CN116492505A - Artificial cornea endothelial transplanting sheet and application thereof - Google Patents
Artificial cornea endothelial transplanting sheet and application thereof Download PDFInfo
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- CN116492505A CN116492505A CN202310529883.4A CN202310529883A CN116492505A CN 116492505 A CN116492505 A CN 116492505A CN 202310529883 A CN202310529883 A CN 202310529883A CN 116492505 A CN116492505 A CN 116492505A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/16—Materials or treatment for tissue regeneration for reconstruction of eye parts, e.g. intraocular lens, cornea
Abstract
The invention provides an artificial cornea endothelial graft, which is treated by plasma technology, and oxygen-containing functional groups are introduced into the surface of the artificial cornea endothelial graft, so that not only is the adhesiveness of the artificial cornea endothelial graft obviously improved, but also the artificial cornea endothelial graft has a certain selective permeability function, and the oxygen permeability coefficient is improved. The plasma treatment artificial endothelial graft provided by the invention can greatly relieve the current situation of lack of cornea donors.
Description
Technical Field
The invention relates to an artificial cornea endothelial transplanting sheet treated by plasma technology and application thereof in cornea injury and cornea function decompensation, belonging to the field of medical artificial substitution materials.
Background
The corneal endothelial cells are single-layered hexagonal cells located between the posterior elastic layer of the cornea and the aqueous humor, constituting a permeable barrier between the corneal stroma and the aqueous humor. Classical theory holds that corneal endothelial cells regulate the hydration state of the corneal stroma via barrier and pumping functions, thereby maintaining corneal transparency and normal thickness. When injury and loss of human corneal endothelial cells are caused by trauma, inflammation or surgery, the cells can be filled only by expansion and migration of peripheral cells, and when the endothelial cell density is reduced to physiological critical value (about 400-500 pieces/mm) 2 ) When the physiological function is insufficient to maintain the constant water content in the cornea stroma, the water content in the cornea stroma is increased, and the cornea dropsy, epithelial blebs, the cornea is caused,Severe eye pain and severe cases caused vision loss. Corneal transplantation is the only effective means of clinically treating corneal endothelial dysfunction, including penetrating keratoplasty and corneal endothelial grafting. Currently, nearly 100 ten thousand patients with corneal endothelial blindness are in need of developing alternative treatment methods for corneal endothelial cells because of shortage of corneal donor materials, which can not meet clinical demands far.
At present, the substitution treatment of the corneal endothelium is a great problem in the field of cornea disease treatment, auffarth teaches that 2 clinical patients who use artificial corneal endothelium to treat chronic corneal edema have fallen off an implant after operation, and the implant needs to be subjected to re-operation and air injection and reposition, and the endothelial piece needs to depend on a stroma of an area which is not covered by the implant to directly contact aqueous humor so as to ensure that nutrient components enter the stroma, and peripheral corneal stroma edema is easy to occur. Therefore, the adhesiveness and permeability of the artificial cornea endothelium are two major problems to be solved urgently, and the design of a new substitute based on a new barrier and selective permeation mechanism of the cornea endothelial cells is a new direction of research and development.
The plasma is a high-energy substance aggregation state containing a large amount of active particles such as electrons, ions, atoms in an excited state, free radicals and the like. At present, in the field of medical materials, a plasma technology is a research hot spot, and comprises the steps of modifying a synthetic high polymer material or a biomedical material so as to meet the required hydrophilic property and biocompatibility requirements and the like. However, no application report of combining plasma with artificial endothelial sheets exists at present.
Disclosure of Invention
In one aspect, the present invention provides a keratoprosthesis having improved performance over prior treatments by plasma treatment; such properties include, but are not limited to, one or more of adhesion, permselectivity, oxygen permeability, hydrophilicity, surface chemistry (oxygen element ratio).
In the invention, the moisture content of the artificial cornea endothelial graft is 40% +/-3%.
Preferably, the thickness of the artificial cornea endothelial graft is 20-90 μm, more preferably 40-70 μm.
Preferably, the diameter of the artificial cornea endothelial graft is 6.0-9.0mm, more preferably 6.5-7.5mm.
