Method for preparing concentrated platelet plasma product through horizontal separation by combining pre-filled catheter flusher with three-way cock

Abstract

The invention discloses a method for preparing a concentrated platelet plasma product through horizontal separation by combining a pre-filled catheter flusher with a three-way cock. According to the method, horizontal separation is carried out in the upward direction of the pipe cap of the pre-filled flusher, the liquid level of the separated sample is protected to be in a horizontal state, and connection and extraction of a three-way cock are facilitated. The concentrated platelet plasma product prepared by the method is in a liquid state, can be mixed with thrombin powder and calcium chloride to prepare a gel state (PG), retains platelet release growth factors in local tissues by activating fibrinogen to form a network scaffold, and promotes cell migration, differentiation and proliferation to achieve tissue repair and regeneration. The invention aims to provide a platelet extraction device which is convenient, complete and standard and can protect separated and extracted platelets from being activated or damaged in advance by mechanical external force. The whole transferring and extracting process is carried out in a clean workbench, bacterial infection is avoided, and meanwhile the skin or mucous membrane of an operator is protected from being polluted by a sample.

Description

Technical Field

[0001] The invention relates to the technical field of medical devices, and in particular to a method for preparing concentrated platelet plasma products by combining a prefilled catheter flusher with a three-way stopcock for horizontal separation. Background Art

[0002] Platelet concentrate (PC) is a platelet concentrated blood product separated from blood by horizontal centrifugation technology. Its characteristics are that the platelet concentration is much higher than the normal blood level and it is rich in various growth factors and plasma proteins. In the medical field, it is widely used in orthopedics, oral and maxillofacial surgery, sports medicine, plastic surgery, dermatology, repair and reconstruction, traditional Chinese medicine, obstetrics and gynecology and other disciplines. And it is used alone or in combination with other technologies in basic medical fields such as cytology, histology, animal experimentation, microbiology, and tissue engineering. A large number of basic and clinical studies have shown that platelet concentrates can promote the repair of various tissues such as bones, muscles, tendons, ligaments, fat, blood vessels, mucosa, nerve fibers, skin, etc., which is beneficial to the overall repair and regeneration of organs. The core component of the tissue that accelerates the repair is that platelets contain a variety of high-concentration growth factors. When these growth factors are released from high-concentration platelet concentrates to the local damaged tissue, they can significantly promote the proliferation, differentiation and migration of repair cells, accelerate the synthesis of extracellular matrix, promote local vascular regeneration and establish an effective microcirculation system. These components play an important role in promoting tissue repair, accelerating wound healing and improving local inflammatory responses. As a natural hemostatic agent and biological scaffold, fibrin helps to form a gel-like matrix, providing a favorable environment for cell migration and proliferation. Plasma proteins include albumin, globulin, etc., which are involved in maintaining osmotic pressure, transporting nutrients and regulating immune responses.

