Hydrogel composition, use of hydrogel comprising same, and kit for preparing hydrogel or delivering cells in body comprising same

Abstract

The present invention relates to a hydrogel composition, a use of a hydrogel comprising same, and a kit for preparing a hydrogel or delivering cells into the body. The hydrogel composition according to one aspect of the present invention exhibits an appropriate gelation rate and can produce a hydrogel of constant quality by simple mixing alone, thereby having excellent procedural convenience and reproducibility. In addition, the hydrogel of the present invention continuously releases growth factors contained therein at a high concentration, thereby stably providing effects of tissue regeneration, wound healing, and induction of a biological response for a long time. The hydrogel of the present invention has excellent biocompatibility, thereby ensuring safety when applied to the human body, and can be widely applied not only to medical fields including dermatology and surgery, but also to the entire field of aesthetics.

Description

Hydrogel composition, uses of a hydrogel containing the same, and a kit for manufacturing a hydrogel containing the same or for in vivo cell delivery.

[0001] The present invention relates to a hydrogel composition, the use of a hydrogel containing the same, and a kit for manufacturing a hydrogel or delivering cells into the body.

[0002] Hydrogels are gel-like materials in which a polymer network structure maintains a state containing a large amount of water and active substances. Hydrogels possess physical properties similar to soft tissue and are characterized by excellent biocompatibility. Due to these characteristics, hydrogels are utilized in the medical field as tissue fillers, hemostatic agents, wound dressings, and drug delivery systems, and are also widely used in the cosmetic field as auxiliary materials for skin procedures, as well as for skin regeneration and soothing. In particular, they have high commercial value as they can be applied directly to the treatment site to induce biological responses while maintaining a consistent shape.

[0003] Hydrogels have been provided in various forms depending on their intended use and application environment. For example, some hydrogels are available in a form that is manufactured in advance as a gel and supplied as a finished product, while others are known to be prepared by mixing a composition of multiple components immediately before the procedure to form a hydrogel, which is then applied immediately. Although finished hydrogels offer the advantage of ease of use, the physical properties of the gel are determined at the manufacturing stage; therefore, there may be limitations in flexibly adjusting these properties according to the condition of the treatment site, the purpose of the procedure, or differences in the treatment environment. Consequently, the method of mixing a composition of multiple components immediately before the procedure to form a hydrogel and applying it immediately has recently become more preferred in medical and cosmetic treatment settings. This method is highly versatile as it allows the practitioner to adjust the conditions for hydrogel formation according to the application site or purpose of the procedure, and it offers the advantage of adapting to various treatment environments.

[0004] Meanwhile, in methods where a hydrogel is formed immediately before the procedure, various factors such as the gelation reaction, mixing conditions, and characteristics of the composition affect the hydrogel formation results. In particular, since complex manufacturing processes or precise condition control are often difficult in the field, there is a continuously increasing demand for compositions that can form a hydrogel in a consistent state with only simple mixing. Additionally, a generally known technique involves using polymeric materials such as hyaluronic acid and methylcellulose to control the viscosity or shape of the gel, but this method has revealed limitations in terms of low reproducibility and convenience.

[0005] In one aspect, the object of the present invention is to provide a hydrogel composition with an appropriate gelation rate.

[0006] In one aspect, the object of the present invention is to provide a hydrogel with excellent homogeneity and chemical, physical, and biological stability.

[0007] In one aspect, the object of the present invention is to provide a hydrogel capable of continuously releasing growth factors at high concentrations.

[0008] In one aspect, the object of the present invention is to provide a hydrogel with excellent biocompatibility.

[0009] In one aspect, the object of the present invention is to effectively deliver cells to a living organism or target.

[0010] In one aspect, the object of the present invention is to improve the viability of the cell tissue at the site where the hydrogel is delivered, and / or the cell tissue provided with the hydrogel.

[0011] In one aspect, the objective of the present invention is to provide a hydrogel with excellent applicability across medical fields, including dermatology and surgery, and cosmetic fields.

[0012] In one aspect, the object of the present invention is to provide a kit for manufacturing a hydrogel and a method for manufacturing the same, which offers excellent ease of manufacturing and reproducibility when used (e.g., at the site of a procedure).

