Patch for eardrum regeneration

Abstract

The present invention relates to an eardrum regeneration patch. The eardrum regeneration patch of the present invention is thin but has high uniformity and appropriate elongation and tensile strength as an eardrum regeneration patch, and thus has an excellent effect of eardrum regeneration, has excellent adhesion with respect to an affected part, and prevents a patient from feeling discomfort such as that from post-procedure fluttering.

Description

eardrum regeneration patch

[0001] The present invention relates to an eardrum regeneration patch.

[0002]

[0003] Chronic otitis media (COM), a condition characterized by persistent tympanic membrane perforation, is one of the most common causes of hearing loss in otolaryngology. However, most treatment options still rely on surgical procedures such as tympanoplasty. These surgical procedures, such as tympanoplasty, are expensive and complex, require postoperative hospitalization, and have numerous drawbacks, including a recurrence rate of over 10%.

[0004] To overcome the limitations of these surgical methods, various methods have been studied, the oldest of which is the application of a paper patch. This involves cutting a piece of paper and applying it to the perforation, allowing cells at the tympanic membrane perforation margin to grow using the patch as a support. While this method is generally effective for small acute tympanic membrane perforations, it is virtually ineffective for chronic tympanic membrane perforations. Furthermore, the use of bioincompatible materials can lead to inflammation, and the success rate is less than 10%.

[0005] With the development of biocompatible materials, there have been attempts to regenerate eardrums in mice using patches made of biocompatible materials. However, the effects of regenerative therapy were insufficient, and the patches had poor adhesiveness, often falling off during activity. Furthermore, because these studies were not based on clinical trials in humans, they were limited in that they did not adequately consider patients' auditory discomfort.

[0006] Therefore, there is a need to develop an eardrum regeneration patch that has excellent adhesiveness and eardrum regeneration effect and does not cause auditory discomfort to patients after the procedure.

[0007]

[0008] The purpose of the present invention is to provide an eardrum regeneration patch that has an excellent eardrum regeneration effect, does not easily detach from an eardrum perforation, is flexible and thin, yet has excellent tensile strength, and does not cause discomfort such as eardrum flapping after the procedure.

[0009]

[0010] The present invention provides an eardrum regeneration patch comprising an adhesive layer and an tympanic membrane regeneration layer formed on the adhesive layer, wherein the tympanic membrane regeneration layer is made of a biocompatible polymer, a pattern for tympanic membrane cell growth is formed on one side or both sides of the tympanic membrane regeneration layer, the pattern is made of repeating peaks and valleys, and the tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 ㎛ to 40 ㎛.

[0011] It is preferable that the tympanic membrane regeneration layer of the above tympanic membrane regeneration patch exhibits an elongation of 4% to 10%.

[0012] The tympanic membrane regeneration layer of the above tympanic membrane regeneration patch can exhibit an elongation of 500% to 700% depending on the material.

[0013] The pattern of the above tympanic membrane regeneration patch may be such that more than 99% of its area satisfies the uniformity (U) of the following mathematical expression 1:

[0014] [Mathematical Formula 1]

[0015] U = |H P1 - H P2 |≤ 0.1H n

[0016] In mathematical expression 1, P1 and P2 are each a point on the floor or a point on the valley, respectively, and H P1 and H P2 represents the height at points P1 and P2, respectively, and H n represents the height difference between the peak and valley of a normal pattern.

[0017] The present invention provides a method for manufacturing an eardrum regeneration patch of the present invention, comprising: a step of manufacturing a sheet-shaped semi-cured article by applying a polymer solution containing a biocompatible polymer onto a polyurethane acrylate mold; a step of positioning the semi-cured article between a polyurethane acrylate mold and a polydimethylsiloxane mold and then applying pressure with upper and lower plates, respectively, to manufacture a patterned semi-cured article; and a step of drying the patterned semi-cured article; and a step of forming an adhesive layer on the eardrum regeneration layer manufactured as described above.

[0018] In the above manufacturing method, the concentration of the biocompatible polymer in the polymer solution may be 15% (w / w) to 24% (w / w).

[0019] In the above manufacturing method, drying can be performed at 60°C to 80°C.

[0020] In the above manufacturing method, unevenness for pattern formation may be formed on a polyurethane acrylate mold or a polydimethylsiloxane mold.

[0021] In the above manufacturing method, the upper plate may be provided with a pressure rod and a guide rod.

[0022] In the above manufacturing method, the pressure can be applied at 0.40 Pa to 0.70 Pa.

[0023]

[0024] The eardrum regeneration patch of the present invention has an excellent eardrum regeneration effect.

[0025] The eardrum regeneration patch of the present invention has excellent adhesion to the affected area and is not easily detached from the eardrum perforation.

[0026] The eardrum regeneration patch of the present invention does not cause the eardrum to flutter when the patient moves or talks after the procedure, so the patient does not feel any discomfort after the procedure.

[0027] The eardrum regeneration patch of the present invention is flexible and thin, yet has excellent tensile strength, so that it can vibrate appropriately in response to sound (sound waves) in the air and maintain adhesion in eardrum environments of various structures or shapes.

[0028] The eardrum regeneration patch of the present invention is flexible and thin, yet has excellent tensile strength, making it easy for a doctor to handle.

[0029] The eardrum regeneration patch of the present invention is flexible and thin, yet has excellent tensile strength and is not easily torn.

[0030] The eardrum regeneration patch of the present invention has excellent pattern uniformity, which is advantageous for eardrum cell regeneration.

[0031] The eardrum regeneration patch of the present invention does not cause any discomfort, such as eardrum flapping, in daily life after the procedure.

[0032]

[0033] Figure 1 is a conceptual diagram of a manufacturing process of an eardrum regeneration patch according to an embodiment of the present invention.

[0034] Figure 2 shows a method for measuring the uniformity (U) of the mathematical formula 1 of the present invention.

[0035] Figures 3a to 3e are surface SEM photographs of the eardrum regeneration patches of Examples 1 to 22 and Comparative Examples 1 and 2.

[0036] Figure 4 is a photograph comparing the uniformity (area %) of the eardrum regeneration patches of Example 4 and Comparative Example 1. The eardrum regeneration patch of the present invention (Example 4) has excellent uniformity in almost all areas, whereas the area of ​​Comparative Example 1 with excellent uniformity is only less than 70% of the total area.

[0037] Figure 5 shows the SEM and FR-IR analysis results of the eardrum regeneration patches of Comparative Example 3 and Example 4.

[0038] Figures 6a and 6b are comparative data of tensile strength and elongation, and comparative data of adhesive strength (adhesion strength) of the eardrum regeneration patches of Comparative Example 3 and Example 4 (Flat: Comparative Example 3, Nano: Example 4).

[0039] Figure 6c is a photograph showing the flexibility of the tympanic membrane regeneration layer of Example 23.

[0040] Figures 7 and 8 show the fibroblast proliferation and migration morphology of the eardrum regeneration patches of Comparative Example 3 and Example 4 (Flat: Comparative Example 3, Nano: Example 4).

[0041] Figure 9 shows the results of quantification by analyzing the degree of cell attachment and proliferation after culturing fibroblasts in the eardrum regeneration patches of Comparative Example 3 and Example 4 (TCPS: TCPS cell culture vessel, Flat: Comparative Example 3, Nano: Example 4).

[0042] Figures 10 and 11 show the results of comparing the cell migration distance, migration speed, and wound healing range of the eardrum regeneration patches of Comparative Example 3 and Example 4 in a wound healing model (Flat: Comparative Example 3, Nano: Example 4).

[0043] Figure 12 shows a method for conducting an animal experiment using an animal model of tympanic membrane perforation.

[0044] Figure 13 shows the results of animal experiments using the existing paper patch and the eardrum regeneration patch of Example 4.

[0045] Figure 14 shows the results of an animal experiment using the tympanic membrane regeneration patch of Example 23.

[0046] Figures 15 and 16 show the results of a clinical trial using the eardrum regeneration patch of Example 4.

[0047] Fig. 17 is a perspective view of a device for manufacturing an eardrum regeneration patch according to one embodiment of the present invention.

[0048] Fig. 18 is a front view of a device for manufacturing an eardrum regeneration patch according to one embodiment of the present invention.

[0049] Fig. 19 is a top view of a device for manufacturing an eardrum regeneration patch according to one embodiment of the present invention.

[0050] Fig. 20 is a cross-sectional view taken along A1-A2 of Fig. 19.

[0051] FIG. 21 is a block diagram illustrating a device for manufacturing an eardrum regeneration patch according to an embodiment of the present invention.

[0052] Fig. 22 is a plan view showing the shape of a pressurizing plate of a pressurizing unit according to an embodiment of the present invention.

[0053] FIG. 23 is a plan view showing another shape of a pressurizing plate of a pressurizing unit according to an embodiment of the present invention.

[0054] FIG. 24 is a plan view showing another shape of a pressurizing plate of a pressurizing unit according to an embodiment of the present invention.

[0055] FIG. 25 is a flowchart illustrating a method for manufacturing an eardrum regeneration patch using an eardrum regeneration patch manufacturing device according to one embodiment of the present invention.

[0056] Figures 26 to 28 are flowcharts of a detailed method for manufacturing an eardrum regeneration patch according to each step of Figure 25.

[0057] FIGS. 29 to 34 are process diagrams illustrating a method for manufacturing an eardrum regeneration patch using an eardrum regeneration patch manufacturing device according to one embodiment of the present invention.

[0058] Figure 35 is a schematic diagram of a tympanic membrane treatment method using the tympanic membrane regeneration patch of the present invention.

[0059]

[0060] The present invention provides the following eardrum regeneration patch.

[0061] eardrum regeneration patch

[0062] The present invention provides an eardrum regeneration patch comprising an adhesive layer and an tympanic membrane regeneration layer formed on the adhesive layer, wherein the tympanic membrane regeneration layer is made of a biocompatible polymer, a pattern for tympanic membrane cell growth is formed on one side or both sides of the tympanic membrane regeneration layer, the pattern is made of repeating peaks and valleys, and the tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 ㎛ to 40 ㎛.

[0063] The tympanic membrane regeneration layer of the tympanic membrane regeneration patch of the present invention has a tensile strength of 3 to 20 MPa, a thickness of 10 µm to 40 µm, and can exhibit an elongation of 4% to 10%.

[0064] The adhesive layer is made of a material that can properly attach the tympanic membrane regeneration layer to the affected area of ​​the tympanic membrane. For example, a hydrocolloid material used as a wound moistening patch can be used.

[0065] When a hydrocolloid layer is used as an adhesive layer, it can exhibit anti-inflammatory activity.

[0066] The adhesive layer should have a larger cross-sectional area than the tympanic membrane regeneration layer for better adhesion. For example, if the tympanic membrane regeneration layer is coin-shaped, the adhesive layer may be coin-shaped with a larger radius than the tympanic membrane regeneration layer.

[0067] The shape of the adhesive layer and the eardrum regeneration layer can be appropriately determined according to the shape of the perforation, the patch attachment environment (humidity, curvature, inclination, etc.), etc.

[0068] The eardrum regeneration patch of the present invention includes a eardrum regeneration layer made of a biocompatible polymer.

[0069] Biocompatible polymers include any polymer known to be usable in living organisms, without limitation. Examples include polycaprolactone (PCL), polylactide-co-glycolide (PLGA), polyethylene glycol (PEG), polyethylene oxide (PEO), polylactic acid (PLA), and polyglycolic acid (PGA).

[0070] In terms of pattern uniformity and the physical properties (elongation, tensile strength, etc.) of the eardrum regeneration layer, it is preferable to use PLGA or PCL as a biocompatible polymer. Among PLGA, a molar ratio of lactide to glycolide of 65-85 to 15-35 is more preferable.

[0071] A pattern for tympanic membrane cell growth is formed on a cross-section or both sides of the tympanic membrane regeneration layer of the present invention.

[0072] When a tympanic membrane regeneration patch is attached to the affected area, tympanic membrane cells surrounding the patch grow along the grooves of the pattern, inducing regeneration of the damaged tympanic membrane.

[0073] Patterns can be formed on either one side or both sides. To facilitate differentiation between the upper and lower surfaces of the eardrum regeneration patch, patterns can be formed only on the upper or lower surface. Furthermore, if the eardrum regeneration patch is small, patterns can be formed on both sides to eliminate the distinction between the upper and lower surfaces.

