Platelet-containing composition and method for producing same

Abstract

To provide a platelet-containing composition having advantages that it can be stored at a room temperature, without adding additional complexes; it can be used without heating, it can be prepared in any shape and thickness of high-purity platelets-containing composition, and it has improved convenience and safety in use and has optimal growth factor controlled release results, and a preparation process.SOLUTION: The present disclosure provides a platelet-containing composition and a preparation process thereof. The platelet-containing composition comprises a quantified and activated platelet-rich plasma (PRP) and a platelet poor plasma (PPP) mixed to the PRP. The preparation process of the platelet-containing composition conducts purification and quantification of platelets from collected whole blood to produce the platelet-containing composition with high purity and optimal growth factor controlled release results.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition and method produced by mixing platelet-rich plasma (PRP) and platelet-poor plasma (PPP) followed by freeze-drying and sterilization, a platelet-containing composition produced, and a method for producing the same. [Background technology]

[0002] Conventional platelet products are prepared by centrifuging whole blood and then compressing it with a manual or automatic compressor to obtain platelet-rich plasma (PRP). However, the platelet count cannot be estimated because the amount of PRP is not quantified. Conventional methods for producing platelet compositions involve adding other components, such as collagen and calcium ions, to PRP to prepare it in various forms, such as liquid, powder, or gel, but this does not eliminate concerns about allergens and toxicity. Furthermore, the gelation process for conventional platelet gels requires heating (usually at 37°C), which is inconvenient and can destroy platelet activity. Furthermore, conventional platelet gels are gelled by forming complexes with other substances, such as gelatin, chitosan, and hyaluronic acid, but platelet activity may be destroyed or reduced by the added substances.

[0003] The applicant searched using keywords such as "platelet lysate", "gel", "~patent / US20120183519A1", "~patent / US20190160104A1", "~patent / US20120183519A1" "sterile", "~patent / CN110876815A" "platelet lysate", "~patent / CN110876815A" "platelets", "gel", etc., and found the following eight prior art.

[0004] Prior art 1, a U.S. patent entitled "Treatment of erectile dysfunction using platelet-rich plasma" (Patent No. US20120183519A1), provides a method for treating erectile dysfunction in a subject, which involves administering a platelet-rich plasma-containing composition to the cavernous tissue, ischemic tissue in the cavernous tissue, or the vicinity thereof. However, because treatment is performed by direct injection of PRP, there is a possibility that it will be consumed soon after injection and it does not have the ability to sustain the release of growth factors. Furthermore, since it cannot be sterilized, there is a risk of infection.

[0005] Prior art 2, a US patent titled "Platelet lysate gel" (Patent No. US20190160104A1), contains human platelet lysate produced by repeating four freeze-thaw cycles on a group of human platelets. Each freeze-thaw cycle involves freezing at temperatures below -190°C. Furthermore, a polymer is added after PRP production, but the polymer may be toxic.

[0006] Prior art 3, a Chinese patent entitled "Hydrogel with Platelet-Rich Plasma and Antibacterial Peptides, and Its Manufacturing Method and Use" (Patent No.: CN110876815A), is produced by mixing PRP with antibacterial peptides and synthetic polymers. The polymers are produced through chemical synthesis, dialysis, and freeze-drying, and according to their Material Safety Data Sheet, they cannot be used on wounds due to their irritating properties.

[0007] Prior art 4, a Chinese patent entitled "Platelet lysate composition, its use and manufacturing method" (Patent No.: CN103702674B), involves extruding the liquid and centrifuging it, discarding the gel and leaving only the treated liquid, which means it cannot be effectively applied to a wider range of wound types.

[0008] Prior art 5, a Chinese patent entitled "Virus-inactivated platelet extract, its use and preparation" (patent number: CN108714155A), uses an organic solvent for sterilization, but the solvent needs to be removed, making the process complicated and potentially leaving residual organic solvent.

