suitable processing method including, but not limited to, milling, grinding, chopping, freeze drying and spray drying. The AP is hydrated by addition of saline ranging from about 1 % to 25% by weight of the AP, about 1 % to 15% by weight of AP, or about 1-5% by weight of the AP to form a concentrated slurry. In an embodiment, additional components are added to the slurry to increase viscosity of the slurry. In an embodiment, the slurry is dispersed in the concavity of a conformer (CR), followed by drying, e.g., in a vacuum oven or incubator, to evaporate liquid from the slurry. The evaporation typically occurs in a few hours, but could be sped up by controlling the temperature and/or pressure of the drying process. As the slurry dries, the AP particles stick together to form a film in the concavity of the conformer. This device with the film of AP on the conformer is called an “Amniotic Conformer or AC”.

[0042] In an embodiment, the mass of AP deposited in a conformer is 2-5 mg to be comparable to the mass of a single layer of amniotic membrane covering the cornea. In an embodiment, the mass of AP is increased to 5-50 mg to provide additional mass, which may be useful for severe injuries. In an embodiment, the mass of AP is reduced to 0.1-2 mg, or to 0.01 - 0.1 mg, which may be sufficient for less severe injuries, particularly because the AP and its components are retained on the cornea for extended periods by the conformer.

[0043] In an embodiment, the AM is converted to a dispersion by adding water or saline followed by sonicating to reduce the particle size to tens to several hundred nanometers. Sonication may be carried out, for example, at 700 W for 5 minutes with intermittent cycles of 30 s sonication followed by 1 min of no-sonication. Stabilizers, such as surfactants or polymers, can be added to the AM dispersion as well. Typically, the amount of water or saline added for sonication will be such that the mass of AM is about 1-10% of the total weight. In other embodiments, 10-25% AM can be added to the water or saline prior to sonication. Sonicated dispersions, produced with this approach, remain stable for extended periods ranging from 1 hour to 15 months. In an embodiment, the hydrophilic polymer carboxyl methyl cellulose (CMC) is added at 1-10% (w/w) to the sonicated AM to produce a viscous AM suspension or hydrogel. In an embodiment, other hydrophilic polymers such as hydroxy propyl methyl cellulose (HPMC), agarose, poly vinyl alcohol (PVA), poly ethylene glycol (PEG), and the like are added to the sonicated AM to form a viscous AM suspension. In an embodiment, the polymer loading in the suspension is 10- 25% w/w. In an embodiment, the polymer is added to the mixture of water/saline and AM prior to sonication. In an embodiment, stem cells are mixed with amniotic particles and/or hydrophilic polymer and applied to a conformer without undergoing sonication.

[0044] FIG. 1 shows side-by-side photographs of a conformer containing amniotic powder (“AP”) and the same conformer containing sonicated powder dispersed in a hydroxy propyl methyl cellulose (HPMC) film. The hydrogel film is transparent due to the nanosize of the amnion particles while the amniotic powder film is opaque due to the larger size of the particles.

[0045] FIG. 2 shows a convex surface of a conformer blank. FIG. 3 shows a concave surface of the conformer blank of FIG. 2. FIG. 4 shows a conformer containing amniotic powder (“AP”). FIGS. 5, 6, and 7 show various angles of a conformer containing amnion powder dispersed in a saline film, according to an embodiment.

[0046] In an embodiment, the suspension of nanoparticles containing 1-10% nanoparticles is freeze dried to manufacture powder. Alternatively, spray drying can be used to form the powder. The powder is then converted to a slurry by addition of a small amount, typically 1-5% (w/w), water or saline or a physiological buffer. In an embodiment, the slurry is placed in the concavity of the conformer and/or on the convex surface of the conformer, followed by placing the AP-loaded conformer into a vacuum oven or incubator to evaporate the liquid from the slurry. The evaporation typically occurs in a few hours but could be sped up by controlling the temperature and/or pressure while drying. As the slurry dries, the AP particles stick together to form a film.

