CN103989553B - Method for manufacturing and storing corneal injury scar-free repairing device - Google Patents
Method for manufacturing and storing corneal injury scar-free repairing device Download PDFInfo
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- CN103989553B CN103989553B CN201410256997.7A CN201410256997A CN103989553B CN 103989553 B CN103989553 B CN 103989553B CN 201410256997 A CN201410256997 A CN 201410256997A CN 103989553 B CN103989553 B CN 103989553B
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Abstract
The invention relates to a method for manufacturing and storing a corneal injury scar-free repairing device. Specially-prepared amnion elements are settled or bonded in or on the surfaces of various biological or synthetic attaching devices such as the invisible contact lens through the novel carrier controlled release technology and the biological tissue engineering technology so as to form the corneal or conjunctiva injury scar-free repairing device. At the early stage of the corneal or conjunctiva injury, the affected corneal tissue or the affected conjunctiva tissue is covered with the amnion elements of the corneal or conjunctiva injury scar-free repairing device, the amnion elements, such as the necessary cell growth factors, the tissue repair factors and protease inhibitors, of the corneal or conjunctiva injury scar-free repairing device are diffused to the affected corneal tissue or the affected conjunctiva tissue due to the special superfine structure of an amnion basilar membrane and the stem cell characteristic of amniotic epithelial cells, and therefore growth of epithelial cells of the corneal or the conjunctiva is promoted, the affected corneal tissue or the affected conjunctiva tissue is stimulated, regulated and controlled to heal orderly, and less scars are formed on the affected corneal tissue or the affected conjunctiva tissue.
Description
Technical Field
The invention relates to a brand-new biomedical product and a novel ophthalmic treatment material developed by utilizing a novel carrier controlled release technology and a tissue engineering technology, in particular to the field of scar-free repair of corneal injury, and has great improvement and innovation on the existing method for clinically preventing and treating the scar formed by corneal trauma. In particular to a corneal injury scar-free repair device and application thereof.
Background
According to the latest published statistics of the World Health Organization (WHO), 700 million people become blind people every year in the world, and at present, 4500 million blind people are shared in the world. China is the country with the most blind people in the world, and has about 1200 million people with visual disabilities, wherein the blind people have more than 700 million people and account for about 1/6 of the total number of the blind people in the world. The keratopathy accounts for 15.4% of the cause of blindness in China.
The human eye is a very powerful focusing device, and the focusing elements of the eye are made up of two parts, the cornea and the crystalline lens, the crystalline lens making up about 20% (12 diopters) and the cornea making up 80% (48 diopters) of the eye's focusing power. The cornea is divided into five layers which are sequentially from front to back: epithelial cell layer, anterior elastic layer (also known as Bowman's membrane), stromal layer, posterior elastic layer (also known as Descemet's membrane), and endothelial cell layer. One of the important factors of corneal transparency is the absence of blood vessels in the corneal tissue, which terminate at the limbus, forming a network of blood vessels from which the nutrients diffuse into the cornea. The sensory nerve of the cornea is abundant and reaches the cornea mainly from the eye branch of the trigeminal nerve via the ciliary nerve.
In view of the specific role of the cornea, corneal epithelial cells have the ability to continually self-renew and rapidly heal wounds. In this process, the overall synergy of proliferation, migration, differentiation, death, etc. of corneal epithelial cells is important. Epithelial cells in the middle of the cornea were injured 5 hours later and epithelial cells around the injured cells slid horizontally to cover the denuded part thereof. The surrounding epithelial cells continue to provide the cells needed for the site of injury until complete recovery. However, this rapid proliferation of corneal epithelial cells often leads to the development of scar tissue, which in turn leads to corneal haze. In addition, the middle and front elastic layers, the stroma layer and the back elastic layer of the cornea cannot be regenerated after being damaged and can only be repaired and filled by opaque scar tissues, so that the vision disorder is caused by the formation of scars in different degrees. Studies have shown that maintenance of the ocular surface (the entire mucosal epithelium between the upper and lower eyelid margins, including the epithelium of the conjunctiva and cornea) stable and intact can prevent corneal disease, not only providing a good microenvironment for corneal and conjunctival reconstructive surgery, but also being the basis and prerequisite for preventing corneal scarring.
