Separation device and preparation method of injectable concentrated growth factor

Abstract

The invention relates to a separation device and a preparation method of injectable concentrated growth factors, which make up the defects of the prior art in the preparation of injectable CGF, such as incapability of separation caused by blood coagulation in the preparation process, and incapability of slow release of growth factors due to inhibition of formation of a fibrin scaffold by using an anticoagulant. After blood sampling, blood is injected into the lower cavity of the separation device, PPP, CGF and RBC are obtained through a specific centrifugal program, the first piston assembly is pushed to enable the PPP on the upper layer to enter the upper cavity, the upper cover body is moved downwards to enable the third barrel to be communicated with the lower cavity, the first piston assembly is moved upwards to enable the CGF to enter the third barrel, and separation can be completed. During separation, only the piston needs to be pushed up and the upper cover body needs to be moved downwards, operation is simple, samples do not need to be transferred, the separation process is isolated from the outside, and safety is high; the prepared CGF can continuously and slowly release various autologous growth factors, and is beneficial to playing a continuous role in a long-term tissue repair process.

Description

Technical Field

[0001] The invention relates to the technical field of medical experimental equipment, in particular to a separation device and a preparation method of injectable concentrated growth factors. Background Art

[0002] Concentrated growth factor was first reported by Sacco in 2005. It is a blood concentrate containing multiple growth factors prepared from autologous peripheral blood by differential centrifugation. No anticoagulant is added during the preparation process. After centrifugation, autologous fibrin gel rich in growth factors is directly obtained. It has the advantages of high biological activity, no immune rejection, and long-term effect. It plays an important role in tissue repair and tissue regeneration. After a specific differential centrifugation, the blood is divided into three layers. The top layer is liquid platelet-poor serum (PPP), the middle layer is gel-state concentrated growth factor (CGF), and the bottom layer is red blood cell clot (RBC). During the differential centrifugation process, the programmed switching between acceleration and deceleration can promote the conversion of fibrinogen into fibrin, forming a high-strength fiber network to prevent the growth factors from being hydrolyzed by proteins. The alternating centrifugal force provides more opportunities for collision with the centrifuge tube, resulting in more platelet rupture and the release of growth factors. Compared with platelet-rich plasma (PRP), CGF has more fully activated platelets, and the fibrin network can maintain and slowly release growth factors, which is closer to the natural process of tissue healing. CGF has a wide range of applications, and can be used in many different fields such as oral implants, plastic surgery, dermatology, etc., and has broad application prospects.

[0003] However, the existing concentrated growth factor preparation technology has the following technical problems: CGF is gel-like after centrifugation and cannot be used for injection therapy; vacuum blood collection tubes are often used as blood containers during the preparation process, and the tube caps need to be opened after centrifugation to remove the blocky CGF with tweezers, which has a high risk of contamination. In the existing preparation technology, anticoagulants need to be added and centrifuged to obtain injectable CGF, which not only increases the risk of immune rejection, but also inhibits fibrin polymerization. This type of CGF cannot form a scaffold in the body after injection and cannot play the role of maintaining and slowly releasing growth factors.

[0004] Chinese invention patent CN202410037415.X discloses a CGF separation and extraction device and an extraction method thereof, including a CGF collecting part, a sealed upper cover, a preparation cavity, a separation piston, a push-up support part, a lower cover, and a push-up spiral member. The preparation cavity includes an upper separation chamber and a lower blood receiving chamber, the separation chamber is connected to the blood receiving chamber through a chamber channel part, the sealed upper cover is installed on the separation chamber, the CGF collecting part is installed in the sealed upper cover, the push-up support part is installed in combination below the separation piston, the separation piston and the push-up support part are sealed and slidably installed in the blood receiving chamber, the lower cover is installed at the lower end opening of the blood receiving chamber, the push-up spiral member is installed in the push-up guide hole on the lower end surface of the lower cover through a threaded connection, and the upper end of the push-up spiral member cooperates with the lower end of the push-up support part.

[0005] The above scheme has the following problems during use. Specifically, since there is no anticoagulant added, blood is easily coagulated during centrifugation, and CGF cannot be transferred and injected after centrifugation. During operation, not only the CGF collection part needs to be raised and lowered back and forth, but also the sealing cover needs to be raised and lowered at the same time to connect the blood receiving chamber and the cavity in the CGF collection part, or to create a gap between the chamber channel part and the CGF collection part and the sealing cover. The operation is cumbersome, time-consuming and labor-intensive. Summary of the invention

[0006] 1. Technical issues to be solved

[0007] In order to solve the above problems, the present invention provides a separation device for injectable concentrated growth factors, which does not require moving the upper cover body and the second cylinder body, is simpler and more convenient to operate, and saves time and effort.

