WO2005072753A1 - 多血小板血漿からなる組成物 - Google Patents
多血小板血漿からなる組成物 Download PDFInfo
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- WO2005072753A1 WO2005072753A1 PCT/JP2005/001231 JP2005001231W WO2005072753A1 WO 2005072753 A1 WO2005072753 A1 WO 2005072753A1 JP 2005001231 W JP2005001231 W JP 2005001231W WO 2005072753 A1 WO2005072753 A1 WO 2005072753A1
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- rich plasma
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/16—Blood plasma; Blood serum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- Composition comprising platelet-rich plasma
- the present invention relates to platelet-rich plasma used in the medical field and a method for preparing the same.
- Non-Patent Document 1 Platelet-rich plasma for clinical testing is collected from the median vein of the forearm, and 4.5 mL of whole blood is added to a plastic tube containing 1.5 mL of 3.1% w / v sodium citrate and mixed by inversion 500 g for 15 minutes at 22 ° C. It is obtained from the supernatant centrifuged at C (Non-Patent Document 1).
- the operation is complicated, and even if red blood cells can be sedimented, it is inevitable that blood cells such as platelets and plasma proteins are damaged during the operation.
- the platelet-rich plasma obtained by the conventional method is referred to as “PRP” and is distinguished from the platelet-rich plasma of the present invention.
- Patent Document 2 In order to separate red blood cells from whole blood to obtain white blood cell-rich plasma (LRP), a method of adding a coinage-forming coagulant and an enhancer to whole blood has been disclosed (Patent Document 2).
- dextran, hespan, pentaspan, ficoll and the like are exemplified as the coin-forming coagulant
- oxalic acid salts, malonic acid salts, mannitol and sucrose are exemplified as the enhancers. I have.
- Platelet-rich plasma used as an agent for promoting postoperative hemostasis and wound healing includes not only platelet components but also blood components including plasma proteins such as fibrinogen and leukocytes in the same state as they exist in the body as much as possible. There is a need for such highly active multiplatelet plasma which is desired to contain a large amount.
- PRP is obtained by centrifugation or sedimentation of blood cells from blood, at least platelet activity has already been enhanced, and PRP is used as a tissue and / or organ repair promoter. Required the use of a coagulation reaction inducer such as thrombin.
- Patent Document 1 Japanese Unexamined Patent Publication No. 5-203639
- Patent Document 2 Published Japanese Translation of PCT International Publication No. 8-510322
- Non-Patent Document 1 Clinical Laboratory Method Proposal, 31st Edition, p.400
- An object of the present invention is to use a platelet-rich plasma as a tissue and / or organ repair promoter without necessarily using a coagulation reaction inducer such as thrombin.
- An object of the present invention is to provide a highly active platelet-rich plasma easily and inexpensively. Means for solving the problem
- the present inventors have found that a platelet-rich plasma obtained by adding a polymer having an organic phosphate compound residue to whole blood to aggregate and sediment erythrocytes in a selective promoting manner is obtained.
- the present inventors have found that the present invention includes intact platelets, which are close to platelets in the body, and can be used as a tissue and / or organ repair promoter without using a coagulation reaction inducer such as thrombin, thereby completing the present invention.
- the present invention includes the following.
- a method for preparing platelet-rich platelet plasma by adding a polymer having an organophosphate compound residue to blood 1.
- a method for preparing platelet-rich plasma comprising a step of adding a polymer having a residue of an organophosphate compound to blood to selectively precipitate erythrocytes.
- R1, R2 and R3 are each independently a straight-chain or branched alkyl group having 118 carbon atoms, and a is an integer of 112) in the side chain. 3.
- X is -H or -CH
- Rl, R2 and R3 are each independently a linear or branched alkyl group having 18 carbon atoms
- b is an integer of 1 4,000
- c is 1 6 3.
- a tissue and / or organ repair promoter comprising the platelet-rich plasma according to item 8 above.
- tissue and / or organ repair promoter containing platelet-rich plasma described in 8 above an additive for bone formation around a dental implant, an additive for implanting bone or artificial bone into a bone defect, a wound Healing promoter, post-treatment or post-treatment tissue healing promoter for the purpose of cosmetic and / or cosmetic treatment, therapeutic agent for skin disease, therapeutic agent for skin ulcer, repair agent for nerve tissue, and / or tissue repair agent after surgery.