A great deal of corneal injury caused blindness is caused by scar tissue, and the common causes of corneal injury are: 1. trauma (Corneal injures): perforation, contusion, explosion, chemical burn, hot scald, etc. of cornea; 2. infectious Corneal lesions (Corneal Infections): including keratitis and corneal ulcer caused by bacteria, fungi and viruses; 3. refractive surgery (reflctive surgery): since the first excimer laser refractive surgery was first developed in the early 90 s, refractive surgery techniques have become widespread worldwide, with over 100 million people doing the surgery in the united states alone each year, and nearly 100 million people doing the surgery in china each year. Currently, refractive surgery is technically divided into two major directions, namely PRK and LASIK, and PRK laser surgery has been recommended in the united states from the last year more than LASIK laser surgery because PRK can bring better vision to patients. The biggest obstacle preventing the development of PRK refractive surgery is post-operative scarring. 3. Allergic Corneal injury (Corneal allorgies): such as bleb keratitis and the like; 4. degenerative or dystrophic corneal injuries (Degenerative or Nutritional disorders): such as corneal arcus senilis, corneal zonal degeneration, lattice dystrophy, corneal softening, etc.; 5. cicatricial Corneal injury (Corneal Scars): corneal clouding, leukoplakia, adhesive blood spots, corneal staphyloma, etc.
Corneal diseases are now common diseases and frequently encountered diseases in ophthalmology, and are also one of important blindness-causing eye diseases. The cornea is positioned at the forefront of the eyeball and is directly contacted with the external environment, so that the cornea is damaged frequently, and the healed scar after the injury can have serious influence on the visual function of a patient. Therefore, reasonable treatment of the cornea will greatly help the patients to recover visual function.
In 1999, Ming X.Wang et al in the United states treated CORNEAL injury FOR the first time with an amniotic CONTACT LENS, the amniotic CONTACT LENS contacted with the amniotic membrane component to deposit the amniotic membrane component on the surface of the amniotic CONTACT LENS or permeate into the interior of the amniotic CONTACT LENS FOR reducing or eliminating the scar formed after the CORNEAL injury and preventing CORNEAL opacity (patent: BIOCHEMICAI CONTACT LENS FOR TREATINGINJURED CORNEAL TISSUE), but the amniotic CONTACT LENS did not consider the individual difference of patients, and the size and shape of the amniotic CONTACT LENS used were the same. When a patient uses the amniotic contact lens, the damaged part of the patient is easily damaged secondarily, and the comfort level of the patient is poor. Meanwhile, the amniotic membrane contact lens directly covers the cornea, so that the sealing property is stronger, the oxygen permeability is insufficient, and the cornea is easy to lack of oxygen. The clinical application still has certain limitations.
In terms of drug therapy: topical steroids have been used to reduce corneal haze and, for refractive outcomes, while glucocorticoids maintain the transition from hyperopia, their effects are reversed after treatment is discontinued and the use of such drugs as a long-term treatment is not possible. In fact, many diseases associated with corneal haze are not directly caused by pathogens or trauma, but rather by scar tissue, such as: cicatricial pemphigoid and Steve-Johnson syndrome (SJS).
Amnion transplantation (AMT) can successfully promote growth and healing of ocular surface tissues (cornea and conjunctival epithelium) after injury, and inhibit scar formation of the eye surface and the cornea. But also has the defects that the method cannot overcome: 1) the transplanted amnion can be dissolved and absorbed within days or weeks after operation, and sometimes needs to be operated for many times, which brings great pain to patients; 2) the amnion transplantation operation can be implemented by only a few famous ophthalmologists in complete-equipment ophthalmology hospitals worldwide; 3) the sewing process of the amnion inevitably causes certain damage to the eye tissue; 4) amniotic membrane transplantation surgery presents a risk of infection and corneal perforation during the acute phase of corneal trauma; 5) AMT has great limitations for the treatment of severe eye diseases and corneal wounds.
Corneal transplantation is a main method for treating corneal scar blindness in the world, and is difficult to popularize in the world due to the limitation of corneal sources, surgical equipment, technical requirements, immunological rejection and the like.
In view of the lack of the current treatment means for the keratopathy, the haze/scar of the cornea caused by the existing laser ablation operation of the corneal tissue and the failure of the ideal treatment effect of the drug treatment, an effective treatment means is urgently needed.
Disclosure of Invention
The invention aims to provide a corneal injury scar-free repair device and application thereof, which are used for extracting amnion or recombining into amnion from amnion tissue and placenta tissue. The device introduces the individual technical innovation concept into the manufacturing field, is customized according to the shape of the cornea wound, effectively protects the wound part of the cornea of the eye, and has better biocompatibility. Meanwhile, the oxygen permeable hole is designed in the device, so that the throughput of oxygen can be greatly increased, the cornea can freely breathe, and the cornea is kept in the optimal state. The device should be stored in amniotic fluid stock immediately after preparation. The device can also fix the medicine for treating the corneal injury on the amniotic membrane to treat the injured corneal tissue.