[0013] The object of the present invention is not limited to the technical problem as described above, and other technical problems may be derived from the following description.

[0014] To achieve the above objectives, the present invention provides, in one aspect, a hydrogel composition comprising a platelet extract and thrombin of greater than 25 Unit / ml and less than 100 Unit / ml.

[0015] In addition, the present invention provides a hydrogel composition in which the platelet extract and thrombin are included in a volume ratio of 1:0.1 to 10 in the above aspects.

[0016] In addition, the present invention provides a hydrogel composition in which the concentration of thrombin is 30 to 70 Unit / ml, in addition to the above aspects.

[0017] In addition, the present invention provides a hydrogel composition having a gelation time of 5 to 15 seconds, in the above aspects.

[0018] In addition, in the above aspects, the present invention provides a hydrogel composition comprising a platelet extract obtained by freezing platelets in an environment of -100 to -60 degrees Celsius for 90 to 150 minutes and then thawing them.

[0019] In addition, the present invention provides a hydrogel composition comprising a platelet extract that has undergone freezing and thawing 1 to 5 times, in addition to the above aspects.

[0020] In addition, the present invention provides, in the above aspects, a hydrogel composition comprising a composition for cosmetic or medical use.

[0021] In addition, the present invention provides a hydrogel composition that includes, in the above aspects, a composition for hemostasis, skin tissue regeneration, skin soothing after dermatological procedures, or restoration of depressed tissue.

[0022] In addition, the present invention, in the above aspects, provides a kit for manufacturing a hydrogel, wherein the kit comprises a platelet extract and thrombin of greater than 25 Unit / ml and less than 100 Unit / ml.

[0023] In addition, the present invention provides a kit for manufacturing a hydrogel or for in vivo delivery, wherein the kit further comprises instructions, and the instructions comprise the platelet extract and thrombin in a volume ratio of 1:1 to 10.

[0024] In addition, in the above aspects, the present invention provides a kit for manufacturing a hydrogel or for in vivo delivery that further comprises cells.

[0025] In addition, the present invention provides a kit for manufacturing a hydrogel or delivering cells in vivo, wherein the cell comprises one or more selected from the group consisting of stem cells, immune cells, progenitor cells, and skin or tissue regeneration cells.

[0026] In addition, in the above aspects, the present invention provides a kit for manufacturing a hydrogel or for in vivo delivery comprising peripheral blood mononuclear cells (PBMC).

[0027] A hydrogel composition according to one aspect of the present invention exhibits an appropriate gelation rate and allows for the production of a hydrogel of consistent quality with only simple mixing, thereby offering excellent convenience and reproducibility for procedures. Furthermore, the structural and mechanical properties of the hydrogel are stably maintained even after gel formation, so that no degradation of the gel occurs in a biological environment. Additionally, the hydrogel of the present invention can stably provide tissue regeneration, wound healing, and biological response induction effects for a long period of time by continuously releasing growth factors contained therein at a high concentration. Accordingly, the hydrogel of the present invention has excellent biocompatibility, ensuring safety when applied to the human body, and can be widely applied across the entire field of beauty as well as in medical fields including dermatology and surgery.

[0028] The various and beneficial advantages and effects of the present invention are not limited to those described above and may be more easily understood in the process of explaining specific embodiments of the present invention.

[0029] Figure 1 is the result of measuring the release concentration of PDGF-BB according to the freezing time and the number of Freeze-Thaw cycles when preparing a platelet extract included in a hydrogel, which is one aspect of the present invention.

[0030] Figure 2 shows the results of measuring the release concentration of VEGF according to the freezing time and the number of Freeze-Thaw cycles when preparing a platelet extract included in a hydrogel, which is one aspect of the present invention.

[0031] Figure 3 shows the results of measuring the release concentration of EGF according to the freezing time and the number of Freeze-Thaw cycles when preparing a platelet extract included in a hydrogel, which is one aspect of the present invention.

[0032] The present invention is capable of various modifications and may have various embodiments, and specific embodiments are illustrated and described in the drawings. However, this is not intended to limit the invention to specific embodiments, and it should be understood that the invention includes all modifications, equivalents, and substitutions that fall within the spirit and scope of the invention.