- a reagent or reagent kit for preparing platelet-rich plasma comprising the polymer according to item 12 above.
- red blood cells are selectively aggregated and sedimented than other blood cells, and platelet-rich plasma containing platelets in an intact state with little damage is provided.
- platelet-rich plasma obtained by this method, it is possible to reduce the amount of a coagulation reaction inducer such as thrombin or to use no such agent in some cases.
- FIG. 1 is a view showing a sedimentation state of red blood cells when a polymer is added to blood and allowed to stand for 20 minutes and 30 minutes. (Example 7)
- FIG. 2 is a graph showing the platelet recovery rates of platelet-rich plasma of the present invention and PRP obtained by a conventional method, centrifugation. (Example 8)
- FIG. 3 is a graph showing the amount of fibrinogen in platelet-rich plasma of the present invention and PRP obtained by a conventional centrifugation method. (Example 8)
- FIG. 4 is a graph showing the platelet activation ratio (CD62P) in the platelet-rich plasma of the present invention and PRP obtained by a conventional centrifugation method. (Example 8)
- Platelets adhere to and aggregate with injured tissue to form thrombi, and play a very important role in hemostasis and in the storage and release of substances that migrate and differentiate other cells. Fibrin is produced by the action of thrombin on fibrinogen in plasma and is involved in the final stage of blood coagulation. Furthermore, it is also important as a scaffold for cell infiltration and cell differentiation for tissue repair.
- leukocytes have the function of preventing invasion of bacteria and harmful microorganisms, and have an immune function and a bactericidal function.In particular, among leukocytes, monocytes / macrophages also play an important role in tissue repair. It has been known.
- Platelets rapidly lose their function over time after stimulation or blood collection. For this reason, it was not possible to obtain platelets with a certain degree of function by a method other than centrifugation, which can be separated in a short time with little stimulation on platelets. Erythrocytes sediment gradually when whole blood is allowed to stand, but this rate is usually very slow, so that plasma containing platelets can be obtained even if left for several hours or more, except for pathological blood such as during severe inflammatory reactions. Not known.
- erythrocytes can be selectively precipitated from whole blood in a promoting manner. It is possible to obtain each of them without damaging them while retaining their respective activities.
- the platelet-rich plasma obtained according to the present invention contains platelets in an intact state where almost no activated state is found. Intact platelets have the characteristics of high hemostasis and clotting effect as well as high growth factor release Such platelet-rich plasma has excellent quality because performance is maintained.
- erythrocytes can be agglutinated and sedimented in a selective promotion manner, and all erythrocytes can be aggregated and precipitated within 3 hours, preferably within 2 hours, more preferably within 1 hour, more preferably within 30 minutes after the start of the treatment.
- Erythrocytes contained in blood are sedimented by aggregating at least 80%, preferably at least 90%, to obtain platelet-rich plasma of at least 15% of the total blood volume.
- Whole blood used for preparing the platelet-rich plasma of the present invention may be obtained from a human or an animal other than a human according to a normal blood sampling method.
- an anticoagulant in advance after blood collection.
- Any anticoagulant that is commonly used and does not show toxicity to living organisms may be used, for example, sodium citrate, anticoagulant dextrose (ACD), EDTA, heparin.
- ACD anticoagulant dextrose
- EDTA heparin.
- Any commonly used materials such as phosphorus, low-molecular-weight heparin, fusan, hirudin, and argatroban may be used.
- the amount of the treated blood for preparing the platelet-rich plasma of the present invention is not particularly limited, and varies depending on its use. An appropriate amount can be selected according to the purpose of use and the amount used.
- the method of the present invention can be applied to, for example, blood collected from one's own blood, and can also be applied to blood obtained by blood donation or the like. Since the present invention is a method for easily obtaining platelets having such high activity, it can be used for preparing platelet preparations.
- a polymer having a residue of an organophosphate compound can be added to whole blood.
- a polymer having a residue of an organic phosphate compound a compound represented by the formula (I):
- R1, R2 and R3 are each independently a linear or branched alkyl group having 118 carbon atoms, and a is an integer of 112).
