Tympanic membrane regenerative patch

Abstract

This invention relates to a tympanic membrane regeneration patch. Although the tympanic membrane regeneration patch of this invention has a thin thickness, it possesses high uniformity and appropriate elongation and tensile strength for a tympanic membrane regeneration patch, resulting in excellent tympanic membrane regeneration effect and excellent adhesion to the affected area. Furthermore, it ensures that postoperative patients do not experience discomfort such as tremors.

Description

Technical Field

[0001] This invention relates to a tympanic membrane regeneration patch. Background Technology

[0002] Chronic otitis media is a disease characterized by persistent tympanic membrane perforation and is one of the most common causes of hearing loss in the field of otolaryngology. However, most treatments still rely on surgical methods such as tympanic membrane regeneration. These surgical methods, including tympanic membrane regeneration, have many problems, such as high cost, complex procedures, the need for postoperative hospitalization, and a recurrence rate of over 10%.

[0003] To overcome the problems of these surgical methods, various approaches have been investigated, the oldest of which is the paper patch method. This involves cutting a paper patch and attaching it to the perforation site, allowing cells at the edge of the tympanic membrane perforation to grow using the paper patch as a scaffold. This paper patch method is generally effective for small, acute tympanic membrane perforations, but almost ineffective for chronic perforations. Furthermore, the use of non-biocompatible materials can induce inflammation, and the success rate is often less than 10%.

[0004] While there have been attempts to use biocompatible materials to create patches for tympanic membrane regeneration in mice, the regeneration results have been unsatisfactory, and the patches have shown poor adhesion, often detaching during activity. Furthermore, because these attempts were not based on clinical trials on human subjects, they have limitations in fully considering patients' hearing discomfort.

[0005] Therefore, there is an urgent need to develop a tympanic membrane regeneration patch that has excellent adhesion and tympanic membrane regeneration effect and ensures that patients do not experience hearing discomfort after surgery. Summary of the Invention

[0006] Problems to be solved

[0007] The present invention aims to provide a tympanic membrane regeneration patch that has excellent tympanic membrane regeneration effect, is not easy to fall off from the tympanic membrane perforation, is flexible and thin, and has excellent tensile strength, so that there is no discomfort such as tympanic membrane tremor after surgery.

[0008] Problem Solving Methods

[0009] The present invention provides a tympanic membrane regeneration patch, comprising an adhesive layer and a tympanic membrane regeneration layer formed on the adhesive layer, wherein the tympanic membrane regeneration layer is composed of a biocompatible polymer, and a pattern for tympanic membrane cell growth is formed on one or both sides of the tympanic membrane regeneration layer. The pattern consists of repeated peaks and valleys. The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

[0010] Preferably, the tympanic membrane regeneration layer of the above-mentioned tympanic membrane regeneration patch exhibits an elongation of 4% to 10%.

[0011] The tympanic membrane regeneration layer of the above-mentioned tympanic membrane regeneration patch can have an elongation of 500% to 700% depending on the material.

[0012] The pattern of the aforementioned tympanic membrane regeneration patch has a uniformity (U) of over 99% of its area, as shown in Equation 1 below:

[0013]

Mathematical Formula 1

[0014] U=|H P1 -H P2 |≤0.1H n

[0015] In mathematical formula 1, P1 and P2 are points on the peak or on the valley, respectively, and H... P1 and H P2 H represents the height at points P1 and P2, respectively. n This represents the height difference between the peaks and valleys in a normal pattern.

[0016] This invention provides a method for manufacturing a tympanic membrane regeneration patch, the method comprising: a step of manufacturing a tympanic membrane regeneration layer, the step of coating a polymer solution containing a biocompatible polymer onto a polyurethane acrylate mold to manufacture a sheet-like semi-cured material; a step of placing the semi-cured material between a polyurethane acrylate mold and a polydimethylsiloxane mold, and then applying pressure using an upper plate and a lower plate respectively to manufacture a patterned semi-cured material; a step of drying the patterned semi-cured material; and a step of forming an adhesive layer on the tympanic membrane regeneration layer manufactured as above.

[0017] In the above manufacturing method, the concentration of biocompatible polymer in the polymer solution can be from 15% (w / w) to 24% (w / w).

[0018] In the above manufacturing method, drying can be carried out at 60°C to 80°C.

[0019] In the above manufacturing method, the polyurethane acrylate mold or the polydimethylsiloxane mold may be formed with concave and convex shapes for forming patterns.

[0020] In the above manufacturing method, the upper plate may have a pressure bar and a guide bar.

[0021] In the above manufacturing method, pressure can be applied in the range of 0.40 Pa to 0.70 Pa.

[0022] Invention Effects

[0023] The tympanic membrane regeneration patch of the present invention has excellent tympanic membrane regeneration effect.

[0024] The tympanic membrane regeneration patch of the present invention has excellent adhesion to the affected area, thus making it less likely to fall off from the tympanic membrane perforation.

[0025] The tympanic membrane regeneration patch of the present invention will not cause tympanic membrane vibration when the patient moves or talks after surgery, so the patient will not feel discomfort after surgery.

[0026] The tympanic membrane regeneration patch of the present invention is flexible, thin and light, and has excellent tensile strength, so that it can make appropriate vibration response to sound (sound waves) in the air, and maintain its adhesion even in tympanic membrane environments of various structures or shapes.

[0027] The tympanic membrane regeneration patch of the present invention is flexible, thin, and lightweight, while having excellent tensile strength, thus making it easy for doctors to operate.

[0028] The tympanic membrane regeneration patch of the present invention is flexible, thin, and lightweight, while having excellent tensile strength, making it less prone to tearing.

[0029] The tympanic membrane regeneration patch of the present invention has excellent uniformity of pattern, which is beneficial to tympanic membrane cell regeneration.

[0030] The tympanic membrane regeneration patch of the present invention does not cause discomfort such as tympanic membrane tremor in daily life after surgery. Attached Figure Description

[0031] Figure 1 This is a conceptual diagram of the manufacturing process of a tympanic membrane regeneration patch according to an embodiment of the present invention.

[0032] Figure 2 A method for measuring the uniformity (U) of mathematical formula 1 of this application is shown.

[0033] Figures 3a to 3e These are surface SEM images of the tympanic membrane regeneration patches of Examples 1 to 22 and Comparative Examples 1 and 2.

[0034] Figure 4 These are comparative photographs showing the uniformity (area %) of the tympanic membrane regeneration patches of Example 4 and Comparative Example 1. The tympanic membrane regeneration patch of the present invention (Example 4) has excellent area uniformity for almost the entire area, while in Comparative Example 1, the area with excellent uniformity is less than 70% of the total area.

[0035] Figure 5 The results are SEM and FR-IR analyses of the tympanic membrane regeneration patches of Comparative Example 3 and Example 4.

[0036] Figure 6a and Figure 6b Comparative data on tensile strength and elongation, as well as adhesive strength (attachment strength) of the tympanic membrane regeneration patches of Comparative Example 3 and Example 4 (Flat: Comparative Example 3, Nano: Example 4).

[0037] Figure 6c This is a photograph showing the flexibility of the tympanic membrane regeneration layer in Example 23.

[0038] Figure 7 and Figure 8 The fibroblast proliferation and migration morphology of the tympanic membrane regeneration patches of Comparative Example 3 and Example 4 are shown (Flat: Comparative Example 3, Nano: Example 4).

[0039] Figure 9 The results are obtained by culturing fibroblasts in the tympanic membrane regeneration patches of Comparative Example 3 and Example 4, analyzing and quantifying the degree of cell adhesion and proliferation (TCPS: TCPS cell culture container, Flat: Comparative Example 3, Nano: Example 4).

[0040] Figure 10 and Figure 11 The results are obtained by comparing the cell migration distance, migration speed and wound healing extent of the tympanic membrane regeneration patches of Comparative Example 3 and Example 4 in a wound healing model (Flat: Comparative Example 3, Nano: Example 4).

[0041] Figure 12 This illustrates the animal experimental methods using a tympanic membrane perforation animal model.

[0042] Figure 13 Animal experiment results are shown using existing paper patches and the tympanic membrane regeneration patch of Example 4.

[0043] Figure 14 Animal experiment results using the tympanic membrane regeneration patch of Example 23 are shown.

[0044] Figure 15 and Figure 16 The results of a clinical trial using the tympanic membrane regeneration patch of Example 4 are shown.

[0045] Figure 17 This is a perspective view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0046] Figure 18 This is a front view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0047] Figure 19 This is a top view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0048] Figure 20 It is along Figure 19 A cross-sectional view of line A1-A2.

[0049] Figure 21This is a block diagram illustrating a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0050] Figure 22 This is a top view showing the shape of the pressure plate of the pressurizing unit according to an embodiment of the present invention.

[0051] Figure 23 This is a top view showing another shape of the pressure plate of the pressurizing unit according to an embodiment of the present invention.

[0052] Figure 24 This is a top view showing another shape of the pressure plate of the pressurizing unit according to an embodiment of the present invention.

[0053] Figure 25 This is a flowchart illustrating a method for manufacturing a tympanic membrane regeneration patch using a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0054] Figures 26 to 28 It is based on Figure 25 A detailed flowchart of the manufacturing process for each step of the tympanic membrane regeneration patch.

[0055] Figures 29 to 34 This is a process diagram illustrating a method for manufacturing a tympanic membrane regeneration patch using a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0056] Figure 35 This is a schematic diagram of a tympanic membrane treatment method using the tympanic membrane regeneration patch of the present invention.

[0057] Symbol explanation:

[0058] 10: Tympanic membrane regeneration patch manufacturing device; 100: Support;

[0059] 110: Support body, 120: Workbench,

[0060] 200: Frame unit, 210: Frame body

[0061] 212: First through hole, 215: Second through hole

[0062] 213: First cylinder guide component; 216: Second cylinder guide component.

[0063] 220: Frame support section; 300: Conveying unit;

[0064] 310: Conveyor rail; 320: Carrier.

[0065] 322: Sliding part, 328: Mounting surface,

[0066] 330: Transmission drive unit; 400: Pressurization unit;

[0067] 410: Main body of the pressurization unit; 420: Pressure bar;

[0068] 430: Pressure plate; 432: Below the pressure plate.

[0069] 437: Above the pressure plate; 439: Placement section.

[0070] 440: Guide rod, 450: Universal joint section

[0071] 470: Pressure sensor; 500: Molding unit.

[0072] 510: Bottom board; 512: Below the bottom board.

[0073] 517: Above the lower plate, 520: Upper plate,

[0074] 522: Pressure surface of the upper plate, 527: Joint surface of the upper plate,

[0075] 550: Pressure distribution section, 600: Input section,

[0076] 700: Display section, 800: Lower mold,

[0077] 810: Second pattern surface, 860: Second support surface

[0078] 900: Upper mold, 910: First pattern surface,

[0079] 960: First support surface; 1000: Regenerated support body.

[0080] 1000a: Biocompatible polymer solution; 1000b: Semi-solidified product.

[0081] 1010: First pattern, 1015: First side.

[0082] 1020: Second pattern, 1025: Second side,

[0083] 1050: Pattern support section; 1051: First pattern surface;

[0084] 1052: Second pattern surface, 1100: First mold,

[0085] 1200: Second mold; SA: Surrounding area.