[0074] Since the tympanic membrane regeneration layer is formed on the adhesive layer, a pattern must be formed on the side of the tympanic membrane regeneration layer that is not in contact with the adhesive layer for tympanic membrane cell growth.

[0075] Patterns formed on the tympanic membrane regeneration layer can influence not only tympanic cell growth but also the material properties of the tympanic membrane regeneration layer. Patterns formed on both sides of the tympanic membrane regeneration layer can improve tensile strength and elongation compared to patterns formed only on one side.

[0076] The pattern is not limited to a specific shape, as long as it has repeating peaks and valleys. For example, the pattern can be a straight or curved shape with repeating peaks and valleys. The shape of the pattern can be a straight line parallel to one edge of the eardrum regeneration patch, a diagonal line at a predetermined angle to one edge, or a wavy curve.

[0077] The vertex refers to the relatively protruding part, and the trough refers to the relatively depressed part between the vertices. The tympanic membrane cells grow along the trough between the vertices.

[0078] The width between the floors can be designed in various ways, for example, 600 to 1000 nm, 700 to 1000 nm, 800 to 1000 nm, 600 to 900 nm, 600 to 800 nm, 600 to 700 nm, etc.

[0079] The width between the grooves can also be designed to be the same as the width between the floors.

[0080] The height difference between the peak and the valley (the height of the peak relative to the valley) can be designed in various ways, such as 600 to 1000 nm, 700 to 1000 nm, 800 to 1000 nm, 600 to 900 nm, 600 to 800 nm, and 600 to 700 nm.

[0081] The distance between the peaks of the pattern, the distance between the valleys, and the height difference between the peaks and valleys may be the same or different. When they are different, the difference may be 100 nm or less, 90 nm or less, 80 nm or less, 70 nm or less, 60 nm or less, or 50 nm or less, respectively.

[0082] For example, the distance between crests, the distance between valleys, and the height difference between crests and valleys can be equal to 800 nm, 700 nm, or 600 nm, respectively. In addition, the distance between crests and the distance between valleys can be 800 nm, and the height difference between crests and valleys can be 750 nm or 850 nm. In addition, the distance between crests and the distance between valleys can be 700 nm, and the height difference between crests and valleys can be 650 nm or 750 nm.

[0083] The distance between the peaks of the pattern, the distance between the valleys, and the height difference between the peaks and valleys can be changed by adjusting the size and spacing of the pattern formed in the mold used to manufacture the eardrum regeneration patch.

[0084] The pattern may have an area satisfying the uniformity (U) of the following mathematical expression 1 of 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 99.9% or more of the total area.

[0085] [Mathematical Formula 1]

[0086] U = |H P1 - H P2 |≤ 0.1H n

[0087] In mathematical expression 1, P1 and P2 are each a point on the floor or a point on the valley, respectively, and H P1 and H P2 represents the height at points P1 and P2, respectively, and H n refers to the height difference between the peak and valley of a normal pattern.

[0088] P1 and P2 can be any points on the floor of the pattern. P1 and P2 can be any points on the valley of the pattern.

[0089] For example, P1 may be a point on a floor, and P2 may be a point on another floor adjacent to that floor, that is, across a valley to the left or right of that floor. P1 may be a point on a floor, and P2 may be a point on another floor spaced apart from that floor, that is, across multiple valleys to the left or right of that floor.

[0090] H P1 is the height at P1. The height at P1 means the height from the valley to any point on the crest, P1. If the crest is formed at the intended height, H P1is the same height as the pattern formed in the mold, and H is the difference in height between the peak and valley of the normal pattern. n is the same as H. If the pattern formation is poor, P1 The height of the pattern formed in the silver mold is different.

[0091] H P2 refers to the height at P2. H P1 It is the same as .

[0092] The tympanic membrane regeneration layer is the height difference between the peak and valley of the normal pattern (H) at P1 and P2. n ) is 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less, 2% or less, 1% or less, 0.5% or less, or 0.1% or less.

[0093] If the pattern of the tympanic membrane regeneration layer is highly uniform, tympanic membrane cells will proliferate smoothly along the uniform pattern, resulting in excellent tympanic membrane regeneration. If the pattern is not uniform and there are partially damaged areas, cell proliferation may be halted in the damaged areas.

[0094] When P1 and P2 are points on the same floor, when measuring the uniformity of the pattern, the height difference between P3 and P4, which are arbitrary points on the valley of the pattern, can optionally be additionally considered.

[0095] P3 and P4 can be points on the same bone. P3 can be a point on a bone, and P4 can be a point on another bone adjacent to that bone, that is, across a crest to the left or right of that bone. P3 can also be a point on a bone, and P4 can be a point on another bone separate from that bone, that is, across multiple crests to the left or right of that bone.

[0096] When P1 and P2 are arbitrary points on the floor and P3 and P4 are arbitrary points on the valley, U1 = |H P1 - HP2 |≤ 0.1H n and U2= |H P3 - H P4 |≤ 0.1H n (H P3 is the height at P3, H P4 refers to the height at P4 and H n The area satisfying the height difference between the peak and valley of the normal pattern may be 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, or 99.9% or more of the total area.

[0097] The thickness of the tympanic membrane regeneration layer ranges from 10 to 40 μm. If it is thinner than 10 μm, the mechanical strength (tensile strength and compressive strength) is low, making the patch or the nano-pattern formed on the patch susceptible to damage. Furthermore, the patch may flap after the procedure, causing discomfort to the patient. If it is thicker than 40 μm, the adhesive strength is poor, making it difficult for the patch to remain attached to the tympanic membrane for a sufficient amount of time for tympanic membrane regeneration.

[0098] If the thickness is thicker than 40㎛, the flexibility of the eardrum regeneration patch may be reduced, making it difficult to vibrate appropriately in response to sound (sound waves), or the adhesion may not be maintained in eardrum environments with various structures or shapes.

[0099] The optimal thickness of the tympanic membrane regeneration layer may vary depending on the material. For example, if the tympanic membrane regeneration layer is made of PLGA material, the thickness is preferably 10 µm to 30 µm, and more preferably 15 µm to 20 µm. For example, if the tympanic membrane regeneration layer is made of PCL material, the thickness is preferably 18 µm to 35 µm, and more preferably 25 µm to 30 µm.

[0100] The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa. This tensile strength allows the tympanic membrane regeneration layer to maintain adhesion to tympanic membrane environments of various structures and shapes, despite its flexibility and thinness. It also vibrates appropriately in response to airborne sound (sound waves). Furthermore, its small size makes it easy for doctors to handle during procedures.

[0101] The optimal tensile strength of the tympanic membrane regeneration layer may vary depending on the material and thickness. For example, the tensile strength of the tympanic membrane regeneration layer made of PLGA material and having a thickness of 10 μm to 20 μm may be 3 MPa to 8 MPa. For example, the tensile strength of the tympanic membrane regeneration layer made of PLGA material and having a thickness of 20 μm to 30 μm may be 15 MPa to 20 MPa. For example, the tensile strength of the tympanic membrane regeneration layer made of PCL material and having a thickness of 18 μm to 35 μm may be 4 MPa to 6 MPa.

[0102] The tympanic membrane regenerative layer can have an elongation of 4% to 10%. Elongation exceeding 10% can easily deform and be difficult to handle, and the flap of the patch after the procedure can cause discomfort to the patient. Elongation below 4% can be prone to breakage and, due to insufficient flexibility, may not properly adhere to the tympanic membrane.

[0103] It is desirable that the elongation of the tympanic membrane be 5% to 7%.

[0104] When the eardrum regeneration layer is made of PCL material, it can have an elongation of 500% to 700%. When the tensile strength and thickness are satisfied, the eardrum regeneration effect of the present invention can be expected. However, due to the high elongation, the adhesive strength may be reduced depending on the eardrum attachment environment.

[0105] It is desirable that the elongation of the tympanic membrane be 5% to 7%.

[0106] The adhesion strength of the tympanic membrane regeneration layer may be 0.2 to 0.5 N / ㎠, and is preferably 0.3 to 0.4 N / ㎠.

[0107] The adhesive layer and non-adhesive surface of the tympanic membrane regeneration layer may be coated with any one selected from the group consisting of collagen, fibrin, growth factors, stem cells, exosomes, and therapeutic drugs, which helps promote adhesion or cell growth.

[0108] For example, fibrin applied to the edge of the tympanic membrane regeneration layer can form a fibrous blood clot and act as a bridge for cell movement, thereby facilitating cell movement and proliferation at the tympanic membrane perforation site, thereby improving the speed of tympanic membrane regeneration.

[0109] When attaching the eardrum regeneration layer to the adhesive layer, an adhesive such as medical glue may be used as needed.

[0110] The present invention provides a method for manufacturing the following eardrum regeneration patch.

[0111]

[0112] Method for manufacturing an eardrum regeneration patch

[0113] The present invention provides a method for manufacturing a tympanic membrane regeneration patch, comprising: (1) a step of manufacturing a tympanic membrane regeneration layer by a predetermined method; and (2) a step of forming an adhesive layer on the tympanic membrane regeneration layer manufactured in step (1).

[0114] (1) The steps can be structured as follows:

[0115] (1-1) A step of manufacturing a sheet-shaped semi-cured film by applying a polymer solution containing a biocompatible polymer onto a polyurethane acrylate (PUA) mold, (1-2) a step of manufacturing a patterned semi-cured film by placing the semi-cured film between a polyurethane acrylate (PUA) mold and a polydimethylsiloxane (PDMS) mold and then applying pressure with upper and lower plates, respectively, and (1-3) a step of manufacturing a tympanic membrane regeneration layer including a step of drying the patterned semi-cured film.

[0116] In the above manufacturing method, the concentration of the biocompatible polymer in the polymer solution may be 15% (w / w) to 24% (w / w).

[0117] In the above manufacturing method, drying can be performed at 60°C to 80°C.

[0118] In the above manufacturing method, unevenness for pattern formation may be formed on a polyurethane acrylate mold or a polydimethylsiloxane mold.

[0119] In the above manufacturing method, the upper plate may be provided with a pressure rod and a guide rod.

[0120] In the above manufacturing method, the pressure can be applied at 0.40 Pa to 0.70 Pa.

[0121] More specifically, the tympanic membrane regeneration layer can be manufactured by a method including a semi-hardened material forming step (S100), a semi-hardened material fixing step (S200), a transporting step (S300), a pressurizing step (S400), a drying step (S500), and a separation step (S600).

[0122] Referring to FIGS. 26 and 29(A), the semi-hardened material forming step (S100) may include a step (S110) of placing an upper mold (900) on a base surface. The base may be the ground, a structure with a flat surface, or a lower plate (510). When the base surface is the lower plate (510), the lower plate (510) may be placed on the loading area (LA). In this embodiment, a case where the base is the lower plate (510) will be illustrated and described.

[0123] The upper mold (900) is a polyurethane acrylate (PUA) mold.

[0124] The upper mold (900) may include a first pattern surface (910) on which a pattern is formed, and a first support surface (960) disposed on the opposite surface of the first pattern surface (910). The first support surface (960) may be placed on the base surface.

[0125] The first pattern surface (910) may include a pattern of a predetermined shape. The pattern is not limited to a specific shape as long as it has repeating peaks and valleys. For example, the pattern may be a straight or curved shape with repeating peaks and valleys parallel to each other. The shape of the pattern may be a straight line parallel to one edge of the eardrum regeneration patch, a diagonal line forming a predetermined angle with one edge, a wavy curve, etc.

[0126] The width between the floors can be designed in various ways to suit the pattern of the eardrum regeneration patch to be manufactured, such as 600 to 1000 nm, 700 to 1000 nm, 800 to 1000 nm, 600 to 900 nm, 600 to 800 nm, 600 to 700 nm, etc.

[0127] The width between the grooves can also be designed to be the same as the width between the floors.

[0128] The height difference between the peak and the valley (the height of the peak relative to the valley) can be designed in various ways to suit the pattern of the eardrum regeneration patch to be manufactured, such as 600 to 1000 nm, 700 to 1000 nm, 800 to 1000 nm, 600 to 900 nm, 600 to 800 nm, and 600 to 700 nm.

[0129] The distance between the peaks of the pattern, the distance between the valleys, and the height difference between the peaks and valleys may be the same or different. When they are different, the difference may be 100 nm or less, 90 nm or less, 80 nm or less, 70 nm or less, 60 nm or less, or 50 nm or less, respectively.

