Leukocyte-rich PRP extraction method based on leukocyte-rich PRP extraction device

Abstract

The invention discloses a leukocyte-rich PRP extraction method based on a leukocyte-rich PRP extraction device. The extraction device comprises a centrifugal tube body and a tube cap which are buckled with each other, the centrifugal tube body sequentially comprises a first cavity, a second cavity and a third cavity from top to bottom, the first cavity, the second cavity and the third cavity are communicated in sequence, and the tube inner diameter of the second cavity is smaller than that of the third cavity; one end of the piston is fixedly connected to the base; one end of the second cavity is embedded in the first cavity, the other end of the second cavity is embedded in the third cavity, the base covers an opening of the third cavity through threaded connection, the volume of a collected blood sample is defined as Lml, the minimum volume of the third cavity reaching the top limit position is 0.30-0.35 * L, and the volume of the second cavity is 0.2-0.3 * L; the separating device comprises a screw rod which sequentially penetrates through the base and the piston and extends to the third cavity, and a plug and a knob cap which are connected to the two ends of the screw rod; and the plug is used for blocking the second cavity.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a leukocyte-rich PRP extraction method based on a leukocyte-rich PRP extraction device. Background Art

[0002] Platelet-rich plasma (PRP) is widely used in modern medical fields in tissue repair, regenerative medicine, and cosmetic surgery. PRP promotes tissue healing and regeneration through its high concentration of platelets and growth factors. However, different types of PRP have different effects due to their different components and application purposes. Leukocyte-rich platelet-rich plasma (LR-PRP) can provide stronger immunity and repair functions due to its high concentration of white blood cells and platelets. It has unique advantages in anti-infection, inflammation control, and tissue repair, such as:

[0003] Chronic wounds: including diabetic foot ulcers, bedsores, etc. The high white blood cell content in LR-PRP helps to remove necrotic tissue and fight infection.

[0004] Acute injuries: such as sports injuries, acute muscle tears, and ligament injuries, LR-PRP initiates a stronger healing process by increasing the inflammatory response.

[0005] Post-operative repair: Post-operative wound healing and tissue repair, LR-PRP can promote tissue regeneration and reduce the risk of infection.

[0006] Bone Healing: Used in fracture healing and bone transplantation, the white blood cells in LR-PRP aid in bone regeneration and repair.

[0007] Therefore, LR-PRP shows significant effects in the treatment of chronic wounds, acute injuries and postoperative repair.

[0008] Currently, most PRP extraction devices on the market focus on extracting ordinary PRP or low-leukocyte PRP, and it is difficult to effectively extract leukocyte-rich platelet-rich plasma. Existing technical means usually require multiple centrifugation and manual operation to separate white blood cells and platelets, which is not only complicated and time-consuming, but also easy to introduce contamination, reduce extraction efficiency and sample purity. In addition, the existing devices do not handle blood samples gently during operation, which may lead to reduced activity of platelets and white blood cells, affecting the final treatment effect. Summary of the invention

[0009] In view of the problems existing in the above-mentioned prior art, the present invention provides a leukocyte-rich PRP extraction method based on a leukocyte-rich PRP extraction device, which can effectively solve the problems existing in the above-mentioned prior art.

[0010] The technical solution of the present invention is:

[0011] According to one aspect of the present invention, the extraction device comprises: a centrifugal tube body and a tube cap that are interlocked, wherein the centrifugal tube body comprises, from top to bottom, a first cavity, a second cavity, and a third cavity that are sequentially connected, and the inner diameter of the second cavity is smaller than the inner diameter of the third cavity;

[0012] It also includes a piston and a base, one end of the piston is fixedly connected to the base; the other end is embedded in the third cavity, the base is threadedly connected to the opening of the third cavity, the volume of the collected blood sample is defined as Lml, the minimum volume of the third cavity at the top limit position is 0.30-0.35*L, and the volume of the second cavity is 0.2-0.3*L;

[0013] A separation device, comprising a screw rod that sequentially passes through the base and the piston and extends to the third cavity, and a plug and a knob cap connected to both ends of the screw rod; the plug rod is used to plug the second cavity, and the plug rod can move along the length direction of the second cavity under the drive of the screw rod to connect or close the second cavity and the third cavity;

