CN113018508A - Surface-modified artificial lens and preparation method thereof - Google Patents
Surface-modified artificial lens and preparation method thereof Download PDFInfo
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- CN113018508A CN113018508A CN202110273832.0A CN202110273832A CN113018508A CN 113018508 A CN113018508 A CN 113018508A CN 202110273832 A CN202110273832 A CN 202110273832A CN 113018508 A CN113018508 A CN 113018508A
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- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/14—Eye parts, e.g. lenses, corneal implants; Implanting instruments specially adapted therefor; Artificial eyes
- A61F2/16—Intraocular lenses
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- 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
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- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
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Abstract
The application discloses a surface-modified intraocular lens and a preparation method thereof, and relates to the technical field of medical implant materials. The surface modified artificial lens is prepared by loading a TGF-beta 2 receptor aptamer coated by lactic acid chitosan nano-particles on the surface of the artificial lens. The preparation method comprises the following steps: preparing lactic acid chitosan nano-particles; constructing a TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid; and (3) carrying out surface modification on the artificial lens by using the TGF-beta 2 receptor nucleic acid aptamer coated by the lactic acid chitosan nano-plasmid. The present application provides surface-modified intraocular lenses that solve the problems of low loading of TGF- β 2 antibodies directly loaded onto the surface of an intraocular lens, inability to maintain effective therapeutic concentrations for extended periods of time, and difficulty in maintaining the bioactivity of TGF- β 2 antibodies.
Description
Technical Field
The application relates to the technical field of medical implant materials, in particular to a surface-modified intraocular lens and a preparation method thereof.
Background
Posterior Capsular Opacification (PCO) of the crystalline lens is a main complication affecting the long-term vision recovery of patients after cataract surgery, and generally the opacified posterior capsular sac is cut by adopting laser of the anterior segment of the eye to treat. However, recent studies show that lenticular capsular opacification is also a wound repair reaction spontaneously generated by the body of a patient, a plurality of growth factors, cytokines and inflammatory mediators participate in the process, and epithelial cells (LECs) of residual lenses mainly regulate hyperplasia, migration and epithelial mesenchymal transdifferentiation of the residual lenses through autocrine generated growth factors (such as TGF-beta 2), and play a central role in the generation and development of the lenticular capsular opacification. Meanwhile, in vitro cytology experiments prove that the TGF-beta 2 factor is the key of the induced epithelial cell transdifferentiation effect, and the TGF-beta 2 factor can activate a downstream channel after being combined with a receptor thereof so as to cause the occurrence of lenticular posterior capsular opacification, so that the TGF-beta 2 factor can be neutralized by a TGF-beta 2 receptor aptamer to inhibit the occurrence of the lenticular posterior capsular opacification.
Currently, intraocular lenses (IOLs) are based on the properties of being intraocular lenses as capsular bag implants and capable of being applied to the delivery of drugs or biologically active molecules, thereby inhibiting the development of posterior capsular opacification of the lens by loading the surface of the IOL with a TGF-beta 2 receptor aptamer. The TGF-beta 2 receptor aptamer loaded on the surface of the artificial lens not only can directly act on TGF-beta 2 factors generated by adjacent epithelial cells, but also can form and maintain a microenvironment with effective treatment concentration in the capsular bag, and is not easy to generate toxic injury to normal tissues in eyes of a patient, so that the problems that the conventional eye administration mode (such as eye drop, anterior chamber perfusion during operation or capsular bag injection) is difficult to continuously maintain the effective treatment concentration in the capsular bag, and the side effects of easily generating toxic injury to the normal tissues in the eyes of the patient after the medicine concentration is increased and the like can be effectively avoided. In addition, the loading of the TGF-beta 2 receptor aptamer on the surface of the artificial lens is convenient and easy without adding redundant treatment steps.
However, TGF- β 2 antibody is loaded on the surface of the intraocular lens at present, and is mainly directly attached to the surface of the intraocular lens in a physical adsorption manner, and since the TGF- β 2 antibody and the intraocular lens are non-specifically adsorbed, the loading amount of the TGF- β 2 antibody on the surface of the intraocular lens is low, and the TGF- β 2 antibody is suddenly released, so that the effective treatment concentration cannot be maintained for a long time, and the TGF- β 2 antibody may be directly hydrolyzed by protease, nuclease and the like in vivo to be ineffective, and thus the bioactivity of the TGF- β 2 antibody is difficult to maintain.
Disclosure of Invention
The embodiment of the application solves the technical problems that the loading capacity of TGF-beta 2 antibodies is small, effective treatment concentration cannot be maintained for a long time and the bioactivity of the TGF-beta 2 antibodies is difficult to maintain due to the fact that the TGF-beta 2 antibodies are directly loaded on the surface of the intraocular lens.