[0003] Platelet concentrates originated from the concept of transfusion medicine. In the early days, the term for platelet concentrates was mostly platelet-rich plasma (PRP). This term has been used to this day. Before the 1980s, all research on PRP was focused on hematology fields such as blood diseases or coagulation systems. Before the discovery of platelet-secreting growth factors, platelets were believed to only play a role related to coagulation in the blood system. In 1974, Ross found through a controlled study that using the supernatant of activated platelets for the culture of arterial smooth muscle can accelerate cell mitosis and promote cell proliferation, and pointed out that cell proliferation may be related to certain factors released after platelets are activated by thrombin. In 1978, Witte named this growth factor released from platelets and capable of promoting cell proliferation PDGF (platelet-derived growth factor). In the following 20 years, a variety of growth factors in platelets were discovered, such as TGF-β (transforming growth factor-β) in 1983, IGF-1 (insulin-like growth factor-1) in 1989, bFGF (basic fibroblast growth factor) in 1993, and VEGF (vascular endothelial growth factor) in 1998. At the same time, from the 1970s to the 1990s, the number of articles on PRP increased year by year, and more and more growth factors were discovered, providing a theoretical basis for the subsequent use of PRP in clinical tissue repair. Concentrated platelet plasma products refer to the development history and conceptual connotation of the three generations of platelet-rich plasma, including PRP, PRF, and CGF. Initial application exploration: After the concept of PRP was proposed in the 1950s, Harke et al. tried to separate and extract PRP in 1977 to avoid the loss of platelet function and severe blood loss after surgery caused by CBP during cardiac surgery, and successfully applied it to cardiac surgery; the repair effect was discovered: in the 1970s, it was a viscous coagulant that was quickly produced when PRP was mixed with calcium ions and thrombin, also known as platelet rich plasma gel (PRG). In 1975, some researchers used it as a human corneal adhesive; in 1979, Fishcher used it for nerve anastomosis. In 1980, an article published by Brandstedt's experimental research institute confirmed that fibrin can promote the migration of fibroblasts, collagen deposition, and lead to the formation of granulation tissue. In 1986, Knighton first reported PDWHF (platelet-derived wound healing factor) and activated platelet supernatant, and then began to use it widely in the treatment of chronic wounds. In 1995, Gaudric et al. first demonstrated that PRP can significantly improve the success rate of macular hole surgery. In 1997, Whitman et al. applied PRG to oral and maxillofacial surgery.In 2000, Man et al. prepared concentrated platelets into autologous PG, and added autologous fibrin glue to patients after plastic surgery, and found that it had the effect of reducing bleeding and helping wound healing. 21st century to date: In 2003, Sanchez et al. used plasma rich in growth factors under arthroscopy to enhance the healing and reconstruction after anterior cruciate ligament transplantation. In 2007, Alio et al. successfully used autologous PRP to treat corneal ulcers for the first time and found that it can promote healing and help reduce pain and inflammation. In 2013, Callejo et al. removed bilateral ovarian lesions in patients. At present, more than ten clinical expert consensuses have been formulated by experts in the medical field in my country, covering various clinical fields. Expert Consensus on Autologous Platelet-Rich Plasma for the Treatment of Thin Endometrium (2023 Edition); Expert Consensus on Platelet-Rich Plasma for the Treatment of Talus Osteochondral Injury (2023 Edition); Expert Consensus on Platelet-Rich Plasma for the Treatment of Lateral Epicondylitis of the Humerus (2022 Edition); Expert Consensus on the Clinical Application of Platelet Concentrates in Hair Regeneration (2022 Edition); Expert Consensus on Autologous Platelet-Rich Plasma Intraarticular Injection for the Treatment of Knee Osteoarthritis (2022 Edition); Expert Consensus on Technical Specifications for the Preparation of Autologous Platelet-Rich Plasma (Chinese Pain Medicine Expert Consensus) (2022 Edition); Expert Consensus on Autologous Platelet-Rich Fibrin (PRF) for the Treatment of Uterine Endometrium (2022 Edition); Expert Consensus on Autologous Platelet-Rich Plasma for the Treatment of Uterine Endometrium ... Expert Consensus on Platelet-Rich Plasma Preparation Technology (2021 Edition); Expert Consensus on the Application of Platelet-Rich Plasma in Chronic Musculoskeletal Pain Diseases (2021 Edition); Expert