A new type of drug-loaded 3D printed amnioscope

Abstract

The present invention relates to an ophthalmic therapeutic device. The purpose is to provide a new type of drug-loaded 3D printed amnionoscope, which should have the characteristics of large drug loading, no invasive operation required, and good therapeutic effect. The technical solution is: a new type of drug-loaded 3D printed amnionoscope, characterized in that: the amnionoscope includes an amnion sheet and an upper ring and a lower ring that are clamped and riveted to fix the amnion sheet after being overlapped up and down; the surfaces of the upper ring and the lower ring are respectively prepared with a thin layer of tacrolimus liposomes; the axial projections of the upper ring and the lower ring are both the same elliptical ring, the inner ellipse in the middle part of the elliptical ring is a through hole, and the remaining parts are all arc surfaces suitable for the ocular surface; the amnion sheet is flattened and fixed on the amnionoscope and covers the entire through hole.

Description

Technical Field

[0001] The present invention relates to an ophthalmic therapeutic device, in particular to a novel 3D printed amniotic membrane mirror capable of being loaded with medicine. Background Art

[0002] For eye diseases (such as thermal burns / chemical injuries to the ocular surface, severe dry eyes, etc.), the conventional treatment methods are:

[0003] 1. Drug treatment: glucocorticoids, anti-inflammatory drugs, anti-VEGF drugs, immunosuppressants, tyrosine kinase inhibitors and other drugs are used for treatment. Currently, commonly used drug treatments such as glucocorticoids cannot be used for a long time, and will cause problems such as increased intraocular pressure and cataracts. Anti-vascular endothelial growth factor also has problems such as non-healing of corneal epithelium in the later stage of treatment and high prices. In addition, eye diseases often require frequent use of eye drops and a combination of multiple drugs for treatment, and the treatment effect depends on the patient's compliance.

[0004] 2. Amniotic membrane transplantation: Traditional amniotic membrane transplantation covering surgery requires the use of sutures, which is an invasive operation with high requirements for equipment. It is prone to complications such as intraoperative pain, bleeding, and amniotic membrane slippage, as well as suture reactions, iatrogenic injuries, and other problems.

[0005] 3. Amniotic membrane fixation ring: It can only fix the amniotic membrane and does not have the function of a drug sustained-release carrier. Eye drops need to be used frequently during treatment, which can still lead to problems such as poor patient compliance.

[0006] 4. Surgical treatment: large trauma, high treatment cost, poor patient tolerance and other problems. Summary of the invention

[0007] The purpose of the present invention is to overcome the shortcomings of the above-mentioned background technology and provide a new type of drug-loaded 3D printed amnionoscope, which should have the characteristics of large drug loading capacity, no need for invasive operation, and good therapeutic effect.

[0008] The technical solution provided by the present invention is:

[0009] A novel drug-loaded 3D printed amnionoscope, characterized in that: the amnionoscope comprises an amnion sheet and an upper ring and a lower ring which are clamped and riveted to fix the amnion sheet after being overlapped; a thin layer of tacrolimus liposomes is prepared on the surface of the upper ring and the lower ring respectively;

[0010] The axial projections of the upper ring and the lower ring are both the same elliptical ring, the inner ellipse in the middle of the elliptical ring is a through hole, and the rest of the parts are arc surfaces that fit the ocular surface; the amniotic membrane is flattened and fixed on the amnioscope and covers the entire through hole.

[0011] The lower surface of the upper ring is provided with a plurality of rivets protruding toward the lower ring, and these rivets are arranged along the edge of the through hole; the lower ring is provided with a plurality of rivet holes corresponding to the positions of the rivets, which can be riveted one by one with the rivets; after the rivets are inserted through the amniotic membrane, they are riveted to the corresponding rivet holes.

[0012] The preparation process of the tacrolimus liposome thin layer is as follows:

[0013] 1) After washing and drying, the upper ring and the lower ring are light-cured and then cleaned in PBS solution and 75% ethanol in an ultrasonic water bath, respectively, and the cleaning is repeated several times, and then dried;

[0014] 2) completing the upper ring and the lower ring of step 1), placing them in a sub-atmospheric glow discharge plasma treatment device for treatment, so that negatively charged hydroxyl groups are generated on the surfaces of the upper ring and the lower ring;

[0015] 3) The upper ring and the lower ring of step 2) are immersed in a cationic tacrolimus liposome solution, immersed in ultrapure water, immersed in a negatively charged sodium alginate solution, and immersed in ultrapure water in sequence, and the above liposome-ultrapure water-sodium alginate solution-ultrapure water immersion process is repeated several times; after completion, the rings are taken out and stored in a 4°C refrigerator away from light for later use.