[0009] Prior art 6, a Chinese patent entitled "Method for producing human platelet mixed lysate, human platelet mixed lysate, and its use in treating nervous system diseases" (Patent No.: CN110891582A), requires the platelet lysate to be treated at a higher temperature for a longer period of time, and also needs to be cooled before use, making the usage procedure complicated.

[0010] Prior art 7, the Chinese patent "Closed high-speed platelet lysate manufacturing device" (patent number: CN204446735U), is a high-speed platelet lysate manufacturing device, but the manufactured platelet lysate cannot be stored and must be manufactured from scratch when used.

[0011] Prior art 8, the Chinese patent "Composite Platelet Gel and Its Manufacturing Method" (Patent No.: CN105030826A), adds chitosan and glucose, so more additives may cause more safety issues.

[0012] In view of the above-mentioned problems of the prior art, a main object of the present invention is to provide a platelet-containing composition, which comprises a fixed amount of activated platelet-rich plasma (PRP) and platelet-poor plasma (PPP) mixed with the platelet-rich plasma.

[0013] Another object of the present invention is to provide a method for producing a platelet-containing composition, which comprises the following steps: collecting whole blood, separating the collected whole blood by centrifugation into two liquids, platelet-rich plasma (PRP) and platelet-poor plasma (PPP), quantifying and activating the platelet-rich plasma (PRP) into highly concentrated platelet plasma (HCPP), mixing the quantified and activated highly concentrated platelet plasma (HCPP) with the platelet-poor plasma (PPP), freeze-drying, and sterilizing to complete the production of the platelet-containing composition.

[0014] Another object of the present invention is to provide a method for producing a platelet-containing composition, which can be stored at room temperature, does not require the addition of a complex, and can be produced into a highly pure platelet composition of any shape and thickness without the need for heating.

[0015] Another object of the present invention is to provide a platelet-containing composition and a method for preparing the same, which improves the convenience and safety of use and has optimal sustained growth factor release results. Summary of the Invention [Problem to be solved by the invention]

[0016] Conventional platelet gels are applied externally to a patient's wound and only have the effect of "protecting" the wound. Furthermore, because they are not extracted from the patient's own blood, they cannot avoid rejection reactions, allergens, and toxicity. Furthermore, they cannot quickly heal the wound and there is a risk of infection.

[0017] Furthermore, in conventional platelet composition production, other components such as collagen and calcium ions are added to platelet-rich plasma (PRP) to form a liquid, powder, gel, or other form, but considerations regarding allergens, toxicity, etc. cannot be eliminated. Furthermore, the gelation process of conventional platelet gel requires heating (usually to 37°C), which is inconvenient for use and has the problem that platelet activity is destroyed by heating.

[0018] Furthermore, conventional platelet gels gel by forming complexes with other substances such as gelatin, chitosan, and hyaluronic acid, which may destroy or reduce platelet activity.

[0019] Prior art 1, "Treatment of erectile dysfunction using platelet-rich plasma" (Patent No. US20120183519A1), involves direct injection of PRP, which may disappear soon after injection and does not have the ability to sustainably release growth factors.

[0020] Furthermore, since it cannot be sterilized, there is a risk of infection. Prior art 2, "Platelet lysate gel" (Patent No.: US20190160104A1), is produced by adding a polymer after PRP is produced, but the polymer may be toxic.

[0021] Prior art 3, "Hydrogel with Platelet-Rich Plasma and Antibacterial Peptides, and Its Manufacturing Method and Use" (Patent No. CN110876815A), is produced by mixing PRP with antibacterial peptides and synthetic polymers. The polymers are produced by chemical synthesis, dialysis, and freeze-drying, and according to the Material Safety Data Sheet, they are irritating and cannot be used on wounds.

[0022] Prior art 4, "Platelet lysate composition, its use and manufacturing method" (Patent No.: CN103702674B), discards the gel and therefore cannot be effectively applied to more types of wounds.

[0023] Prior art 5, "Virus-inactivated platelet extract, its use and preparation" (Patent No.: CN108714155A), uses an organic solvent for sterilization, but the solvent must be removed, making the process complicated and potentially leaving behind residual organic solvent.