[0086] LA: Loading area, PA: Molding area Detailed Implementation

[0087] This invention provides the following tympanic membrane regeneration patch.

[0088] Tympanic membrane regeneration patch

[0089] The present invention provides a tympanic membrane regeneration patch, comprising an adhesive layer and a tympanic membrane regeneration layer formed on the adhesive layer. The tympanic membrane regeneration layer is composed of a biocompatible polymer. Patterns for tympanic membrane cell growth are formed on one or both sides of the tympanic membrane regeneration layer. The patterns consist of repeated peaks and valleys. The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

[0090] The tympanic membrane regeneration patch of the present invention has a tympanic membrane regeneration layer with a tensile strength of 3 to 20 MPa, a thickness of 10 μm to 40 μm, and an elongation of 4% to 10%.

[0091] The adhesive layer is made of a material that allows the tympanic membrane regeneration layer to be properly adhered to the affected area of ​​the tympanic membrane. For example, a hydrocolloid material used as a wound moistening patch can be used.

[0092] When hydrocolloid layers are used as adhesive layers, they can exhibit anti-inflammatory activity.

[0093] The adhesive layer has a larger cross-sectional area than the tympanic membrane regeneration layer, which facilitates adhesion. For example, when the tympanic membrane regeneration layer is coin-shaped, the adhesive layer can also be coin-shaped with a larger radius than the tympanic membrane regeneration layer.

[0094] The shapes of the adhesive layer and the tympanic membrane regeneration layer can be appropriately determined based on the perforation shape, the patch application environment (humidity, curvature, tilt, etc.).

[0095] The tympanic membrane regeneration patch of the present invention includes a tympanic membrane regeneration layer composed of a biocompatible polymer.

[0096] Biocompatible polymers can be any type of polymer known to be usable in living organisms. Examples include polycaprolactone (PCL), lactide-glycolic acid copolymer (PLGA), polyethylene glycol (PEG), polyethylene oxide (PEO), polylactic acid (PLA), and polyglycolic acid (PGA).

[0097] Regarding the uniformity of the pattern and the physical properties (elongation, tensile strength, etc.) of the tympanic membrane regeneration layer, PLGA or PCL is preferred as a biocompatible polymer. More preferably, in PLGA, the molar ratio of lactide to glycolide is 65-85:15-35.

[0098] Patterns for tympanic membrane cell growth are formed on one or both sides of the tympanic membrane regeneration layer of the present invention.

[0099] When the tympanic membrane regeneration patch is applied to the affected area, the tympanic membrane cells present around the patch grow along the valleys of the pattern, thereby inducing the regeneration of the damaged tympanic membrane.

[0100] Patterns can be formed on one or both sides. To easily distinguish the top and bottom surfaces of the tympanic membrane regeneration patch, the pattern can be formed only on the top or bottom surface. Furthermore, when the tympanic membrane regeneration patch is small, the pattern can be formed on both sides, thus eliminating the distinction between top and bottom.

[0101] Since the tympanic membrane regeneration layer is formed on the adhesive layer, patterns must be formed on the surfaces of the tympanic membrane regeneration layer that are not in contact with the adhesive layer for tympanic membrane cell growth.

[0102] The patterns formed on the tympanic membrane regeneration layer not only affect tympanic membrane cell growth but also the physical properties of the tympanic membrane regeneration layer. If patterns are formed on both sides of the tympanic membrane regeneration layer, the tensile strength and elongation can be improved compared to patterns formed on only one side.

[0103] As long as the shape is a repetitive pattern of peaks and valleys, the pattern is not limited to a specific shape. For example, the pattern can be a straight line or a curve with peaks and valleys side by side and repeating. The shape of the pattern can be formed as a straight line parallel to one edge of the tympanic membrane regeneration patch, a diagonal line at a predetermined angle to one edge, a wavy curve, etc.

[0104] A peak is a relatively prominent part, and a valley is a relatively concave part that exists between peaks. Tympanic membrane cells grow along the valleys between peaks.

[0105] The width between peaks 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, 600 to 700 nm, etc.

[0106] The width between valleys can also be designed to be the same as the width between peaks.

[0107] The height difference between the peak and the valley (the height of the peak when the valley is taken as the reference) 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, 600 to 700 nm, etc.

[0108] The distances between peaks, the distances between valleys, and the height differences between peaks and valleys in the pattern can be the same or different. When they are different, the differences can be below 100 nm, below 90 nm, below 80 nm, below 70 nm, below 60 nm, or below 50 nm, respectively.

[0109] For example, the distances between peaks, the distances between valleys, and the height differences between peaks and valleys can all be the same, such as 800 nm, 700 nm, or 600 nm. Alternatively, the distances between peaks and valleys can be 800 nm, and the height differences between peaks and valleys can be 750 nm or 850 nm. Alternatively, the distances between peaks and valleys can be 700 nm, and the height differences between peaks and valleys can be 650 nm or 750 nm.

[0110] The distance between peaks, the distance between valleys, and the height difference between peaks and valleys can be changed by adjusting the size and spacing of the pattern formed on the mold used in the manufacture of tympanic membrane regeneration patches.

[0111] The area of ​​the pattern that satisfies the uniformity (U) of the following mathematical formula 1 can be more than 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% of the total area.

[0112]

Mathematical Formula 1

[0113] U=|H P1 -H P2 |≤0.1H n

[0114] In mathematical formula 1, P1 and P2 are points on the peak or on the valley, respectively, and H... P1 and H P2 H represents the height at points P1 and P2, respectively. n This represents the height difference between the peaks and valleys in a normal pattern.

[0115] P1 and P2 can be any points on the peaks of the pattern. P1 and P2 can also be any points on the valleys of the pattern.

[0116] For example, P1 could be a point on a certain peak, and P2 could be a point on a peak adjacent to that peak, that is, a point on another peak after crossing a valley located to the left or right of that peak. Alternatively, P1 could be a point on a certain peak, and P2 could be a point on a peak separated from that peak, that is, a point on another peak after crossing multiple valleys located to the left or right of that peak.

[0117] HP1 is the height at point P1. The height at P1 refers to the height from the valley to a point P1 that exists as a peak. If the peak forms the expected height, then HP1 is the same as the height of the pattern formed on the mold, and the difference H between the heights of the peaks and valleys as a normal pattern is... n Same. When the pattern is poorly formed, the height of HP1 differs from that of the pattern formed on the mold.

[0118] HP2 refers to the height at point P2. It is the same as HP1.

[0119] In the tympanic membrane regeneration layer, the height difference between points P1 and P2 is the height difference between the peaks and valleys of the normal pattern (H). n The percentages are below 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%.

[0120] If the pattern of the tympanic membrane regeneration layer is highly uniform, the cell proliferation of tympanic membrane cells will proceed smoothly along the uniform pattern, resulting in excellent tympanic membrane regeneration. If the pattern is uneven and partially damaged, cell proliferation may cease in the damaged areas.

[0121] When P1 and P2 are points on the same peak, 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 be selectively considered further.

[0122] P3 and P4 can be points on the same valley. Alternatively, P3 can be a point on one valley, and P4 can be a point on a valley adjacent to that valley, i.e., a point on another valley after crossing a peak located to the left or right of that valley. Or, P3 can be a point on one valley, and P4 can be a point on a valley separated from that valley, i.e., a point on another valley after crossing multiple peaks located to the left or right of that valley.

[0123] When P1 and P2 are arbitrary points on the peak, and P3 and P4 are arbitrary points on the valley, U1=|H P1 -H P2 |≤0.1H n and U2=|H P3 -H P4 |≤0.1H n (H) P3 H refers to the height at point P3. P4 The area of ​​the height at P4 (where Hn is the height difference between the peaks and valleys of the normal pattern) can be more than 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% of the total area.

[0124] The thickness of the tympanic membrane regeneration layer is 10 to 40 μm. When the thickness is less than 10 μm, the mechanical strength (tensile and compressive strength) is low, making the patch or the nanopatterns formed on the patch easily damaged, and patients may experience discomfort due to patch tremor after surgery. When the thickness is greater than 40 μm, the adhesion is reduced, making it difficult for the patch to maintain adhesion to the tympanic membrane for the sufficient time required for tympanic membrane regeneration.

[0125] When the thickness exceeds 40μm, the flexibility of the tympanic membrane regeneration patch decreases, making it difficult to respond appropriately to sound (sound waves) vibrations, or it may not be able to maintain adhesion in tympanic membrane environments of various structures or shapes.

[0126] The optimal thickness of the tympanic membrane regeneration layer can vary depending on the material. For example, when the tympanic membrane regeneration layer is made of PLGA material, its thickness is preferably 10µm to 30µm, more preferably 15µm to 20µm. For example, when the tympanic membrane regeneration layer is made of PCL material, its thickness is preferably 18µm to 35µm, more preferably 25µm to 30µm.

[0127] The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa. Because of this tensile strength, the tympanic membrane regeneration layer can maintain adhesion to tympanic membranes of various structures or shapes, even when it is soft and thin. It also provides an appropriate vibrational response to sound waves in the air, making it easy for doctors to manipulate during procedures, even with its small size.

[0128] The optimal tensile strength of the tympanic membrane regeneration layer varies depending on the material and thickness. For example, when the tympanic membrane regeneration layer is made of PLGA material with a thickness of 10µm to 20µm, the tensile strength can be 3MPa to 8MPa. For example, when the tympanic membrane regeneration layer is made of PLGA material with a thickness of 20µm to 30µm, the tensile strength can be 15MPa to 20MPa. For example, when the tympanic membrane regeneration layer is made of PCL material with a thickness of 18µm to 35µm, the tensile strength can be 4MPa to 6MPa.

[0129] The tympanic membrane regeneration layer can have an elongation rate of 4% to 10%. When the elongation rate exceeds 10%, it is prone to deformation and difficult to handle, and patients may experience discomfort due to patch vibration after surgery. When the elongation rate is less than 4%, it is prone to breakage and lacks flexibility, which may lead to problems with proper adhesion to the tympanic membrane.

[0130] The elongation of the tympanic membrane regeneration layer is preferably 5% to 7%.

[0131] When the tympanic membrane 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 tympanic membrane regeneration effect of the present invention can be expected; however, due to the high elongation, the adhesion may be reduced depending on the tympanic membrane adhesion environment.

[0132] The elongation of the tympanic membrane regeneration layer is preferably 5% to 7%.

[0133] The adhesion strength of the tympanic membrane regeneration layer can be 0.2 to 0.5 N / cm², preferably 0.3 to 0.4 N / cm².

[0134] The surface of the tympanic membrane regeneration layer that is not bonded to the adhesive layer can be coated with any one of the following: collagen, fibrin, growth factors, stem cells, exosomes, and therapeutic drugs. This helps with adhesion or promotes cell growth.

[0135] For example, fibrin coated on the edge of the tympanic membrane regeneration layer can form fibrous blood clots, which act as a bridge for cell migration, thereby enabling cells in the tympanic membrane perforation site to migrate and proliferate smoothly, thus improving the speed of tympanic membrane regeneration.

[0136] When attaching the tympanic membrane regeneration layer to the adhesive layer, medical adhesives or other adhesives may be used as needed.

[0137] This invention provides a method for manufacturing the following tympanic membrane regeneration patch.