[0130] For example, the distance between crests, the distance between valleys, and the height difference between crests and valleys can be equal to 800 nm, 700 nm, or 600 nm, respectively. In addition, the distance between crests and the distance between valleys can be 800 nm, and the height difference between crests and valleys can be 750 nm or 850 nm. In addition, the distance between crests and the distance between valleys can be 700 nm, and the height difference between crests and valleys can be 650 nm or 750 nm.

[0131] The size and spacing of the pattern of the tympanic membrane regeneration patch manufactured can be changed by controlling the size and spacing of the pattern formed in the mold.

[0132] Referring to FIG. 26 and FIG. 29(B), the step (S100) of forming a semi-hardened article (1000b) may include a step (S120) of applying a biocompatible polymer solution (1000a) to the first pattern surface (910) of the upper mold (900). By applying the biocompatible polymer solution (1000a) to the PUA mold (upper mold, 900), a sheet-shaped semi-hardened article (1000b) can be formed.

[0133] A biocompatible polymer solution can be prepared by mixing a biocompatible polymer and an organic solvent, and the organic solvent can be, for example, a halomethane solvent such as chloroform or dichloroform.

[0134] The suitable concentration of the biocompatible polymer solution may vary depending on the type of the biocompatible polymer, and for example, the concentration of the biocompatible polymer in the biocompatible polymer solution may be 15% (w / w) to 24% (w / w). When the biocompatible polymer is PLGA, the concentration may be 15% (w / w) to 21% (w / w), and when the biocompatible polymer is PCL, the concentration may be 18% (w / w) to 24% (w / w).

[0135] In the process of applying the biocompatible polymer solution (1000a), some of the solvent evaporates at room temperature to form a sheet-like semi-cured article (1000b), which prevents the semi-cured article from flowing out even when the upper mold (900) is turned over.

[0136] The upper mold (900) on which a semi-hardened material (1000b) is placed on the first pattern surface (910) is referred to as a first mold (1100).

[0137] Since the first mold (1100) is placed on the upper mold (900) with a semi-hardened material (1000b), the first mold (1100) must be turned over later and brought into contact with the lower mold (800).

[0138] Therefore, in order to bring the surface of the exposed semi-hardened material (1000b) into contact with the lower mold (800) by turning over the first mold (1100), a high viscosity biocompatible polymer can be used or a high volatility solvent can be used to quickly evaporate the solvent, thereby increasing the viscosity of the semi-hardened material (1000b) and preventing it from flowing down.

[0139] Referring to Fig. 25, the method for manufacturing a tympanic membrane regeneration layer may include a step (S200) of fixing a semi-hardened material (1000b). Fixing the semi-hardened material (1000b) is a step of flipping over the first mold (1100) and bringing the exposed semi-hardened material (1000b) into contact with the lower mold (800). The mold formed by contacting the lower mold (800) with the first mold (1100) is referred to as a second mold (1200).

[0140] The lower mold (800) is a polydimethylsiloxane (PDMS) mold.

[0141] Referring to FIG. 27 and FIG. 30(A), the step (S200) of fixing the semi-cargo (1000b) may include a step (S210) of placing the carrier (320) on the loading area (LA).

[0142] A lower plate (510) may be placed on a carrier (320). For example, the lower plate lower surface (512) may be placed on the carrier mounting surface (328). The lower plate (510) may be placed on the loading area (LA) such that the upper surface (517) of the lower plate is exposed.

[0143] Referring to FIG. 27 and FIG. 30(B), the step (S200) of fixing the semi-hardened material (1000b) may include a step (S220) of placing the lower mold (800) on the lower plate (510). For example, the lower mold (800) may be placed on the upper surface (517) of the lower plate on the loading area (LA).

[0144] The lower mold (800) may include a flat surface without a pattern and a second support surface (860) formed on the opposite surface of the flat surface. The second support surface (860) may be placed on the upper surface (517) of the lower plate. Therefore, when the lower mold (800) is placed on the lower plate (510) in the loading area (LA), the flat surface may be exposed to the outside. The lower mold (800) having a flat surface without a pattern is used when manufacturing a cross-sectional nano-patterned eardrum regeneration patch.

[0145] The lower mold (800) may include a second pattern surface (810) and a second support surface (860) formed on an opposite side to the second pattern surface (810). The second pattern surface (810) may be a flat surface without a pattern or a surface with a pattern formed thereon. If it is a flat surface without a pattern, it is used when manufacturing a single-sided patterned eardrum regeneration patch, and if it is a surface with a pattern formed thereon, it is used when manufacturing a double-sided patterned eardrum regeneration patch. The second support surface (860) may be placed on the upper surface (517) of the lower plate. Therefore, when the lower mold (800) is placed on the lower plate (510) on the loading area (LA), the second pattern surface (810) may be exposed to the outside.

[0146] Referring to FIG. 27 and FIG. 30(C), the step (S200) of fixing the semi-hardened material (1000b) may include a step (S230) of contacting the first mold (1100) with the lower mold (800). For example, the semi-hardened material (1000b) may be fixed by placing the first mold (1100) so that it contacts the second pattern surface (810).

[0147] The step (S200) of fixing the semi-conductor (1000b) can expose the first support surface (960) of the upper mold (900) to the outside on the loading area (LA).

[0148] Referring to FIGS. 25 and 31, the method for manufacturing a tympanic membrane regeneration layer may include a transfer step (S300) of transferring a second mold (1200).

[0149] The transfer step (S300) can move the second mold (1200) placed on the loading area (LA) to the molding area (PA).

[0150] A pressing plate (430) of a pressing unit (400) may be placed on the upper portion of the forming area (PA). An upper plate (520) may be coupled to the pressing plate (430). Accordingly, the second mold (1200) may be placed on the lower portion of the upper plate (520).

[0151] For example, the pressure plate (430) may include a pressure plate lower surface (432) and a pressure plate upper surface (437). An upper plate (520) may be coupled to the pressure plate lower surface (432).

[0152] The upper plate (520) may include an upper plate pressure surface (522) and an upper plate coupling surface (527). A pressure plate lower surface (432) may be coupled to the upper plate coupling surface (527). In addition, a pressure distribution member (550) may be bonded to the upper plate pressure surface (522).

[0153] At the bottom of the forming area (PA), a second mold (1200) may be moved, and a transfer unit (300) for moving the second mold (1200) and a lower plate (510) placed on the transfer unit (300) may be placed.

[0154] For example, the transport unit (300) may include a transport rail (310) and a carrier (320). A lower plate (510) may be placed on the carrier (320). A second mold (1200) may be placed on the lower plate (510). Accordingly, the second mold (1200) may be placed in the forming area (PA).

[0155] The transport unit (300) may further include a transport drive unit (330) capable of moving the carrier (320). By driving the transport drive unit (330), the carrier (320) may be moved from the loading area (LA) to the forming area (PA).

[0156] Accordingly, a second mold (1200) may be placed in the lower direction of the pressure distribution unit (550) in the forming area (PA). For example, in the transfer step (S300), the pressure distribution unit (550) and the first support surface (960), which is the exposed surface of the second mold (1200), may be placed in the forming area (PA) so as to face each other.

[0157] Referring to FIG. 25, a method for manufacturing a tympanic membrane regeneration layer according to one embodiment may include a pressurizing step (S400) of pressurizing a fixed semi-hardened material (1000b).

[0158] For example, referring to FIG. 26 and FIG. 32(A), the pressurizing step (S400) can be performed in the forming area (PA).

[0159] The pressurizing step (S400) of pressurizing the fixed semi-hardened material (1000b) may include a step (S410) of contacting the upper plate (520) with the second mold (1200). For example, the upper plate pressurizing surface (522) of the upper plate (520) may be contacted with the first support surface (960) of the upper mold (900). Specifically, the step (S410) of contacting the upper plate (520) may contact the pressure distribution unit (550) with the first support surface (960).

[0160] In the step (S410) of bringing the upper plate (520) into contact with the second mold (1200), a step of operating the pressurizing unit (400) may be included. For example, the pressurizing unit (400) may operate an actuator disposed in the pressurizing unit body (410). A pressurizing rod (420) may be connected to the actuator. The pressurizing rod (420) may be moved toward the pressurizing plate (430). The pressurizing rod (420) may be mounted on a mounting portion (439) disposed on one surface of the pressurizing plate (430).

[0161] The pressurizing rod (420) can move in the vertical direction. When the pressurizing rod (420) moves downward, the upper plate (520) coupled to the pressurizing plate (430) can move downward. Accordingly, the upper plate (520) can move toward the lower plate (510).

[0162] The upper plate (520) can be moved to bring the upper plate pressure surface (522) into contact with the second mold (1200) placed on the lower plate (510). More specifically, the pressure distribution unit (550) can be brought into contact with the first support surface (960) of the upper mold (900) exposed in the second mold (1200).

[0163] In the step (S410) of contacting the upper plate (520) with the second mold (1200), the thickness of some areas of the second mold (1200) may be different from that of other areas.

[0164] For example, during the application process of the biocompatible polymer solution (1000a), the thickness difference may occur due to the biocompatible polymer solution (1000a) being unevenly distributed. Alternatively, the thickness difference may occur due to the formation thickness of the upper mold (900) and the lower mold (800).

[0165] If a difference in thickness occurs, the upper plate (520) may tilt, and the front surface (Whole surface) of the pressure distribution unit (550) and the front surface (Whole surface) of the first support surface (960) may not make surface contact.

[0166] Referring to FIG. 26 and FIG. 32 (B), the pressurizing step (S400) may include a step (S420) of adjusting the inclination of the upper plate (520).

[0167] The step of adjusting the inclination (S420) may allow the upper plate (520) and the fixed semi-hardened material (1200) to be in surface contact. For example, the step of adjusting the inclination of the upper plate (520) (S420) may allow the front surface (Whole surface) of the pressure distribution unit (550) and the front surface (Whole surface) of the first support surface (960) to be in surface contact.

[0168] After the above two configurations are brought into contact, a step of leveling the upper plate (520) can be performed. That is, the upper plate (520) can be rotated (the inclination adjusted).

[0169] The inclination of the upper plate (520) can be adjusted through the gimbal unit (450). The gimbal unit (450) can be placed on the upper surface of the pressure plate (430). For example, the gimbal unit (450) can be placed between the guide rod (440) formed in the pressure unit (400) and the pressure plate (430). In addition, the gimbal unit (450) can be placed between the mounting unit (439) and the pressure plate (430).

[0170] If the inclination of the upper plate (520) is not horizontal, the gimbal unit (450) can rotate the upper plate (520) to level the upper plate (520).

[0171] Accordingly, the gimbal unit (450) can provide the load of the upper plate (520) to the area where the thickness difference of the second mold (1200) occurs. For example, the gimbal unit (450) can provide the load to the area where the thickness difference occurs by rotating (tilting) the upper plate (520) to level it. Here, since the upper plate (520) is coupled to the pressure plate (430), the loads of the upper plate (520) and the pressure plate (430) can be provided to the second mold (1200).

[0172] The load of the upper plate (520) and the pressure plate (430) provided to the second mold (1200) can provide pressure to the fixed semi-hardened body (1200). Therefore, the load of the upper plate (520) and the pressure plate (430) can pressurize the fixed semi-hardened body (1000b) in the area where the thickness difference occurs.

[0173] When the level of the upper plate (520) is formed, the gimbal part (450) can stop providing a load to the second mold (1200).

[0174] Referring to FIG. 26 and FIG. 33, a step (S430) of maintaining a pressing force on the pressing plate (430) may be included.

[0175] The step of maintaining the pressure (S430) can be performed in a state where the pressure distribution unit (550) and the first support surface (960) are in surface contact and the upper plate (520) is horizontal.

[0176] In the aforementioned state, the target pressing force can be maintained on the pressing plate (430). The step of maintaining the pressing force (S430) can provide the pressing force of 0.40 Pa to 0.70 Pa, preferably 0.45 to 0.65 MPa, to the pressing plate (430). The time for providing the pressing force can vary depending on the magnitude of the pressing force, and can be provided for a holding time of, for example, 30 to 50 minutes.