[0024] Prior art 6, "Method for producing human platelet mixed lysate, human platelet mixed lysate, and use in treating nervous system diseases" (Patent No.: CN110891582A), requires high-temperature treatment and cooling before use, making the usage procedure complicated.

[0025] Prior art 7, the "closed high-speed platelet lysate manufacturing device" (Patent No.: CN204446735U), does not allow the manufactured lysate to be stored, and must be manufactured from scratch when used.

[0026] Prior art 8, "Composite platelet gel and its manufacturing method" (Patent No.: CN105030826A), requires the addition of chitosan and glucose, and more additives may cause more safety issues. [Means for solving the problem]

[0027] In order to solve the above problems, the present invention provides a platelet-containing composition comprising a fixed amount of activated platelet-rich plasma (PRP) and a platelet-poor plasma (PPP) mixed with the fixed amount of activated platelet-rich plasma, and the platelet-containing composition is freeze-dried and sterilized. The activation is carried out by adding an activator or by ultrasonic vibration, and the activator is adenosine diphosphate (ADP). , The platelet-rich plasma was quantified by centrifuging the plasma and adding water for injection to adjust the platelet concentration to 10 9 Adjust to / mL Platelet-containing compositions are provided.

[0028] The platelet-containing composition is made into a gel or liquid form by adding additives according to the needs of use.

[0029] In order to solve the above problems, the present invention provides a method for producing a platelet-containing composition, which comprises the following steps (1) to (7):

[0030] (1) Take blood from a bag Process. (2) A process in which the collected whole blood is separated into two liquids, platelet-rich plasma (PRP) and platelet-poor plasma (PPP), by centrifugation. (3) Quantify and activate platelet rich plasma (PRP) into highly concentrated platelet plasma (HCPP) The activation is carried out by adding an activator or by ultrasonic vibration, and the activator is adenosine diphosphate (ADP). The platelet-rich plasma is quantified by centrifuging the plasma and adding water for injection to adjust the platelet concentration to 10. 9 / mL. (4) Mixing a fixed amount of activated highly concentrated platelet plasma (HCPP) with platelet poor plasma (PPP). (5) Freeze-drying. (6) Sterilization process. (7) Completing the production of the platelet-containing composition.

[0031] In the step (7) of the present invention, which is the step of completing the production of the platelet-containing composition, Ca 2+ Contains Adding additives to form a gel or by adding water for injection to form a platelet composition liquid. can be done.

[0032] The platelet-containing composition of the present invention Ca 2+ Contains Additives are The aforementioned Constructing the gel Ca 2+ The concentration is 0.01% to 10% .

[0033] Preferably, Added Ca 2+ The concentration ranges from 0.05% to 9.75%.

[0034] Preferably, Added Ca2+ concentration is 0.1% to 9%.

[0035] Preferably, Added Ca 2+ of concentration are 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0. 29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.5 9%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.7%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.8%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0. 89%, 0.9%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%.

[0036] The gel formed by adding additives to the platelet-containing composition in step (7) of the present invention can be formed into any shape and thickness required in practice, and therefore can be used to accommodate irregular wounds or to repair or protect bones.

[0037] The present invention (7) Process of the platelet-containing composition In the process of completing the manufacturing , Depending on your needs Add a solvent (e.g., water for injection) To make into a liquid form Configure.

[0038] The sterilization method in step (6) of the present invention utilizes gamma rays or the like.

[0039] The platelet-rich plasma in step (3) of the present invention is quantified by centrifuging the plasma and adding water for injection to adjust the platelet concentration to 10 9 Adjust to / mL.

[0040] The water for injection added to quantify the platelet-rich plasma of the present invention can be replaced with sterile saline, 0.45% NaCl, 4.5% hypertonic saline, sterile spring water, or isotonic saline.

[0041] The activation of the platelet-rich plasma in step (3) of the present invention is carried out by adding an activator or by ultrasonic vibration.