[0138] Manufacturing method of tympanic membrane regeneration patch

[0139] The present invention provides a method for manufacturing a tympanic membrane regeneration patch, the method comprising: (1) manufacturing a tympanic membrane regeneration layer by a predetermined method; and (2) forming an adhesive layer on the tympanic membrane regeneration layer manufactured in step (1).

[0140] Step (1) can be constructed as follows.

[0141] The manufacturing steps may include: (1-1) coating a polymer solution containing a biocompatible polymer onto a polyurethane acrylate (PUA) mold to manufacture a sheet-like semi-cured material; (1-2) positioning the semi-cured material between a polyurethane acrylate (PUA) mold and a polydimethylsiloxane (PDMS) mold, and then applying pressure using an upper plate and a lower plate respectively to manufacture a patterned semi-cured material; and (1-3) drying the patterned semi-cured material.

[0142] In the above manufacturing method, the concentration of biocompatible polymer in the polymer solution can be from 15% (w / w) to 24% (w / w).

[0143] In the above manufacturing method, drying can be carried out at 60°C to 80°C.

[0144] In the above manufacturing method, the polyurethane acrylate mold or the polydimethylsiloxane mold may be formed with concave and convex shapes for forming patterns.

[0145] In the above manufacturing method, the upper plate may have a pressure bar and a guide bar.

[0146] In the above manufacturing method, the pressure can be applied from 0.40 Pa to 0.70 Pa.

[0147] More specifically, the tympanic membrane regeneration layer can be manufactured by a method including a semi-cured material forming step S100, a semi-cured material fixing step S200, a conveying step S300, a pressurizing step S400, a drying step S500, and a separation step S600.

[0148] refer to Figure 26 and Figure 29 (A) The semi-cured material forming step S100 may include step S110 of setting the upper mold 900 on the surface of the substrate. The substrate may be the ground, a structure with a flat surface, or a lower plate 510. When the substrate surface is the lower plate 510, the lower plate 510 may be set on the loading area LA. In this embodiment, the case where the substrate is the lower plate 510 will be illustrated.

[0149] The upper mold 900 is a polyurethane acrylate (PUA) mold.

[0150] The upper mold 900 may include a first patterned surface 910 on which a pattern is formed and a first support surface 960 disposed on the opposite side of the first patterned surface 910. The first support surface 960 may be placed on the substrate surface.

[0151] The first pattern surface 910 may include a pattern with a predetermined shape. The pattern is not limited to a specific shape, as long as it is a repeating peak and valley shape. For example, the pattern may be a straight line or a curve with side-by-side peaks and valleys repeating. The shape of the pattern may be a straight line parallel to one edge of the tympanic membrane regeneration patch, a diagonal line at a predetermined angle to one edge, a wavy curve, etc.

[0152] The width between peaks can be designed in various ways to match the pattern of the tympanic membrane 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.

[0153] The width between valleys can also be designed to be the same as the width between peaks.

[0154] The height difference between the peak and the valley (the height of the peak when the valley is used as a reference) can be designed in various ways to match the pattern of the tympanic membrane 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.

[0155] The distances between peaks, the distances between valleys, and the height differences between peaks and valleys in the pattern can be the same or different. When they are different, the differences can be below 100 nm, below 90 nm, below 80 nm, below 70 nm, below 60 nm, or below 50 nm, respectively.

[0156] For example, the distances between peaks, the distances between valleys, and the height differences between peaks and valleys can all be the same, such as 800 nm, 700 nm, or 600 nm. Alternatively, the distances between peaks and valleys can be 800 nm, and the height differences between peaks and valleys can be 750 nm or 850 nm. Alternatively, the distances between peaks and valleys can be 700 nm, and the height differences between peaks and valleys can be 650 nm or 750 nm.

[0157] The size and spacing of the pattern in the manufactured tympanic membrane regeneration patch can be changed by adjusting the size and spacing of the pattern formed on the mold.

[0158] refer to Figure 26 and Figure 29 (B) Step S100 of forming the semi-cured material 1000b may include step S120 of coating the biocompatible polymer solution 1000a onto the first patterned surface 910 of the upper mold 900. By coating the biocompatible polymer solution 1000a onto the PUA mold (upper mold, 900), a sheet-like semi-cured material 1000b can be formed.

[0159] Biocompatible polymer solutions can be prepared by mixing biocompatible polymers with organic solvents. For example, halomethane solvents such as chloroform or dichloromethane can be used as organic solvents.

[0160] The appropriate concentration of a biocompatible polymer solution can vary depending on the type of biocompatible polymer. For example, the concentration of biocompatible polymer in a biocompatible polymer solution can range from 15% (w / w) to 24% (w / w). When the biocompatible polymer is PLGA, the concentration can range from 15% (w / w) to 21% (w / w), and when the biocompatible polymer is PCL, the concentration can range from 18% (w / w) to 24% (w / w).

[0161] During the coating process of biocompatible polymer solution 1000a, some of the solvent evaporates at room temperature, forming a sheet-like semi-cured material 1000b. Therefore, even if the upper mold is flipped 900, the semi-cured material will not flow downwards.

[0162] The upper mold 900 on which the semi-cured material 1000b is provided on the first pattern surface 910 is called the first mold 1100.

[0163] Since the first mold 1100 has a semi-cured material 1000b on the upper mold 900, the first mold 1100 must be flipped over to contact the lower mold 800.

[0164] Therefore, in order to bring the surface of the semi-cured material 1000b exposed by flipping the first mold 1100 into contact with the lower mold 800, a high-viscosity biocompatible polymer can be used, or a highly volatile solvent can be used to make the solvent evaporate quickly, thereby increasing the viscosity of the semi-cured material 1000b and preventing it from flowing downward.

[0165] refer to Figure 25 The method for manufacturing the tympanic membrane regeneration layer may include step S200 of fixing the semi-cured material 1000b. Fixing the semi-cured material 1000b is a step of contacting the semi-cured material 1000b exposed by flipping the first mold 1100 with the lower mold 800. The mold formed by contacting the lower mold 800 with the first mold 1100 is called the second mold 1200.

[0166] The lower mold 800 is a polydimethylsiloxane (PDMS) mold.

[0167] refer to Figure 27 and Figure 30 (A) Step S200 of fixing the semi-cured material 1000b may include step S210 of placing the carrier 320 on the loading area LA.

[0168] The lower plate 510 can be disposed on the carrier 320. For example, the lower plate underside 512 can be placed on the carrier mounting surface 328. In the loading area LA, the lower plate 510 can be configured to expose the upper plate surface 517.

[0169] refer to Figure 27 and Figure 30 (B) Step S200 of fixing the semi-cured material 1000b may include step S220 of setting the lower mold 800 on the lower plate 510. For example, in the loading area LA, the lower mold 800 may be set on the upper plate 517.

[0170] The lower mold 800 may include a flat, unpatterned surface and a second support surface 860 formed on the opposite side of the flat surface. The second support surface 860 may be mounted on the lower plate 517. Therefore, when the lower mold 800 is mounted on the lower plate 510 in the loading area LA, the flat surface may be exposed to the outside. The lower mold 800 with the unpatterned flat surface is used in the manufacture of single-sided nano-patterned tympanic membrane regeneration patches.

[0171] The lower mold 800 may include a second patterned surface 810 and a second support surface 860 formed on the opposite side of the second patterned surface 810. The second patterned surface 810 may be a flat surface without a pattern or a patterned surface. When it is a flat surface without a pattern, it is used in the manufacture of single-sided patterned tympanic membrane regeneration patches; when it is a patterned surface, it is used in the manufacture of double-sided patterned tympanic membrane regeneration patches. The second support surface 860 may be mounted on the lower plate 517. Therefore, when the lower mold 800 is mounted on the lower plate 510 in the loading area LA, the second patterned surface 810 may be exposed to the outside.

[0172] refer to Figure 27 and Figure 30 (C) Step S200 of fixing the semi-cured material 1000b may include step S230 of contacting the first mold 1100 with the lower mold 800. For example, the semi-cured material can be fixed by setting the first mold 1100 so that the semi-cured material 1000b contacts the second patterned surface 810.

[0173] Step S200 of fixing the semi-cured material 1000b can expose the first support surface 960 of the upper mold 900 to the outside on the loading area LA.

[0174] refer to Figure 25 and Figure 31 The method for manufacturing the tympanic membrane regeneration layer may include a conveying step S300 of conveying the second mold 1200.

[0175] In the transfer step S300, the second mold 1200, which is set on the loading area LA, can be moved to the molding area PA.

[0176] The pressure plate 430 of the pressure unit 400 can be disposed above the molding area PA. The upper plate 520 can be combined with the pressure plate 430. Therefore, the second mold 1200 can be disposed below the upper plate 520.

[0177] For example, pressure plate 430 may include a lower pressure plate 432 and a upper pressure plate 437. Upper plate 520 may be combined with lower pressure plate 432.

[0178] The upper plate 520 may include an upper plate pressure surface 522 and an upper plate mating surface 527. The lower plate 432 may be mated with the upper plate mating surface 527. Furthermore, the pressure dispersing part 550 may be adhered to the upper plate pressure surface 522.

[0179] A conveying unit 300 that moves the second mold 1200 via the second mold 1200 and a lower plate 510 disposed on the conveying unit 300 may be provided at the lower part of the molding area PA.

[0180] For example, the conveying unit 300 may include a conveying guide rail 310 and a carrier 320. A lower plate 510 may be disposed on the carrier 320. A second mold 1200 may be disposed on the lower plate 510. Therefore, the second mold 1200 may be disposed in the molding area PA.

[0181] The transfer unit 300 may also include a transfer drive unit 330 capable of moving the carrier 320. By driving the transfer drive unit 330, the carrier 320 can be moved from the loading area LA to the molding area PA.

[0182] Therefore, the second mold 1200 can be disposed in the direction below the pressure dispersion section 550 in the molding area PA. For example, in the transfer step S300, the pressure dispersion section 550 and the first support surface 960, which is the exposed surface of the second mold 1200, can be disposed opposite to each other in the molding area PA.

[0183] refer to Figure 25 A method for manufacturing a tympanic membrane regeneration layer according to one embodiment may include a pressurization step S400 of pressurizing a fixed semi-cured material 1000b.

[0184] For example, refer to Figure 26 and Figure 32 (A) The pressurization step S400 can be performed in the molding area PA.

[0185] The pressurizing step S400, which pressurizes the fixed semi-cured material 1000b, may include a step S410 of contacting the upper plate 520 with the second mold 1200. For example, the upper plate pressing surface 522 of the upper plate 520 may be in contact with the first support surface 960 of the upper mold 900. Specifically, step S410, which involves contacting the upper plate 520, may involve contacting the pressure dispersion portion 550 with the first support surface 960.

[0186] In step S410, which brings the upper plate 520 into contact with the second mold 1200, the step of activating the pressure unit 400 may be included. For example, the pressure unit 400 may activate an actuator provided on the pressure unit body 410. The pressure rod 420 may be connected to the actuator. The pressure rod 420 may move toward the pressure plate 430. The pressure rod 420 may be placed on a mounting portion 439 provided on a surface of the pressure plate 430.

[0187] The pressure rod 420 can move in the up and down direction. When the pressure rod 420 moves in the downward direction, the upper plate 520, which is connected to the pressure plate 430, can also move in the downward direction. Therefore, the upper plate 520 can move towards the lower plate 510.