The specific technical scheme is as follows:
a method for manufacturing a scar-free cornea injury repair device, further, bonding an amnion and an adnexal device together by adopting the following method:
the amnion component is bonded on the surface of the attaching device through a covalent bond;
or,
bonding the amniotic membrane component and the epitype device by attractive forces induced by hydrogen bonding, ionic bonding, van der waals forces, and the like;
or,
an adhesive is used to bond the amniotic membrane component and the casting device.
Further, the binder comprises collagen, fibroin, keratin, casein, bovine albumin, laminin, matrigel, polylysine or polyornithine, a positively charged matrix, dextran, chitosan, a polyvinyl acetate film, paraformaldehyde, polyvinyl alcohol, egg white, a hydrogel, biotin-streptavidin, gelatin-alum-formaldehyde, serum, hyaluronic acid, fibrin glue, pre-coated gelatin, or an antigen-antibody.
Further, the attachment device is a contact lens, other biological attachment device or synthetic attachment device.
Further, the amniotic membrane is pretreated, and the method comprises the following steps:
a. performing blunt separation on the placenta amnion, wherein phosphate buffer solution PBS contains penicillin, streptomycin and amphotericin B;
b. laminating to the nitrocellulose membrane such that the filter with the mesenchymal side of the amniotic membrane facing forms a double membrane;
c. the double film is cut into loops of a certain diameter.
Further, the air conditioner is provided with a fan,
step a also includes the following steps: after placenta tissues of infected people are screened conventionally, cesarean section and normal pregnancy are immediately selected; and/or the presence of a gas in the gas,
in step a, 1000 units/ml of penicillin, 20 mg/ml of streptomycin and 2.5 micrograms/ml of amphotericin B are added; and/or the presence of a gas in the gas,
the method also comprises the following steps between the steps a and b: washing amnion with PBS solution for several times;
and/or the presence of a gas in the gas,
the method also comprises the following steps between the steps b and c: the other double membranes were washed with PBS solution;
and/or the presence of a gas in the gas,
the double film was cut into 9 mm diameter circles in step c and stored in 100% 4 ℃ glycerol.
Further, the amniotic membrane is pretreated, and the method comprises the following steps:
(1) blunt separating amnion from chorion of placenta, and attaching onto paper sheet;
(2) the pieces of amniotic membrane attached paper were stored.
Further, the air conditioner is provided with a fan,
the method also comprises the following steps before the step (1): immediately washing blood stain of placenta obtained by selective cesarean section by using normal saline containing 50mg/L of penicillin, 50mg/L of streptomycin and 215 mg/L of amphotericin B under aseptic condition;
and/or the presence of a gas in the gas,
detecting that the placenta donor hepatitis B surface antigen, AIDS antigen, chlamydia, syphilis, cytomegalovirus and the like are negative before delivery, and has no history of other malignant tumors and infectious diseases;
and/or the presence of a gas in the gas,
in the step (1), the separated paper is laid on a paper sheet of an operation pasting paper towel, and the upper leather surface is upward;
and/or the presence of a gas in the gas,
in the step (2), the paper sheet attached with the amnion is cut into pieces with the size of 8cm multiplied by 8cm, and is placed at the temperature of minus 80 ℃ 1: 1, glycerol BDMEM.
According to the storage method of the product of the manufacturing method of the cornea injury scar-free repair device, the device is immediately stored in the amnion storage solution after being prepared; or, the device is freeze-dried and stored after being prepared; or the amnion in the device is freeze-dried after being prepared, and then is adhered and stored with the attaching device.
Further, after lyophilization, the amniotic membrane may be stored at room temperature or stored in amniotic membrane stock solution.
Further, the amniotic membrane is stored in an amniotic fluid stock immediately after being attached to the contact lens, and/or the amniotic fluid stock contains an inflammation-inhibiting agent.
