WO2021195587A1

WO2021195587A1 - Artificial cornea with double-side microtextured phema hydrogel

Info

Publication number: WO2021195587A1
Application number: PCT/US2021/024521
Authority: WO
Inventors: Yiwen Xi; Chang-Hwan Choi; Junfeng Liang; Xiaojun Yu
Original assignee: The Trustees Of The Stevens Institute Of Technology
Priority date: 2020-03-26
Filing date: 2021-03-26
Publication date: 2021-09-30

Abstract

A microtextured artificial cornea made of pHEMA hydrogel and an associated manufacturing method are disclosed. The artificial cornea has two sides, each of which has an associated microstructure. In an embodiment, microlines can be provided on an anterior side, and a posterior side can have micropores. Both the geometry of the microstructures and their dimensions can be customized for an individual patient. The geometry of the artificial cornea itself and its dimensions can also be customized as such. In addition, the lifetime of the artificial cornea can be significantly enhanced by adding co-polymer(s) into the hydrogel to strengthen its mechanical properties. Patient recovery can be aided by adding peptides into the artificial cornea surfaces to improve cell growth post-operation.

Description

Artificial Cornea with Double-Side Microtextured pHEMA Hydrogel

Cross-Reference to Related Applications

This application claims the benefit of and priority to U.S. Provisional Application No. 63/000,422 filed March 26, 2020 and U.S. Patent Application No. 16/879,641, filed May 20, 2020. The entire disclosures of each of the aforesaid applications are incorporated herein by reference.

Field of the Invention

The present invention pertains to the production and use of a novel artificial cornea material. Specifically, it pertains to a biomaterial that integrates effectively with the native ocular cells.

Background of the Invention

As stated by World Health Organization (WHO), over 10 million people suffer from corneal blindness globally and 285 million people are visually impaired; however, only 1/50 of the patients obtained corneal transplants each year worldwide due to lack of donor tissue. In addition, functional defects and heterogeneity of the transplanted cornea commonly occur within a few years of the surgery because of the prevalent use of tissue from older cornea donors. Rejection of donated tissue constitutes another substantive problem in the art.

Overall, full corneal translation from a donor is inefficient and inaccessible for many. A rising geriatric population and the increasing incidence of eye diseases worldwide are expected to be the prime factors driving the demand worldwide for keratoprosthesis (i.e., artificial cornea) treatment.

Summary of the Invention

The present invention relates to an artificial cornea made of Poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogel. The pHEMA is microtextured on both sides to facilitate integration of the artificial cornea with the native tissue of a patient. Specifically, the artificial cornea is designed so that it is biocompatible, driving rapid proliferation of corneal epithelial cells while also enhancing the adhesion of corneal fibroblasts. By fostering its integration with the epithelium as such, the artificial cornea limits the chances of infection, inflammation or extrusion. By integrating with the stroma, the stability of the material within the eye improves. These factors ultimately expedite recovery from surgical implantation. In addition, no donor tissue is required, and the risk of rejection is low. The current artificial cornea products on the market use the stiff materials like PolyCmethyl methacrylate) (PMMA), of which Young’s Modulus ranges from 2 to 3 GPa. Stiff materials are likely to increase the risks of tissue extrusion and inflammation. The pHEMA hydrogel is soft but durable with the Young’s Modulus ranging from 1 to 10 MPa, making it ideal for permanent wearing while decreasing discomfort.

Brief Description of the Figures

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

For a more complete understanding of the invention disclosed herein, reference is made to the following detailed description of an exemplary embodiment considered in conjunction with the accompanying drawings, in which:

Fig. 1A is a flow diagram of the production process of the microtextured pHEMA, with an associated legend;

Fig. IB shows a top view of a schematic of replicated micropores on one- side-textured pHEMA made via the pattern transfer processes of soft lithography;

Fig. 1C shows a top view of a schematic of replicated microlines on one-side-textured pHEMA made via the pattern transfer processes of soft lithography; Fig. ID shows a side view of a schematic of replicated micropores and microlines on one- side-textured pHEMA made via the pattern transfer processes of soft lithography;

Fig. 2A is a schematic diagram showing the preparation of a viscous pHEMA mixture prior to injection into PDMS molds; Fig. 2B is a schematic diagram of the use of PDMS molds in the microtexturing process;

Fig. 3 is a schematic diagram of a molding process to achieve desired curvature of the artificial cornea.