[0033] Furthermore, terms or words used in the specification and claims of the present invention are not limited to their ordinary or dictionary meanings, and must be interpreted in a meaning and concept consistent with the technical spirit of the present invention, based on the principle that the inventor can appropriately define the concept of the terms to best describe his invention.

[0034] Terms including ordinal numbers, such as second, first, etc., may be used to describe various components, but said components are not limited by said terms. Such terms are used solely for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the second component may be named the first component, and similarly, the first component may be named the second component. The term "and / or" includes a combination of a plurality of related described items or any of a plurality of related described items.

[0035] Furthermore, the terms used in this application are used merely to describe specific embodiments and are not intended to limit the invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this application, terms such as “comprising” or “having” are intended to specify the existence of the features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, and should be understood as not precluding the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof.

[0036] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this specification.

[0037] The present invention will be described in detail below.

[0038] In one aspect, the present invention is a hydrogel composition.

[0039] In this specification, the term “hydrogel composition” means a composition used for forming or manufacturing a hydrogel.

[0040] In one aspect, the hydrogel composition may include a platelet extract and thrombin of more than 25 Unit / ml and less than 100 Unit / ml.

[0041] In one aspect, the present invention is used to produce platelet extract and thrombin hydrogen.

[0042] In addition, in one aspect, the present invention is a method for preparing a hydrogel using platelet extract and thrombin.

[0043] In this specification, the term “platelet extract” refers to a substance obtained by destroying or extracting platelets using physical or chemical methods, etc. The platelet extract is used as a synonym for platelet lysate.

[0044] In one aspect, the concentration of the thrombin may be greater than 25 Unit / mL (U / mL), or 27 Unit / mL or more, 29 Unit / mL or more, 31 Unit / mL or more, 33 Unit / mL or more, 35 Unit / mL or more, 37 Unit / mL or more, 39 Unit / mL or more, 41 Unit / mL or more, 43 Unit / mL or more, 45 Unit / mL or more, 47 Unit / mL or more, 49 Unit / mL or more, or 100 Unit / mL or less, 95 Unit / mL or less, 90 Unit / mL or less, 85 Unit / mL or less, 80 Unit / mL or less, 75 Unit / mL or less, 70 Unit / mL or less, 69 Unit / mL or less, 67 Unit / mL or less, 65 Unit / mL or less, 63 Unit / mL or less, 61 Unit / mL or less, 59 Unit / mL or less, 57 Unit / mL or less, 55 It may be Unit / mL or less, 53 Unit / mL or less, or 51 Unit / mL or less.

[0045] In one aspect, the concentration of the thrombin may be greater than 25 Unit / mL to 100 Unit / mL, or 30 to 100 Unit / mL, or 30 to 90 Unit / mL, or 30 to 85 Unit / mL, or 35 to 85 Unit / mL, or 35 to 80 Unit / mL, or 35 to 75 Unit / mL, or 35 to 70 Unit / mL, or 35 to 65 Unit / mL, or 37 to 65 Unit / mL, or 37 to 63 Unit / mL, or 39 to 63 Unit / mL, or 40 to 62 Unit / mL, or 41 to 60 Unit / mL, or 41 to 59 Unit / mL, or 43 to 57 Unit / mL, or 45 to 55 Unit / mL.

[0046] In one aspect, the platelet extract and thrombin may be included in a volume ratio of 1:0.1 to 10. Specifically, the platelet extract and thrombin may be included in a volume ratio of 1:0.1 to 10, in order, or in a volume ratio of 1:0.1 to 9, or in a volume ratio of 1:0.1 to 8, or in a volume ratio of 1:0.3 to 7, or in a volume ratio of 1:0.4 to 6, or in a volume ratio of 1:0.5 to 5, or in a volume ratio of 1:0.6 to 4, or in a volume ratio of 1:0.7 to 3, or in a volume ratio of 1:0.8 to 1.2.

[0047] In one aspect, when the thrombin and platelets are included in the range as described above, the highest quality hydrogel can be obtained.

[0048] In addition, in one aspect, the gelation time of the above composition may be 5 to 15 seconds.