Compared with the prior art, the cornea injury scar-free repair device has the advantages that the amniotic membrane components are deposited in the amniotic membrane contact lens or are connected with the amniotic membrane components on the surface of the amniotic membrane contact lens, the amniotic membrane contact lens is in contact with injured corneal tissues, and necessary tissue growth factors are diffused from the amniotic membrane contact lens, so that the injured corneal tissues are stimulated to heal, and scar formation of the injured corneal tissues is reduced. Inflammation and fibrovascular ingrowth can be greatly reduced by affecting basic molecules and wound healing processes such as cell activation and apoptosis. The device can be customized according to individual needs by the shape of the cornea wound, and effectively protects the wound part of the cornea of the eye. Meanwhile, the oxygen permeable hole is designed in the device, so that the throughput of oxygen can be greatly increased, the cornea can freely breathe, and the cornea is kept in the optimal state.
Specifically, the method comprises the following steps:
1. the device makes the amniotic membrane contact lens contact with injured corneal tissue, necessary tissue growth factors are diffused from the amniotic membrane contact lens and released into the injured corneal tissue along with the lapse of time, so that the injured corneal tissue is stimulated to heal, and the scar formation of the injured corneal tissue is reduced. And because the amniotic membrane component of the device may penetrate into other regions of the eye during corneal treatment, the amniotic membrane component is effective in preventing the formation of corneal scarring in tissues other than or adjacent to the injured region of the eye.
2. The device can be designed into devices with different shapes according to different corneal curvatures and corneal wounds of patients according to personal needs, so that the wound part of the cornea of the eye is effectively protected, and secondary injury is effectively avoided.
3. The oxygen permeable hole is designed in the device, so that the throughput of oxygen can be greatly increased, the cornea can freely breathe, and the cornea can be kept in the optimal state. Not only improves the oxygen permeability, but also ensures the firmness of the material, and has good wettability and anti-precipitation performance.
4. The device can also fix and attach the medicine for treating corneal injury or other intraocular diseases on the amniotic membrane by a special method, and treat the corneal injury or other intraocular diseases such as glaucoma, retinopathy and other diseases by contacting the injured corneal tissue for slow release of the medicine.
5. The device is convenient to use, can implant and take out very easily, can cooperate the amnion eye drops to use simultaneously, and it is convenient, durable to maintain.
Drawings
FIG. 1 shows an amniotic membrane contact lens of the present invention
FIG. 2 is a schematic diagram of an amniotic membrane contact lens of the present invention
Detailed Description
The invention is described in detail below with reference to the attached drawing, which is a preferred example of various embodiments of the invention.
The first embodiment is as follows:
1. pretreatment of amnion
There are many methods for amniotic membrane pretreatment, and this example adopts: immediately washing blood stain of placenta obtained by selective cesarean section under aseptic condition with physiological saline containing 50mg/L penicillin, 50mg/L streptomycin and 215 mg/amphotericin B, blunt-separating amnion from chorion of placenta, spreading on paper sheet of operation sticking paper towel (epithelial surface facing upwards) "and cutting the paper sheet with amnion into pieces of 8cm × 8cm, and placing at-80 deg.C 1: 1, glycerol BDMEM. The prenatal detection of the placenta donor hepatitis B surface antigen, AIDS antigen, chlamydia, syphilis, cytomegalovirus, etc. shows negative results, and has no history of other malignant tumors and infectious diseases.
2. Production of contact lenses
A special biological material is precisely processed into the invisible contact lens by a numerical control lathe, and a plurality of micro small holes are punched on the invisible contact lens to manufacture the oxygen permeable contact lens.
3. Bonding of amniotic membrane to contact lens
The bonding method of the amnion and the contact lens is various and comprises physical, chemical and morphological methods and the like; for example, the amnion component is bonded to the surface of the contact lens by a covalent bond, and the amnion component is bonded by an attractive force due to a hydrogen bond, an ionic bond, a van der waals force, or the like, and may be bonded by an adhesive such as collagen (collagen) or Matrigel (Matrigel). In this embodiment, the amnion component is covalently bonded to the surface of the contact lens.
4. Preservation of corneal injury after scar-free repair device preparation
In the embodiment, the amnion is adhered to the contact lens and then stored in an amnion stock solution containing specific inflammation inhibiting agents, such as antibiotics, to ensure that the tissue repair factors and growth factors of the amnion in the corneal injury scar-free repair device are embedded on the contact lens.
Typically, corneal lesions are best treated between 24-48 hours.