Preferably, the radius of curvature of the artificial cornea endothelial graft is 6.0-9.0mm, and more preferably 6.5-7.5mm.
In the invention, the surface of the artificial cornea endothelial graft is treated by plasma technology, including single-sided or double-sided.
Preferably, the power of the plasma treatment is 700-1100W, more preferably 800-1000W.
Preferably, the voltage of the plasma treatment is 220-400V.
Preferably, the plasma treatment time is 3s to 10s, more preferably 5s to 8s per wafer.
Preferably, the plasma treatment is carried out under the condition that air is used as a medium under the atmospheric pressure (100-110 KPa).
Preferably, the sterilization mode of the artificial cornea endothelial graft after plasma treatment is gamma ray irradiation sterilization or ethylene oxide sterilization.
Preferably, the transmittance of the artificial cornea endothelial graft after plasma treatment is 75% -85%.
Preferably, the oxygen permeability coefficient of the plasma-treated artificial cornea endothelial graft is (6.0-9.0). Times.10 -11 (cm 2 /s) [mLO 2 /(mL·hPa)]。
Preferably, the contact angle of the artificial cornea endothelial graft after plasma treatment is 45-55 degrees.
Preferably, the number of fluorescent microspheres adhered to the artificial cornea endothelial graft after the plasma treatment is 3000-3500/mm 2 (after incubation with fluorescent polystyrene particles (10. Mu.L, 2.5% (w/v), diameter 10. Mu.m, QDSpher) for 5 minutes at 25 ℃, a large flow of Tris-HCl (0.01M L-1) was rinsed for 30 seconds).
Preferably, the elements of the artificial cornea endothelial graft after plasma treatment account for 60% -70% of carbon elements and 30% -40% of oxygen elements.
In another aspect, the invention provides an application of the artificial cornea endothelial graft treated by the plasma technology in replacing or repairing cornea endothelium, thereby relieving or treating diseases such as cornea endothelial injury, cornea endothelial cell decompensation, cornea endothelial cell dysfunction and the like, and symptoms such as cornea thickness abnormality, cornea transparency reduction, cornea edema, vision degradation or loss, eye pain and the like caused by the diseases.
The invention also provides application of the artificial cornea endothelial transplantation treated by the plasma technology in medical equipment for relieving or treating corneal endothelial injury, corneal endothelial cell function decompensation and corneal endothelial cell dysfunction. The invention also provides application of the artificial cornea endothelial transplantation treated by the plasma technology in medical equipment for relieving or treating abnormal cornea thickness, reduced cornea transparency, corneal edema, vision degradation or loss and eye pain of patients suffering from corneal endothelial cell dysfunction.
Preferably, the medical device is a implant, patch or device case.
The invention also provides application of the plasma technology in improving the performance of the artificial cornea endothelial graft. The properties include one or more of adhesion, permselectivity, oxygen permeability, surface hydrophilicity, surface chemistry.
The beneficial effects are that:
the invention provides an artificial cornea endothelial graft treated by plasma technology, which uses non-polymeric inorganic gas (O) 2 、N 2 、H 2 Etc.), oxygen-containing functional groups are introduced on the surface of the artificial endothelial graft, and the invention surprisingly found that the adhesiveness of the artificial endothelial graft is obviously improved, and the artificial endothelial graft also has a certain selective permeability function and an improved oxygen permeability coefficient after the plasma treatment. Animal experiments show that after the artificial endothelial sheet is transplanted after plasma treatment, two problems of poor adhesion and permeability are solved, and the artificial endothelial sheet can be transplanted in a simulated donor sheet and a stripped endothelial scope. The artificial endothelial graft after plasma treatment provided by the invention can greatly relieve the angleCurrent status of membrane donor deficiency.
Drawings
FIG. 1 is a schematic diagram of the mechanism of an artificial endothelial graft.
Fig. 2, contact angle test patterns of artificial endothelial grafts obtained in example 1 and example 2.
FIG. 3, fluorescent microsphere adhesion test charts of artificial endothelial grafts obtained in example 1 and example 2.