In order to achieve the technical purpose, the embodiment of the application adopts the following technical scheme:
in a first aspect, embodiments of the present application provide a surface-modified intraocular lens having a surface loaded with a lactated chitosan nanoparticle coated TGF- β 2 receptor aptamer.
In an alternative of the embodiments of the present application in combination with the first aspect, the intraocular lens is pre-treated to have a surface carrying a negative charge.
In an alternative to the first aspect, in an embodiment of the present application, the intraocular lens is a hydrophobic acrylate.
In an alternative of this embodiment of the application, in combination with the first aspect, the lactosylated chitosan nanoparticles constitute a globular encapsulation of the TGF- β 2 receptor aptamer.
In a second aspect, embodiments of the present application also provide a method for preparing a surface modified intraocular lens, comprising:
preparing lactic acid chitosan nano-particles;
constructing a TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid; and
and (3) carrying out surface modification on the artificial lens by using the TGF-beta 2 receptor nucleic acid aptamer coated by the lactic acid chitosan nano-plasmid.
In combination with the second aspect, in an alternative of embodiments of the present application, the preparing the lactosylated chitosan nanoparticles includes:
dissolving the lactylated chitosan to obtain a lactylated chitosan solution;
dissolving sodium tripolyphosphate to obtain a sodium tripolyphosphate solution;
dropwise adding the sodium tripolyphosphate solution into the lactic acid chitosan solution, and continuously stirring for several hours at room temperature; and
and (3) carrying out centrifugal separation on the stirred mixed solution, and cleaning the separated product to obtain the lactic acid chitosan nano-particles.
In an alternative of the embodiments of the present application, the constructing a lactated chitosan nanoparticle coated TGF- β 2 receptor aptamer comprises:
dissolving the lactic acid chitosan nano-particles to obtain a lactic acid chitosan nano-particle solution;
preparing a sodium tripolyphosphate/pDNA mixed solution;
dropwise adding the sodium tripolyphosphate/pDNA mixed solution into the lactic acid chitosan nanoparticle solution, and continuously stirring for several hours at room temperature; and
and (3) centrifugally separating the stirred mixed solution, and cleaning the separated product to obtain the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid.
In an alternative of the embodiments of the present application, the surface modification of the intraocular lens with the TGF- β 2 receptor aptamer coated with a lactonized chitosan nanoparticle includes:
carrying out ultrasonic cleaning and soaking in absolute ethyl alcohol on the artificial lens, then placing the artificial lens in a surfactant solution for soaking activation and vacuum drying to obtain an activated artificial lens;
carrying out low-temperature plasma double-sided injection treatment on the activated intraocular lens so as to enable the intraocular lens to carry negative charges;
carrying out matched impregnation on the artificial lens subjected to low-temperature plasma double-side injection treatment and the TGF-beta 2 receptor aptamer suspension coated by the lactylated chitosan nano-particles to obtain a crude artificial lens with a modified surface;
and drying the crude product of the surface-modified intraocular lens under the nitrogen atmosphere, and packaging to obtain the surface-modified intraocular lens.
In an alternative of the embodiments of the present application, in combination with the second aspect, the performing a low temperature plasma double-sided implantation process on the activated intraocular lens comprises:
carrying out plasma double-sided implantation treatment on the activated intraocular lens by adopting a low-temperature ion generator under the condition that the treatment energy is 70 keV;
the ion dose of the double-sided plasma implantation is 5 multiplied by 1013ions/cm2-5×1015ions/cm2。
In a third aspect, embodiments of the present application further provide applications of the surface modified intraocular lens in cataract treatment based on the above studies.