[0049] In this specification, the term “gelation time” means the time required for a liquid or fluid substance to transition into a gel state after being mixed and reacted.

[0050] In one aspect, the gelation time of the above composition may be 5 to 15 seconds, or 5 to 13 seconds, or 5 to 11 seconds, or 6 to 10 seconds.

[0051] In one aspect, the platelet extract may be prepared by a freeze / thaw method.

[0052] Specifically, the platelet extract may include platelets or a platelet concentrate (concentrate) that have been frozen and then thawed in an environment of -100 to -60 degrees Celsius for 90 to 150 minutes.

[0053] In one aspect, the freezing temperature may be -95 to -65 degrees Celsius, or -90 to -65 degrees Celsius, or -90 to -70 degrees Celsius, or -90 to -75 degrees Celsius, or -85 to -75 degrees Celsius, or -83 to -77 degrees Celsius.

[0054] In addition, the freezing time may be 90 to 150 minutes, or 100 to 150 minutes, or 100 to 140 minutes, or 105 to 140 minutes, or 105 to 135 minutes, or 110 to 135 minutes, or 110 to 130 minutes, or 115 to 130 minutes, or 115 to 125 minutes.

[0055] In one aspect, the freezing and thawing may be performed 1 to 5 times. Specifically, the freezing and thawing may be performed 1 or more times, or 2 or more times, or 3 or more times, or 4 or more times, or 5 times or less, or 4 or less times, or 3 or less times, or 2 or less times, or 1 time under the above conditions.

[0056] In one aspect, if the above freezing is performed 2 to 4 times at approximately -90 to -70 degrees, a platelet extract containing a high concentration of growth factors can be obtained.

[0057] In addition, platelet extracts are generally obtained by performing a process of freezing for 8 hours or overnight followed by thawing about 3 to 7 times, and the platelet extract of the present invention is prepared by the above method, and has high manufacturing efficiency.

[0058] In one aspect, the growth factors may include, but are not limited to, PDGF (Platelet-Derived Growth Factor), EGF (Epidermal Growth Factor), FGF (Fibroblast Growth Factor), VEGF (Vascular Endothelial Growth Factor), TGF-α (Transforming Growth Factor-α), IGF-1 (Insulin-Like Growth Factor-1), HGF (Hepatocyte Growth Factor).

[0059] In one aspect, the above composition or the hydrogel containing the above composition may be for cosmetic or medical use.

[0060] In one aspect, the composition or the hydrogel comprising the composition may be used for hemostasis, skin tissue regeneration, skin soothing after dermatological procedures, or restoration of depressed tissue, but is not limited thereto.

[0061] The hydrogel of the present invention has excellent biocompatibility and can be formed immediately before the procedure and maintain its gel structure at the application site, so it can be utilized for various medical and cosmetic uses applied directly to the human body. The medical and cosmetic uses are not limited to specific procedures or functions, but can be understood as encompassing all forms of use that come into contact with human tissue or are applied to the surface or inside the human body.

[0062] Furthermore, the hydrogel of the present invention can continuously release high concentrations of growth factors. Accordingly, cells delivered to the body or a target together with the hydrogel of the present invention are protected within a stable microenvironment and can maintain a high survival rate even in the initial environment after in vivo injection. In addition, continuous stimulation by growth factors contributes to the functional activity and maintenance of biological quality of cells, and the cell-friendly microenvironment formed by the hydrogel can promote cell adhesion, proliferation, and functional expression. That is, the hydrogel of the present invention is advantageous for improving the efficiency and reproducibility of cell-based treatments or cosmetic procedures.

[0063] When applied for medical purposes, the hydrogel of the present invention may be used for the purposes of tissue protection, tissue support, assistance in tissue recovery, assistance in procedures, or induction of biological responses. For example, it may protect the application site during surgical or dermatological procedures, or function as a physical support when applied to tissue defects or damaged areas; if it contains bioactive components, it may be utilized as a medical material to assist in improving the tissue environment or recovery. Such medical applications are not limited to specific medical departments or procedures and can be applied to skin areas throughout the human body, including various skin parts such as the face, scalp, neck, hands, arms, and legs.