[0177] The target pressing force provided to the pressing plate (430) can be transmitted to the upper plate (520). The pressing force transmitted to the upper plate (520) can be provided to the second mold (1200) through the upper plate pressing surface (522). The pressing force provided to the second mold (1200) can be transmitted to the fixed semi-hardened material (1200).

[0178] The pattern shape formed on the first pattern surface (910) and the second pattern surface (810) can be transferred to the semi-hardened material (1000b) through the pressure applied to the second mold (1200).

[0179] Referring to FIG. 25 and FIG. 34, a drying step (S500) for drying a semi-hardened material (1000b) may be included.

[0180] The drying step (S500) can provide target drying heat to the second mold (1200) through the lower plate (510). Alternatively, the second mold (1200) can be dried in a dry oven or hot plate, thereby drying the semi-hardened article (1000b).

[0181] When drying the second mold (1200) through the lower plate (510), a heating plate may be provided inside the lower plate (510). The heating plate may transmit thermal energy to the upper surface (517) of the lower plate to dry the second mold (1200). The thermal energy transmitted to the second mold (1200) may be transmitted to the semi-hardened article (1000b) to dry the semi-hardened article (1000b).

[0182] Meanwhile, when drying the second mold (1200) in a dry oven, the carrier (320) can be moved from the forming area (PA) to the loading area (LA), and the second mold (1200) loaded on the lower plate (510) can be unloaded from the loading area (LA). The second mold (1200) can be unloaded and loaded into the dry oven (DO), and drying heat can be provided to the second mold (1200).

[0183] The drying step (S500) here can provide thermal energy of 15°C to 45°C to the second mold (1200) for 6 to 10 hours.

[0184] Alternatively, the drying step (S500) may be performed by placing the second mold (1200) on a hot plate to dry it. When using a hot plate, a steel plate weighing 0.5 kg to 2 kg may be laminated on the second mold (1200), and the hot plate may be heated to 60°C to 80°C to dry the second mold (1200). The second mold (1200) may be dried by maintaining the temperature above for 4 to 8 hours.

[0185] Here, the steel plate can prevent the upper mold (900) or the lower mold (800) from being separated from the semi-hardened material (1000b) during the drying process.

[0186] If the upper mold (900) or the lower mold (800) is separated during the drying process, the surface pattern of the semi-hardened article (1000b) may be damaged. Therefore, the drying process may be performed by placing a steel plate on the second mold (1200) to prevent damage to the surface pattern of the semi-hardened article (1000b).

[0187] In this way, the second mold (1200) can be dried to form a hardened eardrum regeneration patch (1000) between the upper mold (900) and the lower mold (800).

[0188] Referring to FIG. 25 and FIG. 34, a step (S600) of separating the eardrum regeneration patch (1000) from the second mold (1200) can be performed.

[0189] The eardrum regeneration patch (1000) formed between the lower mold (800) and the upper mold (900) constituting the second mold (1200) can be separated from the lower mold (800) and the upper mold (900).

[0190] After the separation step (S600), the biosupport (1000) can be immersed in 70% ethanol for 10 seconds and washed with purified water.

[0191] The manufacturing step of the tympanic membrane regeneration layer can be performed using the following tympanic membrane regeneration patch manufacturing device (10).

[0192] Below, the drawings are provided and explained in detail.

[0193] FIG. 17 is a perspective view of an tympanic membrane regeneration patch manufacturing device according to one embodiment of the present invention, FIG. 18 is a front view of an tympanic membrane regeneration patch manufacturing device according to one embodiment of the present invention, FIG. 19 is a top view of an tympanic membrane regeneration patch manufacturing device according to one embodiment of the present invention, and FIG. 20 is a cross-sectional view taken along line A1-A2 of FIG. 19.

[0194] Referring to FIGS. 17 to 20, the eardrum regeneration patch manufacturing device (10) may include a stand (100).

[0195] The stand (100) may include a stand body (110) and a table (120).

[0196] The table (120) may be formed as a plate having a flat surface. A plurality of components may be arranged on the table (120). The flat surface may provide a stable seating surface for the components.

[0197] The stand body (110) may be equipped with a plurality of legs. The plurality of legs may be positioned at the corner areas of the table (120). The plurality of legs may be positioned with different or identical set lengths. The plurality of legs with adjusted set lengths may form a flat surface of the table (120).

[0198] Additionally, multiple legs can be arranged with preset lengths that correspond to the worker's working height. For example, multiple legs can be arranged at the worker's expected working height. Multiple legs with adjustable heights can improve the worker's workability.

[0199] The eardrum regeneration patch manufacturing device (10) may include a frame unit (200). The frame unit (200) may be placed on a table (120). The frame unit (200) may include a frame support (220) and a frame body (210).

[0200] The frame support (220) can support the frame body (210). The frame support (220) can be placed on the upper surface of the table (120). The frame support (220) can be installed on the table (120) to fix the frame body (210). The frame support (220) and the table (120) can be fixed through a joint structure such as a fixing screw, but the fixing means is not limited to the fixing screw. In addition, the frame support (220) and the frame body (210) can also be fixed by a similar fixing means.

[0201] The frame body (210) may be placed on the table (120). For example, the frame body (210) may be positioned on the upper surface of the table (120) and supported by a frame support (220). The frame body (210) may be placed in a plate shape. Accordingly, the frame body (210) may be placed in a direction parallel to the flat surface of the table (120).

[0202] The frame body (210) may have a plurality of through holes (212, 215). The frame body (210) may have cylinder guides (213, 216) respectively arranged on the plurality of through holes (212, 215).

[0203] The plurality of through holes (212, 215) may include a first through hole (212) and a second through hole (215). A guide rod (440) may be placed in the first through hole (212). A pressure rod (420) may be placed in the second through hole (215).

[0204] A plurality of first through holes (212) may be arranged around the second through hole (215). The first through holes (212) may be arranged symmetrically around the second through hole (215).

[0205] In this embodiment, the first through-holes (212) are arranged symmetrically two around the second through-hole (215). However, the arrangement of the first through-holes (212) is not limited to this, and three to eight first through-holes (212) may be arranged symmetrically around the second through-hole (215).

[0206] The guide rod (440) can distribute the concentrated pressure provided from the pressure rod (420) in the surface direction of the pressure plate (430). Therefore, since the number of first through holes (212) is related to the number of guide rods (440), it can play a role in forming a distributed pressure of the pressure plate (430). The pressure plate (430) on which the distributed pressure is formed can help form a uniform surface pressure in the forming unit (500).

[0207] The eardrum regeneration patch manufacturing device (10) may include a transport unit (300).

[0208] Here, for easy explanation, in the table (120), the area housed by the frame support (220) and the frame body (210) is defined as the forming area (PA), and the area other than the forming area (PA) is defined as the peripheral area (SA). Among the peripheral areas (SA), the area where the transfer unit (300) is placed is defined as the loading area (LA).

[0209] The transport unit (300) may include a transport rail (310), a carrier (320), and a transport drive unit (330).

[0210] The transport rail (310) may be arranged on the upper surface of the table (120). The transport rail (310) may be arranged from the loading area (LA) to the forming area (PA). The transport rail (310) may form a rail portion with a pair of rails arranged in the direction from the loading area (LA) to the forming area (PA). At least one rail portion may be arranged. For example, when the transport rail (310) has multiple rail portions formed in the direction from the loading area (LA) to the forming area (PA), the loading area (LA) may form multiple work windows.

[0211] A carrier (320) can be placed on a transport rail (310). The carrier (320) can move along the transport rail (310). Accordingly, the carrier (320) can be placed in a loading area (LA) and a forming area (PA).

[0212] The carrier (320) can be movably coupled to the transport rail (310). For example, the carrier (320) can be slidably coupled to the transport rail (310), but is not limited thereto, and the carrier (320) and the transport rail (310) can be coupled in any structure as long as the means is movable.

[0213] The carrier (320) may include a sliding portion (322) having a sliding structure and a mounting surface (328) arranged on the opposite side of the sliding portion (322). The sliding portion (322) may be coupled to a transport rail (310). A lower plate (510) may be mounted on the mounting surface (328). The mounting surface (328) may face the lower plate (510).

[0214] The carrier (320) can be connected to a transport drive unit (330). The transport drive unit (330) can be connected to a thickness surface formed between the sliding unit (322) and the seating surface (328).

[0215] The transport drive unit (330) may be positioned in the peripheral area (SA). The transport drive unit (330) may be configured with an actuator to move the carrier (320) to the loading area (LA) and the forming area (PA). In the present embodiment, a structure in which the actuator moves forward and backward is illustrated and described, but in some cases, the carrier (320) may be moved through a structure in which the actuator moves left and right.

[0216] The eardrum regeneration patch manufacturing device (10) may include a pressurizing unit (400). The pressurizing unit (400) may be placed on a molding area (PA). The pressurizing unit (400) may include a pressurizing unit body (410), a pressurizing rod (420), a pressurizing plate (430), and a guide rod (440).

[0217] The pressurized unit body (410) may be placed on the upper surface of the frame body (210). The pressurized unit body (410) may include an actuator. The actuator may move the pressurized rod (420) in the up-and-down direction.

[0218] The pressurizing rod (420) can be placed in the second through hole (215) that penetrates the upper and lower surfaces of the frame body (210). For example, the pressurizing rod (420) can be fitted into the second cylinder guide (216) placed in the second through hole (215).

[0219] The pressure rod (420) can move up and down. The pressure rod (420) can move to contact the pressure plate (430). The pressure rod (420) can move the pressure plate (430) downward. The pressure rod (420) can apply pressure to the pressure plate (430) when it faces the molding object in the target area.

[0220] The pressurizing unit (400) may include a pressurizing plate (430) placed between the frame body (210) and the table (120).

[0221] The pressure plate (430) may include a pressure plate upper surface (437) with which the pressure rod (420) comes into contact. A mounting portion (439) may be provided on the pressure plate upper surface (437). The pressure rod (420) may be mounted on the mounting portion (439).

[0222] The mounting portion (439) may be positioned in an area including the center of gravity of the pressure plate (430). For example, if the pressure plate (430) is positioned in a square shape, the center of gravity of the pressure plate (430) may be the center area of ​​the square shape. Accordingly, the mounting portion (439) may be positioned on the center area.

[0223] One end of the pressure rod (420) can be mounted on the mounting portion (439). The mounting portion (439) can be formed of a material capable of withstanding the pressure force, such as rubber or silicone, on the upper surface (438) of the pressure plate.

[0224] In this way, the pressure rod (420) can move downward and the pressure rod (420) can move the pressure plate (430) downward. And, when the pressure plate (430) moves to the target location, the pressure rod (420) can provide a pressure force to the center of gravity of the pressure plate (430).

[0225] A guide rod (440) may be placed around the pressure rod (420). The guide rod (440) may be placed between the frame body (210) and the pressure plate (430).

[0226] One end of the guide rod (440) can be coupled to the pressure plate (430). The other end of the guide rod (440) can be inserted into a first through hole (212) that penetrates a portion of the frame body (210). For example, a first cylinder guide (213) can be placed in the first through hole (212) formed in the frame body (210). The guide rod (440) can be fitted into the first cylinder guide (213).

[0227] As described above, a plurality of first through holes (212) may be symmetrically arranged around the second through hole (215). A guide rod (440) may be arranged in each of the plurality of first through holes (212).

[0228] Accordingly, a plurality of guide rods (440) can be symmetrically arranged around the pressure rod (420). The plurality of guide rods (440) can serve to distribute the pressure so that the forming unit (500) can form a uniform surface pressure. The plurality of guide rods (440) can form a uniform pressure on the entire surface of the pressure plate (430).

[0229] Meanwhile, a gimbal member (450) may be placed between the guide rod (440) and the pressure plate (430). For example, the gimbal member (450) may connect the lower surface of the guide rod (440) and the upper surface of the pressure plate (430). In addition, the gimbal member (450) may also be selectively placed between the mounting member (439) and the pressure plate (430). The gimbal member (450) may connect the lower surface of the mounting member (439) and the upper surface of the pressure plate (430).

[0230] The gimbal unit (450) can be symmetrically arranged around the mounting unit (439) with the mounting unit (439) as the center. In other words, as a plurality of guide rods (440) are arranged around an area including the center of gravity of the pressing plate (430) on the upper surface of the pressing plate (430), the gimbal unit (450) arranged on each of the plurality of guide rods (440) can also be arranged in the same arrangement structure.