Compared with the prior art, the beneficial effects or advantages of the embodiment of the application include:
according to the surface-modified intraocular lens provided by the embodiment of the application, by loading the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid on the surface of the intraocular lens, the problems that the loading amount is small when the TGF-beta 2 antibody is directly loaded on the surface of the intraocular lens, the effective treatment concentration cannot be maintained for a long time, and the bioactivity of the TGF-beta 2 antibody is difficult to maintain are solved. Specifically, the present embodiment uses the chitosan lactate nanoparticle as a slow release carrier of the TGF-beta 2 receptor aptamer, on one hand, because the chitosan lactate nanoparticle can encapsulate the TGF-beta 2 receptor aptamer therein through the bonding effect of chemical bonds, thereby leading the TGF-beta 2 aptamer to accurately target the TGF-beta 2 receptor so as to block the TGF-beta 2 effect inside and outside epithelial cells, effectively prevent TGF-beta 2 aptamer from being hydrolyzed by protease, nuclease and the like in vivo before being combined with TGF-beta 2 receptor on the surface of epithelial cells, realize the lasting and stable maintenance of the bioactivity of the TGF-beta 2 receptor aptamer, and TGF-beta 2 aptamer is encapsulated by the lactic acid chitosan nano-plasmid to delay the release, thereby effectively preventing the side effects of toxic damage and the like of the normal tissues in the eyes of the patient caused by overhigh medicine concentration; on the other hand, the surface of the artificial lens after low-temperature plasma injection treatment carries negative charges, and the lactic acid chitosan nano-particles carrying the positive charges can be tightly crosslinked with the artificial lens carrying the negative charges through charge adsorption, so that a stable slow release coating of the lactic acid chitosan nano-particles coated with the TGF-beta 2 receptor aptamer is formed on the surface of the artificial lens, and the loading capacity of the TGF-beta 2 aptamer on the surface of the artificial lens is increased. In addition, the surface-modified intraocular lens provided by the application has high adhesion with the lens capsular bag, so that the space for epithelial cell migration and proliferation can be eliminated, and the sustained-release coating of the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nanoparticle can stably, durably, targetedly and low-toxicity release the TGF-beta 2 receptor aptamer for inhibiting epithelial cell proliferation, adhesion and transformation, so that the generation of capsular opacification behind the lens can be safely, conveniently and effectively inhibited.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings used in the description of the embodiments will be briefly introduced below. It is obvious that the drawings in the following description are only some of the embodiments described in the present application, and that other drawings can be derived from these drawings by a person skilled in the art without inventive effort.
FIG. 1 is a flow chart of a process for preparing a surface modified intraocular lens provided in accordance with an embodiment of the present application;
FIG. 2 is a flow chart of a process for preparing a lactonized chitosan nanoparticle provided in an example of the present application;
FIG. 3 is a process flow diagram for constructing a lactated chitosan nanoparticle coated TGF-beta 2 receptor aptamer provided in the embodiments of the present application;
FIG. 4 is a process flow diagram for surface modification of an intraocular lens using a lactonized chitosan nanoparticle coated TGF- β 2 receptor aptamer provided in an embodiment of the present application;
fig. 5 is an electron microscope morphology characterization diagram of the lactonized chitosan nanoparticles provided in the examples of the present application.
Detailed Description
In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. It should be apparent that the described embodiments are only some of the embodiments of the present application, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
In order to solve the technical problems that the loading capacity of loading TGF-beta 2 antibody on the surface of an artificial lens is small, effective treatment concentration cannot be maintained for a long time, and the bioactivity of the TGF-beta 2 antibody is difficult to maintain, the embodiment of the application provides the artificial lens with a modified surface and a preparation method thereof.
This example provides a surface-modified intraocular lens having a surface carrying a TGF- β 2 receptor aptamer coated with a lactonized chitosan nanoparticle. In this embodiment, the TGF- β 2 receptor aptamer is cross-linked and coated by the lactic acid chitosan nanoparticle and then loaded on the surface of the artificial lens, so that the lactic acid chitosan nanoparticle can be used as a slow release carrier of the TGF- β 2 receptor aptamer, and the release amount/release speed of the TGF- β 2 receptor aptamer in the capsular bag is regulated, so as to prevent the toxic damage and other side effects of the overhigh drug concentration on the normal tissues in the eyes of the patient; the lactic acid chitosan nano-plasmid can form bonding encapsulation on the TGF-beta 2 receptor aptamer, so that the TGF-beta 2 aptamer can accurately target the TGF-beta 2 receptor to block TGF-beta 2 effect inside and outside epithelial cells, and simultaneously effectively prevent the TGF-beta 2 aptamer from being hydrolyzed by protease, nuclease and the like in vivo before being combined with the TGF-beta 2 receptor on the surface of the epithelial cells, so that the biological activity of the TGF-beta 2 receptor aptamer can be durably and stably maintained; the lactic acid chitosan nano-plasmid can be used as an intermediate medium (connecting carrier) between the TGF-beta 2 receptor aptamer and the artificial lens, so that the TGF-beta 2 receptor aptamer and the artificial lens can be tightly crosslinked through charge adsorption, on one hand, a stable lactic acid chitosan coated TGF-beta 2 receptor aptamer load coating can be formed on the surface of the artificial lens, and on the other hand, the load of the TGF-beta 2 aptamer on the surface of the artificial lens can be increased. In addition, the TGF-beta 2 receptor aptamer coated by the lactylated chitosan is tightly crosslinked with the intraocular lens through the charge adsorption effect, so that the surface-modified intraocular lens has high adhesive force with the lens capsular bag, the space for epithelial cell migration and proliferation can be eliminated, and the generation of capsular opacification after lens can be safely, conveniently and effectively inhibited.