- Compounds containing as a chain can be included in the structural unit.
- the polymer used in the present invention may be a homopolymer having only one compound selected from the formula (II) as a constituent unit, or may be a homopolymer having two or more compounds selected from the formula (II). Make up the compound It may be a heteropolymer containing as a unit. Furthermore, a heteropolymer containing another structural unit may be used. Specifically, equation (m):
- Z is - ⁇ , -OH, -COOH, -COONa, -COOK, -SO H
- x is an integer of 1-5
- [R4 is an aliphatic hydrocarbon, the number of hydrocarbons is 0 20] or -0_R5 [R5 is an aromatic hydrocarbon group], e is an integer of 1 to 4000, and f is an integer of 0 to 20) It may contain the metathalylate compound shown.
- polymer used in the present invention include a methacrylic polymer, for example, a compound represented by the formula (IV): H
- (2-methacryloyl oxyethyl phosphorylcholine (MPC)) or a derivative thereof may be included.
- heteropolymer used in the present invention include the following formulas (V) and / or (VI):
- PMSB contains 2-methacryloyloxyshethylphosphorylcholine (MPC), potassium 3-methacryloyloxypropinolesnorefonate (Potassium 3-Methacryloyloxypropyl sulfonate (PMPS)) and n-butyl metharylate (BMA) Can be appropriately selected, for example, 25:20:55, 45: 5: 50, 41: 4: 55, and the like.
- PMSB25 means that the composition ratio of MPC: PMPS: BMA is 25:20:55
- PMSB45 means MPC: PMPS: BMA Means 45: 5: 50.
- the average molecular weight of the polymer that can be used in the present invention is preferably in the range of 10 to 5,000,000, more preferably 1000 to 1,000,000.
- the amount of the polymer having the residue of the organophosphate compound is 0.0015 to 150 mg, preferably 0.15 to 45 mg, more preferably to 1.5 mL of the blood volume containing the anticoagulant. 1.5-4.5mg can be added. When converted, it becomes about 0.0001-10 w / v%, preferably 0.01-3 w / v%, more preferably 0.1-0.3 w / v%.
- a polymer having a residue of an organophosphate compound in the same ratio as described above.
- the polymer having a residue of the organophosphate compound can be added to a container for storing collected blood, or can be directly added to a syringe for collecting blood.
- An amount of a polymer selected from the above range is added to blood containing an anticoagulant, mixed gently so that the polymer spreads throughout the blood, and allowed to stand still to promote selection.
- sedimentation of erythrocytes is observed, and platelet-rich plasma containing not only platelets but also plasma components and blood components such as leukocytes can be obtained in the supernatant. Sedimentation of erythrocytes is observed at the latest 3 hours, preferably within 2 hours, as early as about 10 minutes after the start of the treatment, and about 30 to 40 minutes, the platelet-rich plasma of the present invention can be obtained. .
- centrifugation at 142 G for 5 minutes or less can be performed.
- the platelet-rich plasma prepared by the method of the present invention may be a wound healing agent, an additive for bone formation around a dental implant, an additive for implanting bone or artificial bone into a bone defect, a wound healing promoter, Skin disease treatment agent, skin ulcer treatment agent, tissue healing promoter after treatment or treatment for the purpose of formation or beauty, tissue repair agent after surgery, post-operative tissue repair agent in orthopedic field, nerve tissue repair It can be applied as an agent for promoting healing in any tissue or organ such as an agent. That is, the platelet-rich plasma of the present invention causes Or damage to the skin or tissue.
- the platelet-rich plasma of the present invention can be applied not only to humans but also to mammals other than humans.
- mammals include animals that in particular live on the ground, and can be applied to dogs, cats, hamsters, and the like that are generally kept as pets. It can also be applied to animals used in sports, such as racehorses and bullfighters.
- Platelet-rich plasma prepared from autologous blood by the method of the present invention can be used for the purpose of the above-mentioned therapy or treatment. If the blood type is compatible, it is of course possible to use it even if it is not derived from own blood.
- an administration method such as applying or injecting a required amount of the platelet-rich plasma of the present invention to a wound site can be applied.
