Membrane for periodontal tissue regeneration and method for preparing the same

Abstract

Disclosed is a membrane for periodontal tissue regeneration and a method of preparing the same. The method of preparing a membrane for periodontal tissue regeneration according to an embodiment includes the steps of: preparing a membrane using a biodegradable polymer; cross-linking the membrane; dyeing a portion of the surface of the membrane; and freeze-drying the membrane, wherein the biodegradable polymer includes collagen.

Description

Technical Field

[0001] This disclosure relates to a method for manufacturing a membrane capable of regenerating periodontal tissue and the membrane manufactured by the method. Background Technology

[0002] Typically, membranes for periodontal tissue regeneration are used during tooth extractions or dental treatments to increase alveolar bone around the teeth and promote periodontal tissue regeneration. For example, if a membrane for periodontal tissue regeneration is attached to a site where bone regeneration is needed, space for bone regeneration is ensured and soft tissue is prevented from shifting downwards, thereby aiding in periodontal tissue regeneration. Relatedly, Korean Patent Publication No. 10-2019-0007373 discloses a technique for membranes that facilitate periodontal tissue regeneration.

[0003] Regarding membranes used for periodontal tissue regeneration, the thicker the membrane, the greater its durability. However, if the thickness increases, the membrane's flexibility may decrease, potentially causing gum pain in patients undergoing membrane surgery.

[0004] In previous membranes used for periodontal tissue regeneration, because one surface of the membrane was the same color as the other, it was difficult for surgeons to distinguish them with the naked eye when growth factors that promote periodontal tissue regeneration were only contained on a specific surface of the membrane. Summary of the Invention

[0005] The technical concept disclosed herein is intended to solve the aforementioned technical problems, and its purpose is to provide a technique that can improve the durability and flexibility of membranes used for periodontal tissue regeneration.

[0006] Another objective of the technical concept disclosed herein is to provide a technique that allows for the visual differentiation of one surface of a membrane used for periodontal tissue regeneration from another surface.

[0007] Another objective of the technical concept disclosed herein is to provide a method for manufacturing membranes for periodontal tissue regeneration.

[0008] Another objective of the technical concept disclosed herein is to provide a membrane for periodontal tissue regeneration.

[0009] The technical problems to be solved by this disclosure are not limited to the aforementioned technical problems. Those skilled in the art to which this disclosure pertains can clearly understand other technical problems not mentioned by the following description.

[0010] To achieve this objective, as an embodiment of this disclosure, a method for preparing a membrane for periodontal tissue regeneration includes the following steps: a membrane preparation step, using a biodegradable polymer to prepare the membrane; a crosslinking step, crosslinking the membrane; a staining step, staining a portion of the surface of the membrane; and a freeze-drying step, freeze-drying the membrane, wherein the biodegradable polymer includes collagen.

[0011] In one embodiment, the method for preparing the membrane for periodontal tissue regeneration may further include the following steps: preparing a solution containing the biodegradable polymer.

[0012] In one embodiment, the method for preparing a membrane for periodontal tissue regeneration may further include the following step: an extrusion step, in which the membrane is extruded.

[0013] In one embodiment, the method for preparing a membrane for periodontal tissue regeneration may further include the following step: a washing step, in which the membrane is washed.

[0014] In one embodiment, the method for preparing the membrane for periodontal tissue regeneration may further include the following step: a sterilization step, in which the membrane is sterilized.

[0015] As one embodiment, in the dyeing step, a portion of the membrane can be dyed while a portion of the membrane is immersed in the dyeing solution and the remaining portion of the membrane floats in the dyeing solution.

[0016] In one embodiment, the staining solution may contain growth factors.

[0017] In one embodiment, the dyeing solution may contain the same type of biodegradable polymer as the type of biodegradable polymer used to prepare the membrane.

[0018] In one embodiment, during the freeze-drying step, the membrane can be placed into a freeze dryer to perform the freeze-drying process while a portion of the membrane is immersed in the dyeing solution and the remaining portion of the membrane floats in the dyeing solution.