Fig. 4A displays a pore size of 3 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image; Fig. 4B displays a pore size of 6 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image;

Fig. 4C displays a pore size of 12 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image;

Fig. 4D displays a line gap size of 3 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image;

Fig. 4E displays a line gap size of 6 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image;

Fig. 4F displays a line gap size of 12 pm on the pHEMA hydrogel’s microstructure patterned on silicon substrate as a scanning electron microscope image; Fig. 5A is a perspective view of an artificial cornea made in accordance with an embodiment of the present invention;

Fig. 5B is a side view of the artificial cornea shown in Fig. 5A; Fig. 5C is a schematic diagram, including an anterior and a posterior view, of an embodiment of the artificial cornea of the present invention having a microline texture on one side and a micropore texture on the opposite side, or the same microline or micropore texture on both sides; and Fig. 6 is a schematic diagram of the implanted double-side-microtextured pHEMA hydrogel functioning as an artificial cornea.

Detailed Description of an Exemplary Embodiment

To give the pHEMA hydrogel the desired microstructure of the final artificial cornea, templates (e.g., silicon masters) having the desired microstructure (i.e., micropores or microlines) are first fabricated. Referring now to Fig. 1 A, a UV light lithography process is used to create a microtextured photoresist layer deposited on the top of silicon by shining UV light through a designed micropatterned photomask. The microtextured photoresist layer is used as etch mask in deep reactive ion etching (DRIE) of a silicon wafer. During the DRIE, the photoresist micropattems are transferred on to the silicon wafer, where only the silicon wafer area uncovered by the photoresist is etched by the etching gases. Therefore, the photoresist mask layer and the silicon master should share the same dimensions of microtextures. The depth of the silicon microstructure can be controlled by varying the etching time in the process of DRIE. The photoresist residue is then removed from the silicon wafer.

Once the silicon masters are made, the microstructure patterns are transferred to polydimethylsiloxane (PDMS) via a soft lithography technique to form a pair of molds. To this end, uncured PDMS (e.g., in monomer form with a cure agent in a 10: 1 ratio) is flow-coated onto the silicon masters. Once fully cured, the PDMS, which now constitutes the molds, is peeled off from the silicon master. (Refer to Fig. 1 A)

Once the molds are prepared, liquid pHEMA monomer (2-Hydroxyethyl methacrylate, Sigma-Aldrich, CAS No.: 868-77-9) is first mixed with an initiator (2 -Hydroxy -2- methylpropiophenone 97%, Sigma-Aldrich, CAS No.: 7473-98-5) while being exposed to UV light for a few minutes, resulting in a viscous pHEMA mixture (see Fig. 2A). Additional initiator and crosslinker (Ethylene glycol dimethacrylate, Sigma-Aldrich, CAS No.: 97-90-5) for pHEMA are then added. Once fully mixed and degassed, the pHEMA mixture is slowly poured into a chamber defined by a bottom one of the PDMS molds and non-textured PDMS walls (see FIG. 2B). The other PDMS mold (i.e., the top stamp) is then gently pressed down on top of the pHEMA such that the pHEMA flows into the PDMS microstructures on the top and bottom molds completely, creating an imprint of the PDMS molds’ microstructures on the pHEMA material with high fidelity. Thus, the pHEMA microstructures now replicate the microstructures of the silicon masters. The pHEMA and its surrounding structure is then exposed to UV light until the pHEMA is completely cured (see Fig. 2B). The PDMS molds can be peeled off from the cured pHEMA to obtain the desired hydrogel. Then, the pHEMA is quickly washed with ethanol and then with phosphate-buffered saline (approximate pH of 7.4) to remove unreacted chemical residue from the finished pHEMA material’s surface. Fig. 3 explains one of the possible methods to achieve the desired curvature on the artificial cornea by using a molding process. The curved base is made of quartz glass (or PDMS). After fixing the bended PDMS mold with microtextures onto two bases by clamps, the uncured pHEMA mixture (after pre-UV-light-crosslinking) can be injected into the gap between two molds for final UV-light crosslinking step. Referring now to Fig. 4, exemplary microstructures are shown, demonstrating how pore size can be varied. Figs. 4A, 4B and 4C show micropore diameters of, respectively, 3 pm, 6 pm and 12 pm. Figs. 4D, 4E and 4F show line gap sizes of, respectively, 3 pm, 6 pm and 12 pm.