[0008] (II) Technical solution

[0009] To achieve the above object, the present invention provides the following technical solutions:

[0010] The present invention provides a separation device for injectable concentrated growth factors, comprising:

[0011] A first cylinder, the first cylinder is through-set from top to bottom, the first cylinder is provided with a first nipple protruding upward inside, the first nipple can divide the inside of the first cylinder into an upper cavity located above and a lower cavity located below, the top of the first cylinder is detachably connected to an upper cover body for sealing, and the bottom of the first cylinder is detachably connected to a lower cover body for sealing;

[0012] A second cylinder, the second cylinder is fixedly connected to the bottom of the upper cover and is arranged to pass through from top to bottom, the bottom of the second cylinder is provided with a second nipple which protrudes downward and corresponds to the first nipple, and there is a distance between the second nipple and the first nipple, and the distance can be reduced by moving the upper cover downward;

[0013] A third barrel, the third barrel can be connected to the inside of the second barrel and is arranged to penetrate up and down, the bottom of the third barrel is provided with a third nipple which protrudes downward and corresponds to the first nipple, the outer wall of the third nipple is slidably connected to the inner wall of the second nipple, the third nipple has a first state of abutting against the first nipple and a second state of not abutting against the first nipple, when the third nipple is in the first state, the third barrel is connected to the lower cavity, when the third nipple is in the second state, the third barrel is connected to the upper cavity, the second state is the initial state, and moving the upper cover downward can make the second barrel drive the third barrel to move downward, so that the third nipple is transformed into the first state;

[0014] a first piston assembly, the first piston assembly being sealingly and slidably connected to the interior of the lower cavity and being able to be close to or away from the first nipple;

[0015] The second piston assembly is sealingly and slidably connected to the interior of the third cylinder and can be close to or away from the third nipple.

[0016] Preferably, the first piston assembly comprises:

[0017] A screw rod, the screw rod is vertically arranged and threadedly connected to the middle part of the lower cover body, the screw rod has a first end and a second end which are far away from each other, the first end is located inside the first cylinder body, and the second end is located outside the first cylinder body;

[0018] A support member, wherein the support member is made of a rigid material, and the bottom of the support member is connected to the first end of the screw rod, so that the support member can move up and down as the screw rod rotates;

[0019] The first plug body is made of a flexible material, the bottom of the first plug body is detachably connected to the top of the support member to fix the relative positions of the first plug body and the support member, and the side of the first plug body is sealingly slidably connected to the inner wall of the first cylinder to prevent the liquid in the lower cavity from leaking from below.

[0020] Preferably, an injection hole is provided on the support member, one end of the injection hole abuts against the first plug body, and the other end opposite to the injection hole is offset from the screw rod.

[0021] Preferably, the second piston assembly comprises:

[0022] A core rod, the core rod is vertically arranged and slidably connected to the inner wall of the third cylinder, the core rod has a driving end and a driven end that are far away from each other, the driving end is located outside the third cylinder, and the driven end is located inside the third cylinder;

[0023] The second plug body is made of a flexible material, the top of the second plug body is detachably connected to the driven end of the core rod to fix the relative position of the second plug body and the core rod, and the side of the second plug body is sealingly slidably connected to the inner wall of the third cylinder to prevent the liquid in the third cylinder from leaking from above.

[0024] Preferably, a first hole is provided on the upper cover body, and the first hole connects the upper cavity and the outside world. A breathable membrane is placed at the first hole and fixed by a fixing plug. The fixing plug is provided with a breathable hole corresponding to the position of the first hole, so that the gas in the upper cavity and the gas in the outside world can circulate and exchange through the breathable membrane, and the gas passing through the breathable membrane is filtered, so that the air pressure in the upper cavity and the air pressure in the outside world can be kept consistent, and the gas in the upper cavity and the gas in the outside world can be prevented from being contaminated.

[0025] Preferably, the upper cover body is provided with a second hole, which connects the upper cavity and the outside and is used for inputting fluid into the upper cavity or extracting fluid from the upper cavity. A sealing plug for sealing is detachably connected to the second hole.

[0026] Preferably, an annular protrusion is provided on the top of the first nipple, and there is a distance between the top of the annular protrusion and the bottom of the second nipple. The inner wall of the annular protrusion can be slidably connected to the outer wall of the third nipple to enhance the sealing degree of the third nipple when it is in the first state.

[0027] Preferably, a plug is further included, which is detachably connected to the top of the second cylinder and is used to seal the second cylinder after the third cylinder is taken out.

[0028] Preferably, the first cylinder, the second cylinder and the third cylinder are all made of transparent material to facilitate observation of the liquid separation process.

[0029] Preferably, the first cylinder and the third cylinder are both provided with scales for observing the volume of the liquid.

[0030] The present invention also provides a method for preparing an injectable concentrated growth factor, which uses the above-mentioned injectable concentrated growth factor separation device to prepare the injectable concentrated growth factor. The preparation method comprises the following steps:

[0031] S1, blood collection;

[0032] S2, differential centrifugation;

[0033] S3, concentrated growth factor isolation.

[0034] Preferably, no anticoagulant is added during the blood collection process in step S1.

[0035] Preferably, the procedure of differential centrifugation in step S2 is as follows: the collected blood is injected into the lower cavity in the first cylinder, and variable speed centrifugation is performed at 10°C-20°C, and the centrifugal force reaches 610-650G for 100-140s through physical acceleration for 24-30s, and then the centrifugal force is decelerated to 460-510G for 220-240s, and then accelerated to 630-680G for 200-240s, and then accelerated to 700-750G for 160-200s, and finally decelerated for 35-40s before stopping.