Fig. 4, and the light transmittance test charts of the artificial endothelial grafts obtained in example 1 and example 2.
Fig. 5, and the spectrum test charts of the artificial endothelial grafts obtained in example 1 and example 2.
Fig. 6, OCT scans 1 day after the artificial endothelial sheet implantation in application example 1 and application example 2.
Fig. 7, application example 1 and application example 2 are a general view of cornea and OCT scan 2 weeks after the artificial endothelial sheet implantation and 3 months after the operation.
FIG. 8, application example 1 and application example 2 are proteomic sequencing charts of rabbit cornea tissues 3 months after surgery.
Detailed Description
The invention is further illustrated by the following examples which illustrate the invention, which are intended to be illustrative only and should not be construed as limiting the scope of the invention. Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The source of the artificial cornea endothelial sheet is not particularly limited in the present invention, and the artificial cornea endothelial sheet known in the art may be used. In the embodiment of the invention, the artificial cornea endothelial sheet is self-made by an affiliated ophthalmology hospital of Shandong first medical university, and is made of a hydroxyethyl methacrylate/methyl methacrylate copolymer.
The specific operation of the plasma treatment of the present invention is not limited, and methods, steps, equipment (plasma processor or corona treatment machine), preservation method and rehydration method known in the art may be adopted.
In the invention, the artificial cornea endothelial graft is made of transparent, soft and foldable acrylic ester materials with good biocompatibility, and can be selected from one or more of hydroxyethyl methacrylate/methyl methacrylate copolymer, polymethyl methacrylate, polyhydroxy ethyl methacrylate, acrylic acid hydrogel and methacrylic acid hydrogel.
In the invention, the artificial cornea endothelial graft after plasma treatment can maintain the nutrition source of the cornea stroma by the nutrient substances in the aqueous humor entering the cornea stroma through the selective permeability of the artificial cornea endothelial graft itself, and does not rely on the area not covered by the artificial endothelial sheet for nutrient permeation.
FIG. 1 is a schematic view of the mechanism of a plasma-treated artificial corneal endothelial graft according to the present invention, which may also be referred to as artificial corneal endothelium, artificial endothelial membrane, corneal implant, corneal endothelial patch, and corneal endothelial sheet, and may be used in place of donor corneal endothelium for corneal endothelial grafting. The size of the artificial endothelial graft subjected to plasma treatment is equivalent to that of the stripped endothelial, and the artificial endothelial graft is used as a waterproof barrier, and meanwhile, has better oxygen permeability and adhesion performance, has a certain permeation effect, and can allow part of small molecules to enter the matrix from aqueous humor, so that the normal metabolism and thickness of the cornea matrix are maintained.
In the present invention, plasma technology improves the performance of artificial endothelial grafts, including but not limited to one or more of adhesion, permselectivity, oxygen permeability coefficient, surface wettability, surface hydrophilicity, surface chemistry. As used herein, "improved" means a state of improved performance as compared to the artificial endothelial graft prior to plasma treatment, for example, improving the adhesion (adherence) of the artificial endothelial graft includes promotion of adhesion (adherence) between the post-corneal stroma and the artificial corneal endothelial graft, between corneal endothelial cells and the artificial corneal implant surface, between the artificial corneal implant surface and adjacent molecules or particles; improving the permselectivity of an artificial endothelial graft includes up-regulating glutathione metabolic pathways and/or sulfur metabolic pathways, promoting permeation of small molecule nutrients (including, but not limited to, water molecules, small molecule peptides, glycogen, amino acids, vitamins, lipids, etc.) between the post-corneal stroma and the artificial cornea endothelial graft, between corneal endothelial cells and the artificial cornea implant surface, between the artificial cornea implant surface and adjacent molecules or particles; improving the surface hydrophilicity of the artificial endothelial graft includes reducing the contact angle of the artificial endothelial graft, promoting adhesion (attachment) between the stroma and the artificial cornea endothelial graft after cornea, and leading the artificial endothelial graft to have better biocompatibility in aqueous environment; improving the surface chemistry of the artificial endothelial graft includes increasing the surface oxygen content of the artificial endothelial graft, promoting the incorporation of oxygen-rich groups (including, but not limited to, carboxyl, carbonyl, hydroxyl, etc.) into the tissue of the posterior stroma of the cornea.