Example two:
the angle (node) membrane injury scar-free repair device comprises: the method comprises the steps of pretreatment of the amnion, the structure of various biological or synthetic attached devices including a contact lens, the adhesion method of the amnion and various biological or synthetic attached devices including the contact lens, the preparation method of oxygen permeation and oxygen permeation holes of a scar-free repair device for corneal (nodal) membrane injury, the use method of amnion eye drops/amnion storage solution, and the method for fixing the medicine for treating corneal and conjunctival injury on the amnion. Further: the method comprises the steps of extracting amnion from human or mammal (such as pig, horse or cow) amnion tissue and placenta tissue and biologically recombining to form the amnion, bonding the amnion and various biological or synthetic attached devices including a contact lens and the like together through a novel carrier controlled release technology and a biological tissue engineering technology to prepare an angular (node) membrane injury scarless repair device, contacting the amnion component of the angular (node) membrane injury scarless repair device with injured cornea and conjunctival tissue, and releasing necessary cell growth factors, tissue repair factors, protease inhibitors and the like from the amnion component of the angular (node) membrane injury scarless repair device into the injured cornea and conjunctival tissue, thereby stimulating the injured cornea and conjunctival tissue to heal and reducing scars formed by the injured cornea and conjunctival tissue. The device can be customized according to individual needs by the shape of the cornea wound, and effectively protects the wound part of the cornea of the eye. Meanwhile, the oxygen permeable characteristic and the oxygen permeable hole are designed, so that the oxygen passing amount can be greatly increased, the cornea and the conjunctiva can breathe freely, and the cornea and the conjunctiva are kept in the optimal state. The device can be stored at room temperature after freeze-drying is completed, and the physiological activity is completely kept for more than six months; if the amnion is preserved in the specially prepared amnion storage solution, the curative effect is increased and the time period is prolonged. The device can also fix the medicine for treating the cornea and conjunctival injury on the amniotic membrane to treat the injured cornea and conjunctival tissue.
The injured corneal and conjunctival tissues comprise: 1. trauma: including natural and unnatural wounds such as contusion, explosion, chemical burns, thermal burns, wearing and tearing of common contact lenses and orthokeratology lenses, and damage caused by medical operations on the cornea and conjunctiva (various laser operations for myopia, hyperopia and astigmatism; corneal contact ring for presbyopia; and corneal and conjunctival diseases including corneal transplantation operations, etc.); 2. infectious corneal injury: including keratitis and corneal ulcer caused by bacteria, fungi and viruses; 3. allergic corneal injury: such as bullous keratitis and Steve-Johnson syndrome (SJS); 4. degenerative or dystrophic corneal injury: such as corneal arcus senilis, corneal zonal degeneration, lattice dystrophy, corneal softening, etc.; 5. cicatricial corneal injury: corneal clouding, leukoplakia, adhesive blood spots, corneal staphyloma, etc.
The amnion is obtained by extracting amnion from human or mammal (such as pig, horse or cattle) amnion tissue and/or placenta tissue, and recombining into amnion.
The said attachment includes various biological or synthetic attachments such as contact lenses.
The contact lenses in the attaching device comprise soft contact lenses, hard contact lenses, high oxygen permeability cornea shaping lenses (RGPs) and organic polymer and inorganic polymer mixed gel made of various nano materials.
Various biological or synthetic attachments including contact lenses are made of polyalkyl (poly-acrylonitrile), Polymethacrylate (PMMA), methylmethacrylate, polysilicate, polymethylmethacrylate, polyfluoroacrylate, polyfluorosilicate methacrylate, polymethacrylate, polyacrylate, polyurethane, polysilicate, polymethylene succinate, polycyanoacrylate, polybutylcyanoacrylate, polycyanoacrylate, polyhexamethylene cyanoacrylate, polycyanoacrylate, polyethylene cyanoacrylate, polyisocyanuroacrylate, silicon dioxide (Silica), Silicone (Silicone), polyethylene (polyethylene), polyvinyl pyrrolidone, polyvinyl alcohol, polyvinyl,
polydioxanone (gdm), Glycerol Methacrylate (GMA), hydrogel (hydrogel), hydroxyethyl methacrylate (HEMA), N- (2-hydroxypropyl) methacrylate (HPMA), hillafilcon B, and the like.
The contact lens can introduce various high oxygen permeability keratoplasty lens (RGP) concepts into the contact lens, and the shapes of the middle and the periphery of the contact lens are different, so that the cornea wound part is protected.
The said attached device includes various biological or synthetic attached devices such as contact lens, etc. with auxiliary oxygen-permeable hole and high oxygen permeability.
The amnion component in the repair device is adhered to the inner surface of various biological or synthetic attachments including contact lenses and the like by biological tissue engineering technology.