[0231] The gimbal unit (450) can adjust the inclination of the pressure plate (430). For example, the gimbal unit (450) can adjust the horizontality of the upper plate (520). That is, the gimbal unit (450) can rotate (adjust the inclination) of the upper plate (520).

[0232] If the inclination of the upper plate (520) is not horizontal, the gimbal unit (450) can rotate the upper plate (520) to level the upper plate (520).

[0233] Accordingly, the gimbal unit (450) rotates (tilts) the upper plate (520) to level it, so that the load of the upper plate (520) can be provided to the second mold (1200 of FIG. 32). For example, since the upper plate (520) is coupled to the pressure plate (430), the loads of the upper plate (520) and the pressure plate (430) can be provided to the second mold (1200).

[0234] The gimbal unit (450) can adjust a pressure imbalance that may occur due to a difference in the thickness of the molding object. For example, when the thickness of the molding object is different, the gimbal unit (450) can adjust the inclination of the pressure plate (430) so that the pressure plate (430) corresponds to the whole surface of one surface of the molding object.

[0235] Accordingly, when the thickness of some areas of the molding object is different from that of other areas, the gimbal unit (450) can prevent the pressure plate (430) from concentrating the pressure force on some areas of the molding object or transmitting more pressure force to them.

[0236] In this way, the gimbal unit (450) can improve the molding uniformity of the molding target by adjusting the inclination of the pressure plate (430).

[0237] The eardrum regeneration patch manufacturing device (10) may include a molding unit (500). The molding unit (500) may include a lower plate (510), an upper plate (520), and a pressure distribution unit (550).

[0238] The lower plate (510) may be placed on the upper surface of the carrier (320). For example, the lower plate (510) may be mounted on the mounting surface (328) of the carrier (320). The lower plate (510) may be formed in a shape similar to or smaller than the shape of the carrier (320).

[0239] The lower surface (512) of the lower plate (510) can be mounted on the mounting surface (328) of the carrier (320). The carrier (320) can move between the loading area (LA) and the forming area (PA) along the transport rail (310). Accordingly, the lower plate (510) mounted on the carrier (320) can move between the loading area (LA) and the forming area (PA) via the carrier (320).

[0240] The lower plate (510) may include an upper surface (517) of the lower plate (510) on the opposite side of the lower surface (512) of the lower plate (510). A lower mold (800 of FIG. 29) may be placed on the upper surface (517) of the lower plate (510). The manufacturing method with respect to the lower mold (800) will be described in detail below.

[0241] The upper plate (520) may be placed in the forming area (PA). The upper plate (520) may be coupled to the pressure plate (430). The upper plate (520) may include an upper plate coupling surface (527) coupled to the pressure plate (430). For example, the upper plate coupling surface (527) may be coupled to a lower surface (432) of the pressure plate (430). The lower surface (432) of the pressure plate may be a surface facing the upper surface (437) of the pressure plate on which the mounting portion (439) is arranged.

[0242] The shape of the upper plate (520) may be formed to have a larger area than the shape of the lower plate (510). A molding object may be placed between the upper plate (520) and the lower plate (510). Since the upper plate (520) has a larger area than the shape of the lower plate (510), it can provide uniform pressure to the molding object. Therefore, the size of the molding object may be limited by the shape of the lower plate (510).

[0243] As described above, the eardrum regeneration patch manufacturing device (10) according to the present invention can form a pressure plate (430) in which a uniform pressure is formed by dispersing the pressure, thereby increasing the area of ​​the lower plate (510). Accordingly, a molding object placed on the lower plate (510) can be formed over a large area.

[0244] The upper plate (520) may include an upper plate pressure surface (522) on the opposite side of the upper plate joining surface (527). A pressure distribution unit (550) may be arranged on the upper plate pressure surface (522).

[0245] The pressure distribution unit (550) can be bonded to the upper plate pressure surface (522) using an adhesive. The pressure distribution unit (550) can be formed of a silicone material, but is not limited thereto, and any material capable of transmitting pressure can be used. The pressure distribution unit (550) can evenly distribute the pressure input from the pressure plate (430) across the entire surface of the upper plate (520).

[0246] The eardrum regeneration patch manufacturing device (10) may include an input unit (600) and a display unit (700). The input unit (600) and the display unit (700) may be placed in a peripheral area (SA).

[0247] The eardrum regeneration patch manufacturing device (10) may include an input unit (600) that receives input from a worker. For example, the input unit (600) allows the worker to directly input a setting value.

[0248] The input unit (600) may include input means such as an input button for inputting a setting value. Here, the setting value may include, for example, the pressure of the pressurizing unit (400), the heating temperature of the lower plate (510), the pressure provision time and heating time of the pressurizing unit (400), etc.

[0249] The display unit (700) can display the setting values ​​on the screen. In other words, the display unit (700) can display input information on the screen. For example, the display unit (700) can display the pressure of the pressurizing unit (400), the heating temperature of the lower plate (510), the pressure application time, the heating time, etc. in numerical form.

[0250] In addition, a control unit capable of controlling the setting values ​​of the eardrum regeneration patch manufacturing device (10) may be further placed in the space where the display unit (700) is placed.

[0251] In this way, the eardrum regeneration patch manufacturing device (10) according to the embodiment of the present invention forms a uniform pressure, so that an eardrum regeneration patch with improved uniformity can be manufactured.

[0252] Fig. 21 is a block diagram illustrating a device for manufacturing an eardrum regeneration patch according to an embodiment of the present invention.

[0253] To avoid redundant explanation, Fig. 21 will be described by citing Figs. 17 to 20 for easy explanation.

[0254] The eardrum regeneration patch manufacturing device (10) according to an embodiment of the present invention may include a sensor unit (30) and a control unit (50).

[0255] The control unit (50) can be connected to the display unit (700), the input unit (600), and the sensor unit (30). The control unit (50) can be placed in the area where the display unit (700) is placed.

[0256] The control unit (50) may also be connected to a configuration in which the sensor unit (30) is arranged. For example, the control unit (50) may be connected to a configuration in which the pressure sensor (470) and / or the temperature sensor (515) are arranged.

[0257] For example, if a temperature sensor (515) is disposed on the lower plate (510), the control unit (50) may be connected to the lower plate (510). For another example, if a pressure sensor (470) is disposed on the pressurizing unit (400), the control unit (50) may be connected to the pressurizing unit (400). The sensor unit (30) may include a pressure sensor (470). The pressure sensor (470) may be disposed on the upper surface of the frame body (210).

[0258] In this drawing, a drawing in which a pressure sensor (470) is placed in a pressurizing unit (400) is illustrated and explained. Here, the pressure sensor (470) may also be placed in a forming unit (500).

[0259] The input unit (600) can receive input from the operator. The input unit (600) can input a pressure to be provided to the pressurizing rod (420) of the pressurizing unit (400). The input provided can be a set pressure at which the pressurizing unit (400) operates. The input unit (600) can form a first signal (SG1) composed of the set pressure.

[0260] The input unit (600) can provide a first signal (SG1) to the control unit (50). The control unit (50) can convert the provided first signal (SG1) and transmit it to the display unit (700) and the pressurizing unit (400).

[0261] The control unit (50) can convert the first signal (SG1) and transmit the first-first signal (SG11) to the display unit (700). The display unit (700) that receives the first-first signal (SG11) can display the set pressure on the screen.

[0262] Additionally, the control unit (50) can convert the first signal (SG1) and transmit the first-second signal (SG1)2) to the pressurizing unit (400).

[0263] The pressurizing unit (400) can operate according to the first-second signal (SG12). For example, the pressurizing unit (400) can operate the pressurizing rod (420) according to the first-second signal (SG12). For example, the pressurizing rod (420) can move the pressurizing plate (430) according to the first-second signal (SG12). When the pressurizing plate (430) moves according to the first-second signal (SG12) and comes into contact with the molding object, the pressurizing rod (420) can provide pressure to the pressurizing plate (430).

[0264] The pressure sensor (470) of the sensor unit (30) can sense the pressure formed in the pressurizing unit (400). The sensor unit (30) can form a second signal (SG2) with information obtained by sensing the pressure formed in the pressurizing unit (400). The sensor unit (30) can transmit the second signal (SG2) to the control unit (50).

[0265] The control unit (50) that receives the second signal (SG2) can calculate whether information according to the first-second signal (SG12) is transmitted as pressure through the pressurization unit (400). For example, the control unit (50) can calculate the difference between the pressure set in the pressurization rod (420) and the pressure transmitted from the pressure sensor (470). The control unit (50) can form the third signal (SG3) and the fourth signal (SG4) from the value obtained by calculating the difference in pressure (hereinafter, “calculated value”).

[0266] If the operation value is within the error range compared to the measured data, the control unit (50) can transmit the fourth signal (SG4) to the display unit (700). In addition, the control unit (50) can generate a third signal (SG3) that is identical to the first-second signal (SG12) and provide the third signal (SG3) to the pressurizing unit (400).

[0267] Meanwhile, if the operation value is outside the error range compared to the measured data, the control unit (50) can provide a corrected input value to the pressurization unit (400).

[0268] Here, when the control unit (50) is set manually, a fourth signal (SG4) consisting of an operation value can be transmitted to the display unit (700). The display unit (700) can display the operation value according to the fourth signal (SG4) on the screen.

[0269] The operator can check the operation value displayed on the display unit (700). The operator can input the corrected input value based on the displayed information into the input unit (600).

[0270] The control unit (50) can generate a third signal (SG3) using the corrected input value. The third signal (SG3) formed by the control unit (50) can be transmitted to the pressurizing unit (400). Through the third signal (SG3), the pressurizing unit (400) can provide the corrected pressure value to the pressurizing plate (430). The corrected pressure value can be the pressure provided to the pressurizing plate (430).

[0271] Additionally, the control unit (50) can generate a 4-1 signal (SG4-1) based on information according to the 3rd signal (SG3). The 4-1 signal (SG4-1) can be provided to the display unit (700). The display unit (700) can display information composed of the 4-1 signal (SG4-1) on the screen.

[0272] In this way, the eardrum regeneration patch manufacturing device (10) according to the present invention can control the pressure provided to the pressurizing unit (400) through the sensor unit (30).

[0273] Meanwhile, when the control unit (50) is set to automatic, if the value calculated based on the difference in pressure (hereinafter referred to as “calculated value”) shows a difference, the fourth signal (SG4) may not be generated.

[0274] The pressure sensor (470) can sense the pressure formed in the pressurization unit (400). Through the sensing information, the sensor unit (30) can generate a second-first signal (SG21). The sensor unit (30) can transmit the second-first signal (SG21) to the control unit (50).

[0275] The control unit (50) can determine whether the calculated value is within the error range by comparing the calculated value with the measured data based on the measured data. If it is determined that the calculated value is within the error range, the control unit (50) can generate a third signal (SG3) that is identical to the first-second signal (SG12).

[0276] The third signal (SG3) formed in the control unit (50) can be transmitted to the pressurizing unit (400). Through the third signal (SG3), the pressurizing unit (400) can provide pressure according to the first-second signal (SG12) to the pressurizing plate (430). The pressure according to the first-second signal (SG12) can be the pressure provided to the pressurizing plate (430).

[0277] Additionally, the control unit (50) can generate a fourth signal (SG4) based on information according to the third signal (SG3). The fourth signal (SG4) can be provided to the display unit (700). The display unit (700) can display information composed of the fourth signal (SG4) on the screen.

[0278] In this way, the eardrum regeneration patch manufacturing device (10) according to the present invention can control the pressure provided to the pressurizing unit (400) through the sensor unit (30).

[0279] On the other hand, the control unit (50) can compare the input pressure with the pressure provided to the molding object. For example, the pressurizing unit (400) can provide pressure to the molding unit (500). For example, the pressurizing unit (400) can transmit the pressure formed on the pressurizing plate (430) to the molding unit (500).

[0280] A difference may occur between the pressure formed in the pressurizing unit (400) and the pressure formed in the forming unit (500). The control unit (50) may compare the pressure provided by the pressurizing unit (400) with the pressure output from the forming unit (500). The output pressure may be the pressure provided to the forming object.

[0281] The pressure provided can be provided to the forming unit (500) through the pressure plate (430). The output pressure provided to the forming object can be formed through the forming unit (500).