[0019] To achieve this objective, as another embodiment of this disclosure, the membrane for periodontal tissue regeneration can be prepared by the aforementioned method for preparing a membrane for periodontal tissue regeneration.

[0020] The above-described technical solutions are merely exemplary and should not be construed as limiting this disclosure. In addition to the exemplary embodiments described above, there may be additional embodiments as illustrated in the accompanying drawings and the detailed description of this disclosure.

[0021] As described above, according to various embodiments of the present disclosure, the membrane exhibits excellent durability because its tensile strength is above a predetermined level.

[0022] According to various embodiments of this disclosure, since the membrane has an elongation rate above a predetermined level, it exhibits excellent flexibility, which can help alleviate the pain of patients undergoing membrane surgery.

[0023] According to various embodiments of this disclosure, since specific surfaces of the membrane can be selectively stained, these specific surfaces can be easily distinguished with the naked eye, thereby improving surgical convenience.

[0024] The effects of this disclosure are not limited to those mentioned above, and those skilled in the art can clearly understand other effects not mentioned from the description in the claims. Attached Figure Description

[0025] Figure 1 This is a flowchart schematically illustrating a method for preparing a membrane for periodontal tissue regeneration according to an embodiment of the present disclosure.

[0026] Figure 2 These are photographs taken of one surface and another surface of the membrane according to an embodiment of the present disclosure.

[0027] Figure 3 These are photographs taken of one surface and another surface of the membrane according to another embodiment of the present disclosure. Detailed Implementation

[0028] The preferred embodiments of this disclosure will be described in more detail with reference to the accompanying drawings. For the sake of brevity, well-known technical aspects will be omitted or summarized.

[0029] It should be noted that references to "a" or "an" embodiment of this disclosure in this specification do not necessarily refer to the same embodiment, but rather to at least one.

[0030] In the following embodiments, the terms "first," "second," etc., are not intended to be limiting, but are used to distinguish one constituent element from another.

[0031] In the following embodiments, the singular expression includes the plural expression unless otherwise explicitly stated in the context.

[0032] In the following embodiments, terms such as "comprising" or "having" indicate the presence of the features or constituent elements described in the specification, rather than pre-excluding the possibility of adding more than one other feature or constituent element.

[0033] In the following embodiments, when a membrane, region, constituent element, or other part is located above or on top of other parts, this includes not only the case where it is located immediately above other parts, but also the case where other membranes, regions, constituent elements, or other parts are sandwiched in between.

[0034] Throughout the entire specification of this application, when a unique tolerance for preparation and substance is given for the meaning of the reference, the term "about" used before the numerical value is used to mean that the value is close to or within the range of the numerical value, and is used to prevent unscrupulous infringers from improperly using the disclosure of accurate or absolute numerical values ​​for the purpose of understanding this application.

[0035] Where an embodiment can be implemented in different ways, the specific process sequence may also be performed differently from the described sequence. For example, two processes described consecutively may be performed substantially simultaneously, or they may be performed in the reverse order of the description. That is, unless otherwise stated in the specification or there is a clear conflict in the context, the steps of the methods described herein may be performed in any appropriate order.

[0036] A method for preparing a membrane for periodontal tissue regeneration according to an embodiment may include a solution preparation step, a membrane preparation step, an extrusion step, a crosslinking step, a washing step, a staining step, a freeze-drying step, a hot-pressing step, and a sterilization step. The method for preparing a membrane for periodontal tissue regeneration according to an embodiment... Figure 1 The flowchart shown will be used for explanation, and for convenience, the explanations will be presented in a sequential order.

[0037] 1. Solution preparation steps <s101> In this step, a biodegradable polymer solution can be prepared. In one embodiment, the biodegradable polymer solution, as a solution containing biodegradable polymers, can be a solution mixed with a solvent and biodegradable polymers. For example, the solvent may include at least one of water and physiological saline. The biodegradable polymer may include collagen.