[0042] The present invention provides First, adjust the platelet concentration to the platelet-rich plasma, and then , Adenosine diphosphate (ADP) Add The platelet-rich plasma is activated by The supernatant is removed by centrifugation, and then mixed with the platelet poor plasma (PPP).

[0043] The activation of the platelet-rich plasma in step (3) of the present invention is carried out by ultrasonic vibration or the like. [Effects of the Invention]

[0044] Compared with the prior art, the platelet-containing composition of the present invention comprises a fixed amount of activated platelet-rich plasma (PRP) and a platelet-poor plasma (PPP) mixed with the platelet-rich plasma. The method for producing the platelet-containing composition of the present invention includes the following steps: collecting whole blood, centrifuging the collected whole blood to separate it into two liquids, platelet-rich plasma (PRP) and platelet-poor plasma (PPP), quantifying and activating the platelet-rich plasma (PRP) to form highly concentrated platelet plasma (HCPP), mixing the fixed amount of activated highly concentrated platelet plasma (HCPP) with the platelet-poor plasma (PPP), freeze-drying, and sterilizing the mixture to complete the production of the platelet-containing composition.

[0045] These processes allow the platelet-containing composition to be stored at room temperature, and can be produced into a highly pure platelet composition of any shape and thickness without the need for adding a complex or heating. This improves convenience and safety of use, and provides optimal sustained growth factor release results, thereby greatly expanding industrial applicability and being novel and inventive. [Brief explanation of the drawings]

[0046] [Figure 1] FIG. 1 is a diagram illustrating the constitution of a platelet-containing composition of the present invention. [Figure 2] 1 is a production flowchart of a method for producing a platelet-containing composition of the present invention. [Figure 3] 1 is a flowchart illustrating an embodiment of the platelet gel and platelet composition solution of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0047] Hereinafter, embodiments of the present invention will be described with reference to the drawings, but the dimensions and arrangement of each part in the drawings are shown only as an outline and do not necessarily correspond to the actual dimensions and arrangement. Therefore, the present invention is not limited to the dimensions and arrangement shown in the drawings.

[0048] 1 is a diagram illustrating the constitution of the platelet-containing composition of the present invention. In a preferred embodiment, the platelet-containing composition of the present invention 100 comprises at least platelet-rich plasma (PRP) 1 and platelet-poor plasma (PPP) 2.

[0049] The platelet-rich plasma (PRP) 1 was obtained by centrifuging whole blood and collecting the intermediate layer. The platelet-rich plasma 1 was quantified and activated.

[0050] The platelet-poor plasma (PPP) 2 is obtained by centrifuging whole blood and collecting the upper layer.

[0051] Figure 2 is a production flowchart of the method for producing a platelet-containing composition of the present invention. Figure 3 is a diagram showing examples of the platelet gel and platelet composition solution of the present invention. In a preferred embodiment, the core technology of the present invention (shown in Figure 2) provides a method for producing a platelet-containing composition, which comprises the following steps:

[0052] (1) Step of collecting whole blood (e.g., from a blood bag) (S1) (shown in Figure 2).

[0053] (2) Step (S2) of separating the collected whole blood into two liquids, platelet-rich plasma (PRP) and platelet-poor plasma (PPP), by centrifugation (shown in Figure 2).

[0054] (3) A step (S3) of quantifying and activating platelet-rich plasma (PRP) to produce highly concentrated platelet plasma (HCPP) (shown in FIG. 2).

[0055] (4) A step (S4) of mixing the quantified and activated highly concentrated platelet plasma (HCPP) with platelet poor plasma (PPP) (shown in FIG. 2).

[0056] (5) Freeze-drying step (S5) (shown in Figure 2).

[0057] (6) Sterilization step (S6) (shown in Figure 2).

[0058] (7) Step (S7) of completing the production of the platelet-containing composition 100.

[0059] See Figure 3 In the step (7) of completing the production of the platelet-containing composition, the platelet-containing composition 100 is Ca 2+ Contains Add additive 3 to form a gel Constructing .