Consensus on the Application of Concentrated Platelet Products in Facial Skin and Soft Tissue Rejuvenation (2020 Edition); National Expert Consensus on the Application of Concentrated Platelet Products in Wound Repair (2020 Edition); Clinical Practice Guidelines for Intra-articular Injection of Platelet-Rich Plasma for the Treatment of Knee Osteoarthritis (2018 Edition); Expert Consensus on the Clinical Application of Platelet-Rich Plasma in Orthopedic Surgery (2018 Edition). This technology has attracted more and more attention in research and clinical application at home and abroad. The principle of preparing concentrated platelets is mainly based on the density and specific gravity of different components in the blood. The blood products are obtained after horizontal centrifugation. The top layer of liquid is the brown-yellow layer serum, the middle layer is the white film layer, and the bottom layer is the red blood cell layer. The middle layer of the buffy coat and the 1.5 ml of plasma on the buffy coat are platelet concentrated plasma.Every 10ml of blood containing anticoagulants can prepare 1ml-1.5ml of concentrated platelet plasma. Currently, there are two types of platelet-rich plasma kits on the market: imported and domestic. Imported ones include: REGEN LAB SA; Arthrex, Inc., domestic Shandong Weigao Company, Guangzhou Paili Company, Wekan Medical Technology Company, etc. Most of them are based on high-value consumables, which are expensive and the single blood collection volume is about 50ml. On the one hand, it is not conducive to the widespread application of this technology in clinical practice. On the other hand, due to the high price of consumables, it can only be used by some patients with better economic conditions, which is not conducive to the exploration and development of the technology in a wider range of clinical applications. There are also medical institutions in China that consider using conventional consumables to prepare open laboratories by reducing the value of consumables. Although it reduces the preparation cost and is conducive to the benefit of more patients with this technology, it also has the following disadvantages: 1. Conventional medical consumables are divided into one, two and three categories. If they are not distinguished and carefully identified as Class III medical devices, it is easy to cause medical risks. The document "Summary of the Results of the First Classification and Definition of Medical Devices in 2024" released by the Medical Device Standard Management Center of our National Medical Products Administration on April 9, 2024 clearly stipulates that platelet-rich plasma obtained from disposable vacuum blood collection tubes is only used for in vitro diagnostic testing, not for other purposes such as transfusion and treatment. Class III medical devices must be used to prepare platelet-rich plasma. The "Expert Consensus on Autologous Platelet-Rich Plasma Preparation Technology" also clearly states: medical devices related to the collection and preparation of PRP and clearly belong to Class III; 2. The equipment selected for the preparation of PRP is an ordinary laboratory centrifuge. The operator needs to set the parameters for each patient, and the conventional consumables used are not compatible with the ordinary centrifuge cups. The containers containing blood samples cannot be sealed in the cups. During the operation of the centrifuge, blood samples are prone to leakage and contamination of the centrifugal chamber, resulting in preparation failure. The expert review of "Controversy and Research Progress in the Clinical Application of Platelet-Rich Plasma" also pointed out: Only by using a centrifugation method that matches the equipment can effective PRP be prepared; 3. Experimental 1. The laboratory preparation method is mostly an open environment, and there is a risk of blood contamination during the preparation and transfer of blood products; 2. The ordinary angle rotor centrifuge is used without a closed adapter. The sample liquid surface prepared by the laboratory preparation method is inclined and there is no closed adapter. Manual needle extraction may cause too many red blood cells or fail to effectively extract the white film layer, resulting in insufficient preparation concentration and unsatisfactory treatment effect; 3. The consumables selected for the laboratory preparation method are placed in different directions, which will affect the preparation and cause premature activation of platelets; 4. The non-standard operation steps also lead to the lack of a unified standard to monitor the concentration of the prepared platelet-rich plasma, and the stability of each preparation and extraction cannot be ensured. Summary of the invention