[0282] The pressure sensor (470) of the sensor unit (30) can be placed in each of the pressurizing unit (400) and the forming unit (500). The pressure sensor (470) can sense the supply pressure of the pressurizing unit (400). In addition, the pressure sensor (470) can sense the output pressure of the forming unit (500).

[0283] The sensor unit (30) can generate a second signal consisting of a supply pressure and an output pressure.

[0284] There may be a difference (hereinafter referred to as “pressure difference”) between the input pressure from the pressurizing rod (420) and the output pressure from the forming unit (500). For example, a pressure difference may occur during the process of distributing pressure on the front surface of the pressurizing plate (430) or during the process of distributing pressure in the pressure distributing unit (550).

[0285] In such cases, the probability of molding defects in molded products may increase. Furthermore, power loss from the manufacturing equipment may occur.

[0286] The pressure sensor (470) senses the pressure in each set zone, and the sensor unit (30) can convert the pressure in each set zone into a signal and transmit the second signal (SG2) to the control unit (50).

[0287] The control unit (50) can calculate the pressure difference. Reflecting the calculated value, the setting value of the pressurization unit (400) can be readjusted. Furthermore, the coupling structure between components can be readjusted. Accordingly, the pressure sensor (470) can reduce the probability of defects and reduce power loss resulting from leakage of the manufacturing device.

[0288] FIG. 22 is a plan view illustrating a shape of a press plate of a press unit according to an embodiment, FIG. 23 is a plan view illustrating another shape of a press plate of a press unit according to an embodiment, and FIG. 24 is a plan view illustrating another shape of a press plate of a press unit according to an embodiment.

[0289] To avoid redundant explanation, FIGS. 22 to 24 will be described by citing FIGS. 17 to 20 for easy explanation.

[0290] Referring to FIGS. 22 to 24, the pressurizing unit (400) may include a pressurizing rod (420), a pressurizing plate (430), and a guide rod (440).

[0291] The pressure plate (430) may include a mounting portion (439). The mounting portion (439) may be positioned in an area including the center of gravity of the pressure plate (430). A pressure rod (420) may be positioned at a position corresponding to the mounting portion (439).

[0292] A guide rod (440) may be arranged around the pressure rod (420). The guide rod (440) may be arranged symmetrically around the pressure rod (420). The symmetrically arranged guide rods (440) may form a pressure force distributed on the entire surface of the pressure plate (430).

[0293] Meanwhile, the lower plate (510) of the forming unit (500) may be arranged in a shape with a smaller area than the upper plate (520). And the upper plate (520) may be combined with the pressure plate (430) in the same shape.

[0294] Accordingly, the pressure provided from the pressure plate (430) in which the distributed pressure is formed can be transmitted to the upper plate (520). The upper plate (520) that has received the pressure can pressurize the molding object by facing the lower plate (510).

[0295] Therefore, the pressure plate (430) in which the pressure force distributed over the whole surface is formed can help form a uniform surface pressure on the forming unit (500).

[0296] In FIGS. 17 to 20, the shape of the pressure plate (430) is formed in a rectangular shape, and the guide rods (440) can be arranged symmetrically on both sides of the pressure rod (420) with the pressure rod (420) as the center. Accordingly, two guide rods (440) can be arranged so that they are connected to the pressure plate (430).

[0297] Referring to Fig. 22, the pressure plate (430) has an identical rectangular shape, and the guide rod (440) can be arranged symmetrically at the corner region of the pressure plate (430) with the pressure rod (420) as the center. Accordingly, four guide rods (440) can be arranged so that they are connected to the pressure plate (430).

[0298] In this way, by symmetrically arranging four guide rods (440) on the pressure plate (430) around the pressure rod (420), the distribution of the pressure force can be further improved.

[0299] Referring to Fig. 23, the pressure plate (430) may be formed in a triangular shape. Three guide rods (440) may be arranged on the triangular pressure plate (430). For example, when the pressure plate (430) is triangular in shape, three guide rods (440) may be arranged on the pressure plate (430) so that the guide rods (440) are arranged symmetrically around the pressure rod (420).

[0300] In this way, when the pressure plate (430) has a triangular shape, the pressure distribution can be further improved by arranging three guide rods (440) symmetrically around the pressure rod (420) on the pressure plate (430).

[0301] Referring to Fig. 24, the pressure plate (430) may be formed in a circular shape. Four to eight guide rods (440) may be arranged on the circular pressure plate (430). In the present embodiment, a case where eight guide rods (440) are arranged on the pressure plate (430) is illustrated.

[0302] Specifically, when the pressure plate (430) has a circular shape, 4 to 8 guide rods (440) can be placed on the pressure plate (430) so that the guide rods (440) are symmetrically placed around the pressure rod (420).

[0303] In this way, when the pressure plate (430) has a circular shape, the pressure distribution can be further improved by arranging 4 to 8 guide rods (440) symmetrically around the pressure rod (420) on the pressure plate (430).

[0304] FIG. 25 is a flowchart illustrating a method for manufacturing an eardrum regeneration patch using an eardrum regeneration patch manufacturing device according to an embodiment of the present invention, FIGS. 26 to 28 are flowcharts illustrating detailed methods for manufacturing an eardrum regeneration patch according to each step of FIG. 25, and FIGS. 29 to 34 are process diagrams illustrating a method for manufacturing an eardrum regeneration patch using an eardrum regeneration patch manufacturing device according to an embodiment of the present invention.

[0305] To avoid redundant explanation and for ease of explanation, FIGS. 25 to 34 will be described by citing FIGS. 17 to 20.

[0306] The method for manufacturing a tympanic membrane regeneration layer according to an embodiment of the present invention can form a tympanic membrane regeneration patch using a tympanic membrane regeneration patch manufacturing device (10).

[0307] The tympanic membrane regeneration patch manufacturing device (10) may include a transport unit (300), a forming unit (500), and a pressurizing unit (400). The forming unit (500) may include an upper plate (520) and a lower plate (520). A forming object may be placed between the upper plate (520) and the lower plate (510). The tympanic membrane regeneration patch manufacturing device (10) may provide pressure to the forming object to form a tympanic membrane regeneration patch with improved pattern uniformity on one or both sides.

[0308] The eardrum regeneration layer manufactured as above is positioned on the adhesive layer to manufacture the eardrum regeneration patch of the present invention.

[0309] Hereinafter, the present invention will be described in more detail with examples.

[0310]

[0311] Example

[0312] 1. Preparation of mold and polymer solution for manufacturing eardrum regeneration patches

[0313] (1) PUA mold production

[0314] A 600 μL PUA solution was applied using a micropipette onto a silicon master mold with a uniform nano-pattern measuring 800 nm in length, width, and height. A PET film was placed on the applied area and rolled thoroughly with a urethane roller to prevent bubbles. After confirming that there were no bubbles, the wafer and PET film were placed in a UV curing machine while still combined and UV-cured for 5 minutes. After 5 minutes, the master mold and wafer were removed, and the PET film and PUA mold were UV-cured for an additional 20 hours.

[0315]

[0316] (2) Fabrication of pattern-free PDMS mold

[0317] 27 g of PDMS base and 3 g of curing agent were mixed and dispensed into a petri dish (125 mm × 125 mm × 20 mm). The petri dish was then placed in a vacuum chamber and vacuumed at room temperature and a pressure of 0.1 MPa for 1 hour. After 1 hour, the vacuum was released, and after confirming that all air bubbles in the PDMS had disappeared, it was cured at 70°C for 5 hours. After curing, it was cut to produce a PDMS mold measuring 10 cm × 10 cm.

[0318]

[0319] (3) Fabrication of PDMS mold with nano-patterns

[0320] A mixture solution was prepared by mixing 27 g of PDMS base and 3 g of curing agent. The PUA mold was attached to a petri dish with the pattern facing upward, and the PDMS mixture solution was dispensed into the petri dish (125 mm × 125 mm × 20 mm). The petri dish was placed in a vacuum chamber, and the vacuum was maintained at room temperature and a pressure of 0.1 MPa for 1 hour. After 1 hour, the vacuum was released, and it was confirmed that all air bubbles in the PDMS had disappeared. Then, it was cured at a temperature of 70℃ for 5 hours, and after curing, it was cut to produce a PDMS mold measuring 10 cm × 10 cm. The manufactured PDMS mold had a uniform nano-pattern with a width, length, and height of 800 nm.

[0321]

[0322] (4) Preparation of polymer solution

[0323] PLGA (molar ratio of laclite and glycolide 75:25) and dichloromethane (DCM) were mixed for 8 hours to prepare PLGA solutions of 12% (w / w), 15% (w / w), 18% (w / w), and 21% (w / w).

[0324] PCL (poly(ε-caprolactone)) and dichloromethane (DCM) were mixed for 8 hours to prepare PCL solutions of 15% (w / w), 18% (w / w), 21% (w / w), and 24% (w / w).

[0325]

[0326] 2. Manufacturing of PLGA eardrum regeneration layer

[0327] (1) Manufacturing of the eardrum regeneration layer of Example 1

[0328] After attaching a 2 mm thick PDMS mold to the plate of the manufacturing device of Fig. 17, a 12% (w / w) PLGA solution (3 g) was evenly sprayed on the PUA mold with nano-patterns, and the PDMS mold was placed on top and rolled. Afterwards, it was compressed at a pressure of 0.5 MPa from the top (see Fig. 1). The PDMS mold was removed, and the organic solvent was evaporated in an 80°C drying oven for about 10 hours. The PLGA patch was then separated from the PUA mold to manufacture the tympanic membrane regeneration layer of Example 1.

[0329] The PDMS mold and PUA mold used here are designed to have a pattern spacing (distance between the peaks of the pattern) and a pattern height (height difference between the peaks and valleys of the pattern) of 800 nm each, and one side of the PLGA patch is patterned by the PDMS mold, and the other side is patterned by the PUA mold.

[0330]

[0331] (2) Preparation of the eardrum regeneration layer of Examples 2 to 10 and 23

[0332] The tympanic membrane regeneration layers of Examples 2 to 10 were prepared in the same manner as in Example 1, except that the concentration of the PLGA solution and the drying temperature were as shown in Table 1 below.

[0333] The tympanic membrane regeneration layer of Example 23 was manufactured so that the concentration of the PLGA solution and the drying temperature were the same as those of Example 4, the pattern was formed only on the cross-section, and the thickness was 30 μm.

[0334]

[0335] (3) Manufacturing of eardrum regeneration layer of comparative examples 1 to 3

[0336] The eardrum regeneration patches of Comparative Examples 1 and 2 were manufactured in the same manner as Example 1, except that the concentration of the PLGA solution and the drying temperature were as shown in Table 1 below.

[0337] Comparative Example 3 was manufactured using a non-patterned PDMS mold and a PUA mold. The concentration and drying temperature conditions of the PLGA solution used in the manufacture of the eardrum regeneration patch of Comparative Example 3 are as shown in Table 1 below.

[0338]

[0339] (4) Manufacturing conditions and thickness of the tympanic membrane regeneration layer of Examples 1-10 and 23 and Comparative Example 1-3

[0340] Examples 1-10 and 23 and Comparative Examples 1-3 were manufactured as shown in Table 1 below.

[0341] ClassificationPLGA Concentration % (w / w) Drying Temperature (℃) Pattern Thickness (㎛) Example 11280 Double-sided 14.3 Example 21560 Double-sided 16.9 Example 31570 Double-sided 16.4 Example 41580 Double-sided 15.1 Example 51860 Double-sided 19.1 Example 61870 Double-sided 18.3 Example 71880 Double-sided 17.8 Example 82160 Double-sided 21.1 Example 92170 Double-sided 19.2 Example 102180 Double-sided 18.6 Example 231580 Single-sided 30.0 Comparative Example 11260 Double-sided 16.5 Comparative Example 21270 Double-sided 15.6 Comparative Example 31580 None 15.4

[0342]

[0343] 3. Manufacturing of PCL eardrum regeneration layer

[0344] (1) Preparation of the eardrum regeneration layer of Examples 11 to 22

[0345] 10 g each of 15% (w / w), 18% (w / w), 21% (w / w), and 24% (w / w) PCL solutions were dropped onto tempered glass (10 cm × 10 cm × 1.1 mm) and spin-coated. The thickness of the patch was controlled by varying the rpm of the spin-coating. After spin-coating, the tempered glass was placed on a hot plate set to 70°C with the coated PCL facing up and melted for 5 minutes. A PDMS mold without nano-patterns was placed on the PCL layer and rolled with a urethane roller to prevent air bubbles. After that, [tempered glass, PCL, and PDMS] were placed on the plate of the patch manufacturing device of Fig. 17 set to 70°C and compressed at a pressure of 0.5 MPa for 5 minutes.