[0036] (III) Beneficial effects

[0037] Compared with the prior art, the present invention has the following beneficial effects:

[0038] In actual use, blood is injected into the lower cavity in the first cylinder and centrifuged. After centrifugation, the blood is layered to obtain PPP in the upper layer, CGF in the middle layer, and RBC in the lower layer. Then, the first piston assembly is moved upward to allow the PPP in the lower cavity to enter the upper cavity through the first nipple. Then, the upper cover is moved downward to allow the third nipple to be in a first state of abutting the first nipple. The first piston assembly is continued to be moved upward to allow the target product CGF in the lower cavity to pass through the first nipple and the third nipple in sequence, and finally completely enter the interior of the third cylinder to obtain the concentrated growth factor CGF. Compared with the prior art, since there is a gap between the second nipple on the second cylinder and the first nipple on the first cylinder, during the entire operation, there is no need to move the upper cover and the second cylinder back and forth. Only the upper cover needs to be moved downward to complete the separation of CGF, which simplifies the operation steps and makes the preparation more convenient and quick. In general, by using the separation device for injectable concentrated growth factor, CGF does not need to be transferred to a syringe. After separation, it can be directly injected by connecting the injection needle, which is simpler and more convenient to operate, saving time and effort. The separation process is kept isolated from the outside world without the risk of pollution.

[0039] Furthermore, the present invention significantly slows down the polymerization rate of fibrin in CGF by controlling the centrifugal temperature and adjusting the centrifugal force. Under the heat preservation of the centrifuge, CGF can maintain a liquid state within 30 minutes to 60 minutes, and the liquid CGF can be transferred to the third cylinder of the separation device and directly used for clinical injection. CGF can quickly form a gel scaffold in the body by heating, thereby protecting and slowly releasing the growth factor. This technology solves the technical defects that the gel-like CGF prepared by the prior art cannot be used for injection, and the liquid CGF prepared by adding an anticoagulant cannot form a scaffold after injection and cannot slowly release the growth factor. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0041] Figure 1 A perspective view of a separation device for injectable concentrated growth factors of the present invention is shown;

[0042] Figure 2 Shows Figure 1 A three-dimensional view of a vertical cross section of the separation device for injectable concentrated growth factors when the third nipple is in a first state;

[0043] Figure 3 Shows Figure 2 Plan view at the cross section;

[0044] Figure 4 Shows Figure 1 A three-dimensional view of a vertical cross section of the separation device for injectable concentrated growth factors when the third nipple is in a second state;

[0045] Figure 5 Shows Figure 4 A magnified view of part A in FIG.

[0046] Figure 6 Shows Figure 4 A magnified view of part B in FIG.

[0047] Figure 7 Shows Figure 1 A three-dimensional schematic diagram of the exploded structure of the separation device for injectable concentrated growth factors;

[0048] Figure 8 Shows Figure 7 The enlarged view of the C part in FIG.

[0049] Fig. 9 Shows Figure 7 A three-dimensional schematic diagram from another angle;

[0050] Fig.10 Shows Figure 7 A plan view of a certain side;

[0051] Fig.11 Shows Figure 1 A perspective view of the middle screw and the first handle;

[0052] Fig.12 Shows Figure 1 A perspective view of the middle support member;

[0053] Fig.13 Shows Figure 1 A perspective view of the core rod and the second handle;

[0054] Fig.14 Shows Figure 1 A three-dimensional diagram of the separation device for injectable concentrated growth factor after the third cylinder is removed and the plug is connected;

[0055] Fig.15 Shows Fig.14 A three-dimensional diagram of the middle plug;

[0056] Fig.16 The experimental results of injectable CGF for sustained release of growth factors under physiological conditions are shown.

[0057] In the figure: 1. first cylinder; 101. first nipple; 1011. annular protrusion; 102. upper cavity; 103. lower cavity; 2. second cylinder; 21. second nipple; 3. third cylinder; 31. third nipple; 4. first piston assembly; 41. screw; 411. first end; 412. second end; 42. support member; 421. injection hole; 43. first plug body; 44. first handle; 5. second piston assembly; 51. core rod; 511. driving end; 512. driven end; 52. second plug body; 53. second handle; 6. upper cover body; 7. lower cover body; 8. first hole; 81. breathable membrane; 82. fixing plug; 821. breathable hole; 9. second hole; 91. sealing plug; 10. plug; 11. scale. DETAILED DESCRIPTION

[0058] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0059] Embodiment 1

[0060] See attached Figure 1 -Attached Figure 4The embodiment of the present invention discloses a separation device for injectable concentrated growth factor, comprising a first barrel 1, a second barrel 2, a third barrel 3, a first piston assembly 4, and a second piston assembly 5. The first barrel 1 is arranged to be through-connected from top to bottom, and a first nipple 101 protruding upward is arranged inside the first barrel 1. The first nipple 101 can divide the interior of the first barrel 1 into an upper cavity 102 located above and a lower cavity 103 located below. The top of the first barrel 1 is detachably connected with an upper cover body 6 for sealing, and the bottom of the first barrel 1 is detachably connected with a lower cover body 7 for sealing; the second barrel 2 is fixedly connected to the bottom of the upper cover body 6 and is arranged to be through-connected from top to bottom, and a second nipple 21 protruding downward and corresponding to the first nipple 101 is arranged at the bottom of the second barrel 2, and there is a distance between the second nipple 21 and the first nipple 101, and the distance can be reduced by moving the upper cover body 6 downward; the third barrel 3 can be connected to the first barrel 2, and the second nipple 21 is arranged to be through-connected from top to bottom. The interior of the second cylinder 2 is set to be connected up and down, and the bottom of the third cylinder 3 is provided with a third nipple 31 which protrudes downward and corresponds to the first nipple 101. The outer wall of the third nipple 31 is slidably connected to the inner wall of the second nipple 21. The third nipple 31 has a first state of abutting the first nipple 101 and a second state of not abutting the first nipple 101. When the third nipple 31 is in the first state, the third cylinder 3 is connected to the lower cavity 103. When the third nipple 31 is in the second state, the third cylinder 3 is connected to the upper cavity 102. The second state is the initial state. Moving the upper cover 6 downward can make the second cylinder 2 drive the third cylinder 3 to move downward, thereby changing the third nipple 31 to the first state; the first piston assembly 4 is sealingly slidably connected to the interior of the lower cavity 103 and can be close to or away from the first nipple 101; the second piston assembly 5 is sealingly slidably connected to the interior of the third cylinder 3 and can be close to or away from the third nipple 31.