[0060] The platelet-containing composition 100 Ca 2+ Contains additive 3 teeth , The gel (i.e., platelet gel 31) is formed Ca 2+ The concentration is 0.01% to 10%. Preferably, added Ca 2+ of concentration is 0.05% to 9.75%. More preferably, Added Ca 2+ of concentration is 0.1% to 9%.

[0061] More preferably, Added Ca 2+ of concentrationare 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0. 29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.5 9%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.7%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.8%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0. 89%, 0.9%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%.

[0062] In addition, the platelet-containing composition 100 in the step (7) Ca 2+ Contains The gel formed by adding additive 3 can be formed into any shape and thickness required in practice, and can therefore be used to treat irregular wounds or to repair or protect bones. Additive 3 of the platelet-containing composition 100 forms a platelet composition liquid by adding water for injection or the like.

[0063] Sterilization in the step (6) utilizes gamma rays or the like. The platelet-rich plasma in the step (3) is quantified by centrifuging the plasma and adding water for injection to reduce the platelet concentration to 10. 9 The water for injection added to measure the platelet-rich plasma can be replaced with sterile saline, 0.45% NaCl, 4.5% hypertonic saline, sterile spring water, or isotonic saline.

[0064] The activation of platelet-rich plasma in step (3) is carried out by adding an activator or by ultrasonic vibration (see Figure 3). In step (3), platelet-rich plasma (PRP)1 is activated by using an activator such as adenosine diphosphate (ADP) to maintain platelets in an activated state, and then removing the activator to obtain highly purified platelets and a high content of growth factors.

[0065] In step (3), highly concentrated platelet plasma (HCPP) is activated by ultrasonic vibration, and then mixed with the platelet-poor plasma (PPP) (shown in Figure 3). The platelet-rich plasma of the present invention is activated by adding an activator, centrifuging to remove the supernatant, and then mixed with the platelet-poor plasma (PPP).

[0066] In step (3) of the present invention, the platelet-rich plasma is activated by ultrasonic vibration or the like, and then the platelet-rich plasma is mixed with the platelet-poor plasma (PPP) (as shown in FIG. 3).

[0067] 3 is a flowchart showing the process for producing the platelet gel and platelet composition of the present invention. Taking "platelet gel" as an example, the steps in the process for producing the platelet-containing composition are as follows:

[0068] First, whole blood was collected and centrifuged to prepare platelet-rich plasma (PRP) and platelet-poor plasma (PPP). The platelet-rich plasma (PRP) was further centrifuged to obtain a platelet concentration of 10 9 In the step of measuring, water for injection may be added.

[0069] The measured platelet-rich plasma (PRP) is further activated by ultrasonic vibration or the addition of an activator, resulting in 100% platelet purity and a higher growth factor content. The highly concentrated platelet plasma (HCPP) obtained through the activation process and measurement is centrifuged to remove the supernatant, and the activated highly concentrated platelet plasma (HCPP) from which the supernatant has been removed is mixed with platelet-poor plasma (PPP) obtained by centrifugation, and then freeze-dried and sterilized with gamma rays. Ca 2+ Contains By adding additive 3, Ca constituting the gel 2+ The concentration is 0.01% to 10% Configure.

[0070] 3 is a flowchart showing the process for producing the platelet gel and platelet liquid composition of the present invention. Taking the platelet liquid composition as an example, the steps in the process for producing the platelet-containing composition are as follows:

[0071] First, whole blood was collected and centrifuged to prepare platelet-rich plasma (PRP) and platelet-poor plasma (PPP). The platelet-rich plasma (PRP) was further centrifuged to obtain a platelet concentration of 10 9 The platelet-rich plasma (PRP) is measured at 100% platelet purity and contains a higher amount of growth factors. Water for injection may be added to the measured amount. Activating the measured platelet-rich plasma (PRP) by ultrasonic vibration or by adding an activator results in 100% platelet purity and a higher growth factor content.