[0064] Meanwhile, when applied for cosmetic purposes, the hydrogel of the present invention may be used for the purpose of improving skin condition, post-procedure skin care, skin protection, or skin condition regulation. For example, it may be applied to the skin before or after a cosmetic procedure to alleviate skin irritation, or applied to the skin surface or skin tissue to improve or maintain the appearance of the skin. Such cosmetic applications are also not limited to specific procedures or methods of use and may be applied to various skin areas such as the face, eye area, mouth area, neck, and hands.

[0065] In one aspect, the present invention is a hydrogel comprising the above composition.

[0066] In addition, in one aspect, the present invention is a kit for manufacturing hydrogels, for in vivo delivery to cells, for improving cell viability, or for maintaining cell quality.

[0067] In addition, in one aspect, the present invention is a method for delivering cells into the body using the hydrogel.

[0068] In addition, in one aspect, the present invention is a method for maintaining the quality of cells using the hydrogel.

[0069] In addition, in one aspect, the present invention is a use of the hydrogel for delivering cells into the body.

[0070] In addition, in one aspect, the present invention is a use of the hydrogel for maintaining the quality of cells.

[0071] In this specification, the term “for in vivo delivery of cells” refers to a use for delivering cells from an in vitro environment to an in vivo environment (injection, transplantation, etc.).

[0072] Additionally, in this specification, the term “for improving cell viability” means maintaining or increasing the cell survival rate by inhibiting cell damage or death in an in vitro environment.

[0073] Additionally, in this specification, the term “for maintaining cell quality” means preventing the biological activity of the cell from deteriorating.

[0074] In one aspect, the above kit may be used in the medical or cosmetic fields, but is not limited thereto.

[0075] In one aspect, the kit may comprise a platelet extract, thrombin of more than 25 Unit / mL and less than 100 Unit / mL, and cells for delivery into the body or to a target.

[0076] In one aspect, the subject may include a person or an animal.

[0077] In one aspect, the cell may include one or more selected from the group consisting of stem cells, immune cells, progenitor cells, and skin or tissue regeneration cells, but is not limited thereto.

[0078] For example, the stem cells may include mesenchymal stem cells (MSC), hematopoietic stem cells (HSC), induced pluripotent stem cells (iPSC), or a combination thereof, and the mesenchymal stem cells may include bone marrow-derived mesenchymal stem cells (BM-MSC), adipose-derived stem cells (ADSC), umbilical cord blood-derived stem cells (UCB-MSC), umbilical cord tissue-derived stem cells (UC-MSC), or dental pulp stem cells (DPSC), but are not limited thereto.

[0079] In addition, the immune cells may include peripheral blood mononuclear cells (PBMC), T cells (T lymphocytes), B cells (B lymphocytes), natural killer cells (NK cells), monocytes, macrophages, dendritic cells (DCs), or combinations thereof, but are not limited thereto.

[0080] Additionally, the progenitor cells may include, but are not limited to, endothelial progenitor cells (EPCs), neural progenitor cells (NPCs), muscle progenitor cells (MPCs), osteoprogenitor cells (OPCs), chondrogenic progenitor cells (CPCs).

[0081] Additionally, the skin or tissue regeneration cells may include, but are not limited to, fibroblasts, keratinocytes, melanocytes, chondrocytes, osteoblasts, myoblasts, neurons, or combinations thereof.

[0082] In one aspect, the platelet extract, thrombin, and cells may be provided in freeze-dried or liquid form.

[0083] In one aspect, the platelet extract, thrombin, and cells in the kit may be provided in a separated state. In one aspect, the platelets, thrombin, and cells in the kit may be provided in a separated state within the same container using partitions, etc. Alternatively, the platelets, thrombin, and cells may be provided individually contained in different containers. For example, the platelet extract may be sealed in a first container, the thrombin in a second container, and the cells in a third container.

[0084] In one embodiment, the platelet extract, thrombin, and cells may be provided in the form of a dual syringe, a mixing tip, or a cartridge, but are not limited thereto. In one embodiment, when the kit is provided in the form of a dual syringe, the first syringe may contain the platelet extract or a mixture of the platelet extract and cells, and the second syringe may contain thrombin, but is not limited thereto.