[0061] In actual use, blood is injected into the lower cavity 103 in the first cylinder 1 and centrifuged. After centrifugation, the blood is stratified to obtain PPP in the upper layer, CGF in the middle layer, and RBC in the lower layer. Then, the first piston assembly 4 is moved upward to allow the PPP in the lower cavity 103 to enter the upper cavity 102 through the first nipple 101. Then, the upper cover 6 is moved downward to allow the third nipple 31 to be in the first state of abutting against the first nipple 101. The first piston assembly 4 is continued to be moved upward to allow the target product CGF in the lower cavity 103 to pass through the first nipple 101 and the third nipple 31 in sequence. , and finally completely enters the interior of the third barrel 3 to obtain the concentrated growth factor CGF; compared with the prior art, since there is a distance between the second nipple 21 on the second barrel 2 and the first nipple 101 on the first barrel 1, during the entire operation, there is no need to move the upper cover body 6 and the second barrel 2 back and forth, and only the upper cover body 6 needs to be moved down to complete the separation of CGF, which simplifies the operation steps and makes the preparation more convenient and quick; in general, by using the separation device for injectable concentrated growth factor, there is no need to move the upper cover body 6 and the second barrel 2 back and forth, and the operation is simpler and more convenient, saving time and effort.

[0062] It should be noted that the present application does not limit the manner of detachable connection and sealing between the upper cover body 6 and the first cylinder body 1, and between the lower cover body 7 and the first cylinder body 1, and can be flexibly selected according to actual needs, such as detachable connection through threaded connection, sealing by setting a sealing gasket at the connection between the two components and applying pressure, etc.

[0063] See attached Figure 2 And attached Fig. 9 -Attached Fig.11 In order to allow the first piston assembly 4 to be close to or away from the first nipple 101, the following design is carried out in this embodiment. Specifically, the first piston assembly 4 includes a screw 41, a support member 42, and a first plug body 43. The screw 41 is vertically arranged and threadedly connected to the middle part of the lower cover body 7. The screw 41 has a first end 411 and a second end 412 that are far away from each other. The first end 411 is located inside the first cylinder 1, and the second end 412 is located outside the first cylinder 1; the support member 42 is made of a rigid material, and the bottom of the support member 42 is connected to the first end 411 of the screw 41 so that the support member 42 can move up and down with the rotation of the screw 41; the first plug body 43 is made of a flexible material, and the bottom of the first plug body 43 is detachably connected to the top of the support member 42, which is used to fix the relative positions of the first plug body 43 and the support member 42. The side of the first plug body 43 is sealingly slidably connected to the inner wall of the first cylinder 1 to prevent the liquid in the lower cavity 103 from leaking from below.

[0064] Through the design of the above structure, by rotating the second end 412 of the screw 41, the first end 411 of the screw 41 can drive the support member 42 and the first plug body 43 to move up and down, thereby approaching or moving away from the first nipple 101, so that the first piston assembly 4 can push the liquid in the lower cavity 103 out of the first nipple 101.

[0065] It should be noted that the present embodiment does not limit the connection method between the screw 41 and the support member 42, and can be flexibly selected according to actual needs, such as a fixed connection or a rotating connection, both of which can enable the first plug body 43 to slide stably along the inner wall of the lower cavity 103 without skewing and affecting the sealing effect; in addition, the present embodiment also does not limit the detachable connection method between the support member 42 and the first plug body 43, and can be flexibly designed, such as the common embedded snap-on connection.

[0066] See attached Figure 1 and attached Fig.11 Furthermore, in order to facilitate the rotation of the screw rod 41 , a first handle 44 may be provided at the second end 412 of the screw rod 41 .

[0067] See attached Fig.12 Furthermore, the following design is also carried out in this embodiment. Specifically, an injection hole 421 is provided on the support member 42 , one end of the injection hole 421 abuts against the first plug body 43 , and the other end opposite to the screw rod 41 is staggered.

[0068] Through the design of the above structure, blood can be injected into the lower cavity 103 without pulling out the first piston assembly 4 from the first cylinder 1. Specifically, open the lower cover 7, move the support 42 to a position close to the bottom opening of the lower cavity 103, and then pass the metal injection needle through the injection hole 421 and pierce the first plug 43 made of flexible material, so that the blood in the syringe can be pushed into the lower cavity 103. After the injection is completed, pull out the metal injection needle. Since the first plug 43 is made of flexible material and obviously has a certain thickness, it can play a sealing role to prevent blood from leaking downward from the first plug 43. Then, connect the lower cover 7 to the bottom of the first cylinder 1 to prepare concentrated growth factors.

[0069] Furthermore, the injection hole 421 may be arranged at an angle to facilitate the insertion and withdrawal of the metal needle.