The artificial endothelial graft is subjected to surface modification by a plasma discharge technology, so that the surface of the material is rich in carbonyl, carboxyl and the like, thereby increasing the hydrophilicity of the material, ensuring that the material has good adhesion, and adopting gamma-ray irradiation for sterilization after treatment for standby. In an embodiment, the plasma source has a power of 700-1100W, a voltage of 220-400V, and a corneal substitute treatment time of 3s-10s in air at atmospheric pressure, thereby completing the plasma surface treatment.
For further explanation of the present invention, an artificial endothelial graft and its application will be described in detail below with reference to the accompanying drawings and examples, which should not be construed as limiting the scope of the present invention.
Example 1
The artificial endothelial sheet is prepared by adopting hydroxyethyl methacrylate/methyl methacrylate copolymer, and the parameters of the artificial endothelial graft are as follows: diameter 6.5mm, thickness 50 μm, radius of curvature 7.32mm, water content 38%.
Example 2
The artificial endothelial graft obtained in example 1 was subjected to plasma treatment on both sides by using a corona treatment machine, the power of the plasma source was 800W, the voltage was 400V, and the treatment time of the artificial endothelial graft in air at atmospheric pressure was 5s, to obtain a plasma-treated artificial endothelial graft. The transplanting is put into balanced salt solution for rehydration before the transplanting.
Application example 1
After anesthetizing healthy New Zealand white rabbits (weight 3.0-3.5 Kg), the local analgesia is carried out by using proparacaine hydrochloride eye drops, the endothelium of a region with the thickness of 6.5mm in the central region of the cornea is scraped, the artificial endothelial graft prepared in the example 1 is implanted, anterior chamber air injection fixation is carried out, and the anterior chamber air injection fixation is carried out by applying classical crinis-bone water and eye ointment for 1 time each day within seven days after operation.
Application example 2
The artificial endothelial graft prepared in example 2 was implanted, and the other procedures were the same as in application example 1.
Performance testing
Test example 1
The artificial endothelial grafts obtained in examples 1 and 2 were subjected to oxygen permeability coefficient measurement (see GB/T11417.7-2012, section 7 of ophthalmic optical contact lens: physical and chemical Property test method), and the oxygen permeability coefficient of the artificial endothelial graft obtained in example 1 before treatment was 4.79×10 -11 (cm 2 /s) [mLO 2 /(mL·hPa)]The oxygen permeability coefficient of the artificial endothelial graft after plasma treatment obtained in example 2 was 6.14X10 -11 (cm 2 /s) [mLO 2 /(mL·hPa)]。
Test example 2
The artificial endothelial grafts obtained in examples 1 and 2 were subjected to contact angle measurements using a contact angle analyzer at 25 ℃ using a bubble trapping method, each contact angle measurement being an average of at least five different locations on the sample surface. In FIG. 2, the contact angle of the artificial endothelial graft before treatment obtained in example 1 is 58-59 DEG, and the contact angle of the artificial endothelial graft after plasma treatment obtained in example 2 is 51-52 DEG, and hydrophilicity is enhanced.
Test example 3
Fluorescent microsphere adhesion test was performed at 25 ℃): polystyrene with fluorescenceParticles (10. Mu.L, 2.5% (w/v), diameter 10. Mu.m, QDSpher) were added to the surface of the artificial endothelial grafts obtained in examples 1 and 2. After 5 minutes incubation, the layers were rinsed with a large flow of Tris-HCl (0.01L-1) for 30 seconds. Images of residual microsphere counts (n=3 per group) were captured using confocal microscopy (zeiss LSM 880, carzeiss, germany). As can be seen from FIG. 3, the number of fluorescent microspheres adhered to the artificial endothelial graft obtained in example 2 is 3000-3500/mm 2 The adhesion performance of the artificial endothelial graft obtained in example 1 was superior.