[0072] That is, the high-concentration platelet-rich plasma (HCPP) obtained through the activation process and quantification is centrifuged to remove the supernatant, and the activated high-concentration platelet-rich plasma (HCPP) from which the supernatant has been removed is mixed with platelet-poor plasma (PPP) obtained by centrifugation, and then the mixture is freeze-dried and sterilized with gamma rays. Finally, the mixture is dissolved in a solvent. additive 3 (for example, water for injection) to form the platelet composition 32.

[0073] The above describes the manufacturing process for the platelet-containing composition of the present invention, flowcharts of examples of platelet gel and platelet composition liquid, and the manufacturing processes for the two types of products. Platelet-poor plasma (PPP) 2 in Figure 3 is the upper layer of platelet-rich plasma, and platelet-rich plasma (PRP) 1 in Figure 3 is the middle layer of platelet-rich plasma.

[0074] The platelet-containing composition 100 produced by the above-described production method has the following properties and effects.

[0075] 1. The platelet-containing composition produced is freeze-dried into a powder form, which can be stored at room temperature and does not require special storage conditions. It can be taken and used immediately, eliminating the need for blood collection and preparation every time.

[0076] 2. Platelet-containing compositions can be prepared without the need for additional complexes (eg, anticoagulants).

[0077] 3. The produced platelet gel does not need to be heated during use or production, and can be gelled at room temperature for use. Conventional gels require high-temperature heating or other additional steps for use (the procedure is as follows: first, pressing, squeezing, filtering, ultrasonic vibration, or freezing and thawing cycles are performed before use), so unlike the present invention, they cannot be directly gelled at room temperature for use.

[0078] 4. The shape of the preparation is not limited (for example, conventional blood collection tubes have a capacity limit of 10 cc), and platelet-containing compositions with appropriate shapes and thicknesses can be produced according to demand.

[0079] The advantages and effects of the method for producing a platelet-containing composition of the present invention are verified by experiments. As a result of verification by experiments, the platelet-containing composition produced by the method for producing a platelet-containing composition of the present invention has the following advantages and effects.

[0080] 1. Sterile. 2. No complexes such as gelatin, chitosan, or hyaluronic acid are added. 3. Contains no red blood cells. 4. Does not include separation gel. 5. Contains growth factors. 6. Platelets are quantified. 7. No other substances are added during manufacturing. 8. It can be used as a natural medium for cell culture. 9. The platelet content is 100%. 10. The gelling temperature may be room temperature. 11. There is no need to add growth factors separately. 12. No residual activator.

[0081] Applications of the present invention 1. The benefits of no additives. In the production of the platelet-containing composition of the present invention, no other substances are added, which minimizes the risk of infection to patients and reduces raw material and production costs. 2. Produce a 100% pure platelet composition with ADP. In the present invention, platelets are activated by adding adenosine diphosphate (ADP) to the platelets to maintain them in an activated state, and then removing ADP, thereby achieving 100% platelet purity and a higher growth factor content. 3. Since it is produced from the patient's own blood, rejection and toxicity are avoided. Since the platelet-containing composition of the present invention is produced by collecting blood from the patient's own blood, when the composition is administered to the patient, rejection and toxicity can be avoided, and the composition can be used safely. 4. A manufacturing method that allows large quantities to be produced at once.

[0082] The present invention enables the production of a large amount of platelet-containing composition at one time, and can be used for the treatment of large wounds and injuries, or for mass production. This method solves the problems of conventional blood collection tubes, which have a capacity limit of 10 cc (only blood collection tubes with a certain shape and thickness can be produced), which limits the volume of the platelet gel formed, and makes it difficult for the gel to change shape.

[0083] 5. Can be manufactured into any shape and thickness depending on the actual usage.

[0084] The platelet-containing composition of the present invention can be prepared in a shape and thickness according to actual needs, and therefore can be used to repair or protect irregular wounds and bones. That is, the present invention allows the preparation of a platelet-containing composition having a shape and thickness according to actual needs through the above-mentioned method for producing a large amount of platelet in a single batch, and therefore can be used to repair or protect irregular wounds (e.g., wounds caused by burns, etc.), bones, etc., without the need to combine small pieces of platelet-containing composition or use inappropriate dressing materials.