[0188] The upper plate 520 can be moved so that the pressure surface 522 of the upper plate comes into contact with the second mold 1200 disposed on the lower plate 510. More specifically, the pressure distribution part 550 can come into contact with the first support surface 960 of the upper mold 900 exposed from the second mold 1200.

[0189] In step S410, when the upper plate 520 comes into contact with the second mold 1200, some areas of the second mold 1200 may have a different thickness than other areas.

[0190] For example, during the coating process of the biocompatible polymer solution 1000a, the thickness difference may occur due to the uneven distribution of the biocompatible polymer solution 1000a. Alternatively, the thickness difference may also occur in the forming thickness of the upper mold 900 and the lower mold 800.

[0191] When a thickness difference occurs, the upper plate 520 tilts, which may result in the entire surface of the pressure distribution part 550 not being in contact with the entire surface of the first support surface 960.

[0192] refer to Figure 26 and Figure 32 (B) The pressurization step S400 may include step S420 of adjusting the tilt of the upper plate 520.

[0193] Step S420, which adjusts the tilt angle, allows the upper plate 520 to come into contact with the surface of the fixed semi-cured material 1200. For example, step S420, which adjusts the tilt angle of the upper plate 520, allows the entire surface of the pressure dispersion part 550 to come into contact with the entire surface of the first support surface 960.

[0194] After the two structural surfaces mentioned above come into contact, the leveling step of the upper plate 520 can be performed. That is, the upper plate 520 can be rotated (to adjust the tilt).

[0195] The tilt angle of the upper plate 520 can be adjusted by the universal joint 450. The universal joint 450 can be provided on top of the pressure plate 430. For example, the universal joint 450 can be provided between the guide rod 440 formed on the pressurizing unit 400 and the pressure plate 430. In addition, the universal joint 450 can be provided between the mounting part 439 and the pressure plate 430.

[0196] When the tilt of the upper plate 520 is not horizontal, the universal joint 450 can rotate the upper plate 520 to level it.

[0197] Therefore, the universal joint 450 can provide the load of the upper plate 520 to the thickness difference region of the second mold 1200. For example, for leveling, the universal joint 450 can rotate (tilt) the upper plate 520 to provide the load to the thickness difference region. Here, since the upper plate 520 is combined with the pressure plate 430, the load of the upper plate 520 and the pressure plate 430 can be provided to the second mold 1200.

[0198] The loads provided to the upper plate 520 and pressure plate 430 of the second mold 1200 can apply pressure to the fixed semi-cured material 1200. Therefore, the loads of the upper plate 520 and pressure plate 430 can pressurize the fixed semi-cured material 1200b in the area where the thickness difference occurs.

[0199] When the upper plate 520 is horizontal, the universal joint 450 can stop providing load to the second mold 1200.

[0200] refer to Figure 26 and Figure 33 It may include the step S430 of maintaining pressure on the pressure plate 430.

[0201] The step S430 of maintaining pressure can be performed when the pressure distribution part 550 is in contact with the first support surface 960 and the upper plate 520 is horizontal.

[0202] In the aforementioned state, a target pressure can be maintained on the pressure plate 430. The step S430 of maintaining the pressure can provide a pressure of 0.40 Pa to 0.70 Pa, preferably 0.45 to 0.65 MPa, to the pressure plate 430. The duration of providing the pressure can vary depending on the magnitude of the pressure; for example, a holding time of 30 to 50 minutes can be provided.

[0203] The target pressure applied to the pressure plate 430 can be transmitted to the upper plate 520. The pressure transmitted to the upper plate 520 can be provided to the second mold 1200 through the pressure surface 522 of the upper plate. The pressure provided to the second mold 1200 can be transmitted to the fixed semi-cured material 1200.

[0204] By applying pressure to the second mold 1200, the pattern shape formed on the first pattern surface 910 and the second pattern surface 810 can be transferred to the semi-cured material 1000b.

[0205] refer to Figure 25 and Figure 34 It may include a drying step S500 of drying the semi-cured material 1000b.

[0206] In drying step S500, the target drying heat can be supplied to the second mold 1200 via the lower plate 510. Alternatively, the second mold 1200 can be dried in an oven or on a heating plate, ultimately drying the semi-cured product 1000b.

[0207] When drying the second mold 1200 via the lower plate 510, a heating plate can be prepared inside the lower plate 510. The heating plate can dry the second mold 1200 by transferring heat energy to the upper plate 517. The heat energy transferred to the second mold 1200 can be transferred to the semi-cured material 1000b, thereby drying the semi-cured material 1000b.

[0208] On one hand, when the second mold 1200 is drying in the oven, the carrier 320 can be moved from the molding 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 oven DO, and drying heat can be provided to the second mold 1200.

[0209] Here, the drying step S500 can provide the second mold 1200 with heat energy of 15°C to 45°C for 6 to 10 hours.

[0210] On the other hand, in drying step S500, the second mold 1200 can be placed on a heating plate for drying. When using a heating plate, a steel plate of 0.5 kg to 2 kg can be stacked on the second mold 1200, and the heating plate can be heated to 60°C to 80°C to dry the second mold 1200. The second mold 1200 can be dried at the above heating temperature for 4 to 8 hours.

[0211] Here, the steel plate prevents the upper mold 900 or lower mold 800 from separating from the semi-cured product 1000b during the drying process.

[0212] When the upper mold 900 or the lower mold 800 separates during the drying process, the surface pattern of the semi-cured material 1000b may be damaged. Therefore, in order to prevent damage to the surface pattern of the semi-cured material 1000b, a steel plate can be placed on the second mold 1200 for the drying process.

[0213] In this way, the second mold 1200 can be dried, and a cured diaphragm regeneration patch 1000 can be formed between the upper mold 900 and the lower mold 800.

[0214] refer to Figure 25 and Figure 34 Step S600 can be performed to separate the tympanic membrane regeneration patch 1000 from the second mold 1200.

[0215] The tympanic membrane 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.

[0216] After separation step S600, the biological support 1000 can be immersed in 70% ethanol for 10 seconds and washed with purified water.

[0217] The manufacturing process of the tympanic membrane regeneration layer can be performed using the following tympanic membrane regeneration patch manufacturing apparatus 10.

[0218] The following is a detailed description of the accompanying illustrations.

[0219] Figure 17 This is a perspective view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention. Figure 18 This is a front view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention. Figure 19 This is a top view of a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention. Figure 20 It is along Figure 19 A cross-sectional view of line A1-A2.

[0220] refer to Figures 17 to 20 The tympanic membrane regeneration patch manufacturing apparatus 10 may include a support 100.

[0221] The support 100 may include a support body 110 and a worktable 120.

[0222] The worktable 120 can be formed from a plate with a flat surface. Multiple components can be mounted on the worktable 120. The flat surface provides a stable mounting surface for the components.

[0223] The support body 110 may have multiple legs. These legs may be positioned along the edge of the worktable 120. The legs may be configured with different or identical lengths. The adjusted lengths of the legs can form a flat surface on the worktable 120.

[0224] Furthermore, the outriggers can be set to a predetermined length corresponding to the operator's working height. For example, the outriggers can be set to the operator's expected working height. The adjustable length of the outriggers improves the operator's ease of use.

[0225] The tympanic membrane regeneration patch manufacturing apparatus 10 may include a frame unit 200. The frame unit 200 may be mounted on a worktable 120. The frame unit 200 may include a frame support 220 and a frame body 210.

[0226] The frame support 220 can support the frame body 210. The frame support 220 can be mounted on the worktable 120. The frame support 220 can be installed on the worktable 120 to fix the frame body 210. The frame support 220 and the worktable 120 can be fixed together using screws or similar connecting structures, but the fixing method is not limited to screws. Alternatively, the frame support 220 and the frame body 210 can also be fixed using similar fixing methods.

[0227] The frame body 210 can be mounted on the worktable 120. For example, the frame body 210 can be located on the upper part of the worktable 120 and supported by the frame support 220. The frame body 210 can be plate-shaped. Therefore, the frame body 210 can be mounted in a direction parallel to the flat surface of the worktable 120.

[0228] The frame body 210 may have multiple through holes 212 and 215. The frame body 210 may have cylinder guides 213 and 216 respectively provided on the multiple through holes 212 and 215.

[0229] The multiple through holes 212, 215 may include a first through hole 212 and a second through hole 215. A guide rod 440 may be disposed in the first through hole 212. A pressure rod 420 may be disposed in the second through hole 215.

[0230] Multiple first through holes 212 can be provided around the second through hole 215. The first through holes 212 can be symmetrically arranged around the second through hole 215.

[0231] In this embodiment, the illustration shows two first through holes symmetrically arranged around the second through hole 215. However, the arrangement of the first through holes 212 is not limited to this; three to eight first through holes 212 can be symmetrically arranged around the second through hole 215.

[0232] The guide rod 440 can disperse the concentrated pressure provided by the pressure rod 420 along 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 dispersed pressure on the pressure plate 430. The pressure plate 430, which forms a dispersed pressure, helps to form a uniform surface pressure in the forming unit 500.

[0233] The tympanic membrane regeneration patch manufacturing apparatus 10 may include a conveying unit 300.

[0234] For ease of explanation, the area enclosed by the frame support 220 and the frame body 210 within the workbench 120 is defined as the forming area PA, and the area outside the forming area PA is defined as the peripheral area SA. Within the peripheral area SA, the area where the conveyor unit 300 is located is defined as the loading area LA.

[0235] The transmission unit 300 may include a transmission guide rail 310, a carrier 320, and a transmission drive unit 330.

[0236] The transfer guide rail 310 can be mounted on the worktable 120. The transfer guide rail 310 can be arranged to span from the loading area LA to the molding area PA. The transfer guide rail 310 can be formed by a pair of guide rails arranged in the direction from the loading area LA to the molding area PA. At least one guide rail portion can be provided. For example, when the transfer guide rail 310 consists of multiple guide rail portions formed in the direction from the loading area LA to the molding area PA, the loading area LA can form multiple working windows.

[0237] The carrier 320 can be disposed on the conveyor rail 310. The carrier 320 can move along the conveyor rail 310. Therefore, the carrier 320 can be disposed in the loading area LA and the forming area PA.

[0238] 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. The carrier 320 and the transport rail 310 can be coupled by any structure as long as it is a movable means.

[0239] The carrier 320 may include a sliding portion 322 having a sliding structure and a mounting surface 328 disposed on the opposite side of the sliding portion 322. The sliding portion 322 may be coupled to the conveying guide rail 310. A lower plate 510 may be mounted on the mounting surface 328. The mounting surface 328 may be opposite to the lower plate 510.

[0240] The carrier 320 can be connected to the transfer drive unit 330. The transfer drive unit 330 can be connected to the thickness surface formed between the sliding part 322 and the mounting surface 328.

[0241] The transfer drive unit 330 can be located in the peripheral area SA. The transfer drive unit 330 includes an actuator that can move the carrier 320 to the loading area LA and the molding area PA. In this embodiment, the diagram illustrates a structure where the actuator moves back and forth, but depending on the situation, the carrier 320 can be moved using a structure that moves left and right.