[0085] In one aspect, the above kit may further include instructions.

[0086] In one aspect, the above instructions may include methods for handling and using platelet extracts, thrombin and / or cells, methods and conditions for mixing platelet extracts, thrombin and / or cells, methods and environments for applying the formed hydrogel, dosages according to the purpose of use, etc.

[0087] Specifically, the above instructions may include a description of the order of use, the timing of mixing, or the mixing conditions, in which the platelet extract, thrombin, and / or cells are provided in a separated state so as not to react with each other, and then mixed immediately before the procedure to form a hydrogel. Additionally, it may include guidance on the mixing ratio or range of the platelet extract, thrombin, and / or cells, the time at which gelation begins after mixing, the process of gel formation, and the time range for application after mixing.

[0088] Additionally, if platelet extracts, thrombin, and / or cells are provided in a lyophilized form, the instructions may include instructions on how to reconstitute them before use, and may describe the type or amount of solvent used for reconstitution and the time range for use after reconstitution. For example, the instructions may include a step of resuspending or dissolving the lyophilized platelet extracts, thrombin, or cells in a specified solvent, the type or amount of solvent used for reconstitution, and guidance on the time range for use after reconstitution.

[0089] In addition, the above instructions may include a description of a method for applying, injecting, or placing the formed hydrogel onto the skin or tissue, and may describe examples of application on various skin areas and treatment environments, based on medical or cosmetic applications. Specifically, the above instructions may include a description of the recommended usage amount or application amount depending on the purpose of use of the hydrogel. For example, depending on whether it is used for hemostasis, tissue protection or tissue support, or for post-procedure skin care or cosmetic application, guidance on the range of usage amount considering the area, depth, or treatment environment of the application site may be provided.

[0090] In addition, if the hydrogel manufacturing kit includes a dual syringe, a mixing tip, or a cartridge-type structure, instructions regarding how to combine or operate each component may be included. For example, if the hydrogel manufacturing kit is provided in the form of a dual syringe, the instructions may include a description of a method of use in which platelet extract and thrombin are each contained in the syringe barrels of the dual syringe, and the two components are simultaneously discharged and mixed by operating the plunger. For example, it may include a method of mixing the platelet extract, thrombin, and cells during the discharge process through the mixing tip combined with the dual syringe, or a method of controlling the mixing ratio and the gelation initiation time by operating the plunger at a constant speed.

[0091] In one aspect, the above instructions may include using the platelet extract and thrombin mixed in a volume ratio of 1:1 to 10.

[0092] In addition, in one aspect, the above instructions may include an explanation of the amount of cells used when using the cells. The amount of cells used may vary depending on the purpose of use of the kit, the site of administration, etc. For example, in the case of intramuscular injection, the amount of cells used may be 5 to 15 M, or 8 to 12 M, or 9 to 11 M, and in the case of intralesional injection, the amount of cells used may be 0.1 to 1 M, or 0.2 to 0.8 M, or 0.4 to 0.6 M.

[0093] The present invention will be described in detail below through manufacturing examples and experimental examples. These examples are merely illustrative of the present invention, and therefore the scope of the present invention is not limited by these examples.

[0094]

[0095] [Preparation Example]

[0096] [Preparation Example 1] Preparation of Platelet Extract

[0097] 250 ml of platelet concentrate was aliquoted into 5 ml aliquots in sterile serum vials. Each aliquoted platelet concentrate was frozen at -80°C for 1 hour, 2 hours, 4 hours, and overnight, respectively, and then thawed in 1, 3, 5, and 7 cycles, respectively. Afterward, the concentrations of three growth factors (PDGF-BB, VEGF, EGF) were measured by ELISA.

[0098] As a result, the group frozen for 2 hours had a significantly higher release of growth factors compared to the group frozen for 1 hour, and there was no significant difference compared to the groups frozen for 4 and 8 hours.

[0099] In addition, repeated experiments were conducted on 5 ml vials with freeze-thaw cycles divided into 1, 3, 5, and 7 cycles. As a result, it was confirmed that the increase in growth factor concentration stagnated at 3 or more cycles, and that repeating the cycle 3 times was the optimal condition (Figs. 1 to 3).