[0085] 6. Optimizing the results of sustained release of growth factors.

[0086] The present invention makes it possible to produce a platelet-containing composition with better results in sustained growth factor release without the addition of an anticoagulant, thereby avoiding the risk of excessive anticoagulant use.

[0087] Compared with the prior art, the present invention has the following advantages:

[0088] 1. Conventional platelet gels are applied externally to a patient's wound and only have the effect of "protecting" the wound. The platelet gel of the present invention is produced using blood collected from the patient's own blood, which avoids rejection and toxicity, improves protective effects, and allows the patient's wound to heal more quickly.

[0089] 2. Conventional methods for producing platelet compositions involve adding other components such as collagen and calcium ions to platelet-rich plasma (PRP) to produce different forms such as liquid, powder, or gel, but these methods not only pose concerns about allergens and toxicity, but also make it difficult to estimate platelet count. The present invention produces a 100% pure platelet composition using ADP and quantifies platelets, thereby maintaining platelets in an activated state using ADP (adenosine diphosphate) while removing plasma, thereby obtaining 100% pure platelets.

[0090] 3. The gelation process of conventional platelet gel requires heating (usually 37°C), which is inconvenient to use. The platelet gel produced by the present invention does not require heating during use or production, which avoids the problem of platelet activity being destroyed by heating.

[0091] 4. Conventional platelet gels are gels formed after forming a complex with other substances (e.g., gelatin, chitosan, hyaluronic acid, etc.), which may destroy or reduce platelet activity. The production method of the present invention allows the production of a platelet-containing composition without the need to add a complex separately.

[0092] The features of the present invention are as follows.

[0093] 1. The platelet-containing composition can be stored at room temperature. The prepared platelet-containing composition can be stored at room temperature without being limited to any special storage conditions.

[0094] 2. No need to add complex separately The platelet-containing composition can be prepared without adding a complex such as gelatin, chitosan, or hyaluronic acid.

[0095] 3. No need to heat before use The platelet gel thus produced does not require heating during use or production and can be used in gel form at room temperature.

[0096] 4. Platelet-containing compositions of any shape and thickness can be produced There is no limitation on the shape (for example, conventional blood collection tubes have a volume limit of 10 cc), and platelet-containing compositions having appropriate shapes and thicknesses can be produced according to needs.

[0097] 5. 100% pure platelet composition Activation brings the platelet concentration to 100%.

[0098] The platelet-containing composition 100 of the present invention includes a fixed amount of activated platelet-rich plasma (PRP) 1 and platelet-poor plasma (PPP) 2 to be mixed with the platelet-rich plasma.

[0099] The method for producing the platelet-containing composition of the present invention includes the steps of collecting whole blood, separating the collected whole blood by centrifugation into two liquids, platelet-rich plasma (PRP) 1 and platelet-poor plasma (PPP) 2, quantifying and activating the platelet-rich plasma (PRP) 1, mixing the quantitatively-quantified and activated highly concentrated platelet plasma (HCPP) with the platelet-poor plasma (PPP) 2, freeze-drying, sterilizing, and completing the production of the platelet-containing composition.

[0100] As described above, the platelet-containing composition of the present invention can be stored at room temperature, does not require the addition of a complex, and does not require heating before use. It is possible to produce a highly pure platelet composition of any shape and thickness. Furthermore, the platelet-containing composition of the present invention is more convenient and safe to use, has optimal sustained growth factor release properties, and is thus novel and inventive, greatly expanding its industrial applicability.

[0101] As can be seen from the above, the present invention breaks through prior art, achieves the intended effect, and is not easily conceivable by a person skilled in the art. Therefore, it meets the requirements for a patent, such as novelty and inventive step.