[0038] According to one embodiment, the concentration of the biodegradable polymer in the biodegradable polymer solution can be more than about 1 w / w% and less than about 3 w / w%. For example, the concentration of the biodegradable polymer can be 1 w / w%, 1.1 w / w%, 1.2 w / w%, 1.3 w / w%, 1.4 w / w%, 1.5 w / w%, 1.6 w / w%, 1.7 w / w%, 1.8 w / w%, 1.9 w / w%, 2 w / w%, 2.1 w / w%, 2.2 w / w%, 2.3 w / w%, 2.4 w / w%, 2.5 w / w%, 2.6 w / w%, 2.7 w / w%, 2.8 w / w%, 2.9 w / w%, or 3 w / w. The concentration of the biodegradable polymer in the biodegradable polymer solution can be in the range of more than one of the above values ​​and less than another of the above values.

[0039] For example, the concentration of biodegradable polymers in a biodegradable polymer solution can be above 1 w / w% and below 3 w / w%, above 1 w / w% and below 2 w / w%, or above 2 w / w% and below 3 w / w%.

[0040] In a biodegradable polymer solution, if the concentration of the biodegradable polymer is less than 1 w / w%, the tensile strength of the membrane used for periodontal tissue regeneration may decrease and it may be easily torn, or it may have a negative impact on durability. If the concentration of the biodegradable polymer is greater than 3 w / w%, the membrane thickness may become too thick when the membrane is hydrated, and the membrane's flexibility may decrease, thus posing a risk of pain at the site of membrane surgery.

[0041] 2. Membrane preparation steps <s102>< / s102> In this step, a biodegradable polymer can be used to prepare the membrane. For example, in this step, the biodegradable polymer solution prepared in step S101 can be poured into a mold of a specific shape (as an example, a quadrilateral mold) and freeze-dried to prepare a membrane of a predetermined shape.

[0042] According to one specific example, a biodegradable polymer solution can be frozen at a temperature above about -40°C and below about -30°C (e.g., -40°C, -39°C, -38°C, -37°C, -36°C, -35°C, -34°C, -33°C, -32°C, -31°C, or -30°C) for about 18 hours and below about 24 hours (e.g., 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours), and then at a temperature above 0°C and below 10°C (e.g., 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, or 10°C). Vacuum drying for more than 12 hours and less than 48 hours (e.g., 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours or 48 hours).

[0043] 3. Extrusion Step <s103>< / s103> In this step, the membrane prepared in step S102 can be extruded. For example, in this step, the membrane can be physically extruded by applying pressure at a predetermined pressure. According to one specific example, a pressure of approximately 5 kgf / cm² can be used. 2 Above and approximately 10 kgf / cm 2 The following pressure is applied to the membrane for approximately 5 seconds to approximately 10 seconds.

[0044] 4. Crosslinking step <s104>< / s104> In this step, the membrane can be cross-linked. For example, the membrane can be chemically or non-chemically cross-linked in this step. According to one specific example, a cross-linking agent can be used to cross-link the membrane. The cross-linking agent may include at least one of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC: 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide) and glutaraldehyde. When chemical cross-linking is performed, the membrane is immersed in the cross-linking agent and the cross-linking process is carried out at a temperature of 4°C or higher and 20°C or lower for 4 hours or more and 24 hours or less.

[0045] According to another specific example, non-chemical methods can be used to crosslink the membrane. For example, the membrane can be crosslinked by dehydrogenation heat treatment (DHT). The dehydrogenation heat treatment crosslinking method is a non-chemical crosslinking method that crosslinks the membrane by heating it. It involves placing the membrane in a vacuum oven and reacting it at a temperature of 100°C to 200°C for 10 hours to 48 hours.

[0046] 5. Washing steps <s105>< / s105> In this step, the crosslinked membrane from step S104 can be washed. In one embodiment, the crosslinked membrane is repeatedly washed several times (e.g., 5 to 10 times) with an aqueous ethanol solution of 70 w / w% or more and 80 w / w% or less for a period of 1 to 2 hours. Then, the membrane can be immersed in distilled water for a predetermined time to remove any residual crosslinking agent.