[0100]

[0101] That is, according to the conventional method for preparing platelet extracts, it takes about 36 hours or more (overnight x 3 cycles), but according to the method of the present invention, it takes about 6 hours (2 hours x 3 cycles), so it was confirmed that the time for preparing platelet extracts is significantly shortened.

[0102]

[0103] [Preparation Example 2] Preparation of a hydrogel containing platelet extract and thrombin

[0104] Hydrogels were prepared by mixing 1 ml of platelet extract (Human Platelet Lysate, HPL) with an equal amount of thrombin at different concentrations (10 Unit / ml, 25 Unit / ml, 50 Unit / ml, 100 Unit / ml) at room temperature.

[0105]

[0106] [Experimental Example 1] Verification of Hydrogel Quality According to Thrombin Concentration

[0107] The gelation time and characteristics of hydrogels prepared with different thrombin concentrations were observed.

[0108] As a result, as can be seen in Table 1 below, it was confirmed that using 50 Unit / mL of thrombin resulted in an appropriate gelation time and hydrogel quality.

[0109]

[0110] Thrombin Concentration (U / mL) Gelation Time (sec) Gel Strength (Relative Viscoelasticity) Characteristics 1060 sec or more Incomplete gel formation Unstable liquid form 2525 sec Moderate partial gel formation 508 sec Homogeneous gel formation Gel formation with no microbubbles and excellent elasticity Less than 1003 sec Formation of coagulated brittle gel, cracking

[0111]

[0112] [Experimental Example 2] Confirmation of Growth Factor Release Maintenance Effect

[0113] The concentration of growth factors released from hydrogels prepared with different thrombin concentrations was measured.

[0114] After dropping the prepared hydrogel onto a cell culture plate, PBS was added to submerge the hydrogel, and the plate was incubated in an incubator for 24 hours. Afterward, the supernatant was collected, impurities were removed by centrifugation, and then the concentrations of VEGF, PDGF-BB, and EGF were measured using a sandwich ELISA method with an ELISA kit.

[0115] As a result, the hydrogel prepared using 50 U / mL thrombin maintained a release rate of more than 95% compared to liquid HPL, while the hydrogel prepared using 100 U / mL thrombin had its diffusion and release of growth factors restricted due to reasons such as excessive coagulation and increased gel density, resulting in an average release rate of less than 65%.

Claims

1. A hydrogel composition comprising a platelet extract and thrombin of greater than 25 Unit / ml and less than 100 Unit / ml.

2. In Paragraph 1, A hydrogel composition comprising the above platelet extract and thrombin in a volume ratio of 1:0.1 to 10.

3. In Paragraph 1, A hydrogel composition having a thrombin concentration of 30 to 70 Unit / ml.

4. In Paragraph 1, A hydrogel composition having a gelation time of 5 to 15 seconds.

5. In Paragraph 1, The above platelet extract is, A hydrogel composition comprising platelets frozen at -100 to -60°C for 90 to 150 minutes and then thawed.

6. In Paragraph 5, A hydrogel composition comprising the above platelet extract obtained by performing the above freezing and thawing 1 to 5 times.

7. In any one of paragraphs 1 through 6, The above composition is, A hydrogel composition including one for cosmetic or medical use.

8. In Paragraph 7, The above composition is, A hydrogel composition comprising for hemostasis, skin tissue regeneration, skin soothing after dermatological procedures, or restoration of depressed tissue.

9. As a kit for manufacturing hydrogels, for in vivo delivery of cells, or for improving cell viability, The above kit is, A kit comprising platelet extract; and thrombin of greater than 25 Unit / ml and less than 100 Unit / ml.

10. In Paragraph 9, The above kit is, Includes additional instructions, The above description describes a kit comprising the above platelet extract and thrombin in a volume ratio of 1:1 to 10.

11. In Paragraph 9, The above kit is, A kit containing additional cells.

12. In Paragraph 11, The above cell is, A kit comprising one or more selected from the group consisting of stem cells, immune cells, progenitor cells, and skin or tissue regeneration cells.

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