[0242] The tympanic membrane regeneration patch manufacturing apparatus 10 may include a pressure unit 400. The pressure unit 400 may be disposed on the molding area PA. The pressure unit 400 may include a pressure unit body 410, a pressure rod 420, a pressure plate 430, and a guide rod 440.

[0243] The pressurizing unit body 410 can be mounted on top of the frame body 210. The pressurizing unit body 410 may include an actuator. The actuator can move the pressure rod 420 in the up-down direction.

[0244] The pressure rod 420 can be disposed in the second through holes 215 on the upper and lower parts of the frame body 210. For example, the pressure rod 420 can be inserted into the second cylinder guide 216 disposed in the second through hole 215.

[0245] The pressure bar 420 can move vertically. The pressure bar 420 can move to contact the pressure plate 430. The pressure bar 420 can move the pressure plate 430 downwards. When the pressure bar 420 faces the object being molded in the target area, it can apply pressure to the pressure plate 430.

[0246] The pressurizing unit 400 may include a pressure plate 430 disposed between the frame body 210 and the worktable 120.

[0247] The pressure plate 430 may include a pressure plate top 437 that contacts the pressure rod 420. A mounting portion 439 may be provided on the pressure plate top 437. The pressure rod 420 may be mounted on the mounting portion 439.

[0248] The mounting portion 439 can be provided in the region including the center of gravity of the pressure plate 430. For example, when the pressure plate 430 is square, the center of gravity of the pressure plate 430 can be the central region of the square. Therefore, the mounting portion 439 can be provided in the central region.

[0249] One end of the pressure rod 420 can be placed on the mounting part 439. The mounting part 439 can be made of pressure-resistant materials such as rubber or silicone and formed on the pressure plate 438.

[0250] Thus, the pressure bar 420 can move downwards, causing the pressure plate 430 to move downwards. Moreover, when the pressure plate 430 moves to the target location, the pressure bar 420 can apply pressure to the center of gravity of the pressure plate 430.

[0251] The guide rod 440 can be positioned around the pressure rod 420. The guide rod 440 can be positioned between the frame body 210 and the pressure plate 430.

[0252] One end of the guide rod 440 can be engaged with the pressure plate 430. The other side 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 provided in the first through hole 212 formed on the frame body 210. The guide rod 440 can be inserted into the first cylinder guide 213.

[0253] As mentioned above, multiple first through holes 212 can be symmetrically arranged around the second through hole 215. Guide rods 440 can be respectively arranged in multiple first through holes 212.

[0254] Therefore, multiple guide rods 440 can be symmetrically arranged around the pressure rod 420. The multiple guide rods 440 can disperse the applied pressure to create a uniform surface pressure in the molding unit 500. The multiple guide rods 440 can create a uniform applied pressure across the entire surface of the pressure plate 430.

[0255] On one hand, a gimbal member 450 can be provided between the guide rod 440 and the pressure plate 430. For example, the gimbal member 450 can connect the lower part of the guide rod 440 and the upper part of the pressure plate 430. Alternatively, the gimbal member 450 can be selectively provided between the mounting part 439 and the pressure plate 430. The gimbal member 450 can connect the lower part of the mounting part 439 and the upper part of the pressure plate 430.

[0256] The universal joint portion 450 can be symmetrically arranged around the mounting portion 439, with the mounting portion 439 as the center. In other words, on the top of the pressure plate 430, as multiple guide rods 440 are arranged around the area including the center of gravity of the pressure plate 430, the universal joint portion 450 provided on each of the multiple guide rods 440 can also be provided with the same arrangement structure.

[0257] Universal joint 450 can adjust the tilt of pressure plate 430. For example, universal joint 450 can level the upper plate 520. That is, universal joint 450 can rotate (adjust the tilt) the upper plate 520.

[0258] When the tilt of the upper plate 520 is not horizontal, the universal joint 450 can rotate the upper plate 520 to level it.

[0259] Therefore, since the universal joint 450 rotates (tilts) the upper plate 520 to level it, it is possible to move towards the second mold (reference). Figure 32 The upper plate 520 provides the load to the second mold 1200. For example, since the upper plate 520 is combined with the pressure plate 430, the load of the upper plate 520 and the pressure plate 430 can be provided to the second mold 1200.

[0260] Universal joint 450 can adjust for pressure imbalances that may occur due to differences in the thickness of the molded object. For example, when the thickness of the molded object is different, universal joint 450 can adjust the tilt of pressure plate 430 so that pressure plate 430 corresponds to the whole surface of a surface of the molded object.

[0261] Therefore, when the thickness of one area of ​​the molded object is different from that of another area, the universal joint 450 can prevent the pressure plate 430 from concentrating pressure on one area of ​​the molded object or transmitting more pressure.

[0262] Thus, the universal joint 450 can improve the molding uniformity of the molded object by adjusting the inclination of the pressure plate 430.

[0263] The tympanic membrane regeneration patch manufacturing apparatus 10 may include a molding unit 500. The molding unit 500 may include a lower plate 510, an upper plate 520, and a pressure dispersion section 550.

[0264] The lower plate 510 can be disposed on the carrier 320. For example, the lower plate 510 can be placed on the mounting surface 328 of the carrier 320. The lower plate 510 can be formed into a shape similar to or smaller than that of the carrier 320.

[0265] The lower plate 510's lower surface 512 can be placed on the mounting surface 328 of the carrier 320. The carrier 320 can move along the conveyor guide 310 between the loading area LA and the molding area PA. Therefore, the lower plate 510 placed on the carrier 320 can move between the loading area LA and the molding area PA via the carrier 320.

[0266] The lower plate 510 may be located on the opposite side of its lower surface 512, including its upper surface 517. (Lower mold - reference) Figure 29 The lower mold 800 can be set on top of the lower plate 510 at 517. The manufacturing method for the lower mold 800 is described in detail below.

[0267] The upper plate 520 can be disposed in the molding area PA. The upper plate 520 can be coupled with the pressure plate 430. The upper plate 520 may include an upper plate mating surface 527 that is coupled with the pressure plate 430. For example, the upper plate mating surface 527 may be coupled with the lower surface 432 of the pressure plate 430. The lower surface 432 of the pressure plate may be the surface opposite to the upper surface 437 of the pressure plate where the mounting portion 439 is provided.

[0268] The upper plate 520 can be shaped to have a larger area than the lower plate 510. The molded object can be positioned between the upper plate 520 and the lower plate 510. Because the area of ​​the upper plate 520 is larger than that of the lower plate 510, uniform pressure can be applied to the molded object. Therefore, the size of the molded object can be limited by the shape of the lower plate 510.

[0269] As described above, the tympanic membrane regeneration patch manufacturing apparatus 10 according to the present invention can form a pressure plate 430 with uniform pressure by dispersing pressure, thereby increasing the area of ​​the lower plate 510. Therefore, a molded object disposed on the lower plate 510 can be formed over a large area.

[0270] The upper plate 520 may include an upper plate pressure surface 522 on the opposite side of the upper plate mating surface 527. The pressure dispersion part 550 may be provided on the upper plate pressure surface 522.

[0271] The pressure dispersion section 550 can be bonded to the upper plate pressure surface 522 with an adhesive. The pressure dispersion section 550 can be made of silicone material, but is not limited to it; any material capable of transmitting pressure can be used. The pressure dispersion section 550 can evenly distribute the applied pressure input from the pressure plate 430 to the entire surface of the upper plate 520.

[0272] The tympanic membrane regeneration patch manufacturing apparatus 10 may include an input unit 600 and a display unit 700. The input unit 600 and the display unit 700 may be disposed in the peripheral area SA.

[0273] The tympanic membrane regeneration patch manufacturing apparatus 10 may include an input unit 600 for receiving input from an operator. For example, the input unit 600 allows the operator to directly input set values.

[0274] The input unit 600 may include input means such as input buttons for inputting set values. Here, the set values ​​may include, for example, the pressure of the pressurizing unit 400, the heating temperature of the lower plate 510, the pressure supply time of the pressurizing unit 400, and the heating time.

[0275] The display unit 700 can display the set values ​​on the screen. In other words, the display unit 700 can display the 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 supply time, and the heating time, etc., numerically.

[0276] Furthermore, a control unit capable of controlling the setting values ​​of the tympanic membrane regeneration patch manufacturing apparatus 10 can also be provided in the space where the display unit 700 is located.

[0277] Thus, the tympanic membrane regeneration patch manufacturing apparatus 10 according to an embodiment of the present invention forms uniform pressure, thereby enabling the manufacture of tympanic membrane regeneration patches with improved uniformity.

[0278] Figure 21 This is a block diagram illustrating a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0279] To avoid repetitive descriptions and for ease of explanation, Figure 21 Reference Figures 17 to 20 Please provide an explanation.

[0280] The tympanic membrane regeneration patch manufacturing apparatus 10 according to an embodiment of the present invention may include a sensing unit 30 and a control unit 50.

[0281] 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 installed in the area where the display unit 700 is located.

[0282] The control unit 50 can be connected to a structure equipped with the sensing unit 30. For example, the control unit 50 can be connected to a structure equipped with a pressure sensor 470 and / or a temperature sensor 515.

[0283] Specifically, for example, when a temperature sensor 515 is provided on the lower plate 510, the control unit 50 can be connected to the lower plate 510. As another example, when a pressure sensor 470 is provided on the pressurization unit 400, the control unit 50 can be connected to the pressurization unit 400. The sensing unit 30 may include the pressure sensor 470. The pressure sensor 470 may be disposed on the upper part of the frame body 210.

[0284] This figure illustrates the case where a pressure sensor 470 is provided in the pressurizing unit 400. Alternatively, the pressure sensor 470 can also be provided on the molding unit 500.

[0285] The input unit 600 can receive input from the operator. The input unit 600 can supply pressure to the pressure lever 420 of the pressurizing unit 400. The supplied input can be the set pressure for the operation of the pressurizing unit 400. The input unit 600 can generate a first signal SG1 consisting of the set pressure.

[0286] The input unit 600 can provide a first signal SG1 to the control unit 50. The control unit 50 can convert the received first signal SG1 and transmit it to the display unit 700 and the pressurization unit 400.

[0287] The control unit 50 can convert the first signal SG1 to transmit the first-1 signal SG11 to the display unit 700. Upon receiving the first-1 signal SG11, the display unit 700 can display the set pressure on the screen.

[0288] In addition, the control unit 50 can convert the first signal SG1 and transmit the first-second signal SG1)2 to the pressurization unit 400.

[0289] The pressurizing unit 400 can operate according to the first and second signals SG12. For example, the pressurizing unit 400 can operate the pressure rod 420 according to the first and second signals SG12. For example, the pressure rod 420 can move the pressure plate 430 according to the first and second signals SG12. When the pressure plate 430 moves according to the first and second signals SG12 and comes into contact with the object being molded, the pressure rod 420 can apply pressure to the pressure plate 430.

[0290] The pressure sensor 470 of the sensing unit 30 can sense the pressure generated in the pressurization unit 400. The sensing unit 30 can generate a second signal SG2 as the sensed pressure generated in the pressurization unit 400. The sensing unit 30 can transmit the second signal SG2 to the control unit 50.