The method for bonding the amnion and the contact lens comprises the following steps: including physical, chemical, and morphological methods; such as covalent bonding through the amniotic membrane component to the surface of the contact lens, attractive bonding through hydrogen bonding, ionic bonding, van der waals forces, and the like; it is also possible to use a binder such as collagen, fibroin, keratin, casein, bovine albumin, laminin, matrigel, polylysine or polyornithine, a positively charged matrix (e.g., polylysine), dextran, chitosan, polyvinyl acetate film, paraformaldehyde, polyvinyl alcohol, egg white, hydrogel, biotin-streptavidin, gelatin-alum-formaldehyde, serum, hyaluronic acid, fibrin glue, pre-coated gelatin, antigen-antibody, etc.
The amnion component in the device for repairing damaged cornea (node) without scar is contacted with the damaged corneal tissue.
The amnion part of the device for repairing cornea (node) damage without scar can be stored at room temperature to keep physiological activity by special treatment such as freeze-drying.
The repair device without scar for corneal (nodal) membrane injury has the advantages of increased curative effect and prolonged aging if stored in amniotic fluid storage.
The amnion storage solution contains specific inflammation inhibiting preparations such as antibiotics and the like, and is used together with amnion eye drops.
The scar-free repair device for corneal (nodal) membrane injury can attach a medicine for treating corneal injury to an amniotic membrane by a specific method, and treat the corneal injury by contacting the injured corneal tissue to slowly release the medicine.
The scar-free repair device for corneal (nodal) membrane injury has the best use effect within 24-48 hours of corneal injury.
The amnion component in the device for repairing damaged cornea (node) without scar is contacted with the damaged corneal tissue.
One of the characteristics of the invention is as follows: the pretreatment method of the amnion comprises the steps of immediately selecting caesarean section and normal pregnancy after screening the placenta tissue of the infected human by a conventional method. By blunt placental amniotic membrane separation, Phosphate Buffered Saline (PBS) contained penicillin (1000 units/ml), streptomycin (20 mg/ml) and amphotericin B (2.5. mu.g/ml). The amniotic membrane was washed several times with PBS solution and then laminated to nitrocellulose membrane such that the filter with the mesenchymal side of the amniotic membrane facing it formed a double membrane. The other double membrane was washed with the above-mentioned PBS solution. The double film was cut into 9 mm diameter circles and stored in 100% glycerol at 4 ℃.
The invention has the following two characteristics: the structure of the invisible contact lens is precisely processed by a special biological material through a numerical control lathe, and the structure of the invisible contact lens designs contact lenses with different shapes according to different curvatures of corneas of different patients to form protection on damaged parts; meanwhile, the oxygen permeable contact lens with the auxiliary perforation is designed, so that the throughput of oxygen can be greatly increased, the cornea can freely breathe, the oxygen permeability is improved, the firmness of the material is ensured, and the oxygen permeable contact lens has good wettability and anti-precipitation performance.
Another aspect of the present invention is that the method of adhesion of amniotic membrane to a contact lens, including physical, chemical, and morphological methods, and the like, can be achieved by, for example, covalently bonding the amniotic membrane component to the surface of a contact lens, by bonding through hydrogen bonding, ionic bonding, van der Waals forces, and attractive forces due to such forces, and can also be achieved by collagen, Matrigel, polylysine or polyornithine, positively charged matrices (e.g., fustin), chitosan, Polyvinegar film, triformol, poval, egg white, hydrogel, biotin-streptavidin, Gelatin, Whiteglues, Gelatin-alum-formaldehyde, blood serum, hyaluronic acid, bovine albumin, fibrin gel, Preliminary spread Gelatin, laminin, antigen-antibody reaction, and the like.
The invention has the fourth characteristic that: the application method of the amnion eye drops/amnion storage liquid comprises the following steps: the cornea injury scar-free repair device is prepared and then is stored in amniotic membrane storage liquid; the amnion stock solution contains specific inflammation inhibiting preparation such as antibiotic, and is supplemented with amnion eye drop.
The invention has the five characteristics that: the medicine for treating the corneal injury is fixedly attached to the amniotic membrane by a special method, and the corneal injury is treated by contacting the injured corneal tissue for slowly releasing the medicine.
The invention is characterized in that: the optimal time for implanting the scar-free cornea injury repair device into the cornea is 24-48 hours of cornea injury.