[0346]

[0347] (2) Manufacturing conditions and thickness of the tympanic membrane regeneration layer of Example 11-22

[0348] The PCL tympanic membrane regeneration patches of Examples 11-22 were manufactured as shown in Table 2 below.

[0349] Classification PCL Concentration % (w / w) Spin coating (rpm / concentration) Pattern thickness (㎛) Example 11 152,000 Section 28.9 Example 12 152,500 Section 22.6 Example 13 153,000 Section 18.6 Example 14 182,000 Section 30.4 Example 15 182,500 Section 24.5 Example 16 183,000 Section 20.6 Example 17 212,000 Section 31.8 Example 18 212,500 Section 26.6 Example 19 213,000 Section 21.1 Example 20 242,000 Section 32.9 Example 21 242,500 Section 27.3 Example 22,243,000 sections 22.7

[0350]

[0351] 4. Measurement of nano-pattern uniformity of the tympanic membrane regeneration layer

[0352] In Examples 1 to 23 and Comparative Examples 1 and 2, three points on the peak and two points on the valley were arbitrarily selected, and the height differences between the three points on the peak and the two points on the valley (see Fig. 2) were measured. The average value was substituted into Equation 1 to calculate the uniformity of the nano-pattern. The results are shown in Table 3 below.

[0353] Distinctive Material Pattern|H P1 -H P2 |(nm) Uniformity Evaluation Example 1 PLGA double-sided 9 Suitable Example 2 PLGA double-sided 0.2 Suitable Example 3 PLGA double-sided 0 Suitable Example 4 PLGA double-sided 0 Suitable Example 5 PLGA double-sided 0 Suitable Example 6 PLGA double-sided 0 Suitable Example 7 PLGA double-sided 0 Suitable Example 8 PLGA double-sided 1.1 Suitable Example 9 PLGA double-sided 8.2 Suitable Example 10 PLGA double-sided 5.4 Suitable Example 11 PCL cross-section 0.5 Suitable Example 12 PCL cross-section 0.5 Suitable Example 13 PCL cross-section 0 Suitable Example 14 PCL cross-section 0 Suitable Example 15 PCL cross-section 0 Suitable Example 16 PCL cross-section 0 Suitable Example 17 PCL cross-section 0 Suitable Example 18PCL cross-section 0 Suitable example 19PCL cross-section 0 Suitable example 20PCL cross-section 0.5 Suitable example 21PCL cross-section 0.4 Suitable example 22PCL cross-section 0 Suitable example 23PLGA cross-section 0 Suitable comparison example 1PLGA double-sided 100.2 Unsuitable comparison example 2PLGA double-sided 98.3 Unsuitable comparison example 3PLGA None 0 Unable to measure

[0354] Surface SEM images of the tympanic membrane regeneration layers of Examples 1 to 22 and Comparative Examples 1 and 2 are as shown in FIGS. 3a to 3e. It can be seen that the patterns of the tympanic membrane regeneration layers of Comparative Examples 1 and 2 are partially collapsed (FIG. 3e). The maximum height differences at arbitrary points of the peaks and valleys of the patterns of the tympanic membrane regeneration layers of Comparative Examples 1 and 2 were 100.2 nm and 98.3 nm, respectively. If the nano-pattern is uniform, tympanic membrane cells can proliferate continuously along the pattern, and if the nano-pattern is even partially collapsed, the growth and proliferation of tympanic membrane cells in that part can be stopped.

[0355]

[0356] 5. Measurement of the uniformity of the entire area of ​​the tympanic membrane regeneration layer

[0357] The area satisfying the uniformity of the tympanic membrane regeneration layer of Example 4 and Comparative Example 1 was calculated. According to the following Mathematical Expression 2, the uniformity of Example 4 was calculated to be 99.8% as shown in Fig. 4(A), and that of Comparative Example 1 was less than 70%.

[0358] [Equation 2]

[0359] Uniformity (area %) = (Area satisfying the uniformity of Equation 1 / Total area of ​​the patch)

[0360]

[0361] 6. Analysis of the structural and chemical properties of the tympanic membrane regeneration layer

[0362] The structural and chemical properties of the tympanic membrane regeneration layer of Example 4 and Comparative Example 3 were analyzed. Scanning electron microscopy (SEM) analysis was performed to analyze the surface structure, and FT-IR analysis was performed to confirm the chemical structure.

[0363] Scanning electron microscope (SEM) analysis results showed that the surface of the tympanic membrane regeneration layer of Comparative Example 3 was flat, and that an aligned pattern of 800 nm (800 nm in width, length, and thickness) was well formed on the surface of the tympanic membrane regeneration layer of Example 4. In addition, FT-IR analysis confirmed that the chemical structure of PLGA in the nano-patterned tympanic membrane regeneration layer of Example 4 was maintained the same (Fig. 5).

[0364]

[0365] 7. Analysis of the mechanical properties of the tympanic membrane regeneration layer

[0366] The mechanical properties (tensile strength and elongation) of the tympanic membrane regeneration layer of Example 4 and Comparative Example 3 were measured. The film model samples were cut into 10 mm wide × 50 mm long pieces and measured using a tissue analyzer (Stable Micro Systems Ltd., London, England). The initial grip-to-grip distance was 5 cm, and the crosshead speed was adjusted to 500 mm / min. The experiment was repeated three times to calculate the average and standard deviation. The tensile strength (TS, MPa) was expressed as the maximum tension recorded until the sample broke divided by the initial cross-sectional area of ​​the scaffold (Mathematical Formula 3), and the elongation (E, %) was expressed as the length increased until the sample broke as a percentage of the initial grip distance (Mathematical Formula 4):

[0367] [Equation 3]

[0368] TS(MPa) = F max / A

[0369] (In the equation, F max is the maximum tension (N) recorded until the sample breaks, A is the initial cross-sectional area of ​​the sample (m 2 )lim),

[0370] [Equation 4]

[0371] E(%) = (L / L0) × 100

[0372] (In the equation, L0 represents the initial grip-to-grip distance of the sample (mm), and L represents the length increased until the sample breaks (mm).)

[0373] As a result of the experiment, it was confirmed that both the tensile strength and elongation of Example 4 (Nano) were higher than those of Comparative Example 3 (Flat), as shown in Fig. 6, and it was confirmed that the curve was formed at the top in the strain-stress curve (Fig. 6a).

[0374] By the same method, the tensile strength and elongation of Examples 3, 5-7, 11-19 and 23 and Comparative Example 1-2 were measured, and the results are as shown in Table 4 below.

[0375] Classification Material Pattern Tensile strength (MPa) Elongation (%) Example 3 PLGA double-sided 5.4 6.0 Example 5 PLGA double-sided 7.5 6.9 Example 6 PLGA double-sided 7.1 6.5 Example 7 PLGA double-sided 6.9 6.3 Example 11 PCL cross-section 4.6 5 65 Example 12 PCL cross-section 4.3 5 46 Example 13 PCL cross-section 4.1 5 32 Example 14 PCL cross-section 4.8 5 95 Example 15 PCL cross-section 4.5 5 60 Example 16 PCL cross-section 4.2 5 35 Example 17 PCL cross-section 5.0 6 12 Example 18 PCL cross-section 4.6 5 68 Example 19 PCL cross-section 4.6 5 65 Example 23 PLGA cross-section 17.3 6.7 Comparative example 1PLGA double-sided 2.15.2 Comparative example 2PLGA double-sided 2.25.2

[0376] As the concentration of PLGA decreased, the tensile strength of the manufactured patch tended to weaken. Comparative Examples 1 and 2 were expected to have too low a mechanical strength to be suitable for use in surgery. Comparative Example 3 had a higher curing temperature than Comparative Examples 1 and 2, so the tensile strength increased somewhat, but it was expected that the adhesion would be low and the eardrum regeneration effect would be low because there was no pattern. In the case of the PLGA eardrum regeneration layer, it was confirmed that a relatively excellent uniformity eardrum regeneration layer was manufactured when the PLGA concentration was 15 to 21% (w / w) and the temperature of the drying process was 60-80℃.

[0377] In the case of the PCL tympanic membrane regeneration layer, examples 11 to 13 showed a tendency for the tensile strength to be relatively lower than other examples, and it was judged that it was clinically preferable to use a PCL concentration of 18% (w / w) or more.

[0378]

[0379] 8. Analysis of the adhesive strength of the tympanic membrane regeneration layer

[0380] For the treatment of tympanic membrane perforation, the patch must remain fully attached to the patient's tympanic membrane throughout the regeneration period, requiring an appropriate level of adhesive strength. Therefore, the adhesive strength of the tympanic membrane regeneration layers of Example 4 and Comparative Example 3 was compared.

[0381] The film model samples were cut into circular films with a diameter of 6 mm, and the tissue adhesion was measured using a tensile tester (MCT-1150 tensile tester, A&D Company, Japan). Before measuring the tissue adhesion, the pig dermis layer without contamination was washed three times with distilled water and used for the measurement. A circular tympanic membrane regeneration layer with an area of ​​2.8 cm2 was attached to the surface of the dermis layer by preloading it with ~0.5 N / cm2, and the crosshead speed was adjusted to 100 mm / min, and the adhesion was measured until separation occurred. The adhesion measurement was repeated 10 times for each sample under the same conditions at a relative humidity of 40% and an ambient temperature of 26℃, and the average and standard deviation were calculated.

[0382] [Equation 5]

[0383] Normal adhesion force = F max / A

[0384] (In the equation, F max is the maximum force recorded until the tympanic membrane regeneration patch detaches, and A is the initial area of ​​the tympanic membrane regeneration patch (㎠).

[0385] As a result of the analysis, as shown in Fig. 6b, the adhesive strength (adhesion strength) of Example 4 was superior to that of Comparative Example 3.

[0386] By the same method, the results of measuring the adhesive strength (adhesion strength) of Examples 3, 5-7, 11-19 and 23 and Comparative Example 1-2 are as shown in Table 5 below.

[0387] Classification Material Pattern Adhesion (N / ㎠) Example 3 PLGA double-sided 0.38 Example 5 PLGA double-sided 0.30 Example 6 PLGA double-sided 0.33 Example 7 PLGA double-sided 0.36 Example 11 PCL single-sided 0.32 Example 12 PCL single-sided 0.35 Example 13 PCL single-sided 0.42 Example 14 PCL single-sided 0.31 Example 15 PCL single-sided 0.35 Example 16 PCL single-sided 0.40 Example 17 PCL single-sided 0.28 Example 18 PCL single-sided 0.33 Example 19 PCL single-sided 0.38 Example 23 PLGA single-sided 0.36 Comparative Example 1 PLGA double-sided 0.28 Comparative Example 2 PLGA double-sided 0.29

[0388]

[0389] 9. Analysis of cell proliferation and migration patterns within the nanopattern of the tympanic membrane regeneration layer

[0390] Cells were cultured on the eardrum regeneration patches of Comparative Example 3 and Example 4, and the proliferation and migration patterns of the cells were observed. Fibroblasts, similar to eardrum cells, were used as the cells. In Comparative Example 3 (Flat) without nano-patterns, the fibroblasts tended to spread and proliferate, while in the eardrum regeneration layer (Nano) of Example 4 with nano-patterns, the fibroblasts were confirmed to align along the nano-patterns and proliferate and migrate (Fig. 7).

[0391]

[0392] 10. Evaluation of cell adhesion and function of the tympanic membrane regeneration layer

[0393] Fibroblasts were cultured on the tympanic membrane regeneration layer of Comparative Example 3 and Example 4, and cell attachment and proliferation were observed. Analysis of cell attachment 6 hours after fibroblast culture confirmed that attachment increased in the tympanic membrane regeneration layer (800 nm) of Example 4 compared to Comparative Example 3 (Flat), and this was confirmed to be the same in the proliferation analysis results on the 3rd and 5th days (Fig. 8). In particular, a similar proliferation rate was confirmed compared to the existing cell proliferation-specific cell culture vessel (TCPS) (Figs. 9 and 10, Flat: Comparative Example 3, 800 nm: Example 4).