[0291] Upon receiving the second signal SG2, the control unit 50 can calculate whether pressure has been transmitted through the pressurization unit 400 based on the information from the first and second signals SG12. For example, the control unit 50 can calculate the difference between the pressure set on the pressure rod 420 and the pressure received from the pressure sensor 470. The control unit 50 can generate the third signal SG3 and the fourth signal SG4 from the calculated pressure difference value (hereinafter referred to as the "calculated value").

[0292] If the calculated 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. Additionally, the control unit 50 can generate a third signal SG3 identical to the first and second signals SG12 and provide the third signal SG3 to the pressurization unit 400.

[0293] On the one hand, if the calculated value exceeds the error range compared with the measured data, the control unit 50 can provide the corrected input value to the pressurization unit 400.

[0294] Here, when the control unit 50 is manually set, a fourth signal SG4 consisting of calculated values ​​can be transmitted to the display unit 700. The display unit 700 can then display the calculated values ​​based on the fourth signal SG4 on the screen.

[0295] The operator can confirm the calculated value displayed on the display unit 700. The operator can use the displayed information as a reference to input the correction value into the input unit 600.

[0296] The control unit 50 can generate a third signal SG3 based on the corrected input value. The third signal SG3 generated in the control unit 50 can be transmitted to the pressurization unit 400. The pressurization unit 400 can provide a corrected pressure value to the pressure plate 430 via the third signal SG3. The corrected pressure value can be the pressure provided to the pressure plate 430.

[0297] Furthermore, the control unit 50 can generate a fourth-first signal SG4-1 based on the information from the third signal SG3. The fourth-first signal SG4-1 can be provided to the display unit 700. The display unit 700 can display information composed of the fourth-first signal SG4-1 on the screen.

[0298] Thus, the tympanic membrane regeneration patch manufacturing apparatus 10 according to the present invention can control the pressure supplied to the pressurizing unit 400 by the sensing unit 30.

[0299] On the one hand, when the control unit 50 is automatically set, if there is a difference in the calculated pressure difference value (hereinafter referred to as the "calculated value"), the fourth signal SG4 may not be generated.

[0300] Pressure sensor 470 can sense the pressure generated in pressurization unit 400. Sensing unit 30 can generate second-first signal SG21 based on the sensed information. Sensing unit 30 can transmit second-first signal SG21 to control unit 50.

[0301] The control unit 50 can determine, based on the measured data, whether the calculated value is within the error range compared to the measured data. If it is determined that the value is within the error range, the control unit 50 can generate a third signal SG3 that is the same as the first-second signal SG12.

[0302] The third signal SG3 generated in the control unit 50 can be transmitted to the pressurizing unit 400. The pressurizing unit 400 can provide pressure based on the first-second signal SG12 to the pressure plate 430 via the third signal SG3. The pressure based on the first-second signal SG12 can be the pressure provided to the pressure plate 430.

[0303] Furthermore, the control unit 50 can generate a fourth signal SG4 based on the information from 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.

[0304] Thus, the tympanic membrane regeneration patch manufacturing apparatus 10 according to the present invention can control the pressure supplied to the pressurizing unit 400 by the sensing unit 30.

[0305] On the other hand, the control unit 50 can compare the input pressure with the pressure supplied to the molding object. For example, the pressurizing unit 400 can supply pressure to the molding unit 500. For example, the pressurizing unit 400 can transmit the pressure formed in the pressure plate 430 to the molding unit 500.

[0306] There may be a difference between the pressure formed in the pressurizing unit 400 and the pressure formed in the molding unit 500. The control unit 50 can compare the pressure provided by the pressurizing unit 400 with the pressure output by the molding unit 500. The output pressure may be the pressure provided to the object being molded.

[0307] Pressure can be supplied to the molding unit 500 via the pressure plate 430. The molding unit 500 can generate the output pressure supplied to the object being molded.

[0308] The pressure sensor 470 of the sensing unit 30 can be respectively provided in the pressurizing unit 400 and the molding unit 500. The pressure sensor 470 can sense the pressure provided by the pressurizing unit 400. In addition, the pressure sensor 470 can sense the output pressure of the molding unit 500.

[0309] The sensing unit 30 can generate a second signal consisting of supplied pressure and output pressure.

[0310] There may be a difference (hereinafter referred to as "pressure difference") between the supply pressure input in the pressure bar 420 and the output pressure output in the forming unit 500. For example, a pressure difference may occur during the process of distributing pressure to the entire surface of the pressure plate 430 or during the process of distributing pressure in the pressure dispersion section 550.

[0311] In this situation, the probability of molded products being defective may increase. Furthermore, power loss due to leakage during the use of the manufacturing equipment may occur.

[0312] The pressure sensor 470 can sense the pressure in each set area, and the sensing unit 30 converts the pressure in each set area into a signal and transmits the second signal SG2 to the control unit 50.

[0313] The control unit 50 can calculate the pressure difference. The setpoint of the pressurization unit 400 can be readjusted based on the calculated value. Furthermore, the connection structure between the components can be readjusted. Therefore, the pressure sensor 470 can reduce the probability of malfunction and minimize power loss from leakage in the manufacturing equipment.

[0314] Figure 22 This is a top view showing the shape of the pressure plate of the pressurizing unit according to an embodiment. Figure 23 This is a top view showing another shape of the pressure plate of the pressurizing unit according to an embodiment. Figure 24 This is a top view of another shape of the pressure plate of the pressure unit according to an embodiment.

[0315] To avoid repetitive descriptions and for ease of explanation, Figures 22 to 24 Reference Figures 17 to 20 Please provide an explanation.

[0316] refer to Figures 22 to 24 The pressurizing unit 400 may include a pressure rod 420, a pressure plate 430, and a guide rod 440.

[0317] The pressure plate 430 may include a mounting portion 439. The mounting portion 439 may be disposed in the region containing the center of gravity of the pressure plate 430. The pressure rod 420 may be disposed at a position corresponding to the mounting portion 439.

[0318] Guide rods 440 can be arranged around pressure rods 420. Guide rods 440 can be arranged symmetrically with respect to pressure rods 420. Symmetrically arranged guide rods 440 can form a distributed pressure across the entire surface of pressure plate 430.

[0319] On one hand, the lower plate 510 of the molding unit 500 can be configured to have a smaller area than the upper plate 520. On the other hand, the upper plate 520 can be attached to the pressure plate 430 with the same shape.

[0320] Therefore, the pressure provided by the pressure plate 430 with dispersed pressure can be transmitted to the upper plate 520. The upper plate 520, which receives pressure through transmission, can be positioned opposite the lower plate 510 to pressurize and mold the object.

[0321] Therefore, the pressure plate 430 with dispersed pressure formed on the entire surface can help to form uniform surface pressure in the molding unit 500.

[0322] exist Figures 17 to 20 In this configuration, the pressure plate 430 can be shaped as a quadrilateral, and the guide rods 440 can be symmetrically arranged on both sides of the pressure rod 420 with the pressure rod 420 as the center. Therefore, the guide rods 440 can be configured as two rods connected to the pressure plate 430.

[0323] refer to Figure 22 Alternatively, the pressure plates 430 can be identical quadrilaterals, and the guide rods 440 can be symmetrically arranged around the pressure rods 420 at the corners of the pressure plates 430. Therefore, the guide rods 440 can be configured as four rods connected to the pressure plates 430.

[0324] Thus, since the four guide rods 440 are symmetrically arranged on the pressure plate 430 with the pressure rod 420 as the center, the dispersion of the applied pressure can be further improved.

[0325] refer to Figure 23 The pressure plate 430 can be formed into a triangle. Three guide rods 440 can be provided on the triangular pressure plate 430. For example, when the pressure plate 430 is triangular, three guide rods 440 can be provided on the pressure plate 430 so that the guide rods 440 are symmetrically arranged with the pressure rod 420 as the center.

[0326] Thus, when the pressure plate 430 is triangular, the pressure dispersion can be further improved because the three guide rods 440 are symmetrically arranged on the pressure plate 430 with the pressure rod 420 as the center.

[0327] refer to Figure 24 The pressure plate 430 can be formed in a circular shape. Four to eight guide rods 440 can be provided on the circular pressure plate 430. In this embodiment, eight guide rods 440 are shown arranged on the pressure plate 430.

[0328] Specifically, when the pressure plate 430 is circular, four to eight guide rods 440 can be set on the pressure plate 430 so that the guide rods 440 are symmetrically arranged with the pressure rod 420 as the center.

[0329] Thus, when the pressure plate 430 is circular, since four to eight guide rods 440 are symmetrically arranged on the pressure plate 430 with the pressure rod 420 as the center, the pressure dispersion can be further improved.

[0330] Figure 25This is a flowchart illustrating a method for manufacturing a tympanic membrane regeneration patch using a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention. Figures 26 to 28 It is based on Figure 25 A detailed flowchart of each step in the manufacturing process of the tympanic membrane regeneration patch. Figures 29 to 34 This is a process diagram illustrating a method for manufacturing a tympanic membrane regeneration patch using a tympanic membrane regeneration patch manufacturing apparatus according to an embodiment of the present invention.

[0331] To avoid repetitive descriptions and for ease of explanation, Figures 25 to 34 Reference Figures 17 to 20 Please provide an explanation.

[0332] The method for manufacturing the 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 apparatus 10.

[0333] The tympanic membrane regeneration patch manufacturing apparatus 10 may include a conveying 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 510. The forming object may be disposed between the upper plate 520 and the lower plate 510. The tympanic membrane regeneration patch manufacturing apparatus 10 may apply pressure to the forming object, thereby forming a tympanic membrane regeneration patch with improved pattern uniformity on one or both sides.

[0334] The tympanic membrane regeneration layer manufactured as described above is placed on the adhesive layer to manufacture the tympanic membrane regeneration patch of the present invention.

[0335] The invention will be described in more detail below through examples.

[0336] Example

[0337] 1. Manufacturing of molds and polymer solutions used for manufacturing tympanic membrane regeneration patches.

[0338] (1) Making PUA molds

[0339] Using a micropipette, 600 μl of PUA solution was coated onto a silicon master mold with a uniform nanopattern of 800 nm in length, width, and height. After placing the PET film on the coated area, it was thoroughly rolled with a polyurethane roller to prevent air bubbles. Once the absence of air bubbles was confirmed, the wafer and PET film were placed together in a UV curing machine for 5 minutes. After 5 minutes, the master mold and wafer were removed, and the PET film and PUA mold were then subjected to a further 20 hours of UV curing.

[0340] (2) Fabrication of patternless PDMS molds

[0341] Mix 27g of PDMS base material and 3g of curing agent, and dispense the mixture into Petri dishes (125mm×125mm×20mm). Then, place the Petri dishes in a vacuum chamber and maintain a vacuum at 0.1MPa for 1 hour at room temperature. After 1 hour, release the vacuum, confirm that all air bubbles in the PDMS have disappeared, and then cure at 70℃ for 5 hours. After curing, cut to create a 10cm×10cm PDMS mold.

[0342] (3) Fabrication of PDMS molds with nanopatterns

[0343] 27g of PDMS base material and 3g of curing agent were mixed to prepare a mixed solution. A PUA mold was attached to a Petri culture dish with the pattern facing upwards, and then the PDMS mixed solution was dispensed into the Petri culture dish (125mm × 125mm × 20mm). The Petri culture dish was placed in a vacuum chamber and maintained under vacuum at 0.1MPa for 1 hour at room temperature. After 1 hour, the vacuum was released, and after confirming that all air bubbles in the PDMS had disappeared, it was cured at 70℃ for 5 hours. After curing, it was cut to produce a PDMS mold measuring 10cm × 10cm. The manufactured PDMS mold has a uniform nanopattern with dimensions of 800nm ​​in length, width, and height.