[0047] 6. Staining steps <s106>< / s106> In this step, a portion of the membrane surface can be dyed. For example, in this step, a portion of the membrane can be dyed within a predetermined time while a portion of the membrane is immersed in the dyeing solution and the rest of the membrane floats in the dyeing solution. According to one specific example, if the membrane is placed into a mold having a predetermined shape after the dyeing solution has been added, a portion of the membrane can be immersed in the dyeing solution.

[0048] In one embodiment, the staining solution may include a staining component. The staining solution may also include at least one of a biodegradable polymer and a growth factor. The staining solution may also include at least one of water, distilled water, physiological saline, and phosphate-buffered saline as a solvent. In a specific example, the staining solution may include a staining component, a biodegradable polymer, a growth factor, and a solvent.

[0049] In one embodiment, the staining component, as a component with a specific color, may include at least one of riboflavin and methylene blue. According to one embodiment, riboflavin is yellow and therefore can be used as a staining component. In a specific example, if riboflavin adheres to the surface of the membrane, it can alleviate inflammation at the surgical site and promote periodontal tissue regeneration. Riboflavin has the advantage of low in vivo toxicity compared to other types of staining components.

[0050] In one embodiment, the concentration of the staining component included in the staining solution may be greater than about 0.1 mg / L and less than about 2 mg / L (e.g., 0.1 mg / L, 0.2 mg / L, 0.3 mg / L, 0.4 mg / L, 0.5 mg / L, 0.6 mg / L, 0.7 mg / L, 0.8 mg / L, 0.9 mg / L, 1 mg / L, 1.1 mg / L, 1.2 mg / L, 1.3 mg / L, 1.4 mg / L, 1.5 mg / L, 1.6 mg / L, 1.7 mg / L, 1.8 mg / L, 1.9 mg / L, or 2 mg / L). The amount of staining component added to the staining solution can be appropriately adjusted as needed.

[0051] According to one embodiment, the staining solution may contain growth factors. The growth factors are at least one selected from bone morphogenetic proteins (BMPs), transforming growth factor-β (TGF-β), insulin-like growth factor (IGF), fibroblast growth factor (FGFs), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), tripotassium phosphate (TCP), human growth hormone, and angiopoietin.

[0052] In one embodiment, the concentration of growth factors included in the staining solution can be above about 1 mg / L and below about 2 mg / L. The amount of growth factors added to the staining solution can be adjusted appropriately as needed.

[0053] In one embodiment, the biodegradable polymer included in the staining solution may be of the same type as the biodegradable polymer constituting the membrane. According to one specific example, the biodegradable polymer included in the staining solution may include collagen. In one specific example, with the presence of collagen in the staining solution, the staining components and growth factors can readily adhere to the surface of the collagen-based membrane. The staining solution is viscous due to the predetermined concentration of collagen, therefore, the membrane may not be completely precipitated or immersed in the staining solution, but only partially immersed.

[0054] According to one embodiment, the concentration of the biodegradable polymer contained in the staining solution can be more than about 0.5 w / w% and less than about 1 w / w%. For example, the concentration of the biodegradable polymer contained in the staining solution can be 0.5 w / w%, 0.6 w / w%, 0.7 w / w%, 0.8 w / w%, 0.9 w / w%, or 1 w / w. The concentration of the biodegradable polymer contained in the staining solution can be in the range of more than one of the above values ​​and less than another of the above values.

[0055] For example, the concentration of biodegradable polymers in the dyeing solution can be 0.5 w / w% to 1 w / w, 0.5 w / w% to 0.9 w / w, 0.6 w / w% to 1 w / w, or 0.7 w / w% to 1 w / w. If the concentration of biodegradable polymers in the dyeing solution is less than 0.5 w / w, the membrane durability may be reduced and it may be easily torn. If the concentration of biodegradable polymers exceeds 1 w / w, there are concerns about negatively impacting the membrane's elongation and flexibility.