[0014] The extraction method comprises the following steps:

[0015] S1, the extraction device is arranged vertically in the forward direction, the base is rotated toward the third cavity to the top limit position, and then the plug is moved away from the second cavity, so that the first cavity, the second cavity and the third cavity are connected;

[0016] S2, injecting the collected blood sample into the centrifuge tube;

[0017] S3, starting the centrifuge, controlling the centrifugal force and time of the centrifuge to perform the first centrifugation, so that the blood sample is separated into a red blood cell layer, a white blood cell layer and a plasma layer from bottom to top;

[0018] S4, slowly rotating the base to drive the piston to move downward, so that the red blood cell layer in the second cavity slowly flows into the third cavity, so that the boundary line between the red blood cell layer and the white blood cell layer is located at the bottom of the second cavity;

[0019] S5, slowly turning the knob cap to make the screw drive the plug to move toward the second cavity, so that the plug is blocked in the second cavity, thereby separating the red blood cell layer from the white blood cell layer;

[0020] S6, turning the extraction device 180 degrees, starting the centrifuge, and controlling the centrifugal force and time of the centrifuge to perform a second centrifugation to concentrate the platelets and leukocytes in the bottom layer;

[0021] S7, insert the syringe from the bottom of the tube cap to collect the concentrated platelets, leukocytes and plasma.

[0022] Further, in step S3, the step of controlling the centrifugal force and time of the centrifuge to perform the first centrifugation so that the blood sample is separated from bottom to top into a red blood cell layer, a white blood cell layer and a plasma layer specifically includes:

[0023] The centrifugal force of the centrifuge is controlled to be 380-420 g, and the centrifugal time is 3-10 minutes for the first centrifugation, so that the blood sample is separated into a red blood cell layer, a white blood cell layer and a plasma layer from bottom to top.

[0024] Furthermore, in step S6, the step of turning the extraction device 180°, starting the centrifuge, and controlling the centrifugal force and time of the centrifuge to perform a second centrifugation to concentrate platelets and leukocytes in the bottom layer specifically includes:

[0025] The centrifugal force of the centrifuge is controlled to be 950-1050 g, and the centrifugal time is 5-15 minutes for a second centrifugation to concentrate the platelets and leukocytes in the bottom layer.

[0026] Further, in step S2, the step of injecting the collected blood sample into the centrifuge tube specifically includes:

[0027] Insert the syringe from the top of the tube cap and inject the collected blood sample into the centrifuge tube.

[0028] Further, in step S3, when the boundary between the red blood cell layer and the white blood cell layer is located in the third cavity, a preliminary judgment is made as to a blood problem, and the process ends.

[0029] Furthermore, L is 5 to 100 ml, and the minimum volume of the third cavity when it reaches the top limit position is 0.33 to 0.34*L.

[0030] Furthermore, the inner diameter of the second cavity is 1 to 1.8 cm.

[0031] Further, in step S5, when the knob cap is slowly turned to make the screw drive the plug to move toward the second cavity, the boundary line between the red blood cell layer and the white blood cell layer rises significantly, the base is slowly rotated to drive the piston to move downward, so that the boundary line between the red blood cell layer and the white blood cell layer is always located at the bottom of the second cavity.

[0032] Further, in step S3, when the boundary between the red blood cell layer and the white blood cell layer is located in the first cavity, a blood problem is preliminarily determined and the process ends.

[0033] By adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art:

[0034] This scheme provides an efficient and easy-to-operate leukocyte-rich platelet-rich plasma (LR-PRP) extraction device, which can effectively separate and extract high-concentration platelets and leukocytes in a single operation, simplify the operating steps, reduce the risk of contamination, improve the extraction efficiency and sample activity, and thus better meet clinical needs; the LR-PRP extracted by this device will be able to play a more significant role in the treatment of chronic wounds, acute injuries, postoperative repair, etc., promote tissue regeneration and healing, and improve the treatment effect and quality of life of patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0036] Figure 1 This is a schematic diagram of the vertical cross-sectional structure of the extraction device in the present invention;

[0037] Figure 2 This is a schematic cross-sectional structure diagram of the plug blocking the second cavity in the present invention;