[0047] In an embodiment, the viscous polymer suspension is placed in the concavity of the conformer and/or on a convex surface of the conformer, followed by placing the conformer into a vacuum oven or incubator to evaporate liquid in the polymer suspension. The evaporation typically occurs in about a day but controlling the temperature and/or pressure while drying could speed it up. Drying of the polymer suspension results in formation of a thin film of the hydrophilic polymer with amnion nanoparticles encapsulated within the film. The amnion-loaded film is transparent and the presence of the amnion-loaded film does not impact the transparency of the conformer.

[0048] In an embodiment, 100 microliter (-100 mg) of amnion suspension is placed in the concavity of the conformer. In an embodiment, the entire concavity of the conformer is filled with the suspension. In an embodiment, 50% or less of the conformer is filled with the suspension. [0049] In an embodiment, the AP is sprayed directly onto a surface of the conformer. In an embodiment, the AP is deposited onto the conformer via ink-jet printing, where AP and/or stem cells are a component of the ink. In both methods, the mass of amnion deposited on the conformer is in the range of 2-5 mg, or in the range 5-50 mg to provide additional mass which may be useful for severe injuries.

In an embodiment, the mass is reduced to 0.1-2 mg and to 0.01 - 0.1 mg.

[0050] In an embodiment, the conformer is rotated about its central axis during the drying process to obtain a fine film on the surface of the conformer.

[0051] In an embodiment, the pH of the slurry or suspension of nanoparticles is maintained in the range of 5-7 and osmolarity is maintained at 300 mOs by addition of salt to minimize the potential for discomfort to the patient.

PHARMACEUTICALS

[0052] There are potential benefits from addition of selective pharmaceuticals within the formulations disclosed herein. Corneal scarring / fibrosis is the major sequel to several forms of insults to the cornea, including alkali burn. Loss of corneal transparency is the second highest cause of vision impairment, therefore appropriate management to prevent corneal fibrosis is imperative. There is yet no specific therapy in clinical practice to address corneal fibrosis, but researchers have previously identified pirfenidone nanoformulation drops as a promising therapy to prevent corneal scarring. They have also reported the anti-inflammatory and antifibrotic effects of pirfenidone in prevention of fibrosis leading to proliferative vitreoretinopathy. Thus, inclusion of pirfenidone within the formulations disclosed herein may be beneficial.

[0053] In an embodiment, pirfenidone (1 mg/mL) is added to the suspension of AM nanoparticles containing polymer, followed by one of the processes described above to form a polymeric film containing amnion nanoparticles and dissolved pharmaceutical. In an embodiment, pirfenidone is encapsulated within PLGA nanoparticles or microparticles to allow for extended release of the drug. Drug- loaded PLGA particles can be produced using a double emulsion approach. Overall, the suspension will contain 5-20% polymer, 1-10% amnion nanoparticles, and 1-5% PLGA nanoparticles which contain drug at a loading of 0.1-10%. Many ophthalmic drugs can bind to the surface of the amnion particles resulting in an increase in drug loading and/or release duration.

[0054] In an embodiment, drug-containing PLGA particles are added to the AP and/or stem cell slurry, followed by one of the processes described above to form an amnion film.

[0055] In an embodiment, other pharmaceuticals are incorporated into the device such as antibiotics (levofloxacin, moxifloxacin); steroids such as dexamethasone; analgesics such as ketorolac and lidocaine. In an embodiment, multiple pharmaceuticals can be incorporated into PLGA particles and then into the amnion film. A typical composition will have PLGA particles with 1-10% drug loading, and 1- 20% of the PLGA particles in an AM nanoparticle and/or stem cell suspension.

[0056] In an embodiment, stem cells and stem cell solutions are added to the conformer with the nanoparticle suspensions. The stem cells are added in a concentration of from 100-100,000 cells per 100 pi of phosphate buffered saline

(PBS at a final concentration of 157 mM Na^÷, 140mM C! , 4.45mM K⁺, 10.1 mM HPO4² , 1.78 mM j-½P0₄ ^~ and a pH of 7.3 - 7.96) either alone or in combination with the AM nanoparticles, for example.