[0070] See attached Figure 2 , Attachment Figure 7 and attached Fig.13In order to allow the second piston assembly 5 to be close to or away from the third nipple 31, the following design is carried out in this embodiment. Specifically, the second piston assembly 5 includes a core rod 51 and a second plug body 52. ​​The core rod 51 is vertically arranged and slidably connected to the inner wall of the third cylinder 3. The core rod 51 has a driving end 511 and a driven end 512 that are away from each other. The driving end 511 is located outside the third cylinder 3, and the driven end 512 is located inside the third cylinder 3; the second plug body 52 is made of a flexible material, and the top of the second plug body 52 is detachably connected to the driven end 512 of the core rod 51, which is used to fix the relative positions of the second plug body 52 and the core rod 51. The side of the second plug body 52 is sealingly slidably connected to the inner wall of the third cylinder 3 to prevent the liquid in the third cylinder 3 from leaking from above.

[0071] Through the design of the above structure, by pulling the driving end 511 of the core rod 51 upward or pushing the driving end 511 of the core rod 51 downward, the driven end 512 of the core rod 51 can drive the second plug body 52 to move up and down, so as to approach or move away from the third nipple 31, so that during the process of liquid in the lower cavity 103 being injected into the third cylinder 3, the second plug body 52 can be moved upward by the core rod 51, so that the air pressure in the third cylinder 3 is kept stable, so that after the third cylinder 3 is removed from the second cylinder 2, the second plug body 52 can be moved downward by the core rod 51, so as to push out the liquid in the third cylinder 3.

[0072] See attached Figure 1 and attached Fig.13 Furthermore, in order to facilitate pushing and pulling the core rod 51 , a second handle 53 may be provided at the driving end 511 of the core rod 51 .

[0073] See attached Figure 4 , Attachment Figure 5 and attached Figure 8 Since the separation device for injectable concentrated growth factors includes multiple chambers, the air pressure in these chambers may have an adverse effect on the flow of the liquid during the liquid separation process. Therefore, the following design is carried out in this embodiment. Specifically, a first hole 8 is provided on the upper cover body 6, and the first hole 8 connects the upper cavity 102 and the outside world. A breathable membrane 81 is placed at the first hole 8 and fixed by a fixing plug 82. The fixing plug 82 is provided with a breathable hole 821 corresponding to the position of the first hole 8.

[0074] Through the design of the above-mentioned structure, the gas in the upper cavity 102 and the external gas can be circulated and exchanged through the breathable membrane 81, so that the air pressure in the upper cavity 102 and the external air pressure can be kept consistent, preventing the liquid in the lower cavity 103 from being hindered by the air pressure during the process of being input into the upper cavity 102; and because the breathable membrane 81 is provided, the circulating gas can be filtered to prevent the gas in the upper cavity 102 and the external gas from being contaminated. At the same time, the breathable membrane 81 also has a waterproof function, which can prevent the liquid in the separation device for injectable concentrated growth factors from leaking through the first hole 8.

[0075] See attached Figure 4 , Attachment Figure 6 and attached Figure 8 Furthermore, the following design is carried out in this embodiment. Specifically, a second hole 9 is provided on the upper cover body 6, and the second hole 9 connects the upper cavity 102 and the outside. A sealing plug 91 for sealing is detachably connected to the second hole 9.

[0076] Through the design of the above structure, after opening the sealing plug 91, the needle can be inserted into the upper cavity 102, so as to accurately extract and separate PPP, so that PPP, CGF and RBC can be accurately separated and extracted at the same time; in addition, disinfection gas or disinfection liquid, such as ethylene oxide in the disinfection and sterilization gas, can be injected into each chamber of the separation device for injectable concentrated growth factor through the second hole 9 to disinfect each component, which is more convenient to use; specifically, the ethylene oxide in the disinfection and sterilization gas is injected into the upper cavity 102 of the first barrel 1 through the second hole 9. Since there is a gap between the second nipple 21 and the first nipple 101, no matter whether the third nipple 31 is in the first state or the second state, and no matter whether the third nipple 31 passes through the second nipple 21, the ethylene oxide in the upper cavity 102 can enter the interior of the second barrel 2, and the third barrel 3 is moved to the second state, and the ethylene oxide can also enter the interior of the third barrel 3 and the interior of the lower cavity 103 of the first barrel 1, so as to achieve a more comprehensive sterilization effect.

[0077] See attached Figure 4 After the third nipple 31 and the first nipple 101 are abutted, only one surface is sealed, but the volume of the third nipple 31 and the first nipple 101 is obviously smaller, that is, the abutting area is small, and due to the existence of processing errors, the two may not be able to abut accurately, which will affect the sealing after the two are abutted. In order to solve the above problem, the following design is carried out in this embodiment. Specifically, an annular protrusion 1011 is provided on the top of the first nipple 101, and there is a gap between the top of the annular protrusion 1011 and the bottom of the second nipple 21, and the inner wall of the annular protrusion 1011 can be slidably connected with the outer wall of the third nipple 31.

[0078] Through the design of the above structure, after the third nipple 31 abuts against the first nipple 101, the bottom and side surfaces of the third nipple 31 are sealed, so the sealing effect is significantly better.

[0079] See attached Fig.14 and attached Fig.15 Since the second cylinder 2 is connected to the first cylinder 1, after the third cylinder 3 is removed from the second cylinder 2, the external environment can be indirectly connected to the first cylinder 1 through the second cylinder 2. The external environment contains various impurities such as dust, which makes the first cylinder 1 and the liquid in the first cylinder 1 very easy to be contaminated. In order to solve the above problem, the following design is carried out in this embodiment. Specifically, it also includes a plug 10, which is detachably connected to the top of the second cylinder 2.