[0056] According to one specific example, in this step, one surface of the membrane (i.e., the surface in contact with the periodontal tissue) can be selectively stained. Therefore, the surgeon can easily distinguish one surface of the membrane (the stained surface) from the other (the unstained surface) with the naked eye, and the stained surface of the membrane can be used as the contact surface with the periodontal tissue.

[0057] 7. Freeze-drying step <s107>< / s107> In this step, the membrane can be freeze-dried. For example, in this step, the membrane can be placed into a freeze dryer to perform the freeze-drying process while a portion of the membrane is immersed in the dyeing solution and the rest floats in the dyeing solution. That is, in this step, instead of removing the membrane from the dyeing solution and placing it into the freeze dryer after performing step S106, the membrane can be placed into the freeze dryer while maintaining a portion of the membrane immersed in the dyeing solution of the mold. In one implementation example, the mold containing the dyeing solution and the membrane can be directly placed into the freeze dryer.

[0058] According to one specific example, in this step, the membrane is frozen at a temperature above approximately -100°C and below approximately -30°C (e.g., -100°C, -95°C, -90°C, -85°C, -80°C, -75°C, -70°C, -65°C, -60°C, -55°C, -50°C, -45°C, -40°C, -35°C, or -30°C) for approximately 10 minutes and less than approximately 24 hours (e.g., 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 1...). 9 hours, 20 hours, 21 hours, 22 hours, 23 hours or 24 hours), then vacuum dry at a temperature above 0°C and below 10°C (e.g. 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C or 10°C) for about 10 minutes and about 24 hours (e.g. 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours or 24 hours).

[0059] 8. Hot pressing step <s108>< / s108> In this step, the membrane freeze-dried in step S107 can be extruded by heating. For example, in this step, the membrane can be pressurized at a predetermined temperature and a predetermined pressure. According to one specific example, the membrane can be pressurized at a temperature above about 50°C and below about 90°C, at a pressure of about 5 kgf / cm³. 2 Above and approximately 10 kgf / cm 2 The membrane is pressurized under the following pressure and squeezed for approximately 5 seconds to approximately 15 seconds.

[0060] 9. Sterilization steps <s109>< / s109> In this step, the membrane can be sterilized. For example, sterilization can be performed by irradiating the membrane with radiation or by exposing the membrane to ethylene oxide gas.

[0061] According to one embodiment, sterilization can be performed by irradiating the membrane with gamma rays of 5 kGy or more but less than 30 kGy. According to another embodiment, the membrane can be placed in a sterilization chamber and sterilized by exposing it to ethylene oxide gas under vacuum for a predetermined time period.

[0062] The present disclosure will now be described in more detail through specific embodiments and experimental examples. The embodiments and experimental examples described below are merely illustrative examples to aid in understanding the present disclosure, and therefore the scope of the disclosure is not limited thereto.

[0063] Membrane preparation <Examples 1-3 and Comparative Examples 1-2> A biodegradable polymer solution was prepared by mixing collagen (derived from bovine tendon) with physiological saline. The collagen concentration was set according to the values ​​listed in Table 1 below. 20g of the prepared collagen solution was poured into a quadrilateral mold and frozen at -30℃ to -40℃ for 18 to 24 hours, then vacuum-dried at 0℃ to 10℃ for 12 to 48 hours. The vacuum-dried membrane was then subjected to a flow rate of 5 kgf / cm². 2 ~10kgf / cm 2 Apply pressure for 5 to 10 seconds. Immerse the membrane in a crosslinking agent containing 5 mM EDC in 95 w / w% ethanol and crosslink at 4°C to 20°C for 4 to 24 hours. Wash the crosslinked membrane with a 70 w / w% ethanol aqueous solution for 1 to 2 hours, and then immerse the membrane in distilled water for 1 hour to remove residual crosslinking agent.