[0038] Figure 3 This is a structural schematic diagram of the syringe inserted into the first cavity when the extraction device is rotated 180° and the extraction device is vertically reversed in the present invention;

[0039] In the figure: centrifuge tube body-1, first cavity-11, third cavity-12, second cavity-13, tube cap-2, slope surface-21, bracket-3, piston-4, limiting groove-41, base-5, connecting column-51, limiting convex edge-52, separation device-6, plug-61, screw-62, knob cap-63, syringe-A. DETAILED DESCRIPTION

[0040] The present invention will be further described in detail below in conjunction with the accompanying drawings and examples. It is particularly noted that the following examples are only used to illustrate the present invention, but are not intended to limit the scope of the present invention. Similarly, the following examples are only partial embodiments of the present invention rather than all embodiments, and all other embodiments obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present invention.

[0041] like Figures 1 to 3 As shown, the present scheme provides a leukocyte-rich PRP extraction method based on a leukocyte-rich PRP extraction device.

[0042] See also Figure 1 and Figure 2 The extraction device comprises: a centrifugal tube body 1 and a tube cap 2 that are interlocked, and in this embodiment, a bracket 3 is further included, the bracket 3 is fixedly sleeved outside the tube cap 2, and the bracket 3 is threadedly connected to the first cavity 11. The tube cap 2 is made of silicone; it is convenient for the syringe A to be inserted into the tube cap 2 to collect platelets, white blood cells and plasma.

[0043] See also Figure 1 and Figure 2 , further comprising a base 5 and a piston 4, the piston 4 being embedded in the third cavity 12 so as to be movable up and down, one end of the piston 4 being fixedly connected to the base 5, and the base 5 being covered at the opening of the third cavity 12 by a threaded connection. The initial state of the base 5 is to rotate to the limit position toward the third cavity 12. Specifically, the internal thread of the base 5 is cooperatively connected with the external thread of the third cavity 12. Preferably, a connecting column 51 is provided at one end of the base 5 extending toward the third cavity 12, the connecting column 51 is provided with a limiting convex edge 52, and the piston 4 is correspondingly provided with a limiting groove 41, and the limiting convex edge 52 is engaged with the limiting groove 41.

[0044] See also Figure 1 and Figure 2 , the centrifugal tube body 1 includes, from top to bottom, a first cavity 11, a second cavity 13 and a third cavity 12 which are connected in sequence, and the inner diameter of the second cavity 13 is smaller than the inner diameter of the third cavity 12. The volume of the collected blood sample is defined as Lml, wherein L can be 5-100ml, in which many embodiments, L is 10-30ml, and in this embodiment, L is about 20ml. Since the red blood cell content in normal human whole blood is between 35% and 55%. In order to make the boundary between the red blood cell layer and the white blood cell layer located in the second cavity 13 in the subsequent separation process, preferably, the minimum volume of the third cavity 12 at the top limit position should be 0.30-0.35*L, more preferably, the minimum volume of the third cavity 12 at the top limit position should be 0.33-0.34*L, in this embodiment, the minimum volume of the third cavity 12 at the top limit position is about 6.8ml. The maximum volume of the third cavity 12 at the bottom limit position is not limited, and preferably can be about 0.5*L. In this embodiment, the maximum volume of the third cavity 12 when it reaches the bottom limit position is about 10 ml.

[0045] The volume of the second cavity 13 is preferably 0.2 to 0.3*L. The advantage is that when about 35% of the red blood cells fill the third cavity 12 at the top limit position during the first centrifugal separation, the remaining red blood cells will be located in the second cavity 13, so that the boundary between red blood cells and white blood cells is also located in the second cavity 13, which facilitates the subsequent separation of the red blood cell layer and the white blood cell layer. In this embodiment, the volume of the second cavity 13 is preferably 4ml to 6ml, and preferably, the volume of the second cavity 13 is preferably 5ml.