[0344] (4) Preparation of polymer solutions

[0345] PLGA (lactide to glycolide molar ratio of 75:25) was mixed with dichloromethane (DCM) for 8 hours to prepare PLGA solutions of 12% (w / w), 15% (w / w), 18% (w / w) and 21% (w / w).

[0346] PCL (poly(ε-caprolactone)) was mixed with dichloromethane (DCM) for 8 hours to prepare PCL solutions of 15% (w / w), 18% (w / w), 21% (w / w), and 24% (w / w).

[0347] 2. Manufacturing of PLGA tympanic membrane regeneration layer

[0348] (1) Manufacturing of the tympanic membrane regeneration layer in Example 1

[0349] Attach a 2mm thick PDMS mold to Figure 17 After the fabrication apparatus is mounted on the plate, a 12% (w / w) PLGA solution (3g) is uniformly sprayed onto a PUA mold with a nano-pattern, and then a PDMS mold is placed on top for rolling. Subsequently, compression is performed on the top at a pressure of 0.5MPa (reference). Figure 1After removing the PDMS mold and evaporating the organic solvent in an oven at 80°C for about 10 hours, the PLGA patch was separated from the PUA mold to manufacture the tympanic membrane regeneration layer of Example 1.

[0350] The pattern spacing (distance between peaks of the pattern) and pattern height (height difference between peaks and valleys of the pattern) of the PDMS mold and PUA mold used here are designed to be 800nm ​​respectively. One side of the PLGA patch is patterned by the PDMS mold and the other side is patterned by the PUA mold.

[0351] (2) Manufacturing of the tympanic membrane regeneration layer in Examples 2 to 10 and 23

[0352] Except for the concentration of the PLGA solution and the drying temperature shown in Table 1 below, the tympanic membrane regeneration layers of Examples 2 to 10 were manufactured by the same method as in Example 1.

[0353] The concentration and drying temperature of the PLGA solution used to prepare the tympanic membrane regeneration layer in Example 23 were the same as those in Example 4. The pattern was formed only on one side and the thickness was 30 μm.

[0354] (3) Manufacturing of the tympanic membrane regeneration layer in Comparative Examples 1 to 3

[0355] Except for the concentration of the PLGA solution and the drying temperature shown in Table 1 below, the tympanic membrane regeneration patches of Comparative Examples 1 and 2 were manufactured by the same method as in Example 1.

[0356] Comparative Example 3 was manufactured using a PDMS mold and a PUA mold without a pattern. The concentration of the PLGA solution and the drying temperature conditions used in the manufacture of the tympanic membrane regeneration patch of Comparative Example 3 are shown in Table 1 below.

[0357] (4) Manufacturing conditions and thickness of the tympanic membrane regeneration layer in Examples 1-10 and 23 and Comparative Examples 1-3

[0358] The manufacturing processes of Examples 1-10 and 23 and Comparative Examples 1-3 are shown in Table 1 below.

[0359] Table 1

[0360]

[0361] 3. Manufacturing of PCL tympanic membrane regeneration layer

[0362] (1) Manufacturing of the tympanic membrane regeneration layer in Examples 11 to 22

[0363] 10g each of 15% (w / w), 18% (w / w), 21% (w / w), and 24% (w / w) PCL solutions were added dropwise to tempered glass (10cm × 10cm × 1.1mm) and spin-coated. The thickness of the patch was adjusted by changing the spin-coating speed (rpm). After spin-coating, the tempered glass was placed on a hot plate set to 70°C with the coated PCL facing upwards and melted for 5 minutes. A PDMS mold without nano-patterns was placed on the PCL layer and rolled using a polyurethane roller to prevent air bubbles. Subsequently, [tempered glass, PCL, and PDMS] were placed on a hot plate set to 70°C. Figure 17 On the plate of the patch manufacturing device, it is compressed at a pressure of 0.5 MPa for 5 minutes.

[0364] (2) Manufacturing conditions and thickness of the tympanic membrane regeneration layer in Examples 11 to 22

[0365] The manufacturing process of the PCL tympanic membrane regeneration patches in Examples 11 to 22 is shown in Table 2 below.

[0366] Table 2

[0367]

[0368] 4. Measurement of the uniformity of nanopatterns in the tympanic membrane regeneration layer

[0369] Three points on the peak and two points on the trough of the tympanic membrane regeneration layer of Examples 1 to 23 and Comparative Examples 1 and 2 were randomly selected, and the height difference between the three points on the peak and the height difference between the two points on the trough were measured (refer to...). Figure 2 The average value was substituted into Equation 1 to calculate the uniformity of the nanopatterns. The results are shown in Table 3 below.

[0370] Table 3

[0371]

[0372]

[0373] SEM images of the surface of the tympanic membrane regeneration layer in Examples 1 to 22 and Comparative Examples 1 and 2 are shown below. Figures 3a to 3e As shown. It can be seen that the pattern of the tympanic membrane regeneration layer in Comparative Examples 1 and 2 partially collapsed ( Figure 3e The maximum height difference between any point on the peaks and valleys of the tympanic membrane regeneration layer in Comparative Examples 1 and 2 was 100.2 nm and 98.3 nm, respectively. If the nanopattern is uniform, tympanic membrane cells can proliferate continuously along the pattern; if the nanopattern collapses even partially, tympanic membrane cells may stop growing and proliferating in that part.

[0374] 5. Measurement of the uniformity of the total area of ​​the tympanic membrane regeneration layer

[0375] The areas in the tympanic membrane regeneration layers of Example 4 and Comparative Example 1 that satisfy the uniformity of Mathematical Formula 1 are calculated. According to Mathematical Formula 2, the uniformity of Example 4 is calculated as follows: Figure 4 As shown in (A), it is 99.8%, while Comparative Example 1 is less than 70%.

[0376]

Mathematical Formula 2

[0377] Uniformity (area %) = (Area satisfying uniformity in equation 1 / Total area of ​​the patch)

[0378] 6. Structural and chemical properties analysis of the tympanic membrane regeneration layer

[0379] The structure and chemical properties of the tympanic membrane regeneration layers of Example 4 and Comparative Example 3 were analyzed. Scanning electron microscopy (SEM) was performed to analyze the surface structure, and FT-IR analysis was performed to confirm the chemical structure.

[0380] Scanning electron microscopy (SEM) analysis showed that the tympanic membrane regeneration layer surface of Comparative Example 3 was flat, while the tympanic membrane regeneration layer of Example 4 had a neatly arranged pattern of 800 nm structures (length, width, and thickness). Furthermore, FT-IR analysis confirmed that the chemical structure of PLGA in the nanopatterned tympanic membrane regeneration layer of Example 4 remained the same. Figure 5 ).

[0381] 7. Mechanical property analysis of the tympanic membrane regeneration layer

[0382] The mechanical properties (tensile strength and elongation) of the tympanic membrane regeneration layers of Example 4 and Comparative Example 3 were measured. Thin 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 clamping distance was adjusted to 5 cm, and the crosshead speed to 500 mm / min. The experiment was repeated three times, and the mean and standard deviation were calculated. Tensile strength (TS, MPa) was expressed as the maximum tension recorded before sample breakage divided by the initial cross-sectional area of ​​the support (Equation 3), and elongation (E, %) was expressed as the percentage increase in length before sample breakage relative to the initial clamping distance (Equation 4).

[0383]

Mathematical Expression 3

[0384] TS(MPa)=F max / A

[0385] (In the formula, F) max , where is the maximum tension (N) recorded before the sample breaks, and A is the initial cross-sectional area of ​​the sample (m²).

[0386]

Mathematical Expression 4

[0387] E(%) = (L / L0) × 100

[0388] (In the formula, L0 is the initial clamping distance of the sample (mm)), and L is the length increase before the sample breaks (mm).

[0389] The experimental results are shown in Figure 6, confirming that the tensile strength and elongation of Example 4 (Nano) are both higher than those of Comparative Example 3 (Flat), and confirming that the stress-strain curve forms on the upper side (…). Figure 6a ).

[0390] The results of measuring the tensile strength and elongation of Examples 3, 5-7, 11-19 and 23 and Comparative Examples 1-2 using the same method are shown in Table 4 below.

[0391] Table 4

[0392]

[0393] As the PLGA concentration decreases, the tensile strength of the manufactured patches tends to weaken. Comparative Examples 1 and 2 are predicted to be unsuitable for surgery due to their excessively low mechanical strength. Although Comparative Example 3 has a slightly improved tensile strength due to its higher curing temperature compared to Comparative Examples 1 and 2, it is predicted to have poor adhesion and reduced tympanic membrane regeneration effect due to the lack of a pattern. For PLGA tympanic membrane regeneration layers, it was confirmed that a relatively uniform tympanic membrane regeneration layer was manufactured when the PLGA concentration was 15% to 21% (w / w) and the drying process temperature was 60 to 80°C.

[0394] For the PCL tympanic membrane regeneration layer, Examples 11 to 13 showed a trend of relatively lower tensile strength compared with other examples. Therefore, it was determined that a PCL concentration of 18% (w / w) or higher is preferred for clinical use.

[0395] 8. Adhesion analysis of the tympanic membrane regeneration layer

[0396] For the treatment of tympanic membrane perforation, the patch needs to adhere completely to the patient's tympanic membrane during regeneration; therefore, appropriate adhesion strength is required. Thus, the adhesion strength of the tympanic membrane regeneration layers of Example 4 and Comparative Example 3 was compared.

[0397] After cutting the film model sample into circular films with a diameter of 6 mm, the tissue adhesion force was measured using a tensile testing machine (MCT-1150 tensile tester, A&D Company, Japan). Before measuring the tissue adhesion force, uncontaminated porcine dermal layers were washed three times with triple-distilled water. A circular tympanic membrane regeneration layer with an area of ​​2.8 cm² was pre-pressed onto the dermal surface with a pressure of ~0.5 N / cm², and the crosshead speed was adjusted to 100 mm / min. The adhesion force was measured until separation occurred. The adhesion force was measured by repeating the experiment 10 times for each sample under the same conditions at 40% relative humidity and an ambient temperature of 26°C, and the mean and standard deviation were calculated.

[0398]

Mathematical Expression 5

[0399] Normal adhesion force = F max / A

[0400] (In the formula, F) max The maximum force recorded until the tympanic membrane regeneration patch separates is A, where A is the initial area (cm²) of the tympanic membrane regeneration patch.

[0401] Analysis results as follows Figure 6b As shown, the adhesive strength (adhesion strength) of Example 4 is superior to that of Comparative Example 3.

[0402] Using the same method, the adhesive force (adhesion strength) of Examples 3, 5-7, 11-19 and 23 and Comparative Examples 1-2 was measured, and the results are shown in Table 5 below.