[0004] In view of the problems of the existing laboratory preparation method, the purpose of the present invention is to provide a method for preparing concentrated platelet plasma products by combining a prefilled catheter flusher with a three-way stopcock for horizontal separation, which is convenient, standardized, and can prevent premature platelet activation or the risk of blood product transfer contamination, protect the safety of operators, and maintain the stability of preparation. In order to solve the above problems, the present invention adopts the following technical scheme: a method for preparing concentrated platelet plasma products by combining a prefilled catheter flusher with a three-way stopcock for horizontal separation, the preparation method comprising a prefilled catheter flusher, a three-way stopcock, a medical centrifuge, an adapter with the same shape as the prefilled catheter flusher, and a clean workbench.

[0005] The prefilled catheter flusher is of 10 ml specification and is composed of a tube cover of a Luer connector, an outer sleeve, a piston and a detachable core rod.

[0006] The three-way cock is an upper longitudinal pipe and a lower longitudinal pipe, and a transverse pipe connected between the upper longitudinal pipe and the lower longitudinal pipe and communicated with the upper longitudinal pipe; and is controlled by a directional valve.

[0007] The medical centrifuge includes: a main machine (equipped with a variable frequency motor); a rotating shaft, which is rotatably arranged in the centrifugal chamber of the main machine and rotated after being driven by the main machine; and there are four U-shaped connecting grooves on the rotating shaft, which are used to fix four adapters respectively. The adapter (currently patented, authorized patent number: CN220737990U) is an adapter cap, a sealing ring, and an adapter column. The internal structure of the adapter column is consistent with the shape of the prefilled catheter flusher.

[0008] The clean workbench can provide local dust-free cleanliness, and the sterile working environment is a box-type air purification of the local operating environment of Class 100, which is used for environmental control during blood transfer operations.

[0009] The specific steps of the method for preparing concentrated platelet plasma products (taking PRP as an example) by combining a prefilled catheter flusher with a three-way stopcock for horizontal separation are as follows: before preparation, use 75% alcohol to wipe the clean bench, put in spare consumables (three-way, prefilled catheter flusher, tube rack, 20ml disposable sterile syringe, waste recycling bin, marker pen, treatment tray, sterile drape, sterile gauze), turn on the ultraviolet lamp for disinfection for at least 30 minutes, then turn off the ultraviolet lamp, turn on the fluorescent lamp and the clean bench and continue to blow air for standby use.

[0010] Use a 20ml syringe to first draw out 2ml of 3.8% citrate anticoagulant (or 4% citrate anticoagulant is also acceptable), then draw out 18ml of the patient's blood (the ratio of anticoagulant to blood is 1:9), and store it on a tray covered with a sterile drape.

[0011] After thoroughly mixing the venous blood and the anticoagulant, take about 0.2 ml of blood sample for platelet count testing.

[0012] The operator wears clean gloves, transfers the anticoagulated blood sample to the operating table, opens the prefilled catheter flusher package, carefully unscrews the tube cap, taking care to avoid touching the interface end, and places the cap in a clean and untouchable location.

[0013] After draining the saline in the original tube of the prefilled catheter flusher, move the 20ml syringe to the two 10ml prefilled catheter flushers through the three-way stopcock. Pay attention to the aseptic operation during the transfer process and inject blood at a low speed as much as possible to avoid bubbles. After the transfer is completed, cover the prefilled catheter tube cap and tighten it, rotate to remove the piston core rod, place the prefilled catheter tube cap vertically in the adapter with the upward direction, and cover the adapter cap.

[0014] Put two adapters with prefilled catheter flushers into the "U" slot on the rotor of the medical centrifuge and fix them symmetrically. Click on the PRP mode on the centrifuge screen to perform the first horizontal centrifugation, with a speed of 1600rpm, a temperature of 18°, a speed increase of 5 gears, a speed decrease of 2 gears, and a centrifugation time of 10 minutes and 30 seconds. The blood is divided into three layers, from top to bottom, the plasma layer, the buffy coat layer, and the red blood cell layer.