CONFORMER DESIGN

[0057] The conformer is shaped to have an anterior side that faces the eyelids and the posterior side that faces the cornea. The radius of curvature of the posterior side must be steeper than that of the cornea so that placing the conformer on the cornea leads to formation of a vault in the central cornea with the edges of the conformer resting beyond the limbus, on the conjunctiva, to minimize the potential for discomfort. The central section of the conformer should preferably be semi- spherical to create a pocket when the conformer is placed on the eye, and the central section is surrounded by a skirt of flatter curvature to conform to the shape of the sclera. When the conformer is placed on the cornea, the central portion creates a vault or pocket and the periphery rests flush against the sclera. The thickness of the vault, i.e., the spacing between the posterior surface of the conformer and the cornea should be at least 50 microns, and preferably 50-400 microns to accommodate the amniotic film therebetween. A thickness of larger than 400 microns could lead to reduced oxygen permeation that may inhibit the healing process. A diameter of the conformer should be at least 6 mm and preferably 6-16 mm. These dimensions are only guides and the actual dimensions may depend on the specific would and shape of the patient’s cornea. The anterior surface of the conformer must be smooth to allow gliding of the eyelid without discomfort. In an embodiment, the conformer is a rigid gas permeable contact lens. In an embodiment, the conformer is soft allowing the vault or pocket to be compressed, resulting in intimate contact of the amnion film with the cornea. In an embodiment, the conformer is a soft silicone-hydrogel contact lens. In an embodiment, the pharmaceuticals are incorporated into the soft contact lenses prior to forming the amnion film in the concavity.

[0058] In an embodiment, the amnion nanoparticles are incorporated directly inside the soft contact lens by soaking the contact lens in the amnion nanoparticle suspension for about 7 days. In an embodiment, dried amnion nanoparticles are dispersed in an organic liquid such as ethanol, followed by soaking the soft contact lens. The lens expands considerably in the ethanol allowing diffusion of nanoparticles into the lens. In another embodiment, amnion nanoparticles are added to a monomer mixture followed by polymerization to form a soft contact lens having amnion nanoparticles dispersed therein. For example, hydroxymethyl methacrylate (HEMA) is added to the amnion nanoparticle suspension at 20-50% w/w, along with 2% crosslinker and 0.1% initiator, followed by polymerization in a mold to form a contact lens loaded with amnion nanoparticles. Upon placing the lens in the eye, the amnion diffuses out of the contact lens to the cornea. In an embodiment, the contact lens is made of dissolvable material such as HPMC.

[0059] In an embodiment, vitamin E is loaded into a soft contact lens along with one or more pharmaceuticals and amniotic nanoparticles to increase the release duration of the loaded drugs. Vitamin E diffusion barriers may be incorporated into contact lenses at 0-30% (w/w in dried lens) to increase the drug release duration.

[0060] In an embodiment, vitamin E and drug-loaded lenses are used as a bandage lens over an amniotic membrane to combine the beneficial effects of the amniotic material and the drugs. The device in this case is a package comprising a dehydrated AM and a fully hydrated contact lens loaded with vitamin E and ophthalmic drugs such as levofloxacin, dexamethasone, ketorolac, etc.