[0080] Through the design of the above structure, after the third cylinder 3 is removed from the second cylinder 2, a plug 10 can be connected to the top of the second cylinder 2 for sealing, so that the interiors of the first cylinder 1 and the second cylinder 2 can be kept clean.

[0081] It should be noted that the material of the plug 10 and the manner in which the plug 10 and the second cylinder 2 are detachably connected are not limited in this embodiment, so it can be flexibly designed according to actual needs. For example, the plug 10 can be made of rubber material. In this case, the detachable connection between the plug 10 and the second cylinder 2 can be achieved by interference fit. Specifically, the diameter of the insertion portion of the plug 10 should be slightly larger than the diameter of the inner wall of the top of the second cylinder 2. After the plug 10 is inserted into the second cylinder 2, the rubber plug 10 will spontaneously expand outward under the action of its own elasticity, thereby forming a pressure-tight and damped plug-in detachable connection with the second cylinder 2.

[0082] In order to facilitate observation of the liquid separation process, the following design is carried out in this embodiment. Specifically, the first cylinder 1, the second cylinder 2, and the third cylinder 3 are all made of transparent materials.

[0083] See attached Figure 1 and attached Fig.10 Furthermore, the following design is also carried out in this embodiment. Specifically, scales 11 are provided on the first cylinder 1 and the third cylinder 3.

[0084] Through the design of the above structure, the volume of the liquid can be directly observed, and a specified liquid of a specified volume can be separated more accurately.

[0085] The present invention also provides a method for preparing an injectable concentrated growth factor, which uses the above-mentioned injectable concentrated growth factor separation device to prepare the injectable concentrated growth factor. The preparation method includes the following steps: S1, blood collection; S2, differential centrifugation; S3, concentrated growth factor separation.

[0086] Preferably, no anticoagulant is added during the blood collection process in step S1.

[0087] Preferably, the procedure of differential centrifugation in step S2 is as follows: the collected blood is injected into the lower cavity in the first cylinder, and variable speed centrifugation is performed at 10°C-20°C, and the centrifugal force reaches 610-650G for 100-140s through physical acceleration for 24-30s, and then the centrifugal force is decelerated to 460-510G for 220-240s, and then accelerated to 630-680G for 200-240s, and then accelerated to 700-750G for 160-200s, and finally decelerated for 35-40s before stopping.

[0088] Based on the above scheme, the above-mentioned separation device for injectable concentrated growth factor is taken out after centrifugation, and PPP located in the upper layer, CGF located in the middle layer and RBC located in the lower layer can be obtained, so that the third nipple 31 is in the second state where it does not abut the first nipple 101, the screw 41 is rotated and the first piston assembly 4 is moved upward, so that the PPP in the lower cavity 103 passes through the first nipple 101 and enters the upper cavity 102, and then the upper cover body 6 is moved downward, so that the third nipple 31 is in the first state abutting the first nipple 101, and then the first piston assembly 4 is moved upward, so that the target product CGF in the lower cavity 103 passes through the first nipple 101 and the third nipple 31 in turn, and finally completely enters the interior of the third cylinder 3, so that the concentrated growth factor CGF can be obtained; at this time, the injection needle is connected to the third nipple 31 for injection.

[0089] Based on the above scheme, this embodiment introduces a method for preparing a concentrated growth factor, which specifically includes the following steps:

[0090] Step 1: Draw 15-30 mL of peripheral blood from the blood using a sterile syringe without anticoagulant and inject it into the lower cavity 103 in the first barrel 1;

[0091] Step 2: Place the blood-injected separation device into a centrifuge with a temperature control system, and then select a balance separation device of equal weight and place it into the centrifuge at the same time, and keep the two separation devices symmetrical, and then start differential centrifugation, wherein the centrifugation temperature T is 10-20°C, preferably 15°C. If the temperature is too high, blood coagulation is easy to occur during the centrifugation process; if the temperature is too low, the gelation speed of CGF in the body after injection is slow, and the patient will feel uncomfortable; the specific parameters in the differential centrifugation program are: in the first stage, the acceleration time t1=24-30s, the centrifugal force is increased to 610-650G, and the centrifugal time t2 is 110-140s; specifically, t1 can be 24s, 25s, 28s, 30s, etc., and the centrifugal force can be 61 0G, 624G, 636G, 642G, 650G, etc., t2 can be 110s, 120s, 130s, 140s, etc., without specific limitation; the purpose of centrifugation in this stage is to make the blood cells settle. Too fast an increase in speed, too high a centrifugal speed and too long a time will cause the blood cells to rupture, otherwise the blood cells cannot be completely settled; in the second stage, the centrifugal force is adjusted to 460-510G, and the centrifugal time t3 is 220-240s. Specifically, the centrifugal force in this stage can be 460G, 466G, 475G, 484G, 497G, 503G, 510G, etc., and t3 can be 220s, 225s, 230s, 240s, etc., without specific limitation; in the third stage, the centrifugal force is adjusted to 63 0-680G, centrifugal time t4 is 200-240s, specifically, the centrifugal force in this stage can be 630G, 636G, 642G, 655G, 669G, 680G, etc., t4 can be 200s, 210s, 220s, 230s, 240s, etc., without specific limitation; in the fourth stage, the centrifugal force is increased to 700-750G, and the centrifugal time t5 is 160-200s. Specifically, the centrifugal force in this stage can be 700G, 705G, 718G, 734G, 742G, 750G, etc., t5 can be 160s, 170s, 180s, 190s, 200s, etc., without specific limitation; the purpose of the change of centrifugal force and centrifugal time from the second to the fifth stage is The platelets are aggregated to the middle layer, and the fibrin source in the blood is activated by physical acceleration and deceleration, inducing the platelets to release growth factors. Excessive centrifugal force and too long centrifugal time may cause the platelets to settle to the red blood cell layer, otherwise the platelets cannot fully aggregate, and excessive centrifugal force changes will cause the fibrin source to be overactivated and trigger coagulation. Too gentle centrifugal force changes are not conducive to the aggregation of platelets and fibrin in the middle layer, and the platelet activation degree is low, which is not conducive to the release of growth factors; in the fifth stage, the centrifugal force is reduced to 0, and the deceleration time t6 is 35-40s. Specifically, t6 can be 35s, 36s, 37s, 38s, 39s, and 40s, without specific limitation; the deceleration time in this stage affects the stability of the final centrifugal stratification;