The specially prepared amnion component is deposited or adhered in various biological or synthetic attached devices including contact lenses and the like or forms an angle (knot) membrane injury scar-free repair device on the surface by a novel carrier controlled release technology and a biological tissue engineering technology. In the early stage of corneal (nodal) membrane injury (usually within a time window of 24 hours to 48 hours of injury), injured corneal and conjunctival tissues are covered by amniotic membrane components of the corneal (nodal) membrane injury scar-free repair device, and the amniotic membrane components of the corneal (nodal) membrane injury scar-free repair device are utilized to diffuse into the injured corneal (nodal) membrane tissue from the amniotic membrane components of the corneal (nodal) membrane injury scar-free repair device by utilizing the special ultrastructure of the amniotic membrane basement membrane and the characteristics of amniotic epithelial cell stem cells such as necessary cell growth factors, tissue repair factors, protease inhibitors and the like, so that the corneal (nodal) membrane epithelial cell growth is promoted, the ordered healing of the injured corneal (nodal) membrane tissue is stimulated and regulated, and the scar formation of the injured corneal (nodal) membrane tissue is reduced. The amnion component of the repair device for repairing injured scarless cornea (conjunctiva) membrane affects wound healing processes of excessive secretion of cornea and conjunctival extracellular matrix, excessive deposition of collagen, growth and transformation of fibroblasts and the like through the regulation effect on apoptosis, and obviously reduces inflammatory reaction and fibrovascular ingrowth. The device can be customized according to individual needs by the shape of cornea wound, and effectively protects the wound parts of cornea and conjunctiva of the eye. Meanwhile, the biological and synthetic attached part of the device including the invisible contact lens is processed by a nanotechnology, has high oxygen permeability and is also provided with an oxygen permeation hole, so that the throughput of oxygen can be obviously increased, the cornea can freely breathe, and the cornea (conjunctiva) can be kept in the optimal physiological health state. The device can be stored at room temperature after freeze-drying is completed, and the physiological activity is completely kept for more than six months; if the amnion is preserved in the specially prepared amnion storage solution, the curative effect is increased and the time period is prolonged. The device can also fix the medicine for treating the cornea and conjunctival injury on the amniotic membrane, and carry out inflammation repair treatment on the injured cornea and conjunctival tissue.
The invention has been described in connection with the accompanying drawings, it is to be understood that the invention is not limited to the specific embodiments disclosed, but is intended to cover various modifications, adaptations or uses of the invention, and all such modifications and variations are within the scope of the invention.
Claims (5)
1. A manufacturing method of a cornea injury scar-free repairing device is characterized in that amnion and an attached device are bonded together by adopting the following method:
the amnion component is bonded on the surface of the attaching device through a covalent bond;
alternatively, the amniotic membrane component and the epitype device are bound by hydrogen bonding, ionic bonding, and attractive forces induced by van der waals forces;
alternatively, an adhesive is used to bond the amniotic membrane component and the casting device;
the adhesive comprises collagen, fibroin, keratin, casein, bovine albumin, laminin, matrigel, polylysine or polyornithine, positively charged matrix, dextran, chitosan, polyvinyl acetate film, paraformaldehyde, polyvinyl alcohol, egg white, hydrogel, biotin-streptavidin, gelatin-alum-formaldehyde, serum, hyaluronic acid, fibrin glue, pre-laid gelatin, or antigen-antibody;
the attached device is a contact lens;
the pretreatment of the amnion comprises the following steps:
a. blunt separation of placental amniotic membrane;
b. laminating to the nitrocellulose membrane such that the filter with the mesenchymal side of the amniotic membrane facing forms a double membrane;
c. cutting the double film into circles; the double film was cut into 9 mm diameter circles and stored in 100% glycerol at 4 ℃;
the method also comprises the following steps between the steps a and b: washing amnion with phosphate buffer PBS for several times;
the method also comprises the following steps between the steps b and c: washing the double membranes by phosphate buffer PBS;
phosphate buffered saline PBS contains penicillin, streptomycin and amphotericin B; penicillin is 1000 units/ml, streptomycin is 20 mg/ml, amphotericin B is 2.5 μ g/ml;
or, the amnion is pretreated, comprising the following steps:
(1) blunt separating amnion from chorion of placenta, and attaching onto paper sheet;
(2) the pieces of amniotic membrane attached paper were stored.