[0403] Table 5

[0404]

[0405] 9. Morphological analysis of cell proliferation and migration within the nanopattern of the tympanic membrane regeneration layer

[0406] Cells were cultured in the tympanic membrane regeneration patches of Comparative Example 3 and Example 4, and cell proliferation and migration morphology were observed. Fibroblasts similar to tympanic membrane cells were used. In Comparative Example 3 (Flat) without nanopatterns, a tendency for fibroblast proliferation and diffusion was confirmed. In the tympanic membrane regeneration layer (Nano) of Example 4 with nanopatterns, fibroblasts were confirmed to be neatly arranged along the nanopatterns and to proliferate and migrate. Figure 7 ).

[0407] 10. Evaluation of cell adhesion ability and function of the tympanic membrane regeneration layer

[0408] Fibroblasts were cultured in the tympanic membrane regeneration layers of Comparative Example 3 and Example 4, and cell adhesion and proliferation were observed. Analysis of cell adhesion after 6 hours of fibroblast culture showed enhanced adhesion in the tympanic membrane regeneration layer (800 nm) of Example 4 compared to Comparative Example 3 (Flat), and this was confirmed in proliferation analyses on days 3 and 5. Figure 8 In particular, similar proliferation rates were confirmed even when compared to cell culture containers (TCPS) specifically designed for existing cell proliferation. Figure 9 and Figure 10 Flat: Comparative Example 3, 800nm: Example 4).

[0409] Furthermore, cell migration was analyzed using a wound healing model. After inducing wounds by attaching 1 mm diameter SLABs (a substance made of PDMS) to the tympanic membrane regeneration layer of Examples 4 and Comparative Example 3 (wound portion = SLAB attachment portion), fibroblasts were seeded and cultured for 24 hours. Cell migration analysis at 0, 12, and 24 hours after cell culture showed that the migration distance and speed of cells in Example 4 were higher than those in Comparative Example 3, especially confirming a very wide wound healing range in the wound healing model. Figure 10 and Figure 11 Flat: Comparative Example 3, Nano: Example 4).

[0410] 11. Tympanic membrane regeneration effect of the tympanic membrane regeneration layer (in vivo)

[0411] (1) Tympanic membrane regeneration effect in Example 4

[0412] The tympanic membrane regeneration layer of Example 4 and the existing paper patch were attached to an animal model of tympanic membrane perforation, and the results were analyzed. Figure 12 The process of tympanic membrane perforation and patch application in an animal model is briefly illustrated. Animal experimental results show that, compared with the existing paper patch group, the tympanic membrane regeneration rate is increased by more than 40% in the group using the nano-patterned tympanic membrane regeneration patch of this invention (…). Figure 13 ).

[0413] (2) Tympanic membrane regeneration effect in Example 23

[0414] The tympanic membrane regeneration effect of the tympanic membrane regeneration layer in Example 23 was confirmed as follows. Twenty male Sprague-Dawley rats underwent bilateral tympanic membrane perforation, and the observation period was set for a total of 14 days. The tympanic membrane healing status was evaluated in three stages.

[0415] All Sprague-Dawley rats (8 weeks old) were anesthetized with chloral hydrate (1 g / mL, 10% dilution) and mechanically punctured. After observing the tympanic membrane, when it was normal, a 40-50% perforation was prepared in the anterior part of the tympanic membrane under a Zeiss microscope using a micropick.

[0416] For the efficacy evaluation parameters, the presence of tympanic membrane perforation is confirmed by Zeiss microscopy, and the tympanic membrane perforation is evaluated as cured only when complete healing is achieved.

[0417] Evaluation methods

[0418] 1. Complete healing: A state without perforation.

[0419] 2. Partial healing: The perforation size is reduced by more than 50%.

[0420] 3. No healing: The perforation size remains unchanged or shrinks by less than 50%.

[0421] Evaluation results

[0422] Evaluation results as follows Figure 14 As shown.

[0423] Twenty bilateral SD rats were used as subjects, and the perforation healing rate was measured under a microscope. In the experimental group (NG patch), 14 rats achieved complete healing in the first week, 6 rats achieved complete healing in the second week, and all individuals achieved complete healing within two weeks. In the control group (paper patch), only 3 rats healed within one week, and 4 rats ultimately did not heal completely. Therefore, it can be seen that the tympanic membrane regeneration layer of the present invention has a fast recovery speed and excellent tympanic membrane regeneration effect.

[0424] 12. Clinical trials using tympanic membrane regeneration patches

[0425] (1) Use of the tympanic membrane regeneration layer in Example 23

[0426] At Soonchunhyang University Hospital in South Korea, a total of 27 clinical trials were conducted using the tympanic membrane regeneration layer (a circular patch with a thickness of 30 μm and a diameter of 5 mm) from Example 23.

[0427] Clinical trial results confirmed tympanic membrane regeneration and hearing improvement in over 50% of patients with chronic tympanic membrane perforation. Patient information from four cases is shown in Table 6 below. In particular, in patient number 4, it was demonstrated that even in cases of failed tympanic membrane reconstruction surgery, the application of the tympanic membrane regeneration layer of this invention could achieve tympanic membrane regeneration and hearing improvement. Figure 15 No patients developed tremors after the surgery.

[0428] Table 6

[0429]

[0430] However, in some cases, the tympanic membrane regeneration layer failed to adhere consistently to the tympanic membrane during perforation regeneration, detaching prematurely after approximately 7 days. In cases where the tympanic membrane regeneration layer maintained consistent adhesion, a significant reduction in perforation size was confirmed, indicating tympanic membrane regeneration. Therefore, the following clinical trial was conducted by adding an adhesive layer to the tympanic membrane regeneration layer.

[0431] (2) Use of the tympanic membrane regeneration layer + hydrocolloid layer in Example 4

[0432] The tympanic membrane regeneration layer of Example 4 was evaluated to have physical properties most suitable for clinicians to perform the procedure. The tympanic membrane regeneration layer of Example 4 was laminated onto a hydrocolloid layer to apply the tympanic membrane regeneration patch of the present invention, with enhanced adhesion, to a patient. As a result, the tympanic membrane regeneration layer of Example 24, designed to be formed on the hydrocolloid layer, remained permanently attached to the tympanic membrane perforation site.

[0433] A clinical trial was conducted at Soonchunhyang University Hospital in South Korea using the aforementioned tympanic membrane regeneration patch on 18 patients. The selected patients were those who had suffered tympanic membrane perforation for more than 3 months and had no symptoms of ear discharge, pain, or other inflammation.

[0434] The procedure on the patient was performed through the following steps: ① local anesthesia, ② perforation margin trimming, ③ application of the tympanic membrane regeneration layer of Example 4, ④ application of the hydrocolloid layer, and ⑤ confirmation of the tympanic membrane regeneration effect every 4 weeks.

[0435] The following criteria were used to evaluate the postoperative tympanic membrane regeneration effect.

[0436] - Healing rate = Complete healing rate + Partial healing rate

[0437] - Complete healing: A state without tympanic membrane perforation.

[0438] Partial healing: The size of the tympanic membrane perforation decreases by more than 50% after the application of the patch.

[0439] - No healing: When the size of the tympanic membrane perforation does not change or shrinks by less than 50% after the patch is applied.

[0440] The methods and results of this clinical trial are as follows: Figure 16 As shown.

[0441] In a total of 18 cases, the total healing rate reached an excellent 61%, and the complete healing rate reached a high level of approximately 50%. This indicates that the tympanic membrane regeneration patch of the present invention can be used for non-surgical treatment of patients with tympanic membrane perforation and chronic otitis media.

[0442] Of the 18 cases, one case of pain and one case of otorrhea were observed as complications, both of which were minor. These all improved after medication.

Claims

1. A tympanic membrane regeneration patch, comprising an adhesive layer and a tympanic membrane regeneration layer formed on the adhesive layer, wherein, The tympanic membrane regeneration layer is composed of biocompatible polymers. Patterns for tympanic membrane cell growth are formed on one or both sides of the tympanic membrane regeneration layer. The pattern consists of repeated peaks and valleys. The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

2. The tympanic membrane regeneration patch according to claim 1, wherein, The tympanic membrane regeneration layer has an elongation of 4% to 10%.

3. The tympanic membrane regeneration patch according to claim 1, wherein, The adhesive layer is a hydrocolloid layer.

4. The tympanic membrane regeneration patch according to claim 1, wherein, The biocompatible polymer is a lactide-glycolic acid copolymer consisting of 65 to 85 mol% lactide and 15 to 35 mol% glycolide.

5. The tympanic membrane regeneration patch according to claim 1, wherein, The biocompatible polymer is polycaprolactone.

6. The tympanic membrane regeneration patch according to claim 1, wherein, The area of ​​the pattern satisfies the uniformity of more than 99% of the following mathematical formula: 【Mathematical Formula 1】 U=|H P1 -H P2 |≤0.1H n (In the formula, P1 and P2 are points on the peak or points on the valley, respectively, H) P1 and H P2 H represents the height at points P1 and P2, respectively. n (This represents the height difference between the peaks and valleys in a normal pattern).

7. The tympanic membrane regeneration patch according to claim 1, wherein, The surface of the tympanic membrane regeneration layer that is not bonded to the adhesive layer is coated with any one of the following: collagen, fibrin, growth factors, stem cells, exosomes, and therapeutic drugs.

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

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

10. The tympanic membrane regeneration patch according to claim 1, wherein, The peak and the valley each have a width and height of 700 to 900 nm.

11. A method for manufacturing a tympanic membrane regeneration patch according to any one of claims 1 to 10, the method comprising: The steps of manufacturing a tympanic membrane regeneration layer include: coating a polymer solution containing biocompatible polymers onto a polyurethane acrylate mold to manufacture a sheet-like semi-cured material; positioning the semi-cured material between the polyurethane acrylate mold and a polydimethylsiloxane mold, and then applying pressure using an upper plate and a lower plate respectively to manufacture a patterned semi-cured material; and drying the patterned semi-cured material. The step of forming an adhesive layer on the tympanic membrane regeneration layer manufactured as described above.

12. The method for manufacturing a tympanic membrane regeneration patch according to claim 11, wherein, The concentration of the biocompatible polymer in the polymer solution is 15% (w / w) to 24% (w / w).

13. The method for manufacturing a tympanic membrane regeneration patch according to claim 11, wherein, The drying process is carried out at 60°C to 80°C.

14. The tympanic membrane regeneration patch according to claim 11, wherein, The polyurethane acrylate mold or the polydimethylsiloxane mold has raised and recessed areas for forming the pattern.

15. The tympanic membrane regeneration patch according to claim 11, wherein, The upper plate has a pressure bar and a guide bar.

16. The tympanic membrane regeneration patch according to claim 11, wherein, The pressure is between 0.40 Pa and 0.70 Pa.

17. A method for tympanic membrane regeneration, the method comprising: The steps involved in applying a tympanic membrane regeneration layer to a perforated tympanic membrane; and The step of applying a hydrocolloid adhesive layer onto the tympanic membrane regeneration layer, The tympanic membrane regeneration layer is composed of lactide-glycolic acid copolymer or polycaprolactone. Patterns for tympanic membrane cell growth are formed on one or both sides of the tympanic membrane regeneration layer. The pattern consists of repeated peaks and valleys. The tympanic membrane regeneration layer has a tensile strength of 3 to 20 MPa and a thickness of 10 μm to 40 μm.

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

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

20. The tympanic membrane regeneration method according to claim 17, wherein, The peak and the valley each have a width and height of 700 to 900 nm.

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