It should be noted that in this embodiment, by using the lactylated chitosan as the drug delivery system of the TGF- β 2 receptor aptamer, not only can a targeting system of the lactylated chitosan coated TGF- β 2 receptor aptamer be quickly constructed, but also the constructed drug delivery system of the lactylated chitosan coated TGF- β 2 receptor aptamer has the characteristics of low cost, safety, no toxicity, and protection of the TGF- β 2 receptor aptamer from degradation. The lactic acid chitosan serving as a non-viral gene vector has good biocompatibility, biodegradability, oxidation resistance and anti-inflammatory properties, can promote tissue regeneration, enhance drug and DNA (deoxyribonucleic acid) transfer and enhance calcium absorption, so that a drug loading system of the TGF-beta 2 receptor aptamer is constructed by taking the lactic acid chitosan nano-particles as an intermediate medium, and the lactic acid chitosan has the advantages of high safety, low cost and capability of protecting the TGF-beta 2 receptor aptamer from being directly degraded; meanwhile, the lactic acid chitosan nano-particles can form chemical bond bonding effect with a TGF-beta 2 receptor aptamer to be tightly cross-linked, and a targeting system capable of combining a TGF-beta 2 receptor in an epithelial cell is easier to construct, so that the generation of lenticular posterior capsule opacification is stably inhibited in a low-toxicity and long-acting targeting manner. In addition, the preparation of the lactylated chitosan lactic acid nano plasmid is simple and convenient, the cost is low, and based on the excellent characteristics of the lactylated chitosan such as oxidation resistance, anti-inflammation, tissue regeneration promotion, drug and DNA transmission enhancement, calcium absorption enhancement and the like, a drug loading system is constructed by combining the lactylated chitosan with a TGF-beta 2 receptor aptamer, so that the drug loading system not only can generate a synergistic promotion effect in the treatment of the lenticular posterior capsular opacification and improve the treatment effect, but also can greatly reduce the occurrence of other complications after cataract surgery, and has remarkable superiority in the treatment of the lenticular posterior capsular opacification.
In this example, the surface of the intraocular lens after pretreatment carries negative charges to enable the intraocular lens and the lactating chitosan coated TGF- β 2 receptor aptamer to be tightly cross-linked by charge adsorption. Specifically, the lactic acid chitosan nanoparticles carry positive charges and are applied as an intermediate medium between the intraocular lens and a targeting system, so that the intraocular lens is pretreated to enable the surface of the intraocular lens to carry negative charges, the intraocular lens and the lactic acid chitosan nanoparticles can be tightly crosslinked through charge adsorption, and the adhesion force between the intraocular lens and a drug loading system is enhanced.
In this example, the intraocular lens is selected to be a hydrophobic acrylate posterior chamber intraocular lens to enhance adhesion of the drug loaded system intraocular lens to the capsular bag. Particularly, the hydrophobic acrylate posterior chamber type artificial lens has stronger adhesive force after meeting liquid, can sink immediately after being implanted into the eye of a patient and is closely adsorbed with the capsular bag, and has better stability and neutrality in the capsular bag, thereby effectively reducing the occurrence of capsular opacification behind the crystalline lens. In addition, the hydrophobic acrylate posterior chamber type artificial lens can bear larger laser energy, and the persistence of the surface modified artificial lens is improved.
It should be noted that the intraocular lens of the present embodiment is not limited to the hydrophobic acrylate intraocular lens, and may be an intraocular lens of other materials, such as PMMA intraocular lens, silicone gel intraocular lens, and the like. It is to be understood that hydrophobic acrylate intraocular lenses have a superior advantage over PMMA intraocular lenses, silicone gel intraocular lenses, and the like, and thus hydrophobic acrylate intraocular lenses are preferred in this embodiment.
In the embodiment, the lactylated chitosan nanoparticles form spherical encapsulation on the TGF-beta 2 receptor aptamer, so that the TGF-beta 2 receptor aptamer can be protected from being degraded before being combined with the TGF-beta 2 receptor on the surface of lens epithelial cells, and the release of the TGF-beta 2 receptor aptamer can be delayed through the slow degradation effect of the lactylated chitosan, so that a stable, continuous and effective drug treatment process is obtained.
The present embodiment also provides a method for preparing a surface modified intraocular lens, as shown in fig. 1, comprising steps S101-S103.
S101: preparing the lactic acid chitosan nano-particles.
S102: constructing TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-plasmid.