[0064] Next, 100g of staining solution (containing 1mg / L riboflavin, 1w / w% collagen, and 1mg / L bone morphogenetic protein, with distilled water as the solvent) was poured into a quadrilateral mold. The membrane was then placed into the mold and allowed to stand for 1 second to 30 minutes. While maintaining partial immersion in the staining solution within the mold, the membrane was placed in a freeze dryer and frozen at -100℃ to -30℃ for 10 minutes to 24 hours. The membrane was then vacuum dried at 0℃ to 10℃ for 10 minutes to 24 hours. The freeze-dried membrane was then subjected to a vacuum drying process at 50℃ to 90℃ at a rate of 5kgf / cm². 2 ~10kgf / cm 2 The pressure is applied for 5 to 15 seconds. Then, the membrane is exposed to sterilization for 1 hour at a temperature of 38 to 60°C, a relative humidity of 40% to 80%, and an ethylene oxide concentration of 1000 mg / L, thus completing the membrane preparation.

[0065] [Table 1]

[0066] <Examples 4-6 and Comparative Examples 3-4> Except for setting the concentration of collagen in the staining solution according to the values ​​recorded in Table 2 below, everything else was set the same as in Example 3, thereby preparing the membrane.

[0067] [Table 2]

[0068] <Example 7> Except that the staining solution did not contain growth factors (bone morphogenetic proteins), everything else was set the same as in Example 3, and a membrane was prepared.

[0069] <Example 8> Except for the use of methylene blue at a concentration of 1 mg / L instead of riboflavin as the staining component in the staining solution, everything else was set the same as in Example 3, and a membrane was prepared.

[0070] <Comparative Example 5> 100g of staining solution (in which the concentration of riboflavin is 1mg / L, the concentration of collagen is 1w / w%, and the concentration of bone morphogenetic protein is 1mg / L, with distilled water used as solvent) was poured into a quadrilateral mold. The membrane was then placed into the mold and allowed to stand for 1 second to 30 minutes. The process was then identical to that in Example 3, except that the membrane was removed from the mold and placed in a freeze dryer.

[0071] <Comparative Example 6> A biodegradable polymer solution was prepared by mixing collagen (derived from bovine tendon) with physiological saline, with the collagen concentration set at 3 w / w%. 20 g of the prepared collagen solution was poured into a quadrilateral mold and frozen at -30°C to -40°C for 18 to 24 hours, followed by vacuum drying at 0°C to 10°C for 12 to 48 hours. The vacuum-dried membrane was then subjected to a flow rate of 5 kgf / cm². 2 ~10kgf / cm 2 Apply pressure for 5 to 10 seconds.

[0072] Then, 100g of staining solution (containing 1mg / L riboflavin, 1w / w% collagen, and 1mg / L bone morphogenetic protein, with distilled water as the solvent) was poured into a quadrilateral mold. The membrane was then placed into the mold and allowed to stand for more than 1 second but less than 30 minutes. The membrane was then removed from the mold and placed in a freeze dryer. It was frozen at -100℃ to -30℃ for 10 minutes to 24 hours, and then vacuum dried at 0℃ to 10℃ for 10 minutes to 24 hours. The freeze-dried membrane was then subjected to a vacuum drying process at 50℃ to 90℃ at 5kgf / cm². 2 ~10kgf / cm 2 The pressure is applied for 5 to 15 seconds. Afterward, the membrane is immersed in a crosslinking agent containing 5 mM EDC in 95 w / w ethanol and crosslinked at 4°C to 20°C for 4 to 24 hours. The crosslinked membrane is washed with a 70 w / w ethanol aqueous solution for 1 to 2 hours, and then immersed in distilled water for 1 hour to remove residual crosslinking agent. Subsequently, the membrane is sterilized by exposing it to ethylene oxide at 38°C to 60°C, 40% to 80% relative humidity, and 1000 mg / L for 1 hour, thus completing the membrane preparation.

[0073] Sample preparation The membranes prepared according to the various embodiments and comparative examples were cut into dimensions of 10 cm wide and 20 cm long to prepare samples.