[0102] The above examples are merely for illustrating the technical ideas and features of the present invention, and are intended to enable those skilled in the art to understand and practice the contents of the present invention, and are not intended to limit the scope of the claims of the present invention. Any improvements or modifications to the present invention that do not deviate from the spirit and scope of the present invention should be included in the protection scope of the present invention. [Explanation of symbols]

[0103] 100 Platelet-containing composition 1 Platelet rich plasma (PRP) 2 Platelet poor plasma (PPP) 3 Ca 2+ Contains additives, Solvent Additives 31 Platelet Gel 32 Platelet composition S1 Collect whole blood S2 The collected whole blood is centrifuged to separate it into two liquids: platelet-rich plasma (PRP) and platelet-poor plasma (PPP). S3 Quantify and activate platelet rich plasma (PRP) into highly concentrated platelet plasma (HCPP) S4 Quantitative, activated highly concentrated platelet plasma (HCPP) is mixed with platelet poor plasma (PPP). S5 Freeze-dry S6 Sterilize S7. Complete the manufacture of the platelet-containing composition

Claims

1. a fixed amount of activated platelet-rich plasma (PRP); and a platelet-poor plasma (PPP) mixed with the fixed amount of activated platelet-rich plasma, The platelet-containing composition is completed by freeze-drying and sterilization, and the activation is performed by adding an activating agent or by ultrasonic vibration, and the activating agent is adenosine diphosphate (ADP); The platelet-rich plasma is quantified by centrifuging the plasma and adding water for injection to adjust the platelet concentration to 10 9 / mL. Platelet-containing composition.

2. Adding a Ca2+-containing additive to the platelet-containing composition results in a gel, and adding water for injection to the platelet-containing composition results in a platelet-containing liquid. The platelet-containing composition of claim 1.

3. (1) collecting blood from a blood bag; (2) Separating the collected whole blood into two liquids, platelet-rich plasma (PRP) and platelet-poor plasma (PPP), by centrifugation; (3) quantifying and activating platelet-rich plasma (PRP) into highly concentrated platelet plasma (HCPP), the activation being carried out by adding an activator or by ultrasonic vibration, the activator being adenosine diphosphate (ADP), and quantifying the platelet-rich plasma by centrifuging and adding water for injection to adjust the platelet concentration to 10 9 / mL; (4) mixing a quantified amount of activated highly concentrated platelet plasma (HCPP) with platelet poor plasma (PPP); (5) freeze-drying; (6) a sterilizing step; (7) completing the production of the platelet-containing composition. A method for producing a platelet-containing composition.

4. In the step (7), a Ca 2+ -containing additive is added depending on the needs of use to form a gel, or water for injection is added to form a platelet composition solution. The method for producing the platelet-containing composition according to claim 3.

5. The Ca 2+ -containing additive of the platelet-containing composition has a Ca 2+ concentration of 0.01% to 10%. The method for producing the platelet-containing composition according to claim 4.

6. The concentration of added Ca 2+ is 0.05% to 9.75%. The method for producing the platelet-containing composition according to claim 4.

7. The concentration of added Ca 2+ is 0.1% to 9%. The method for producing the platelet-containing composition according to claim 4.

8. Added Ca ２＋ The concentrations of are 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0 .59%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.7%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.8%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0. 0.89%, 0.9%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%. The method for producing the platelet-containing composition according to claim 4.

9. The gel formed by adding additives to the platelet-containing composition in step (7) can be adapted to a shape and thickness required in practice, and therefore can be used to accommodate irregular wounds or to repair or protect bones. The method for producing the platelet-containing composition according to claim 4.

10. The water for injection added to measure the platelet-rich plasma can be replaced with sterile saline, 0.45% NaCl, 4.5% hypertonic saline, sterile spring water, or isotonic saline. The method for producing the platelet-containing composition according to claim 4.

11. The platelet concentration is adjusted to obtain the platelet-rich plasma, and then the platelet-rich plasma is activated by adding adenosine diphosphate (ADP), and the supernatant is removed by centrifugation, and then the platelet-rich plasma is mixed with the platelet-poor plasma (PPP). The method for producing the platelet-containing composition according to claim 3.

12. The activation of the platelet-rich plasma in the step (3) is carried out by ultrasonic vibration. The method for producing the platelet-containing composition according to claim 3.

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