S103: and (3) carrying out surface modification on the artificial lens by using the TGF-beta 2 receptor nucleic acid aptamer coated by the lactic acid chitosan nano-plasmid.
According to the preparation method of the surface-modified intraocular lens provided by the embodiment of the application, after the lactic acid chitosan and lactic acid nano-plasmid is prepared, the TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-plasmid is constructed, and then the intraocular lens is directly subjected to surface modification through the characteristic that the lactic acid chitosan nano-plasmid carries positive charges, so that the intraocular lens with the surface loaded with the lactic acid chitosan nano-plasmid coated TGF-beta 2 receptor aptamer is obtained. In view of this, the preparation method has the advantages of simplicity, convenience and low cost, and the prepared artificial lens product has the characteristics of stability, long-acting effect, low toxicity and targeting, and inevitably generates good economic and social benefits in the treatment of the posterior capsular opacification of the crystalline lens.
In this example, the preparation of lactonized chitosan nanoparticles is shown in FIG. 2, and specifically includes steps S201-S204.
S201: dissolving the lactylated chitosan to obtain a lactylated chitosan solution.
S202: and dissolving the sodium tripolyphosphate to obtain a sodium tripolyphosphate solution.
S203: and dropwise adding the sodium tripolyphosphate solution into the lactic acid chitosan solution, and continuously stirring for several hours at room temperature.
S204: and (3) carrying out centrifugal separation on the stirred mixed solution, and cleaning the separated product to obtain the lactic acid chitosan nano-particles.
It should be noted that, the lactic acid chitosan/sodium tripolyphosphate is dissolved by the double distilled water, so that impurity ions are effectively prevented from being mixed into the lactic acid chitosan nano-particles, and the impurity ions are prevented from interfering and/or influencing the charge quantity of the lactic acid chitosan nano-particles.
In the embodiment, the lactic acid chitosan nano-particles are prepared by an ion gel method, and specifically, the lactic acid chitosan nano-particles are prepared by performing ion-induced gelation on lactic acid chitosan through sodium tripolyphosphate without toxic and side effects. Intermolecular and intramolecular bonding can be formed between the phosphate group of the sodium tripolyphosphate and the amino group of the lactic acid chitosan, and finally the spherical sodium tripolyphosphate coupled lactic acid chitosan nano plasmid is formed. The preparation method of the lactic acid chitosan nano-particles by the ion gel method has the characteristics of simplicity, rapidness and low cost, and the reaction conditions are mild, so that the lactic acid chitosan nano-particles with firmness, good stability and uniform particle size can be obtained without using an organic solvent. In addition, the drug delivery system of the TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-particles prepared by the ionic gel method is easier to construct. Specifically, the formation of the lactylated chitosan drug-loading system mainly depends on the bonding effect of chemical bonds, wherein the amino group of the lactylated chitosan can be bonded with the phosphate group of sodium tripolyphosphate under a proper condition and wraps the TGF-beta 2 receptor aptamer therein to form the drug-loading system, and the drug-loading system has the advantages of good stability and high encapsulation efficiency.
In this example, the construction of a lactonized chitosan nanoparticle-coated TGF- β 2 receptor aptamer by an ionic gel method specifically includes steps S301 to S304 as shown in fig. 3.
S301: and dissolving the lactic acid chitosan nano-particles to obtain a lactic acid chitosan nano-particle solution.
S302: and preparing a sodium tripolyphosphate/pDNA mixed solution.
S303: and dropwise adding the sodium tripolyphosphate/pDNA mixed solution into the lactic acid chitosan nanoparticle solution, and continuously stirring for a plurality of hours at room temperature.
S304: and (3) centrifugally separating the stirred mixed solution, and cleaning the separated product to obtain the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid.
In the embodiment, a drug loading system of the lactic acid chitosan nano-plasmid coated TGF-beta 2 receptor aptamer is constructed by an ion gel method, so that the drug loading system with good stability and high encapsulation efficiency is obtained. Specifically, the drug-loading system of the lactylated chitosan is formed mainly by the bonding effect of chemical bonds, wherein the amino group of the lactylated chitosan can be bonded with the phosphate group of sodium tripolyphosphate under appropriate conditions and wraps a TGF-beta 2 acceptor aptamer therein to form the drug-loading system.
In this example, the surface modification of the intraocular lens using TGF- β 2 receptor aptamer coated with lactonized chitosan nanoparticles, as shown in fig. 4, specifically includes steps S401 to S404:
s401: and (3) carrying out ultrasonic cleaning on the artificial lens, soaking the artificial lens in absolute ethyl alcohol, then placing the artificial lens in a surfactant solution for soaking activation and vacuum drying to obtain the activated artificial lens.