[0074] Evaluation of the physical properties of membranes Measurement of hydration time Immerse the sample in 200 ml of distilled water at 36°C. Observe the sample absorbing the distilled water while the amount of distilled water decreases. Record the time when the amount of distilled water stops decreasing in Table 3 below.

[0075] Thickness measurement Immerse the sample in 200 ml of distilled water at 36°C. Observe the sample absorbing the distilled water while the amount of distilled water decreases with the naked eye. Measure the film thickness at the moment when the amount of distilled water no longer decreases and record the results in Table 3 below.

[0076] Measurement of absorption capacity The absorbance of the specimen (g / cm) was measured according to EN13726-1. 2 ), and record them in Table 3 below.

[0077] Measurement of tensile strength When the specimen is fixed in a universal testing machine and the crosshead speed is set to 200 mm / min and the specimen is stretched, the tensile force at the moment of specimen fracture is measured and recorded in Table 3 below.

[0078] Measurement of elongation Fix the specimen in a universal testing machine. If the crosshead speed is set to 200 mm / min and the specimen is stretched until it breaks, derive the elongation of the specimen and record it in Table 3 below. The elongation is derived using the following formula.

[0079] [Calculation formula] Elongation (%) = (Length of elongation of the specimen - Initial length of the specimen) / Initial length of the specimen × 100 Visual inspection When the appearance of the samples prepared according to the various embodiments and comparative examples is checked by visual inspection, the samples are classified as good, average, or poor according to their condition, and recorded in Table 3 below. In the appearance inspection results, "good" means that no wrinkles or tears are produced on the surface of the sample, "average" means that the area with poor marketability (areas with wrinkles or tears) is less than 10% of the entire area of ​​the sample, and "poor" means that the area with poor marketability is more than 10% of the entire area of ​​the sample.

[0080] [Table 3]

[0081] Referring to the results in Table 3, it was confirmed that the hydration time of Examples 1 to 3, where the collagen concentration in the biodegradable polymer solution was in the range of 1 w / w% to 3 w / w%, was less than 20 seconds, which was faster than that of Comparative Example 2. This resulted in a membrane thickness within the range of 0.1 mm to 0.3 mm, suitable for surgical procedures on periodontal tissues. If the membrane absorbency is too low, the membrane thickness at hydration may be too small, potentially reducing durability; if the absorbency is too high, the membrane thickness at hydration may be too large, potentially reducing flexibility. However, in Examples 1 to 3, an absorbency of 0.2 g / cm³, suitable for surgical procedures on periodontal tissues, was confirmed. 2 ~0.6g / cm 2 Within a certain range. If the tensile strength of the membrane is too low, the membrane may be easily damaged; if the tensile strength is too high, the flexibility decreases, making surgery difficult and potentially causing pain at the surgical site. However, in Examples 1 to 3, a tensile strength range of 1 MPa to 7 MPa was confirmed as suitable for surgery on periodontal tissue sites. If the elongation of the membrane is too low, the membrane's flexibility decreases, making surgery difficult and potentially causing pain at the surgical site. If the elongation is too high, the membrane may be easily damaged. However, in Examples 1 to 3, an elongation range of 10% to 17% was confirmed as suitable for surgery on periodontal tissue sites.

[0082] It was confirmed that Comparative Example 1, in which the collagen concentration in the biodegradable polymer solution was less than 1 w / w%, had lower tensile strength and elongation compared to Examples 1 to 3, and its appearance was deteriorated. It can be seen that since the thickness of Comparative Example 1 was less than 0.1 mm, it had a negative impact on absorption capacity, tensile strength, elongation, etc.

[0083] Comparative Example 2, which had a collagen concentration greater than 3 w / w% in a biodegradable polymer solution, was found to have a thicker thickness, higher tensile strength, and lower elongation compared to Examples 1 to 3, thus potentially causing pain when attached to the surgical site.