[0016] The preparation process of the tacrolimus liposome thin layer is as follows:

[0017] 1) After washing and drying, the upper ring and the lower ring are subjected to light curing under ultraviolet light, and then ultrasonically cleaned in PBS solution and 75% ethanol for 10 minutes respectively, and the cleaning process is repeated 3 times, and then dried;

[0018] 2) After completing the upper and lower rings of step 1), place them in a sub-atmospheric glow discharge plasma treatment device, set the mode to HI, and the treatment time to 1 min, so that negatively charged hydroxyl groups are generated on the surface of the amnion mirror;

[0019] 3) The upper and lower rings of step 2) were placed in a positively charged 0.3 mg / mL tacrolimus liposome solution for 2 hours, in ultrapure water for 5 minutes (to remove the liposomes loosely adsorbed on the surface of the upper and lower rings), in a negatively charged 1 mg / mL sodium alginate solution for 1 hour, and in ultrapure water for 5 minutes; the above liposome-ultrapure water-sodium alginate solution-ultrapure water soaking process was repeated 10 times; after assembly, the assemblies were taken out and stored in a 4°C refrigerator away from light for later use.

[0020] The beneficial effects of the present invention are:

[0021] 1. The novel drug-loaded 3D printed amniotic mirror provided by the present invention is more non-invasive and convenient than the traditional suture method to fix the amniotic membrane, which saves surgical time and reduces the chance of amniotic membrane slippage. The curvature of the amniotic mirror matches the curvature of the normal human ocular surface. The outer and inner edges of the amniotic mirror are blunt, avoiding abrasion of the ocular surface, and can effectively relieve the patient's pain during and early postoperative period, without damaging the tissue structure of the affected eye, and improving its comfort during and after surgery. When using the device, the ring and the amniotic membrane can follow the movement of the eyeball and are not easy to generate tension. When the amniotic membrane is fixed by the suture method, tension is easily generated at the suture point during frequent eyeball movements such as blinking, causing the suture to break and causing the amniotic membrane to slip. Because there is no intraoperative suture, no bleeding, and no stimulation of the ocular surface by the postoperative suture, the patient's tolerance is improved. During the operation, the use of an amniotic mirror can fix the amniotic membrane more quickly and stably, while also saving operation time.

[0022] 2. The medication of ocular surface diseases often requires frequent medication and the combined use of multiple drugs, and the low patient compliance leads to poor drug treatment effect. The amniotic membrane provided by the present invention has a large surface area and can be used as a drug sustained release reservoir. The 72-hour in vitro sustained release curve shows that the drug release amount can reach 642.22±13.76μg / mL, which significantly increases the drug loading, reduces the number of times the eye drops are used, increases patient compliance, and also significantly increases the effective drug concentration on the ocular surface, increasing the therapeutic effect of the disease.

[0023] 3. Most of the existing solutions currently have only one effect. The present invention can not only be mass-produced, but also the amniotic membrane can be repeatedly replaced according to the patient's condition and disease cycle, and a variety of drugs can be assembled as a drug sustained-release device, killing two birds with one stone. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a main structural schematic diagram of the present invention.

[0025] Figure 2 yes Figure 1 sectional view of .

[0026] Figure 3 It is a schematic diagram of the top structure of the present invention.

[0027] Figure 4 It is a three-dimensional structural schematic diagram of the present invention.

[0028] Figure 5 It is a schematic diagram of the main structure of the upper ring in the present invention.

[0029] Figure 6 It is a schematic diagram of the top structure of the upper ring in the present invention.

[0030] Figure 7 It is a schematic diagram of the structure of the upper ring in the present invention when viewed from above.

[0031] Figure 8 It is a schematic diagram of the main structure of the lower ring in the present invention.

[0032] Fig. 9 It is a schematic diagram of the top view of the lower ring in the present invention.

[0033] Fig.10 It is a 72h in vitro sustained-release curve diagram of the present invention.

[0034] Fig.11 It is an electron microscope photograph of the surface of the amnioscope of the present invention. DETAILED DESCRIPTION

[0035] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0036] The new drug-loaded 3D printed amnioscope shown in the figure includes an amniotic membrane sheet 3, an upper ring 1 and a lower ring 2. The upper ring and the lower ring are overlapped and the amniotic membrane sheet is clamped and riveted. The surfaces of the upper ring and the lower ring are also prepared with a thin layer of tacrolimus liposomes.