S402: the activated intraocular lens is subjected to a low temperature plasma double-sided implantation process to cause the intraocular lens to carry negative charges.
S403: and (3) carrying out matched impregnation on the artificial lens subjected to low-temperature plasma double-side injection treatment and the TGF-beta 2 receptor aptamer suspension coated by the lactic acid chitosan nano-particles to obtain a crude product of the artificial lens with the surface modified.
S404: and drying the crude product of the surface-modified intraocular lens under the nitrogen atmosphere, and packaging to obtain the surface-modified intraocular lens.
According to the embodiment, firstly, the artificial lens with the surface cleaned and activated is obtained by carrying out ultrasonic cleaning, absolute ethyl alcohol soaking and surface activator solution soaking treatment on the artificial lens, so that on one hand, interference of other impurity ions on subsequent plasma double-sided injection is avoided, on the other hand, due to the fact that the surface of the artificial lens is activated, the plasma double-sided injection is facilitated, and the plasma double-sided injection process is accelerated; and then the artificial lens carries negative charges through low-temperature plasma double-sided injection treatment, and the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid carries positive charges, so that the artificial lens and the TGF-beta 2 receptor aptamer can be tightly crosslinked together through charge adsorption after carrying the negative charges, and the adhesion between the artificial lens and a medicine carrying system is improved. Meanwhile, the surface of the TGF-beta 2 receptor aptamer is coated with the lactic acid chitosan nanoparticle coating, and the acidified chitosan nanoparticle is of a spherical structure, so that the specific surface area of the drug carrying system is increased, and the artificial lens can be loaded with more drug carrying systems when being combined with the drug carrying system, namely the loading capacity of the TGF-beta 2 receptor aptamer coated with the lactic acid chitosan nanoparticle on the surface of the artificial lens is increased. In addition, the preparation method has the characteristics of simplicity and rapidness, so that the preparation speed is increased and the cost is reduced. It is understood that drug delivery system refers to a lactonized chitosan nanoparticle coated TGF- β 2 receptor aptamer.
In this embodiment, the low-temperature plasma double-side implantation treatment of the activated intraocular lens specifically includes:
carrying out plasma double-sided implantation treatment on the activated intraocular lens by adopting a low-temperature ion generator under the condition that the treatment energy is 70 keV;
the ion dose of the double-sided plasma implantation is 5 multiplied by 1013ions/cm2-5×1015ions/cm2。
The injected ion dosage can ensure that the injected anions in the same plane keep uniform, so that when the injected anions react with the TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-particles, the TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-particles can be uniformly loaded on the surface of the artificial lens in a large area.
The present embodiment is based on the above research and also provides the use of the surface modified intraocular lens in the treatment of cataract. Specifically, the surface modified artificial lens has higher adhesive force with the lens capsular bag, and can eliminate the space for epithelial cell migration and proliferation; meanwhile, the slow release coating of the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid can stably, durably, targetedly and low-toxicity release the TGF-beta 2 receptor aptamer for inhibiting proliferation, adhesion and transformation of epithelial cells, thereby realizing safe, convenient and effective inhibition of capsular opacification after crystalline lens and being widely applied to cataract treatment.
The technical solution of the present application is further described below with reference to specific examples.
A method of making a surface modified intraocular lens comprising the steps of:
the preparation method of the lactic acid chitosan nano-particles comprises the following specific steps:
dissolving the lactylated chitosan: dissolving 25mg of lactylated chitosan in 10mL of double distilled water, and stirring at room temperature by using a magnetic stirrer to fully dissolve the lactylated chitosan to obtain a lactylated chitosan solution, wherein the rotating speed of the magnetic stirrer is 350rpm/min, and the stirring time is 30 min;
dissolving sodium tripolyphosphate: dissolving 10mg of sodium tripolyphosphate in 10mL of double distilled water, and stirring by a magnetic stirrer at room temperature to fully dissolve the sodium tripolyphosphate to obtain a sodium tripolyphosphate solution, wherein the rotating speed of the magnetic stirrer is 350rpm/min, and the stirring time is 30 min;
and (3) dropwise adding a sodium tripolyphosphate solution into the lactic acid chitosan solution: according to the volume ratio of the sodium tripolyphosphate solution to the lactic acid chitosan solution of 3:1, under the magnetic stirring at the rotating speed of 350rpm/min, absorbing 333uL of the sodium tripolyphosphate solution through a liquid transfer gun, dropwise adding the solution into the lactic acid chitosan solution within 5min, and continuously stirring at room temperature overnight;
centrifuging the mixed solution after stirring, and washing the separated product: and on the second day, transferring the mixed solution stirred overnight into an ultrahigh-speed centrifuge, centrifuging for 25min at the rotating speed of 9500r/min, and cleaning the centrifugal product with double distilled water for 3 times in total to obtain the lactylated chitosan nanoparticles.