[0015] Open the adapter cap, take out the prefilled catheter flusher vertically and place it on the clean bench tube rack for use (Note: In the subsequent operations, the prefilled catheter flusher after centrifugation must always be kept vertical, and never tilt or flip it to protect the separated PRP layer); routine hand disinfection, wear clean gloves. Take out a three-way stopcock and a new prefilled catheter flusher, carefully open the package and unscrew its tube cover, connect it to the three-way, then keep the prefilled catheter containing blood products separated for the first time vertical to the ground, unscrew the tube cover and connect it to the same three-way tube, open the three-way connection valve in an "L" shape, connect the prefilled catheter with blood products and the channel of the new prefilled catheter flusher, slowly and evenly transfer the plasma layer and the white film layer of the sample into the new flusher, and stop when it is about 0.3mm below the red blood cell layer. Close the three-way, carefully unscrew the sample tube containing the remaining red blood cells, and discard it into the waste bucket.

[0016] Replace with another sample and repeat the above

[0015] operation. Recover a new tube of sample and carry out the next step of centrifugation.

[0017] Place a new sample tube into the adapter vertically with the cap facing upward, then place the adapter vertically into the medical centrifuge, place another adapter symmetrically for balancing, select PRP mode for the second horizontal centrifugation, speed 2950rpm, temperature 18°, speed up 3 gears, down 2 gears, and centrifugation time 6 minutes.

[0018] Perform routine hand disinfection and wear clean gloves. Keep the sample vertical when taking it out. The upper layer of the sample is serum, the middle layer is platelet-rich plasma, and the bottom layer is a very small amount of red blood cells. Take out a syringe and a three-way stopcock, separate the syringe needle as before, and connect it in an "L" shape with the three-way stopcock and the prefilled catheter flusher with the separated sample. Drain the excess serum in the upper layer until the remaining 2.5 ml of plasma (as required for injection) remains in the prefilled catheter flusher. After discharge, separate the three-way tube from the syringe, and connect the prefilled catheter flusher containing the prepared platelet-rich plasma to the injection needle.

[0019] After fully mixing the 2.5 ml plasma in

[0018] , take about 0.1 ml plasma sample for platelet number detection after preparation. The platelet detection multiple is between 3-5 times the platelet base number detected in whole blood, which can meet the clinical treatment needs. (Since the specimen required for bacterial culture of adult blood is at least 5-10 ml, the bacterial culture of concentrated platelet plasma prepared by this method is selected to be carried out in animal experiments to ensure that the preparation method meets the requirements of infection control). The method for preparing concentrated platelet plasma products (PRF) by a prefilled catheter flusher combined with a three-way stopcock for horizontal separation is a single centrifugation method, and the method for preparing CGF is a single centrifugation method (this mode is a single centrifugation with four-stage frequency conversion). The blood collection and preparation steps are the same as before, except that the preparation mode of the centrifuge is selected differently, which will not be repeated here.

[0020] In the above-mentioned method for preparing concentrated platelet plasma, the maximum rotation speed of the secondary centrifugation of PRP and the primary centrifugation of PRF and CGF is 3000rpm, and the centrifugation method is horizontal centrifugation, which ensures that the liquid surface is not tilted after sample separation, which is convenient for accurate transfer of the three-way stopcock, and avoids the tilt of the liquid surface after separation by the angle rotor centrifuge or the mixing of a large number of red blood cells, which makes it difficult to extract; and the blood product container throughout the preparation process is carried out in a sealed adapter, and the transfer of samples is also carried out in a clean workbench, without the risk of aerosol contamination during centrifugation and infection caused by contact with air. This method selects medical consumables for preparation, professional sealed adapters, medical centrifuges with preparation modes, and clean workbenches for transfer operations to standardize and monitor the entire operation process. The cost of consumables used in this preparation method is about 30 yuan, which is suitable for the wide promotion and application of this technology in clinical practice.

[0021] The method of the present invention has the advantages that the whole process of standardized operation can be monitored, bacteria infection of samples can be avoided, aerosol contamination can be avoided, the safety of operators can be protected, and extraction is convenient.