[0037] The axial projections of the upper ring and the lower ring are both the same elliptical ring; the inner ellipse in the middle part of the elliptical ring is a through hole, and the rest of the parts are arc surfaces that fit the ocular surface; the amniotic membrane is flattened and fixed on the amnioscope and covers the entire through hole.

[0038] The lower surface of the upper ring is provided with a plurality of rivets protruding toward the lower ring, and these are arranged along the edge of the opening; the lower ring is provided with a plurality of rivet holes corresponding to the positions of the rivets, which can be riveted one by one with the rivets (the figure shows: the upper ring has 8 rivets 1-1, and the lower ring has 8 rivet holes 2-1 at corresponding positions); after the rivets are inserted through the amniotic membrane, they are riveted with the corresponding rivet holes (after riveting, the rivets are flush with the lower surface of the lower ring), so that the two rings can be fixed together, and the amniotic membrane 3 can be clamped between the upper and lower rings (as recommended, small holes corresponding to the rivets are also opened around the amniotic membrane to facilitate the insertion and riveting of the rivets) and worn on the eye surface.

[0039] As a recommendation: the horizontal diameter of the amnioscope is 21mm, the vertical diameter is 20mm, and the height is about 4.2mm; the thickness of the upper and lower rings is about 0.5mm, the horizontal diameter of the central elliptical through hole is about 14.7mm, and the vertical diameter is about 12.8mm; the diameter of the rivet hole on the lower ring is about 1.0mm, the height of the rivet on the upper ring is 0.7mm, and the diameter of the rivet is about 1.1mm (the rivet and the rivet hole are tight fit). The shape and curvature of the ring conform to the shape of the normal human eye surface and will not cause pain when worn.

[0040] As a further recommendation, both the upper and lower rings can be obtained by 3D printing, so that the upper and lower ring shapes that match the ocular surface shapes of different patients can be obtained. The material of the upper and lower rings is preferably a commercial light-curable resin surgicalguide (formlabs, Somerville, MA, USA).

[0041] The preparation process of the tacrolimus liposome thin layer is as follows:

[0042] 1) After washing and drying, the upper ring and the lower ring are subjected to light curing under ultraviolet light, and then ultrasonically cleaned in PBS solution and 75% ethanol for 10 minutes respectively, and the cleaning process is repeated 3 times, and then dried;

[0043] 2) The upper ring and the lower ring of step 1) are placed in the central reaction zone of quartz glass in a sub-atmospheric glow discharge plasma treatment device for treatment, the mode is set to HI, and the treatment time is 1 min, so that negatively charged hydroxyl groups are generated on the surfaces of the upper ring and the lower ring;

[0044] 3) After the upper and lower rings of step 2) are completed, they are placed in a positively charged cationic tacrolimus liposome solution (the tacrolimus content is 0.3 mg / mL, and the solvent is 1X PBS solution) and soaked for 2 hours, soaked in ultrapure water for 5 minutes (to remove the liposomes loosely adsorbed on the surface of the upper and lower rings), soaked in a negatively charged 1 mg / mL sodium alginate solution (the solvent is 1X PBS solution) for 1 hour, and soaked in ultrapure water for 5 minutes; repeat the above liposome-ultrapure water-sodium alginate solution-ultrapure water soaking process 10 times; after the assembly is completed, take out and store in a 4°C refrigerator away from light for later use. The concentration and soaking time of the above tacrolimus liposome solution and the concentration and soaking time of the sodium alginate solution can be selected and determined as needed.

[0045] Fig.11 The surface shapes of the upper ring and the lower ring after the above treatment are shown; wherein, Fig.11 Figure A in the figure shows the surface of the upper and lower rings after self-assembly at a lower magnification. The rough granular substances are the assembled tacrolimus liposomes and sodium alginate. Fig.11 Figure B shows the surface of the upper and lower rings after self-assembly at a higher magnification, and the round granular substances are tacrolimus liposomes.

[0046] The principle of the present invention is: plasma glow discharge technology can generate active substances to attack the surface of the 3D printed amniotic ring, add negatively charged hydroxyl groups to the electrically neutral surface of the amniotic ring, and then use electrostatic layer-by-layer self-assembly technology to assemble multiple tacrolimus liposomes and sodium alginate bilayers on the surface of the upper ring and the lower ring. The type of drug and the number of assembled layers can be adjusted according to the needs of the disease, and the larger surface area of ​​the amniotic ring (upper ring and lower ring) is used as a reservoir for drug release to achieve the purpose of sustained drug release.