- the most characteristic feature of the platelet-rich plasma of the present invention is that a platelet-rich plasma effective for treatment or the like can be easily and easily prepared from its own blood.
- the simple and easy preparation can be achieved by providing a polymer having a residue of the organic phosphate compound used in the preparation method of the present invention as a reagent.
- a device used in the preparation method of the present invention specifically, a syringe or blood collection tube used for blood collection, or a device for adding a polymer having a residue of an organophosphate compound to blood collected, is used. Examples include containers or test tubes made of sterilized plastic.
- the platelet-rich plasma preparation kit of the present invention includes kits of reagents and instruments selected from the reagents and instruments exemplified above. By using this kit, the platelet-rich plasma of the present invention can be easily prepared at the collected bedside or the like.
- a polymer in which a polymer having a residue of the organic phosphate compound of the present invention is previously filled in a syringe or a container used for blood collection.
- Sodium citrate and ACD which are widely used as anticoagulants, contain polyphosphates containing organophosphate compounds. It is also particularly preferable to dissolve the mer and fill it into the above-mentioned syringe or container.
- the present invention also extends to platelet-rich plasma prepared by the above method, and to the polymer specified in the present invention used for the purpose of preparing platelet-rich plasma.
- the purpose of this example is to confirm the sedimentation effect of erythrocytes when various monomers or polymers are added to whole blood (peripheral blood).
- ACD anticoagulated dextrose taenoate
- Sample 2 was a heteropolymer containing MPC and BMA described in Formula VI in a ratio of 8: 2, Poly (MPC-co-BMA) (PMB (8: 2) (molecular weight 100000), sample Homopolymer (molecular weight: 100,000) having only MPC as a constituent unit described in Formula IV was used for 3.
- Polygnoretamic acid used was Sigma (molecular weight: 53,785).
- Sample 2 3 mg of PMB (8: 2) was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 3 3 mg of PMPC was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 4 Polyglutamic acid O.lmg was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 5 0.5 mg of polyglutamic acid was dissolved in 0.15 mL of ACD solution, and mixed with 1.35 mL of blood.
- Sample 6 0.7 mg of polyglutamic acid was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 7 3 mg of polyglutamic acid was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- WBC white blood cells
- RBC red blood cells
- PHT platelets
- the supernatant obtained from Sample 3 showed high leukocyte and platelet counts and values similarly to the supernatant obtained from Sample 4 (Table 1).
- the purpose of the present example is to confirm the sedimentation state of erythrocytes due to the presence or absence of an anticoagulant added with PMSB shown in Formula VII.
- Samples 2-5 were prepared by dissolving each weight of PMSB in ACD solution or Hanks solution (manufactured by Invitrogen) and adding it to blood.
- PMSB25 of Samples 2 and 4 is PMSB (molecular weight 100000) containing MPC, PMPS, and BMA in the ratio of 25:20:55 described in Formula VII
- PMSB45 of Samples 3 and 5 is similarly expressed in Formula VII.
- PMSB molecular weight 100000
- Sample 2 3 mg of PMSB25 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 3 3 mg of PMSB45 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 4 3 mg of PMSB25 was dissolved in 0.15 mL of Hanks' solution and mixed with 1.35 mL of blood.
- Sample 5 3 mg of PMSB45 was dissolved in 0.15 mL of Hanks' solution and mixed with 1.35 mL of blood.
- the purpose of this example is to examine the effect of adding each PMSB.
- Sample 2 3 mg of polyglutamic acid was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 3 3 mg of PMSB25 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 4 3 mg of PMSB45 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- WBC platelet count
- PHT platelet count
- CD62P platelet activity
- the clotting time was as follows: 0.5 mL of the supernatant of each sample in a 1.5 mL eppendorf tube, 0.05 mL of calcium chloride injection (l.lg / 20 mL as CaCl), and no anticoagulant added. Autologous blood (0.05 mL) was added and mixed, the tube was tilted every 5 minutes, and coagulation was observed and measured.
- CD62P is a glycoprotein with a molecular weight of 140 kd that is intrinsic to the secretory granule membrane in platelets, and is expressed as an indicator of platelet activation because once activated, the platelets migrate to the cell membrane surface of platelets and are expressed .