[0394] In addition, cell movement was analyzed using a wound healing model. After attaching a 1 mm diameter SLAB (= material made of PDMS) to the eardrum regeneration layer of Example 4 and Comparative Example 3 to induce a wound (wound area = SLAB attachment area), fibroblasts were seeded and cultured for 24 hours. As a result of analyzing cell movement at 0, 12, and 24 hours after cell culture, both the cell movement distance and speed were higher in Example 4 than in Comparative Example 3, and it was confirmed that the wound healing range was particularly wide in the wound healing model (Figs. 10 and 11, Flat: Comparative Example 3, Nano: Example 4).

[0395]

[0396] 11. Effect of the tympanic membrane on tympanic membrane regeneration (in vivo)

[0397] (1) Effect of eardrum regeneration in Example 4

[0398] The results after attaching the tympanic membrane regeneration layer of Example 4 and the existing paper patch to an animal model of tympanic membrane perforation were analyzed. Figure 12 schematically illustrates the animal model tympanic membrane perforation and patch attachment process. The animal experiment results confirmed that the group applying the nano-patterned tympanic membrane regeneration patch of the present invention achieved a tympanic membrane regeneration rate that was more than 40% faster than the group applying the existing paper patch (Figure 13).

[0399]

[0400] (2) Effect of eardrum regeneration in Example 23

[0401] The tympanic membrane regeneration effect of the tympanic membrane regeneration layer of Example 23 was confirmed as follows. After perforation of both tympanic membranes in 20 male Sprague-Dawley rats, an observation period of a total of 14 days was set, and the tympanic membrane healing status was evaluated in three stages.

[0402] All Sprague-Dawley rats (8 weeks old) were anesthetized with chloral hydrate (1 g / mL, 10% dilution) and mechanical perforation was performed. After observing the tympanic membrane, if it was normal, a 40-50% perforation was created in the anterior portion of the tympanic membrane using a micropick under a Zeiss microscope.

[0403] The validity evaluation variable was the presence or absence of tympanic membrane perforation confirmed using a Zeiss microscope, and the tympanic membrane perforation was evaluated as cured only when complete healing was achieved.

[0404] Evaluation method

[0405] 1. Complete healing: No perforation

[0406] 2. Partial healing: The perforation size is reduced by more than 50%.

[0407] 3. No healing: If there is no change in the perforation size or the perforation size is reduced by less than 50%.

[0408] Evaluation results

[0409] The evaluation results are as shown in Figure 14.

[0410] The perforation healing rate was measured using a microscope in 20 SD rats (both ears). In the test group (NG patch), 14 rats completely healed within the first week, and 6 rats completely healed within the second week, showing complete healing within 2 weeks in all rats. In the control group (paper patch), only 3 rats healed within 1 week, and 4 rats ultimately failed to completely heal. Therefore, it can be seen that the tympanic membrane regeneration layer of the present invention has a fast recovery rate and excellent tympanic membrane regeneration effect.

[0411]

[0412] 12. Clinical trials using eardrum regeneration patches

[0413] (1) Use of the tympanic membrane regeneration layer of Example 23

[0414] A total of 27 clinical trials were conducted at Soonchunhyang University Hospital in Korea using the tympanic membrane regeneration layer of Example 23 (circular patch with a thickness of 30 μm and a diameter of 5 mm).

[0415] Clinical trials confirmed tympanic membrane regeneration and hearing improvement in over 50% of patients with chronic tympanic membrane perforation. Patient information for four of these cases is shown in Table 6 below. In particular, in the case of Patient 4, it was confirmed that even in cases where tympanic membrane reconstruction failed, the application of the tympanic membrane regeneration layer of the present invention resulted in tympanic membrane regeneration and hearing improvement (Figure 15). None of the patients complained of ear fluttering after the procedure.

[0416] Age / Gender / Status Patient 138, female, hearing loss and perforated eardrum Patient 232, female, hearing loss and perforated eardrum Patient 359, female, hearing loss and perforated eardrum Patient 435, female, hearing loss and perforated eardrum (tympanic membrane reconstruction failed)

[0417] However, in some cases, the tympanic membrane did not adhere continuously to the eardrum during the tympanic membrane perforation healing period, and prematurely detached after approximately 7 days. In cases where the tympanic membrane perforation remained adherent, the perforation size was significantly reduced, confirming tympanic membrane regeneration. Accordingly, an adhesive layer was added to the tympanic membrane perforation and the following clinical trial was conducted.

[0418]

[0419] (2) Use of the tympanic membrane regeneration layer + hydrocolloid layer of Example 4

[0420] The tympanic membrane regeneration layer of Example 4 was evaluated to have the most suitable properties for clinical use. The tympanic membrane regeneration layer of Example 4 was laminated onto a hydrocolloid layer to enable the tympanic membrane regeneration patch of the present invention with improved adhesiveness to be applied to a patient. As a result, the tympanic membrane regeneration layer of Example 24 formed on the hydrocolloid layer was designed to remain attached to the tympanic membrane perforation site for a long period of time.

[0421] A clinical trial using the tympanic membrane regeneration patch was conducted at Soonchunhyang University Hospital in Korea on 18 patients. Patients were selected if they had a perforated tympanic membrane for more than 3 months and had no inflammatory symptoms such as otitis media or pain.

[0422] The procedure for the patient was performed according to the following process: ① local anesthesia, ② perforation margin trimming, ③ application of the tympanic membrane regeneration layer of Example 4, ④ application of the hydrocolloid layer, ⑤ confirmation of the tympanic membrane regeneration effect at 4-week intervals.

[0423] The effect of tympanic membrane regeneration after the procedure was evaluated according to the following criteria.

[0424] - Healing rate = complete healing rate + partial healing rate

[0425] - Complete healing: No perforation of the eardrum

[0426] - Partial healing: The size of the tympanic membrane perforation is reduced by more than 50% after patch application.

[0427] - No healing: If the size of the eardrum perforation does not change or is reduced by less than 50% after applying the patch.

[0428] The method and results of this clinical trial are as shown in Fig. 16.

[0429] Among the 18 cases, the total healing rate was excellent at 61%, and the complete healing rate was high at approximately 50%. This indicates that patients with tympanic membrane perforation and chronic otitis media can be treated non-surgically using the tympanic membrane regeneration patch of the present invention.

[0430] Of the 18 cases, only one case of pain and one case of laryngeal ulcers were reported, both of which were minor complications. All of these cases improved after drug treatment.

[0431]

[0432] [Explanation of symbols]

[0433] 10: Eardrum regeneration patch manufacturing device 100: Stand

[0434] 110: Stand body 120: Table

[0435] 200: Frame unit 210: Frame body

[0436] 212: First through hole 215: Second through hole

[0437] 213: 1st cylinder guide 216: 2nd cylinder guide

[0438] 220: Frame support 300: Transport unit

[0439] 310: Transfer rail 320: Carrier

[0440] 322: sliding part 328: seating surface

[0441] 330: Transport drive unit 400: Pressurization unit

[0442] 410: Pressurized unit body 420: Pressurized rod

[0443] 430: Pressing plate 432: Pressing plate bottom

[0444] 437: Upper surface of the pressure plate 439: Mounting part

[0445] 440: Guide rod 450: Gimbal part

[0446] 470: Pressure sensor 500: Molding unit

[0447] 510: Lower plate 512: Lower plate

[0448] 517: Lower plate upper surface 520: Upper plate

[0449] 522: Upper plate pressurized surface 527: Upper plate mating surface

[0450] 550: Pressure distribution section 600: Input section

[0451] 700: Display part 800: Lower mold

[0452] 810: Second pattern surface 860: Second support surface

[0453] 900: Upper mold 910: First pattern surface

[0454] 960: 1st support surface 1000: Regeneration support body

[0455] 1000a: Biocompatible polymer solution 1000b: Semi-hardened

[0456] 1010: 1st pattern 1015: 1st side

[0457] 1020: Second pattern 1025: Second side

[0458] 1050: Pattern support part 1051: First pattern surface

[0459] 1052: Second pattern surface 1100: First mold

[0460] 1200: Second mold SA: Peripheral area

[0461] LA: Loading area PA: Forming area

Claims

1. A patch for eardrum regeneration containing: adhesive layer; and a regenerative layer of the eardrum formed on the adhesive layer, wherein the said regeneration layer of the eardrum is made of a biocompatible polymer, wherein on one surface or on both surfaces of the said regeneration layer of the eardrum a relief is formed for the growth of eardrum cells, wherein said relief consists of repeating protrusions and grooves, and wherein said layer of regeneration of the eardrum has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

2. A patch for regenerating the eardrum according to claim 1, wherein said eardrum regenerating layer has a relative elongation of 4% to 10%.

3. A patch for regenerating the eardrum according to claim 1, wherein the adhesive layer is a hydrocolloid layer.

4. The eardrum regeneration patch of claim 1, wherein the biocompatible polymer is poly(lactide-co-glycolide) containing 65 to 85 mol% lactide and 15 to 35 mol% glycolide.

5. A patch for regenerating the eardrum according to claim 1, wherein the biocompatible polymer is polycaprolactone.

6. The eardrum regeneration patch according to claim 1, wherein 99% or more of the relief area satisfies the uniformity condition (U) according to Equation 1 below: Equation 1 , where P1 and P2 represent, respectively, any point on the projection or any point in the groove, H P1 and H P2 denote the heights at points P1 and P2, respectively, and H n denotes the difference in height between the protrusion and the groove of a normal relief.

7. The eardrum regeneration patch according to claim 1, wherein any one component selected from the group consisting of collagen, fibrin, growth factors, stem cells, exosomes and therapeutic agents is applied to the surface of the eardrum regeneration layer that is not attached to the adhesive layer.

8. The eardrum regeneration patch according to claim 1, wherein the eardrum regeneration layer has a tensile strength of 15 to 20 MPa and a thickness of 10 μm to 30 μm.

9. The eardrum regeneration patch according to claim 1, wherein the eardrum regeneration layer has a tensile strength of 3 to 10 MPa and a thickness of 18 μm to 35 μm.

10. The eardrum regeneration patch according to claim 1, wherein the projections and grooves have a width and height of 700 to 900 nm each.

11. A method for manufacturing a patch for regenerating the eardrum according to any one of paragraphs 1-10, comprising: production of a regenerative layer of the eardrum by: applying a polymer solution containing a biocompatible polymer to a polyurethane acrylate mold to produce a semi-cured sheet-shaped product; placing said semi-cured product between said polyurethane acrylate mold and said polydimethylsiloxane mold and applying pressure using a top plate and a bottom plate to form a semi-cured product with a relief; and drying the said semi-hardened product with relief; and formation of an adhesive layer on the manufactured regeneration layer of the eardrum.

12. A method for producing a patch for regenerating the eardrum according to claim 11, wherein the concentration of the biocompatible polymer in the polymer solution is from 15% w / w to 24% w / w.

13. A method for producing a patch for regenerating the eardrum according to claim 11, wherein drying is performed at a temperature of 60°C to 80°C.

14. A patch for regenerating the eardrum according to claim 11, in which the protrusions and recesses for forming the relief are formed on a polyurethane acrylate mold or a polydimethylsiloxane mold.

15. The eardrum regeneration patch of claim 11, wherein the top plate includes a pressure rod and a guide rod.

16. The eardrum regeneration patch according to claim 11, wherein the pressure is from 0.40 Pa to 0.70 Pa.

17. A method for regenerating the eardrum, comprising: applying a layer of tympanic membrane regeneration to the perforation of the tympanic membrane; and application of a hydrocolloid adhesive layer to the regeneration layer of the eardrum, wherein the said regeneration layer of the eardrum is made of poly(lactide-co-glycolide) or polycaprolactone, wherein on one surface or on both surfaces of the regeneration layer of the eardrum a relief is formed for the growth of eardrum cells, wherein said relief consists of repeating protrusions and grooves, and wherein said layer of regeneration of the eardrum has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

18. The method for regenerating the eardrum according to claim 17, wherein the eardrum regeneration layer has a tensile strength of 15 to 20 MPa and a thickness of 10 μm to 30 μm.

19. The method for regenerating the eardrum according to claim 17, wherein the eardrum regeneration layer has a tensile strength of 3 to 10 MPa and a thickness of 18 μm to 35 μm.

20. The method for regenerating the eardrum according to claim 17, wherein the protrusions and grooves have a width and height of 700 to 900 nm each.