[0022] In summary, the present invention has the following specific advantages: 1. The concentrated platelet plasma product of this method is derived from the patient's own blood and is used by the donor himself, without immune rejection reaction and risk of disease transmission; 2. The operation steps and processes are standardized and the preparation is stable; 3. In this preparation method, 18 ml of whole blood is used to prepare 3 ml of PRP; 27 ml of whole blood is used to prepare 4.5 ml of PRP, and 36 ml of whole blood is used to prepare 6 ml of PRP, which can be selected according to treatment needs, which is very convenient and fast; 3. This preparation method is composed of a special medical centrifuge with a preparation mode and an adapter with a sealing ring, which completely eliminates aerosol pollution in the separation process of an ordinary centrifuge and protects the safety of the operator; 4. The preparation operation is horizontal separation, and the centrifugal force is precisely controllable , the air shear force is minimal, which protects the platelets from being prematurely activated or damaged by external mechanical forces to the greatest extent, and facilitates the complete extraction of the three-way stopcock; 5. The samples are transferred and extracted in a Class 100 clean workbench in accordance with the infection control requirements, avoiding the risk of bacterial infection and protecting the operators from skin or mucous membrane contamination by the samples; 6. The operation steps can be replicated and improved, which is convenient for forming training materials for technical application and training promotion; 7. The concentrated platelet plasma product prepared by this method is liquid, which can be mixed with thrombin powder and calcium chloride to make a gel state (PG), which can retain the platelets in local tissues to release growth factors by activating fibrinogen to form a network scaffold, promote cell migration, differentiation and proliferation, and achieve tissue repair and regeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of the exploded structure of the prefilled catheter flusher;

[0024] Figure 2 Schematic diagram of three-way cock structure;

[0025] Figure 3 Schematic diagram of centrifuge structure;

[0026] Figure 4 Schematic diagram of centrifuge rotor structure;

[0027] Figure 5 Rotor operation structure diagram

[0028] Figure 6 Schematic diagram of the column structure with a sealing ring adapter;

[0029] Figure 7 Schematic diagram of the adapter cap structure with sealing ring

[0030] Figure 8 Schematic diagram of clean workbench;

[0031] Fig. 9 Schematic diagram of the structure of a syringe connected to a three-way connection to transfer blood;

[0032] Fig.10Schematic diagram of the structure of the red blood cells after the first separation using two prefilled catheter flushers combined with a three-way stopcock transfer;

[0033] Fig.11 A schematic diagram of the structure of the prefilled catheter after the second centrifugation of the present invention;

[0034] Fig.12 Schematic diagram of the transfer of a syringe and a prefilled catheter flusher after the second separation;

[0035] In the accompanying drawings, the components represented by the reference numerals are listed as follows: 01. Tube cap; 02. Prefilled catheter flusher; 03. Piston; 04. Core rod; 05. Three-way stopcock; 06. Centrifuge cover; 07. Centrifuge chamber; 08. Centrifuge screen; 09. Rotor; 10. Adapter cover; 11. Adapter with sealing ring; 12. Clean workbench; 13. Syringe; 14. Horizontal centrifuge.

Claims

1. Before preparation, wipe the clean bench 12 with 75% alcohol, put in spare consumables (tee, prefilled catheter flusher, tube rack, 20ml disposable sterile syringe, waste recycling bin, marker pen, treatment tray, sterile drape, sterile gauze), turn on the ultraviolet light for disinfection for at least 30 minutes, then turn off the ultraviolet light, turn on the fluorescent light and clean bench and keep blowing, and set aside.

2. Use a 20ml syringe 13 to first draw out 2ml of 3.8% citrate anticoagulant (or 4% citrate anticoagulant is also acceptable), then draw out 18ml of the patient's blood (the ratio of anticoagulant to blood is 1:9), and store the blood on a tray covered with a sterile drape.

3. After thoroughly mixing the venous blood and anticoagulant, take about 0.2 ml of blood sample for platelet count testing.

4. The operator wears clean gloves, transfers the anticoagulated blood sample to the operating table, opens the package of the prefilled catheter flusher 02, carefully unscrews the tube cap 01, taking care to avoid touching the interface end, and places the cap in a clean and untouchable position.