Fig. 5A is a perspective view of an artificial cornea, while Fig. 5B is a side view of same. Fig. 5C shows an embodiment of the present invention in which microlines (i.e., microgrooves) are present on the anterior side of the artificial cornea, while micropores (i.e., microdimples) are present on the posterior face. By way of example, the diameter of the micropores and the width of the microlines’ ridges can range from a few micrometers to few tens of micrometers (e.g. larger than 1 pm but less than 100 pm). The depth of the micropores should be smaller than the thickness of the artificial cornea, e.g., less than 2000 pm. The depth of the microlines should be further restricted by their aspect ratios (e.g., maximum ratio of depth to ridge width = 3:1) due to the weaker mechanical strength of the pHEMA microlines when the aspect ratios are greater than 3:1. For use with a typical adult, the total thickness of the artificial cornea can lie in a range of from about 500 to about 600 pm, the radius of the artificial cornea can lie in a range of from about 3 to about 5 mm, the radius of its curvature from about 8 mm to about 10 mm, and its Young’s Modulus can lie in a range of from about 1 to about 2 MPa. For use with typical infants or individuals with larger corneas (or even larger animals, such as cows), the total thickness of the artificial cornea can lie in a range of from about 100 to about 2000 pm, the radius of the artificial cornea can lie in a range of from about 1 to about 20 mm, the radius of its curvature from about 2 to about 30 mm, and its Young’s Modulus can lie in a range of from about 0.2 to about 50 MPa.

In manufacturing the artificial cornea, it is desirable to vary its dimensions to suit the recipient. For instance, the thickness of the artificial cornea should be controlled. Another factor to control is the curvature of the artificial cornea so that it suits the patient’s eye.

The design of the pHEMA hydrogel is partially motivated by the expectation that the corneal epithelium will be allowed to proliferate well and fast to form a layer of protection and prevent the eyes from bacterial infection, inflammation, stromal melt, or extrusion. The microstructure outer surface of the artificial cornea can be designed to facilitate this outcome. To a similar end, the microstructure of the inner surface of the hydrogel can be designed to allow for fast adhesion of keratocytes (corneal fibroblasts) to the hydrogel, leading to the stable formation of stroma underneath the artificial cornea, thereby decreasing recovery time following the implantation. Enhanced keratocyte adhesion also stabilizes the artificial cornea itself on the top of the eyeball without unnecessary movement, which in conjunction with the softness of the hydrogel makes it ideal for permanent wearing by decreasing discomfort.

Referring now to Fig. 6, the position of the artificial cornea in the eye can be seen in a situation where the artificial cornea has been inserted into a patient’s eye to replace an old malfunctioned cornea. As illustrated in Fig. 6, after the patient’s cornea frontier (with dead or damaged tissue) is cut off (to create a pocket to insert an implant) by the surgeons, the artificial cornea of the present invention can be embedded into the cornea stroma pocket to replace the removed tissue. In applying the artificial cornea to the patient’s eye, it should be noted that no donor tissue is required for either the manufacture or the implantation of the hydrogel. The pHEMA hydrogel from which the artificial cornea is made is an FDA approved biocompatible material having similar mechanical properties to human tissue. In an embodiment, its water content is larger than 10% by weight.

Further methods are available to improve the physical and chemical properties of the artificial cornea described hereinabove. For example, the incorporation of bioactive molecules (e.g., one or more peptides) can speed up the proliferation of patients’ own epithelial cells to cover the whole top surface of the artificial cornea, efficiently, right after surgery, to further prevent potential infection and inflammation. To incorporate peptides into the hydrogel, they can be added into the uncured 2-hydroxyethyl methacrylate solution (i.e., HEMA solution) before final polymerization processes. Alternatively, peptides can be loaded into the pHEMA hydrogel once the gel is polymerized. Peptides with chemotactic and/or chemokinetic can possess the desired biological activity and offer better control of release than such macromolecules.

To further improve the lifetime of the artificial cornea described above, co-polymers (e.g., methacrylic acid (MMA)) can also be added into the HEMA solution, during fabrication of the hydrogel, for the polymerization of the pHEMA hydrogel. The weight ratio between the added co-polymer to pHEMA can be any value less than 1:2. Addition of these co-polymers improve the strength and other mechanical properties of the artificial cornea. The added co-polymer(s) and pHEMA are both FDA approved to be applied to the human body, thus, the biocompatibility of the artificial cornea can be ensured. The Young’s Modules of the pHEMA with co-polymer(s) can range from a few megapascals to a few hundred megapascals, which will be larger than the Young’s Modulus of the original pHEMA by itself.