Test example 4
The transmittance of the artificial endothelial grafts obtained in example 1 and example 2 was measured using an ultraviolet spectrophotometer (spectromax M2, molecular Devices, sunnyvale, CA, USA) using a spectral region between 300 and 800 nm. The light transmittance is calculated according to the following equation: light transmittance (%) =X 100, where a represents the absorbance measured. As can be seen from FIG. 4, the transmittance of the artificial endothelial graft obtained in example 2 is superior to that of the artificial endothelial graft obtained in example 1, and the transmittance of the artificial endothelial graft treated by the plasma technique is 75% or more.
Test example 5
The artificial endothelial grafts obtained in example 1 and example 2 were subjected to X-ray photoelectron spectroscopy (XPS) measurement to analyze the chemical composition change of the artificial endothelial surface after plasma treatment. The test results are shown in fig. 5, wherein a is the XPS spectrum of the artificial endothelial graft obtained in example 1, b is the XPS spectrum of the artificial endothelial graft obtained in example 2, and c is the comparison of C, O, N elements. It can be seen from fig. 4 that the O element content of the plasma treated artificial endothelial graft provided by the present invention is significantly increased.
Test example 6
On postoperative day 1, optical Coherence Tomography (OCT) was performed on rabbit cornea in application example 1 and application example 2 under the following test conditions: the scan pattern is shown in fig. 6, where a is an OCT scan after the implantation of the artificial endothelial graft (untreated) obtained in application example 1 in example 1, and b is an OCT scan after the implantation of the artificial endothelial graft (plasma treated) obtained in application example 2 in example 2, and it can be seen from the a graph that the adhesion of the implant is poor and the corneal edema is thickened after the implantation of the untreated artificial endothelial graft, and it can be seen from the b graph that the adhesion of the plasma treated artificial endothelial graft provided by the present invention is good and the cornea is not edema.
Test example 7
The cornea after the operation in application example 1 and application example 2 was observed and measured roughly by using a slit-lamp microscope and OCT scanning, and the test results are shown in fig. 7, wherein a is a cornea roughly observation image and OCT scanning image obtained after the operation of implanting the artificial endothelial graft (untreated) obtained in application example 1, and b is a cornea roughly observation image and OCT scanning image obtained after the operation of implanting the artificial endothelial graft (plasma treated) obtained in application example 2. As can be seen from graph a, the corneal stroma becomes significantly thinner 2 weeks after the implantation of the untreated artificial endothelial graft, and the cornea is near-perforated 3 months after the operation; as can be seen from panel b, the plasma treated artificial endothelial graft provided by the present invention has normal thickness of the corneal stroma at 2 weeks and is maintained for 3 months after surgery (normal rabbit cornea thickness is about 340 μm).
Test example 8
Proteome sequencing is carried out on rabbit cornea tissues 3 months after operation in application example 1 and application example 2, and meanwhile, normal rabbit cornea is used as a control, and the testing method comprises the following steps: three groups of cornea samples (n=3) were subjected to protein extraction by SDT (4% (w/v) SDS, 100mM Tris/HCl ph7.6, 0.1M DTT) lysis and protein quantification was performed by BCA method. Marking according to the specification of a TMT marking kit of Thermo company, and adopting LC-MS/MS data acquisition to carry out protein identification and quantitative analysis. The test results are shown in fig. 8, where a is the KEGG pathway that the post-operative thinned cornea of application example 1 was down-regulated compared to the normal cornea, and b is the KEGG pathway that the post-operative non-thinned cornea of application example 2 was up-regulated compared to the post-operative thinned cornea of application example 1. As can be seen from the graph a, after the untreated artificial endothelial piece is implanted, the glutathione metabolic pathway and the sulfur metabolic pathway are obviously down-regulated, and as can be seen from the graph b, after the plasma treated artificial endothelial piece is implanted, the glutathione metabolic pathway and the sulfur metabolic pathway are obviously up-regulated, which indicates that the plasma treated artificial endothelial piece has a certain permeability effect and can allow permeation of micromolecular nutrient substances.
Although the foregoing embodiments have been described in some, but not all embodiments of the invention, other embodiments may be obtained according to the present embodiments without departing from the scope of the invention.