PACKAGING

[0061] An amniotic conformer device may be packaged and then sterilized via ebeam. In an embodiment, a medical professional opens the package, fills the concavity of the device with a separately packaged AM nanoparticle suspension (optionally comprising additional agents), and then places the device on a patient’s eye. In an embodiment, the solution accompanying the amniotic conformer comprises buffered saline. In an embodiment, hydrophilic polymer, such as carboxyl methyl cellulose (CMC), is added to the accompanying solution at 0.1-10% (w/w) to increase the viscosity of the fluid which will assist in lubrication and potentially increasing comfort after placing an amniotic conformer on a patient’s eye. In an embodiment, other hydrophilic polymers, such as hydroxy propyl methyl cellulose (HPMC), agarose, poly vinyl alcohol (PVA), poly ethylene glycol (PEG), etc are added at 0.1-10% w/w to the AM nanoparticle suspension. In another embodiment, the AM nanoparticle suspension contains nanoparticles of amniotic material at 0.1- 10% manufactured by sonication or homogenization of an amniotic powder. In another embodiment, the AM nanoparticle suspension includes amniotic nanoparticles at 0.1-10% and hydrophilic polymers at 0.1-10%. In another embodiment, the AM nanoparticle suspension contains one or more surfactants, such as pluronics, to stabilize the amniotic nanoparticle suspension. In another embodiment, the AM nanoparticle suspension includes drugs, such as pirfenidone, to aid wound healing. In an embodiment, other drugs, such as antibiotics (levofloxacin, moxifloxacin); steroids such as dexamethasone; analgesics such as ketorolac, lidocaine, are added to the AM nanoparticle suspension. In an embodiment, one or more drugs can be incorporated into PLGA particles, which are then added to the AM nanoparticle suspension.

STEM CELLS

[0062] Stem cells can be incorporated into a hydrogel and deposited on the conformer to form a device for delivering stem cells to the cornea. BREAKUP OF THE AMNION FILM TO RELEASE PARTICLES

[0063] The amnion film in the conformer breaks up after exposure to tears to release the amniotic particles. In vitro studies of the amnion film conformer formed by evaporation of the slurry demonstrated that the film falls apart slowly releasing the amnion into the fluid. In this example, the concavity of the conformer loaded with amnion was filled with an aqueous solution, and placed on a vibrating stage to simulate the motion inside the eye. The amnion slurry is then allowed to dry to form the film. The dried device is then filled with the aqueous solution to test in vitro how the deposited film breaks releasing the amniotic material. The film falls apart gradually releasing the material into the fluid. The film starts to break around 40 min and completely disintegrates in a few hours. Experiments were also conducted in vivo. It was discovered that upon insertion of the conformer with the film deposited from slurry, the film falls apart faster than in vitro. The film broke apart into many pieces in about 20-30 min, which improved the patient’s vision considerably. Visual acuity was 20/80 after 2 minutes improving to 20/30 after 35 minutes.

[0064] In vitro exposure of the conformer containing the polymeric film with the amnion nanoparticles resulted in dissolution of the film in 10 to 30 min releasing the nanoparticles into the aqueous media.

REFERENCES

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[0070] Zhong H, Sun G, Lin X, Wu K, Yu M. Evaluation of pirfenidone as a new postoperative antiscarring agent in experimental glaucoma surgery. Invest Ophthalmol Vis Sci. 2011;16:3136-3142. doi: 10.1167/iovs.10-6240.

[0071] Sun G, Lin X, Zhong H. et al. Pharmacokinetics of pirfenidone after topical administration in rabbit eye. Mol Vis. 2011;17:2191-2196.

[0072] Meller D, Pauklin M, Thomasen H, Westekemper H, Steuhl KP: Amniotic membrane transplantation in the human eye. Dtsch Arztebl Int 2011; 108(14): 243- 8. DOI: 10.3238/arztebl.2011.0243.

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STATEMENTS REGARDING INCORPORATION BY REFERENCE AND

VARIATIONS

[0077] All references cited throughout this application, for example patent documents including issued or granted patents or equivalents; patent application publications; and non-patent literature documents or other source material; are hereby incorporated by reference herein in their entireties, as though individually incorporated by reference.

[0078] The terms and expressions which have been employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the invention has been specifically disclosed by preferred embodiments, exemplary embodiments and optional features, modification and variation of the concepts herein disclosed can be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims. The specific embodiments provided herein are examples of useful embodiments of the invention and it will be apparent to one skilled in the art that the invention can be carried out using a large number of variations of the devices, device components, and method steps set forth in the present description. As will be apparent to one of skill in the art, methods and devices useful for the present methods and devices can include a large number of optional composition and processing elements and steps. All art-known functional equivalents of materials and methods are intended to be included in this disclosure.