Another method for improving the lifetime of the artificial cornea implant is altering the concentration of the cross-linker of the pHEMA. It should also be noted that other materials can also be loaded into the microdimples and/or microgrooves, such as tears, to mitigate dryness.

An improved pHEMA-based artificial cornea can be made using a method which combines the aforementioned peptides and co-polymers. The resulting product would have sufficient transparency, as aided by the peptides, while also having the improved mechanical properties occasioned by the addition of the co-polymers. Such a product may better resist tear, thereby sparing patients from a subsequent secondary surgery. Therefore, the improved method described above has the potential to fabricate artificial corneas suitable for lifelong use by a patient.

It will be understood that the embodiments described hereinabove are merely exemplary and that a person skilled in the art may make many variations and modifications, especially to the manufacturing methods described herein, without departing from the spirit and scope of the present invention.

Claims

What is claimed is:

1. An ocular implant, comprising: a body made, at least partially, from a hydrogel and having an anterior side and a posterior side opposite said anterior side; a first microstructure pattern on said anterior side of said body; and a second microstructure pattern on said posterior side of said body.

2. The ocular implant of Claim 1, wherein said body includes one or more co-polymers incorporated into said hydrogel.

3. The ocular implant of Claim 2, wherein said one or more co-polymers comprise methacrylic acid.

4. The ocular implant of Claim 1, wherein said body includes one or more bioactive molecules incorporated into said hydrogel.

5. The ocular implant of Claim 4, wherein said one or more bioactive molecules comprise one or more peptides.

6. The ocular implant of Claim 1, wherein said first microstructure pattern is different from said second microstructure pattern.

7. The ocular implant of Claim 6, wherein said first microstructure pattern includes a plurality of microlines and said second microstructure pattern includes a plurality of micropores.

8. The ocular implant of Claim 6, wherein said first microstructure pattern includes a plurality of micropores and said second microstructure pattern includes a plurality of microlines.

9. The ocular implant of Claim 1, wherein said first microstructure pattern is the same as said second microstructure pattern.

10. The ocular implant of Claim 9, wherein each of said first and second microstructure patterns includes a plurality of microlines.

11. The ocular implant of Claim 9, wherein each of said first and second microstructure patterns includes a plurality of micropores.

12. The ocular implant of Claim 1, wherein said body has a thickness in a range of from about 100 microns to about 2000 microns.

13. The ocular implant of Claim 1, wherein said body has a thickness in a range of from about 500 microns to about 600 microns.

14. The ocular implant of Claim 1, wherein said ocular implant has a water content greater than 10% by weight.

15. A method for making an ocular implant, comprising the steps of: providing a body made, at least partially, from a hydrogel and having an anterior side and a posterior side opposite said anterior side; applying a first microstructure pattern to said anterior side of said body; and applying a second microstructure pattern to said posterior side of said body.

16. The method of Claim 15, further comprising the step of applying a curvature to said body.

17. The method of Claim 15, wherein said first microstructure pattern is different from said second microstructure pattern.

18. The method of Claim 17, wherein said first microstructure pattern includes a plurality of microlines and said second microstructure pattern includes a plurality of micropores.

19. The method of Claim 17, wherein said first microstructure pattern includes a plurality of micropores and said second microstructure pattern includes a plurality of microlines.

20. The method of Claim 15, wherein said first microstructure pattern is the same as said second microstructure pattern.

21. The method of Claim 20, wherein each of said first and second microstructure patterns includes a plurality of microlines.

22. The method of Claim 20, wherein each of said first and second microstructure patterns includes a plurality of micropores.

23. A method of repairing a damaged cornea, comprising the steps of: providing an ocular implant comprising a body made, at least partially, from a hydrogel and having a first microstructure pattern on an anterior side of said body and a second microstructure pattern on a posterior side of said body opposite said anterior side of said body; lesioning the damaged cornea of a patient; removing dead or damaged tissue from said damaged cornea to surgically create a cornea stroma pocket; and embedding said ocular implant into said cornea stroma pocket such that said posterior side of said body interfaces with said stroma pocket.