Constructing a TGF-beta 2 receptor aptamer coated by the lactic acid chitosan nano-plasmid, and the specific process comprises the following steps:
dissolving the lactic acid chitosan nano-particles: dissolving 25mg of lactonized chitosan nanoparticles prepared in S10 in 10mL of double distilled water, and stirring at room temperature by using a magnetic stirrer to fully dissolve the lactonized chitosan nanoparticles to obtain a solution of the lactonized chitosan nanoparticles, wherein the rotating speed of the magnetic stirrer is 350rpm/min, and the stirring time is 30 min;
preparing a sodium tripolyphosphate/pDNA mixed solution; dissolving 10mg of sodium tripolyphosphate and 1mg of pDNA in 10mL of double distilled water, stirring by a magnetic stirrer at room temperature to fully dissolve the sodium tripolyphosphate and the pDNA to obtain a sodium tripolyphosphate/pDNA solution, wherein the rotating speed of the magnetic stirrer is 350rpm/min, and the stirring time is 30 min;
and (2) dropwise adding a sodium tripolyphosphate/pDNA mixed solution into the lactic acid chitosan nano-plasmid solution: according to the volume ratio of the sodium tripolyphosphate/pDNA solution to the lactic acid chitosan nanoparticle solution of 3:1, under the magnetic stirring at the rotating speed of 350rpm/min, 333uL of the sodium tripolyphosphate/pDNA solution is sucked by a liquid-transferring gun, is added into the lactic acid chitosan nanoparticle solution dropwise within 5min, and is continuously stirred at room temperature overnight;
centrifuging the mixed solution after stirring, and washing the separated product: and on the second day, transferring the mixed solution stirred overnight into an ultra-high speed centrifuge, centrifuging for 25min at the rotating speed of 9500r/min, and cleaning the centrifugal product with double distilled water for 3 times in total to obtain the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nanoparticles.
In this example, the morphology and characteristics of the TGF-beta 2 receptor aptamer coated by the chitosan lactate nanoparticles are detected, as shown in FIG. 5.
Before the morphology and characteristics of the TGF- β 2 receptor aptamer coated on the chitosan lactate nanoparticles are detected, the following treatments are performed in the example:
the preparation method comprises the steps of coating a TGF-beta 2 receptor aptamer solution on a lactonized chitosan nano-particle, freeze-drying, diluting by 10 times, measuring the particle size, surface Zeta Potential and Dispersity (PDI) of the nano-particle by a laser particle size analyzer, and sucking 1uL of the solution to be dripped to a copper sheet for electron microscope morphology characterization, wherein the result is shown in figure 5.
The surface modification of the artificial lens is carried out by utilizing a TGF-beta 2 receptor nucleic acid aptamer coated by the lactic acid chitosan nano-plasmid, and the specific process comprises the following steps:
and (3) carrying out ultrasonic cleaning on the artificial lens, soaking the artificial lens in absolute ethyl alcohol, then placing the artificial lens in a surfactant solution for soaking activation and vacuum drying to obtain the activated artificial lens. Specifically, firstly, in the environment of a purification workbench, deionized water is used as a cleaning medium, and the artificial lens is repeatedly cleaned for 3 times by an ultrasonic cleaning machine; secondly, soaking the cleaned artificial lens in analytically pure absolute ethyl alcohol for 24 hours, taking out and drying; finally, soaking the dried artificial lens in a surfactant solution for 24 hours, taking out the artificial lens, drying the artificial lens in a vacuum drier, and storing the artificial lens for later use;
the activated intraocular lens is subjected to a low temperature plasma double-sided implantation process to cause the intraocular lens to carry negative charges. Specifically, the activated intraocular lens was subjected to plasma double-sided implantation treatment by a low-temperature ionizer under the condition that the ionizer treatment energy was 70keV, and the implanted ion dose was 5X 1013ions/cm2-5×1015ions/cm2So that the surface of the artificial lens uniformly carries negative charges;
and (3) carrying out matched impregnation on the artificial lens subjected to low-temperature plasma double-side injection treatment and the TGF-beta 2 receptor aptamer suspension coated by the lactic acid chitosan nano-particles for 48 hours to obtain the artificial lens with the surface modified.
And carrying out ultrasonic cleaning on the surface-modified intraocular lens, drying and packaging to obtain the surface-modified intraocular lens.