Claims (8)
1. An artificial cornea endothelial graft, characterized in that the artificial cornea endothelial graft is treated by a plasma technique, thereby improving the performance thereof compared with the artificial cornea endothelial graft before the treatment; the power of the plasma technology treatment is 700-1100W, the voltage is 220-400V, and the treatment time is 3s-10s; the properties include one or more of adhesion, permselectivity, oxygen permeability, surface hydrophilicity, surface chemistry.
2. The artificial cornea endothelial transplanting sheet is characterized in that the surface of the artificial cornea endothelial transplanting sheet is treated by a plasma technology, the power of the plasma technology treatment is 800-1000W, and the treatment time is 5s-8s; the surface comprises one or both sides.
3. The artificial corneal endothelial graft according to claim 1 or 2, wherein the artificial corneal endothelial graft has a water content of 40% ± 3%, a thickness of 20-90 μm, a diameter of 6.0-9.0mm, a radius of curvature of 6.0-9.0mm, and a light transmittance of 75% -85%.
4. The artificial corneal endothelial graft according to claim 1 or 2, wherein the artificial corneal endothelial graft has an oxygen permeability coefficient of (6.0 to 9.0) ×10 -11 (cm 2 /s) [mLO 2 /(mL·hPa)]The method comprises the steps of carrying out a first treatment on the surface of the The contact angle is 45-55 degrees; the oxygen element accounts for 30% -40%.
5. Use of the artificial cornea endothelial graft according to claims 1-4 for medical devices for alleviating or treating corneal endothelial injury, decompensation of corneal endothelial cell function and dysfunction of corneal endothelial cells.
6. Use of the artificial cornea endothelial graft according to claims 1-4 for the preparation of a medical device for alleviating or treating abnormal cornea thickness, reduced cornea transparency, corneal oedema, reduced or lost vision, ocular pain in a patient suffering from decompensation of the function of the cornea endothelial cells.
7. The use according to claim 5 or 6, wherein the keratoprosthesis in the medical device is in a dry vacuum environment.
8. The use according to claim 5 or 6, wherein the medical device is a implant, patch or device box.
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Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4927676A (en) * | 1988-07-01 | 1990-05-22 | Becton, Dickinson And Company | Method for rapid adherence of endothelial cells onto a surface and surfaces prepared thereby |
| US4961954A (en) * | 1987-04-10 | 1990-10-09 | University Of Florida | Surface modified surgical instruments, devices, implants, contact lenses and the like |
| US5433745A (en) * | 1993-10-13 | 1995-07-18 | Allergan, Inc. | Corneal implants and methods for producing same |
| JPH09182762A (en) * | 1995-12-28 | 1997-07-15 | Menicon Co Ltd | Artificial cornea |
| WO2003030958A1 (en) * | 2001-10-06 | 2003-04-17 | Celltran Limited | Corneal graft |
| US20070092550A1 (en) * | 2003-10-10 | 2007-04-26 | Cellular Bioengineering, Inc. | Methods and compositions for growing corneal endothelial and related cells on biopolymers and creation of artifical corneal transplants |
| CN201160919Y (en) * | 2008-03-05 | 2008-12-10 | 黄一飞 | Novel split type artificial cornea |
| CN101726864A (en) * | 2009-11-30 | 2010-06-09 | 温州医学院眼视光研究院 | Method for modifying hydrophility of plasmons at surface of fluorine and silicon hard corneal contact lens |
| US20150306258A1 (en) * | 2014-04-29 | 2015-10-29 | EP Technologies LLC | Plasma sterilization of corneal tissue |
| US20150374881A1 (en) * | 2012-12-27 | 2015-12-31 | Nitta Gelatin Inc. | Human corneal endothelial cell sheet |