5. After draining the saline in the original tube of the prefilled catheter flusher 02, move the 20ml syringe 13 to the two 10ml prefilled catheter flushers 02 through the three-way stopcock 05. Pay attention to the aseptic operation during the transfer process and inject blood at a low speed as much as possible to avoid bubbles. After the transfer is completed, cover and tighten the prefilled catheter tube cover 01, rotate and remove the piston core rod 04, and place the prefilled catheter tube cover 01 vertically in the adapter 11 with the upper direction facing up, and cover the adapter cover 10; the two adapters 11 equipped with prefilled catheter flushers 02 are fixed in the "U" card slot on the medical centrifuge rotor 09 and placed symmetrically. Click on the centrifuge screen 08 to select the PRP mode for the first horizontal centrifugation 14 treatment, with a speed of 1600rpm, a temperature of 18°, a speed increase of 5 gears, a speed decrease of 2 gears, and a centrifugation time of 10 minutes and 30 seconds. The blood is divided into three layers, from top to bottom, respectively, the plasma layer, the buffy coat layer, and the red blood cell layer.

6. Open the adapter cap 10, take out the prefilled catheter flusher 02 vertically and place it on the clean bench tube rack for use (Note: In the subsequent operations, the prefilled catheter flusher after centrifugation must always be kept vertical, and never tilt or flip it to protect the separated PRP layer); perform routine hand disinfection and wear clean gloves. Take out a three-way stopcock 05 and a new prefilled catheter flusher 02, carefully open the package and unscrew its tube cover 01, and connect it to the three-way stopcock 05. Then keep the prefilled catheter flusher 02 containing blood products separated for the first time vertical to the ground, unscrew the tube cover 01 and connect it to the same three-way stopcock 05, open the three-way connection valve in an "L" shape, connect the prefilled flusher 02 containing blood products and the new prefilled catheter flusher 02, and slowly and evenly transfer the plasma layer and the white film layer of the sample into the new flusher, and stop when it is about 0.3mm below the red blood cell layer. Close the three-way stopcock 05, carefully unscrew the sample tube containing the remaining red blood cells, and discard it into the waste bucket; Replace another sample and repeat the above (8) operation. Recover the sample into a new tube for the next centrifugation step.

7. Place a new sample tube vertically into the adapter 11 with the cap 01 facing upwards, then place the adapter vertically into the medical centrifuge 9, symmetrically place another adapter 11 for balancing, select the PRP mode for the second horizontal centrifugation 14 treatment, with a speed of 2950 rpm, a temperature of 18°, a speed increase of 3 gears, a speed decrease of 2 gears, and a centrifugation time of 6 minutes.

8. Perform routine hand disinfection and wear clean gloves. Keep the sample vertical when taking it out. The upper layer of the sample is serum, the middle layer is platelet-rich plasma, and the bottom layer is a very small amount of red blood cells. Take out a syringe 13 and a three-way stopcock 05, separate the syringe needle as before, and connect them in an "L" shape with the three-way stopcock and the prefilled catheter flusher 02 with the separated sample, and drain the excess serum in the upper layer until there is 2.5 ml of plasma left in the prefilled catheter flusher 02 (as required for injection). After draining, separate the three-way stopcock 05 from the syringe 13, and connect the prefilled catheter flusher 02 containing the prepared platelet-rich plasma to the injection needle.

9. After thoroughly mixing the 2.5 ml plasma in (11) above, take about 0.1 ml of plasma sample for platelet count detection. The platelet count detected is between 3-5 times the platelet count of whole blood, which can meet clinical treatment needs. (Since the sample required for bacterial culture of adult blood is at least 5-10 ml, the bacterial culture of concentrated platelet plasma prepared by this method is selected in animal experiments to ensure that the preparation method meets the requirements of infection control).

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