[0092] Step 3: After the centrifugation, the blood is layered to obtain PPP (platelet-poor plasma) in the upper layer, CGF (concentrated growth factor) in the middle layer, and RBC (red blood cells) in the lower layer. The blood is carefully and steadily taken out of the centrifuge, and the first piston assembly 4 is moved upward to allow the PPP in the lower cavity 103 to enter the upper cavity 102 through the first nipple 101;

[0093] Step 4: Move the upper cover 6 downward to make the third nipple 31 in the first state of abutting the first nipple 101, and then move the first piston assembly 4 upward to make the target product CGF in the lower cavity 103 pass through the first nipple 101 and the third nipple 31 in sequence, and finally completely enter the interior of the third cylinder 3;

[0094] Step 5: Move the third barrel 3 upward, separate the third barrel 3 from the second barrel 2, and then connect an injection needle, such as a No. 7 injection needle, at the third nipple 31 to obtain injectable CGF;

[0095] Step 6: Before injecting CGF, first gently shake the third cylinder 3 to fully mix the CGF in the third cylinder 3, then point the third nipple 31 upward and move the second piston assembly 5 upward to discharge the gas and excess CGF in the third cylinder 3, and then inject the specified volume of CGF into the specified part through the injection needle.

[0096] By preparing CGF through the above method, there is no need to use anticoagulants, no adverse reactions will occur, the formation of fibrin network will not be inhibited due to the presence of anticoagulants, and platelets can be more fully activated to release growth factors; the third cylinder 3 can also be directly taken out for injection without the need to transfer CGF again, which makes the operation simpler and can effectively prevent CGF from being contaminated during multiple transfers.

[0097] It should be noted that the target product CGF prepared by the above steps can maintain injectable performance for 30-60 minutes in a centrifuge insulation environment, so it should be injected as soon as possible or stored in time.

[0098] Embodiment 2

[0099] The difference between this embodiment and the first embodiment is that different centrifugation conditions are used for centrifugation; specifically, the effect of temperature and centrifugation procedure on the state of CGF after separation and the gel speed is evaluated.

[0100] 20 mL of blood was drawn and injected into the separation device for the above-mentioned injectable concentrated growth factor. Centrifugation was performed using the centrifugation conditions in Table 1. The state of CGF after centrifugation was observed and the time for the injectable CGF to change from liquid to gel at 37°C was calculated. The test results are shown in Table 2.

[0101] Table 1 is as follows:

[0102]

[0103] Table 2 is as follows:

[0104]

[0105] Embodiment 3

[0106] The difference between this embodiment and the first embodiment is that an experimental study on the sustained release of growth factor by injectable CGF under physiological environment was conducted. The specific experimental process is as follows:

[0107] 36 volunteers were recruited, and 26 mL of autologous blood was collected from each person, of which 20 mL of blood was used to separate 3 mL of CGF by the preparation method and separation device provided by the present invention, and the remaining 6 mL of blood was used to prepare 3 mL of plasma. 3 mL of CGF and 3 mL of plasma were added to 10 mL of serum-free DMEM culture medium, and incubated at 37 ° C. 200 mL of culture medium supernatant was taken every 24 hours, and 200 mL of fresh culture medium was supplemented, and sampling continued for 12 days; the content of TGF-β, PDGF and VEGF growth factors released from CGF and plasma into the culture medium was detected by ELISA kit, and the average value was taken to draw the concentration curve of growth factors over time; the above three growth factors have the functions of promoting cell proliferation and differentiation, regulating inflammation, and promoting angiogenesis in clinical tissue repair, and play a key role in tissue repair and reconstruction.

[0108] The experimental results are as follows Fig.16 As shown, the three growth factors TGF-β, PDGF and VEGF in CGF showed sustained release behavior within 12 days, and the concentrations were higher than those in plasma and did not decrease significantly, indicating that CGF can slowly release growth factors in a physiological environment and can maintain the activity of growth factors for a long time, which has advantages in the application of long-term tissue reconstruction such as bone repair and articular cartilage treatment.

[0109] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0110] In addition, it should be noted that the use of terms such as "first" and "second" to limit components is only for the convenience of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore cannot be understood as limiting the scope of protection of this application.