- CD62P was measured by flow cytometry using an antibody manufactured by BD Biosciences.
- the purpose of this example is to examine the effect of adding each PMSB.
- Sample 2 3 mg of polyglutamic acid was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 3 3 mg of PMSB25 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- Sample 4 3 mg of PMSB45 was dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood.
- the time required for coagulation of the platelet-rich plasma obtained from Samples 3 and 4 was 10 to 20 minutes, showing superior coagulation ability as compared with the platelet-rich plasma obtained from Sample 2.
- the purpose of this example is to examine the effect when various polymers are added.
- ACD solution containing ACD mixed with 3.13 w / v% in water and control Sample 1 was prepared in the same manner as in Example 1.
- Samples 3 and 4 used a homopolymer (molecular weight 100,000) containing only MPC described in Formula IV as a constituent unit
- Sample 5 used a heteropolymer (PMB) containing MPC and BMA described in Formula VI in a ratio of 8: 2. (8: 2)) (molecular weight 100000).
- PMB heteropolymer
- Various polymers dissolved in water at a concentration of 5 w / v% were used as polymer solutions.
- Sample 2 3 mg of polyglutamic acid dissolved in 0.15 mL of ACD solution and mixed with 1.35 mL of blood
- Sample 3 Add 3 mg of polyglutamic acid to 0.06 mL of polymer solution (PMPC), add 0.09 mL of ACD solution, and add blood 1.35 Mixed with mL.
- PMPC polymer solution
- Sample 4 0.09 mL of ACD solution was added to 0.06 mL of polymer solution (PMPC), and mixed with 1.35 mL of blood.
- Sample 5 Polymer solution (PMB (8: 2) 0.09 mL of ACD solution was added to 0.06 mL, and mixed with 1.35 mL of blood.
- WBC white blood cells
- PHT platelet count
- CD62P platelet activation rate
- CD62P showed high values in platelet-rich plasma obtained from samples 2 and 3 containing polyglutamic acid, but low values in platelet-rich plasma obtained from samples 4 and 5.
- Platelet-rich plasma obtained from Samples 4 and 5 solidified within 10 minutes, whereas platelet-rich plasma obtained from Samples 2 and 3 containing polyglutamic acid was the same as in Samples 4 and 5.
- the amount of time required for coagulation was long even when the amount of calcium salt was reduced.
- the platelet-rich plasma obtained from Samples 4 and 5 was coagulated within 10 minutes using only calcium chloride-added cascine without caloricizing anticoagulated blood (Table 4).
- the purpose of this example is to measure the amount of fiprinogen contained in platelet-rich plasma and PRP obtained by the method of the present invention.
- Samples 1 and 2 were allowed to stand for 30 minutes according to the method of Example 1.
- Sample 3 was prepared by adding blood to 0.85 mL of an aqueous solution mixed with 3.8 w / v% of ACD to make a total of 8.5 mL using PRPkit (Kurasan Co., Ltd.). After centrifugation at 3600 ⁇ m for 15 minutes, the mixture was further centrifuged at 2400 ⁇ m for 10 minutes to obtain a supernatant.
- Sample 3 Conventional method (centrifugal method) The amount of fibrinogen in the supernatant obtained from the above sample was measured. The measurement was performed by the thrombin method using a reagent manufactured by Boehringer Mannheim.
- the purpose of this example is to investigate the coagulation time when the heteropolymer represented by the formula (1) is added.
- Sample 1 was allowed to stand for 20 minutes and 30 minutes according to the method of Example 1 to obtain a supernatant.
- Sample 2 was prepared by adding blood to 0.85 mL of an aqueous solution containing 3.8 w / v% of ACD, making 8.5 mL of the whole solution into a 15 mL conical tube (BD Falcon tube), and centrifuging at 1000 ⁇ m for 10 minutes. The supernatant was obtained.
- Figure 1 shows the erythrocyte sedimentation state in sample 1.
- Sample 1 3 mg of MPC and methacrylic acid copolymer (3: 7) was dissolved in 0.15 mL of ACD solution, and
- the clotting time of the supernatant obtained from the above sample was measured.