In this example, the surface-modified intraocular lens was prepared using a hydrophobic acrylate posterior chamber intraocular lens as a manufacturing platform. The surface modified artificial lens is different from the existing layer-by-layer self-assembly preparation method, after the surface of the artificial lens is subjected to ion injection treatment to enable the surface of the artificial lens to carry negative charges, the TGF-beta 2 receptor aptamer encapsulated by the lactic chitosan nano-plasmid is loaded by utilizing the charge adsorption effect, so that the tight crosslinking between a medicine carrying system formed by the TGF-beta 2 receptor aptamer encapsulated by the lactic chitosan nano-plasmid and the artificial lens is realized while the TGF-beta 2 aptamer accurately targets an epithelial cell TGF-beta 2 receptor to block the TGF-beta 2 effect inside and outside the cell, and the probability of sudden release of the TGF-beta 2 receptor aptamer can be reduced in the release process.
The above embodiments are only used to illustrate the technical solutions of the present application, and not to limit the present application; although the present application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art. The technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications or substitutions do not depart from the spirit and scope of the present disclosure.
Claims (10)
1. A surface-modified intraocular lens, characterized in that the surface of the intraocular lens is loaded with a TGF-beta 2 receptor aptamer coated with a lactonized chitosan nanoparticle.
2. The surface modified intraocular lens of claim 1, wherein the intraocular lens is pre-treated to carry a negative charge.
3. The surface modified intraocular lens of claim 2, wherein the intraocular lens comprises a hydrophobic acrylate.
4. The surface-modified intraocular lens of claim 1, wherein the lactosylated chitosan nanoparticles constitute a globular encapsulation of the TGF- β 2 receptor aptamer.
5. A method of making a surface modified intraocular lens comprising:
preparing lactic acid chitosan nano-particles;
constructing a TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid; and
and (3) carrying out surface modification on the artificial lens by using the TGF-beta 2 receptor nucleic acid aptamer coated by the lactic acid chitosan nano-plasmid.
6. The method of manufacturing a surface-modified intraocular lens according to claim 5, wherein the manufacturing of the lactosylated chitosan nanoparticles comprises:
dissolving the lactylated chitosan to obtain a lactylated chitosan solution;
dissolving sodium tripolyphosphate to obtain a sodium tripolyphosphate solution;
dropwise adding the sodium tripolyphosphate solution into the lactic acid chitosan solution, and continuously stirring for several hours at room temperature; and
and (3) carrying out centrifugal separation on the stirred mixed solution, and cleaning the separated product to obtain the lactic acid chitosan nano-particles.
7. The method of manufacturing a surface-modified intraocular lens according to claim 5, wherein the constructing a lactated chitosan nanoparticle-coated TGF- β 2 receptor aptamer comprises:
dissolving the lactic acid chitosan nano-particles to obtain a lactic acid chitosan nano-particle solution;
preparing a sodium tripolyphosphate/pDNA mixed solution;
dropwise adding the sodium tripolyphosphate/pDNA mixed solution into the lactic acid chitosan nanoparticle solution, and continuously stirring for several hours at room temperature; and
and (3) centrifugally separating the stirred mixed solution, and cleaning the separated product to obtain the TGF-beta 2 receptor aptamer coated by the lactylated chitosan nano-plasmid.
8. The method of manufacturing a surface-modified intraocular lens according to claim 5, wherein the surface modification of an intraocular lens with a lactated chitosan nanoparticle coated TGF- β 2 receptor aptamer comprises:
carrying out ultrasonic cleaning and soaking in absolute ethyl alcohol on the artificial lens, then placing the artificial lens in a surfactant solution for soaking activation and vacuum drying to obtain an activated artificial lens;
carrying out low-temperature plasma double-sided injection treatment on the activated intraocular lens so as to enable the intraocular lens to carry negative charges;
carrying out matched impregnation on the artificial lens subjected to low-temperature plasma double-side injection treatment and the TGF-beta 2 receptor aptamer suspension coated by the lactylated chitosan nano-particles to obtain a crude artificial lens with a modified surface;
and drying and packaging the crude product of the surface-modified intraocular lens in a nitrogen atmosphere to obtain the surface-modified intraocular lens.
9. The method of making a surface modified intraocular lens according to claim 7 wherein said subjecting the activated intraocular lens to a low temperature plasma double-sided implantation process comprises:
carrying out plasma double-sided implantation treatment on the activated intraocular lens by adopting a low-temperature ion generator under the condition that the treatment energy is 70 keV;
the ion dose of the double-sided plasma implantation is 5 multiplied by 1013ions/cm2-5×1015ions/cm2。
10. Use of a surface-modified intraocular lens according to any one of claims 1 to 4 in the treatment of cataracts.
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