[0047] The cationic tacrolimus liposomes detached from the amniotic membrane can combine with the negatively charged mucin layer on the ocular surface, significantly increasing the drug's ocular surface residence time and corneal permeability. The assembled drug tacrolimus is a commonly used macrolide immunosuppressant that can be used to treat severe dry eyes, thermal burns of the eye, and other diseases. Tacrolimus can reduce the expression of inflammatory factors IL-1, IL-6, MMP-2, and MMP-9 by binding to the FKBP12 protein in the cell, and can also inhibit the production of VEGF through the PI3K-AKT and p38 pathways, thereby achieving the purpose of treating ocular surface diseases. At the same time, the amniotic membrane clamped between the amniotic membrane rings can promote the differentiation, proliferation, and migration of adjacent conjunctival and corneal epithelial cells, which is beneficial to ocular surface reconstruction.

[0048] The amniotic membrane provided by the present invention has a large surface area and can be used as a drug sustained-release reservoir. The 72-hour in vitro sustained-release curve shows that the tacrolimus drug release amount can reach 642.22±13.76 μg / mL (see Fig.10 ), significantly increased the drug loading capacity, reduced the number of times eye drops were used, and increased patient compliance. It also significantly increased the effective drug concentration on the ocular surface and increased the therapeutic effect of the disease.

Claims

1. A new type of drug-loaded 3D printed amnioscope, characterized by: The amnionoscope comprises an amnion sheet (3) and an upper ring (1) and a lower ring (2) which are overlapped and clamped and riveted to fix the amnion sheet; a thin layer of tacrolimus liposomes is prepared on the surfaces of the upper ring and the lower ring respectively; The axial projections of the upper ring and the lower ring are both the same elliptical ring, the inner ellipse in the middle of the elliptical ring is a through hole, and the rest of the parts are arc surfaces that fit the ocular surface; the amniotic membrane is flattened and fixed on the amnioscope and covers the entire through hole; The preparation process of the tacrolimus liposome thin layer is as follows: 1) After washing and drying, the upper ring and the lower ring are light-cured and then cleaned in PBS solution and ethanol in an ultrasonic water bath, respectively. The cleaning process is repeated several times and then dried. 2) After the upper ring and the lower ring of step 1) are completed, they are placed in a sub-atmospheric glow discharge plasma treatment device to generate negatively charged hydroxyl groups on the surfaces of the upper ring and the lower ring; 3) After the upper and lower rings in step 2) are completed, they are sequentially immersed in a cationic tacrolimus liposome solution, immersed in ultrapure water, immersed in a negatively charged sodium alginate solution, and immersed in ultrapure water. The above liposome-ultrapure water-sodium alginate solution-ultrapure water immersion process is repeated several times; after completion, the rings are taken out and stored in a 4°C refrigerator away from light for later use.

2. The novel drug-loaded 3D printed amnionoscope according to claim 1 is characterized in that: The lower surface of the upper ring is provided with a plurality of rivets (1-1) protruding toward the lower ring, and the rivets are arranged along the edge of the through hole; the lower ring is provided with a plurality of rivet holes (2-1) corresponding to the positions of the rivets, which can be riveted one by one with the rivets; after the rivets are inserted through the amniotic membrane, they are riveted to the corresponding rivet holes.

3. The novel drug-loaded 3D printed amnioscope according to claim 2 is characterized in that: The preparation process of the tacrolimus liposome thin layer is as follows: 1) After washing and drying, the upper ring and the lower ring were light-cured under ultraviolet light, and then ultrasonically cleaned in PBS solution and 75% ethanol for 10 minutes respectively. The cleaning process was repeated 3 times, and then dried; 2) After the upper and lower rings in step 1) are processed in a sub-atmospheric glow discharge plasma treatment device, the mode is set to HI, and the processing time is 1 min, so that negatively charged hydroxyl groups are generated on the surfaces of the upper and lower rings; 3) After the upper and lower rings of step 2) are completed, they are placed in a positively charged 0.3 mg / mL tacrolimus liposome solution for 2 hours and in ultrapure water for 5 minutes to remove the liposomes loosely adsorbed on the surface of the upper and lower rings, and then immersed in a negatively charged 1 mg / mL sodium alginate solution for 1 hour and in ultrapure water for 5 minutes; the above liposome-ultrapure water-sodium alginate solution-ultrapure water immersion process is repeated 10 times; after assembly, take out and store in a 4°C refrigerator away from light for later use.

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