- the clotting time was as follows: 0.5 mL of the supernatant of each sample was placed in a 1.5 mL eppendorf tube, 0.05 mL of calcium chloride injection solution (as CaCl, l.llg / 20 mL) was placed in a 1.5 mL eppendorf tube, as in Example 3. Further, 0.05 mL of autologous blood without the addition of an anticoagulant was added and mixed, and after 5 minutes from the mixing, the tube was tilted every minute to observe coagulation.
- the purpose of this example is to examine the platelet recovery rate, fibrinogen amount and platelet activation rate (CD62P) when the PMB polymer was added.
- Sample 1 was allowed to stand for 30 minutes according to the method of Example 5 to obtain a supernatant.
- sample 2 blood was added to 0.85 mL of an aqueous solution containing 3.8 w / v% of ACD, and 8.5 mL of the whole solution was added to a 15 mL conical tube (BD Falcon tube), which was then centrifuged at 2400 ⁇ m for 10 minutes. Then, a supernatant containing platelets was collected, and further centrifuged at 3600 ⁇ m for 15 minutes to obtain a supernatant.
- BD Falcon tube BD Falcon tube
- Sample 1 Polymer solution (PMB (8: 2)) 0.09 mL of ACD solution was added to 0.06 mL, and mixed with 1.35 mL of blood.
- the platelet recovery rate was determined according to the following equation.
- CD62P was measured by the method described in Example 3.
- the amount of fibrinogen was measured by the method described in Example 6.
- the platelet recovery rate of the supernatant obtained from each sample is shown in Fig. 2, the fibrinogen amount is shown in Fig. 3, and the platelet activation rate (CD62P) is shown in Fig. 4.
- the platelet-rich plasma and platelet-rich plasma obtained from Sample 1 were equivalent to those obtained from Sample 2 (conventional method).
- the platelet activation rate of platelet-rich plasma obtained from sample 1 was lower than that of sample 1, confirming that the platelet-rich plasma had properties superior to those of the supernatant obtained from sample 2.
- platelet-rich plasma obtained by the preparation method of the present invention is The platelet activity was very high in comparison with PRP obtained by the conventional centrifugation method in terms of the percentage and the amount of fibrinogen. This makes it possible to easily provide platelet-rich plasma having excellent properties.
- platelet-rich plasma obtained by the method of the present invention using its own blood component as a raw material is used as a tissue and Z or organ repair promoter, specifically, an additive for bone formation around a dental implant, a bone defect site, and the like.
- an agent for promoting wound healing for transplantation of bone or artificial bone into bones, an agent for promoting wound healing, an agent for promoting tissue healing after treatment or treatment for the purpose of formation and / or cosmetic treatment, an agent for treating skin diseases, an agent for treating skin ulcer, nerve It can be used in medical practice as a tissue repair agent and / or a tissue repair agent after surgery.
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JP2005517521A JPWO2005072753A1 (ja) | 2004-01-30 | 2005-01-28 | 多血小板血漿からなる組成物 |
US10/597,559 US20070253940A1 (en) | 2004-01-30 | 2005-01-28 | Composition Comprising Platelet-Rich Plasma |
EP05709457A EP1723959A4 (en) | 2004-01-30 | 2005-01-28 | COMPOSITION COMPRISING PLASMA BLOOD PLASMA RICH IN PLATELETS |
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JP2004-024815 | 2004-01-30 | ||
JP2004024815 | 2004-01-30 |
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JP2015112262A (ja) * | 2013-12-11 | 2015-06-22 | 学校法人藤田学園 | 組織修復及び/又は再生促進材 |
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CA2740079A1 (en) * | 2008-10-08 | 2010-04-15 | Thrombodyne, Inc. | Methods of making concentrated fibrinogen- and platelet-containing compositions |
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CN102462652B (zh) * | 2010-11-09 | 2016-05-04 | 中央医疗器材股份有限公司 | 含血小板干粉的美肤促进剂 |
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Also Published As
Publication number | Publication date |
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US20070253940A1 (en) | 2007-11-01 |
EP1723959A1 (en) | 2006-11-22 |
JPWO2005072753A1 (ja) | 2007-10-11 |
EP1723959A4 (en) | 2007-09-26 |
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