[0111] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A separation device for injectable concentrated growth factors, characterized in that: include: A first cylinder, the first cylinder is through-set from top to bottom, the first cylinder is provided with a first nipple protruding upward inside, the first nipple can divide the inside of the first cylinder into an upper cavity located above and a lower cavity located below, the top of the first cylinder is detachably connected to an upper cover body for sealing, and the bottom of the first cylinder is detachably connected to a lower cover body for sealing; A second cylinder, the second cylinder is fixedly connected to the bottom of the upper cover and is arranged to pass through from top to bottom, the bottom of the second cylinder is provided with a second nipple which protrudes downward and corresponds to the first nipple, and there is a distance between the second nipple and the first nipple, and the distance can be reduced by moving the upper cover downward; A third barrel, the third barrel can be connected to the inside of the second barrel and is arranged to penetrate up and down, the bottom of the third barrel is provided with a third nipple which protrudes downward and corresponds to the first nipple, the outer wall of the third nipple is slidably connected to the inner wall of the second nipple, the third nipple has a first state of abutting against the first nipple and a second state of not abutting against the first nipple, when the third nipple is in the first state, the third barrel is connected to the lower cavity, when the third nipple is in the second state, the third barrel is connected to the upper cavity, the second state is the initial state, and moving the upper cover downward can make the second barrel drive the third barrel to move downward, so that the third nipple is transformed into the first state; a first piston assembly, the first piston assembly being sealingly and slidably connected to the interior of the lower cavity and being able to be close to or away from the first nipple; The second piston assembly is sealingly and slidably connected to the interior of the third cylinder and can be close to or away from the third nipple.

2. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: The first piston assembly comprises: A screw rod, the screw rod is vertically arranged and threadedly connected to the middle part of the lower cover body, the screw rod has a first end and a second end which are far away from each other, the first end is located inside the first cylinder body, and the second end is located outside the first cylinder body; A support member, wherein the support member is made of a rigid material, and the bottom of the support member is connected to the first end of the screw rod, so that the support member can move up and down as the screw rod rotates; The first plug body is made of a flexible material, the bottom of the first plug body is detachably connected to the top of the support member to fix the relative positions of the first plug body and the support member, and the side of the first plug body is sealingly slidably connected to the inner wall of the first cylinder to prevent the liquid in the lower cavity from leaking from below.

3. The separation device for injectable concentrated growth factors according to claim 2, characterized in that: The support member is provided with an injection hole, one end of the injection hole abuts against the first plug body, and the other end opposite to the injection hole is staggered with the screw rod.

4. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: The second piston assembly comprises: A core rod, the core rod is vertically arranged and slidably connected to the inner wall of the third cylinder, the core rod has a driving end and a driven end that are far away from each other, the driving end is located outside the third cylinder, and the driven end is located inside the third cylinder; The second plug body is made of a flexible material, the top of the second plug body is detachably connected to the driven end of the core rod to fix the relative position of the second plug body and the core rod, and the side of the second plug body is sealingly slidably connected to the inner wall of the third cylinder to prevent the liquid in the third cylinder from leaking from above.

5. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: The upper cover body is provided with a first hole, which connects the upper cavity and the outside world. A breathable membrane is placed at the first hole and fixed by a fixing plug. The fixing plug is provided with a breathable hole corresponding to the position of the first hole, so that the gas in the upper cavity and the gas in the outside world can be circulated and exchanged through the breathable membrane, and the gas passing through the breathable membrane is filtered, so that the air pressure in the upper cavity and the air pressure in the outside world can be kept consistent, and the gas in the upper cavity and the gas in the outside world can be prevented from being contaminated.

6. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: The upper cover body is provided with a second hole, which connects the upper cavity and the outside and is used for inputting fluid into the upper cavity or extracting fluid from the upper cavity. A sealing plug for sealing is detachably connected to the second hole.

7. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: An annular protrusion is provided at the top of the first nipple, and there is a distance between the top of the annular protrusion and the bottom of the second nipple. The inner wall of the annular protrusion can be slidably connected to the outer wall of the third nipple to enhance the sealing degree of the third nipple when it is in the first state.

8. The separation device for injectable concentrated growth factors according to claim 1, characterized in that: It also includes a plug, which is detachably connected to the top of the second cylinder and is used to seal the second cylinder after the third cylinder is taken out.

9. The separation device for injectable concentrated growth factors according to any one of claims 1 to 8, characterized in that: The first cylinder, the second cylinder and the third cylinder are all made of transparent materials to facilitate observation of the liquid separation process.

10. The separation device for injectable concentrated growth factors according to claim 9, characterized in that: The first cylinder and the third cylinder are both provided with scales for observing the volume of the liquid.

11. A method for preparing an injectable concentrated growth factor, characterized in that: The injectable concentrated growth factor separation device according to any one of claims 1 to 9 is used to prepare the injectable concentrated growth factor, and the preparation method comprises the following steps: S1, blood collection; S2, differential centrifugation; S3, concentrated growth factor isolation.

12. The method for preparing an injectable concentrated growth factor according to claim 11, characterized in that: No anticoagulant is added during the blood collection process in step S1.

13. The method for preparing an injectable concentrated growth factor according to claim 11, characterized in that: The procedure of differential centrifugation in step S2 is as follows: injecting the collected blood into the lower cavity in the first cylinder, performing variable speed centrifugation at 10°C-20°C, physically accelerating for 24-30s, the centrifugal force reaches 610-650G for centrifugation for 100-140s, then decelerating to 460-510G for centrifugation for 220-240s, accelerating to 630-680G for centrifugation for 200-240s, then accelerating to 700-750G for centrifugation for 160-200s, and finally decelerating for 35-40s before stopping.

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