[0084] The membranes of Examples 3 to 6, with collagen concentrations in the staining solution ranging from 0.5 w / w% to 1 w / w%, were confirmed to have excellent durability and flexibility. The tensile strength and elongation of Comparative Example 3, with a collagen concentration in the staining solution less than 0.5 w / w%, were confirmed to be inferior to those of Examples 3 to 6. Furthermore, the elongation of Comparative Example 4, with a collagen concentration in the staining solution greater than 1 w / w%, was confirmed to be lower than that of Examples 3 to 6, potentially causing pain when applied to surgical sites.

[0085] In Comparative Example 5, where the membrane was simply removed after being placed into a mold containing a dyeing solution and then freeze-dried, deterioration of the sample's appearance was confirmed.

[0086] In Comparative Example 6, where the membrane was cross-linked after dyeing and freeze-drying, the appearance of the sample was observed, confirming that both sides of the sample were dyed. In Comparative Example 6, selective dyeing of only one surface of the sample was not achieved.

[0087] Figure 2 These are photographs taken of one surface and the other surface of the membrane according to Embodiment 3 of this disclosure. Figure 2 (a) is the front surface of the membrane. Figure 2 (b) is the back surface of the membrane. (Refer to...) Figure 2 It can be seen that only the back surface of the membrane was stained yellow.

[0088] Figure 3 These are photographs taken of one surface and the other surface of the membrane according to Embodiment 8 of this disclosure. Figure 3 (a) is the front surface of the membrane. Figure 3 (b) is the back surface of the membrane. (Refer to...) Figure 3 It can be seen that only the back surface of the membrane was stained blue.

[0089] As described above, according to various embodiments of the present disclosure, the membrane exhibits excellent durability because its tensile strength is above a predetermined level.

[0090] According to various embodiments of this disclosure, the membrane exhibits excellent flexibility due to its elongation rate exceeding a predetermined level, which can help alleviate the pain of patients undergoing membrane surgery.

[0091] According to various embodiments of this disclosure, since specific surfaces of the membrane can be selectively stained, these surfaces can be easily distinguished with the naked eye, thus improving surgical convenience. For example, if the surface in contact with periodontal tissue is stained yellow or blue, the stained surface of the membrane and the unstained surface can be easily distinguished with the naked eye, thereby improving convenience from the surgeon's perspective.

[0092] As described above, although the present disclosure has been specifically illustrated through embodiments, the above embodiments are only preferred examples of the present disclosure and should not be construed as the present disclosure being limited to the above embodiments. The scope of the claims of the present disclosure should be understood as the claims and their equivalents.

Claims

1. A method for preparing a membrane for periodontal tissue regeneration, characterized in that, Includes the following steps: The steps for preparing the membrane involve using biodegradable polymers to prepare the membrane; The crosslinking step crosslinks the membrane; The dyeing step involves dyeing a portion of the surface of the membrane; and The freeze-drying step involves freeze-drying the membrane. The biodegradable polymers include collagen.

2. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, It also includes the following steps: The step of preparing the solution involves preparing a solution containing the biodegradable polymer.

3. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, It also includes the following steps: The extrusion step involves extruding the membrane.

4. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, It also includes the following steps: The washing step involves washing the membrane.

5. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, It also includes the following steps: The sterilization step involves sterilizing the membrane.

6. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, In the dyeing step, a portion of the membrane is dyed while a portion of the membrane is immersed in the dyeing solution and the remaining portion of the membrane floats in the dyeing solution.

7. The method for preparing the membrane for periodontal tissue regeneration as described in claim 6, characterized in that, The staining solution contains growth factors.

8. The method for preparing the membrane for periodontal tissue regeneration as described in claim 6, characterized in that, The staining solution contains the same type of biodegradable polymer as the biodegradable polymer used to prepare the membrane.

9. The method for preparing the membrane for periodontal tissue regeneration as described in claim 1, characterized in that, In the freeze-drying step, the membrane is placed into a freeze dryer to perform the freeze-drying process while a portion of the membrane is immersed in the dyeing solution and the remaining portion of the membrane floats in the dyeing solution.

10. A membrane for periodontal tissue regeneration, characterized in that, The membrane for periodontal tissue regeneration is prepared according to any one of claims 1 to 9.

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