[0079] When a group of substituents is disclosed herein, it is understood that all individual members of that group and all subgroups are disclosed separately. When a Markush group or other grouping is used herein, all individual members of the group and all combinations and subcombinations possible of the group are intended to be individually included in the disclosure.

[0080] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a pharmaceutical" includes a plurality of such pharmaceuticals and equivalents thereof known to those skilled in the art, and so forth. As well, the terms "a" (or "an"), "one or more" and "at least one" can be used interchangeably herein. It is also to be noted that the terms "comprising", "including", and "having" can be used interchangeably. The expression “of any of claims XX- YY” (wherein XX and YY refer to claim numbers) is intended to provide a multiple dependent claim in the alternative form, and in some embodiments is interchangeable with the expression “as in any one of claims XX- YY.”

[0081] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0082] Whenever a range is given in the specification, for example, a range of integers, a temperature range, a time range, a composition range, or concentration range, all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the disclosure. As used herein, ranges specifically include the values provided as endpoint values of the range and all the integer values of the range. For example, a range of 1 to 100 specifically includes the end point values of 1 and 100. It will be understood that any subranges or individual values in a range or subrange that are included in the description herein can be excluded from the claims herein.

[0083] As used herein, “comprising” is synonymous and can be used interchangeably with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of excludes any element, step, or ingredient not specified in the claim element. As used herein, "consisting essentially of does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim. In each instance herein any of the terms "comprising", "consisting essentially of and "consisting of can be replaced with either of the other two terms. The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations which is/are not specifically disclosed herein.

Claims

CLAIMS What is claimed is:

1. An ocular device comprising: a conformer having a concave surface and a convex surface; and a layer of amniotic particles characterized by an average diameter less than or equal to 1 mm disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface.

2. The ocular device of claim 1 , wherein the conformer is transparent.

3. The ocular device of claim 1 , wherein the concave surface comprises a pocket circumscribed by a skirt, the pocket having a steeper curvature gradient than the skirt.

4. The ocular device of claim 3, wherein the amniotic particles are disposed within the pocket.

5. The ocular device of claim 1, wherein the amniotic particles are characterized by an average diameter between 1 micron and 1 mm.

6. The ocular device of claim 1 , wherein the amniotic particles are nanoparticles characterized by an average diameter of 100 nm or less.

7. The ocular device of claim 1 , wherein the layer of amniotic particles consists of the amniotic particles.

8. The ocular device of claim 1 , wherein the amniotic particles are dispersed within a hydrophilic polymer film.

9. The ocular device of claim 8, wherein a concentration of the amniotic particles in the hydrophilic polymer film is between 10% and 90% by weight.

10. The ocular device of claim 8 further comprising a pharmaceutical agent dispersed within the hydrophilic polymer film.

11. The ocular device of claim 10, wherein the pharmaceutical agent is at least partially encapsulated within a microparticle.

12. The ocular device of claim 1 , wherein the amniotic particles are at least partially encapsulated within the conformer.

13. The ocular device of claim 1 , wherein the conformer is a scleral contact lens, a rigid gas permeable contact lens, or a soft polymer contact lens.

14. The ocular device of claim 1 further comprising stem cells disposed on at least a portion of the concave surface, the convex surface, or both the concave surface and the convex surface.

15. The ocular device of claim 14, wherein the stem cells and the amniotic particles are dispersed within a hydrophilic polymer film.

16. The ocular device of claim 15 further comprising a pharmaceutical agent dispersed within the hydrophilic polymer film.

17. The ocular device of claim 14, wherein the stem cells are disposed within a second layer that is distinct from the first layer.

18. The ocular device of claim 14, wherein the stem cells are disposed within a first hydrophilic polymer and the amniotic particles are disposed within a second hydrophilic polymer that is different from the first hydrophilic polymer.

19. The ocular device of claim 14, wherein the stem cells and the amniotic particles are disposed at physically distinct locations of the conformer relative to one another.

20. The ocular device of claim 18, wherein the amniotic particles and the stem cells are disposed on different surfaces of the concave surface and the convex surface relative to one another.