2. The method for manufacturing a scar-free repair device for corneal injuries as claimed in claim 1,
the method also comprises the following steps before the step (1): immediately washing blood stain of placenta obtained by selective cesarean section by using normal saline containing 50mg/L of penicillin, 50mg/L of streptomycin and 215 mg/amphotericin B under aseptic condition;
detecting that the placenta donor hepatitis B surface antigen, AIDS antigen, chlamydia, syphilis and cytomegalovirus are negative before delivery, and has no history of other malignant tumors and infectious diseases;
in the step (1), the separated paper is laid on a paper sheet of an operation pasting paper towel, and the upper leather surface is upward;
in the step (2), the paper sheet attached with the amnion is cut into pieces with the size of 8cm multiplied by 8cm, and is placed at the temperature of minus 80 ℃ 1: 1, glycerol BDMEM.
3. The method for manufacturing a scar-free repair device for corneal injuries as claimed in claim 2,
the device is immediately preserved in amnion storage solution after the preparation; or, the device is freeze-dried and stored after being prepared; or the amnion in the device is freeze-dried after being prepared, and then is adhered and stored with the attaching device.
4. The method of claim 3, wherein the device is lyophilized and stored at room temperature or stored in an amniotic fluid stock.
5. The method of claim 4, wherein the amniotic membrane is stored in an amniotic membrane stock solution immediately after being adhered to the contact lens, and/or wherein the amniotic membrane stock solution comprises an inflammation-inhibiting formulation.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2822412C1 (en) * | 2023-10-19 | 2024-07-04 | Федеральное государственное бюджетное учреждение "Национальный медицинский исследовательский центр глазных болезней имени Гельмгольца" Министерства здравоохранения Российской Федерации (ФГБУ "НМИЦ ГБ им. Гельмгольца" Минздрава России) | Method for preventing keratotransplant lysis in urgent surgery |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105597145A (en) * | 2016-02-22 | 2016-05-25 | 中山大学中山眼科中心 | Retina cell scaffold biological surgical binder and preparing method thereof |
| EP3435971B1 (en) * | 2016-03-28 | 2024-03-27 | Cetel Scientific LLC | Amniotic or placental preparation and device for ophthalmic use as a dressing to enhance healing |
| CN106214319B (en) * | 2016-08-26 | 2019-03-08 | 张晨明 | Amnioscopic and its preservation method |
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| CN118490870B (en) * | 2024-05-13 | 2025-03-14 | 广州友瀚生物科技有限公司 | A suture-free acellular biological amniotic membrane ocular surface composite material and use method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1652686A (en) * | 2002-03-14 | 2005-08-10 | 组织技术公司 | Amniotic membrane covering for a tissue surface and devices facilitating fastening of membranes |
| CN101088479A (en) * | 2006-08-31 | 2007-12-19 | 中山大学中山眼科中心 | Biological membrane fixing device for eye surface |
| CN101843923A (en) * | 2010-01-22 | 2010-09-29 | 中国海洋大学 | Method for preparing carrier bracket of tissue engineering artificial corneal endothelium by using fresh amniotic membrane |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201193228Y (en) * | 2007-02-13 | 2009-02-11 | 刘青 | Three-dimensional cell-culturing insert, manufacturing equipment thereof and kit |
| US20120071580A1 (en) * | 2008-07-31 | 2012-03-22 | The Board Of Trustees Of The University Of Illinois | Suturable Hybrid Superporous Hydrogel Keratoprosthesis for Cornea |
| CZ302915B6 (en) * | 2010-04-23 | 2012-01-18 | Pegas Nonwovens S.R.O. | Process for producing non-woven fabric with barrier and antistatic finish |
| CN102719391A (en) * | 2012-06-07 | 2012-10-10 | 江阴瑞康健生物医学科技有限公司 | Diphasic porous three-dimensional cell culture scaffold |
-
2014
- 2014-06-10 CN CN201410256997.7A patent/CN103989553B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1652686A (en) * | 2002-03-14 | 2005-08-10 | 组织技术公司 | Amniotic membrane covering for a tissue surface and devices facilitating fastening of membranes |
| CN101088479A (en) * | 2006-08-31 | 2007-12-19 | 中山大学中山眼科中心 | Biological membrane fixing device for eye surface |
| CN101843923A (en) * | 2010-01-22 | 2010-09-29 | 中国海洋大学 | Method for preparing carrier bracket of tissue engineering artificial corneal endothelium by using fresh amniotic membrane |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2822412C1 (en) * | 2023-10-19 | 2024-07-04 | Федеральное государственное бюджетное учреждение "Национальный медицинский исследовательский центр глазных болезней имени Гельмгольца" Министерства здравоохранения Российской Федерации (ФГБУ "НМИЦ ГБ им. Гельмгольца" Минздрава России) | Method for preventing keratotransplant lysis in urgent surgery |
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