| KR101655333B1 (en) * | 2015-03-17 | 2016-09-07 | 전북대학교산학협력단 | Surface-modified Silk Fibroin Implant for Biological Film and Preparation Method Thereof |
| CN108852615A (en) * | 2018-05-09 | 2018-11-23 | 山东省眼科医院 | A kind of plant piece and corneal stroma pouch for corneal stroma implantation |
| CN111320779A (en) * | 2020-02-28 | 2020-06-23 | 广州洁特生物过滤股份有限公司 | Method for modifying surface of substrate and cell culture apparatus |
| CN111565675A (en) * | 2018-01-08 | 2020-08-21 | 爱拥医疗有限公司 | Treatment for improving adhesion of corneal implants |
| CN113934016A (en) * | 2021-08-24 | 2022-01-14 | 天津世纪康泰生物医学工程有限公司 | Orthokeratology mirror based on artificial intelligence optimization algorithm |
| WO2023059933A1 (en) * | 2021-10-08 | 2023-04-13 | Purdue Research Foundation | Ocular drug delivery device and related methods |
-
2023
- 2023-05-11 CN CN202310529883.4A patent/CN116492505B/en active Active
Patent Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4961954A (en) * | 1987-04-10 | 1990-10-09 | University Of Florida | Surface modified surgical instruments, devices, implants, contact lenses and the like |
| US4927676A (en) * | 1988-07-01 | 1990-05-22 | Becton, Dickinson And Company | Method for rapid adherence of endothelial cells onto a surface and surfaces prepared thereby |
| US5433745A (en) * | 1993-10-13 | 1995-07-18 | Allergan, Inc. | Corneal implants and methods for producing same |
| JPH09182762A (en) * | 1995-12-28 | 1997-07-15 | Menicon Co Ltd | Artificial cornea |
| WO2003030958A1 (en) * | 2001-10-06 | 2003-04-17 | Celltran Limited | Corneal graft |
| US20070092550A1 (en) * | 2003-10-10 | 2007-04-26 | Cellular Bioengineering, Inc. | Methods and compositions for growing corneal endothelial and related cells on biopolymers and creation of artifical corneal transplants |
| CN201160919Y (en) * | 2008-03-05 | 2008-12-10 | 黄一飞 | Novel split type artificial cornea |
| CN101726864A (en) * | 2009-11-30 | 2010-06-09 | 温州医学院眼视光研究院 | Method for modifying hydrophility of plasmons at surface of fluorine and silicon hard corneal contact lens |
| US20150374881A1 (en) * | 2012-12-27 | 2015-12-31 | Nitta Gelatin Inc. | Human corneal endothelial cell sheet |
| US20150306258A1 (en) * | 2014-04-29 | 2015-10-29 | EP Technologies LLC | Plasma sterilization of corneal tissue |
| KR101655333B1 (en) * | 2015-03-17 | 2016-09-07 | 전북대학교산학협력단 | Surface-modified Silk Fibroin Implant for Biological Film and Preparation Method Thereof |
| CN111565675A (en) * | 2018-01-08 | 2020-08-21 | 爱拥医疗有限公司 | Treatment for improving adhesion of corneal implants |
| CN108852615A (en) * | 2018-05-09 | 2018-11-23 | 山东省眼科医院 | A kind of plant piece and corneal stroma pouch for corneal stroma implantation |
| CN111320779A (en) * | 2020-02-28 | 2020-06-23 | 广州洁特生物过滤股份有限公司 | Method for modifying surface of substrate and cell culture apparatus |
| CN113934016A (en) * | 2021-08-24 | 2022-01-14 | 天津世纪康泰生物医学工程有限公司 | Orthokeratology mirror based on artificial intelligence optimization algorithm |
| WO2023059933A1 (en) * | 2021-10-08 | 2023-04-13 | Purdue Research Foundation | Ocular drug delivery device and related methods |
Non-Patent Citations (5)
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| Publication number | Publication date |
|---|---|
| CN116492505B (en) | 2023-12-08 |
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Effective date of registration: 20240415 Address after: 200333, Room 1009, No. 1777 Zhongshan North Road, Putuo District, Shanghai Patentee after: Shanghai Weishi Medical Equipment Co.,Ltd. Country or region after: China Address before: 250021, No. four, No. 372, Ji'nan, Shandong Patentee before: Affiliated ophthalmic hospital of Shandong First Medical University (Shandong ophthalmic Hospital) Country or region before: China |