[0046] In this embodiment, further, in order to make the boundary between red blood cells and white blood cells more obvious, under the premise that the volume of the second cavity 13 remains unchanged, the inner diameter of the second cavity 13 can be further reduced to increase the height of the second cavity 13. However, as the inner diameter decreases, the stratification of red blood cells and white blood cells will not be complete (as the inner diameter decreases, the centrifugal force also decreases, F = mω2r, where F represents centrifugal force, m represents mass, ω represents angular velocity, and r represents radius). Experiments have shown that when the inner diameter of the second cavity 13 is less than 1 cm, it is difficult to effectively separate red blood cells and white blood cells when the centrifugal force of the first centrifugation is 400g (g is the acceleration of gravity). Taking the volume of the second cavity 13 as 5ml as an example (V = π*R 2 *h, V represents volume, R represents radius, and h represents height), preferably, the inner diameter of the second cavity 13 is 1 cm to 1.8 cm, and the corresponding height of the second cavity 13 is 6.37 to 1.97 cm; more preferably, the inner diameter of the second cavity 13 is 1.2 cm to 1.6 cm, and the corresponding height of the second cavity 13 is 4.42 to 2.49 cm.

[0047] See also Figures 1 to 3 , also includes a separation device 6 passed through the third cavity 12, the separation device 6 moves along the length direction of the second cavity 13 to connect or close the second cavity 13 and the third cavity 12; preferably, the separation device 6 includes a plug 61, a screw 62 and a knob cap 63, the screw 62 passes through the base 5 and the third cavity 12, and the screw 62 is threadedly connected to the base 5; the plug 61 can move along the length direction of the second cavity 13 driven by the screw 62 to connect or close the second cavity 13 and the third cavity 12; one end of the screw 62 is fixed with a plug 61 for blocking the second cavity 13, and the other end is fixed with a knob cap 63.

[0048] See also Figure 1 and Figure 2 One end of the tube cap 2 is provided with an annular slope surface 21 extending toward the inner circumference of the first cavity 11. Therefore, when the extraction device is reversed and vertical, the space from the junction of the first cavity 11 and the tube cap 2 to the upper end surface of the first cavity 11 is wide at the top and narrow at the bottom, which is convenient for observing the collection of platelets, white blood cells and plasma.

[0049] The extraction method comprises the following steps:

[0050] S1, the extraction device is set vertically in the forward direction, the base 5 is rotated to the top limit position toward the third cavity 12, and then the plug 61 is moved away from the second cavity 13, so that the first cavity 11, the second cavity 13 and the third cavity 12 are connected;

[0051] S2, injecting the collected blood sample into the centrifuge tube body 1; the step of injecting the collected blood sample into the centrifuge tube body 1 specifically includes: inserting the syringe A from the top of the tube cap 2 and injecting the collected blood sample into the centrifuge tube body 1.

[0052] S3, start the centrifuge, control the centrifugal force and time of the centrifuge to perform the first centrifugation, so that the blood sample is separated from the bottom to the top into a red blood cell layer, a white blood cell layer and a plasma layer; preferably, the step of controlling the centrifugal force and time of the centrifuge to perform the first centrifugation, so that the blood sample is separated from the bottom to the top into a red blood cell layer, a white blood cell layer and a plasma layer specifically includes: controlling the centrifugal force of the centrifuge to be 380-420g and the centrifugation time to be 3-10 minutes for the first centrifugation, so that the blood sample is separated from the bottom to the top into a red blood cell layer, a white blood cell layer and a plasma layer.

[0053] When the boundary line between the red blood cell layer and the white blood cell layer is located in the third cavity 12, the blood problem is initially determined and the procedure ends. When the boundary line between the red blood cell layer and the white blood cell layer is located in the third cavity 12, it indicates that the red blood cell content of the whole blood is between 35% and 55%.

[0054] When the boundary line between the red blood cell layer and the white blood cell layer is located in the first cavity 11, the blood problem is initially determined and the procedure ends. When the boundary line between the red blood cell layer and the white blood cell layer is located in the first cavity 11, it means that the red blood cell content of the whole blood exceeds 55%.

[0055] S4, slowly rotate the base 5 to drive the piston 4 to move downward, so that the red blood cell layer in the second cavity 13 slowly flows into the third cavity 12, so that the boundary between the red blood cell layer and the white blood cell layer is located at the bottom of the second cavity 13;

[0056] S5, slowly turn the knob cap 63 so that the screw 62 drives the plug 61 to move toward the second cavity 13, so that the plug 61 is blocked in the second cavity 13, thereby separating the red blood cell layer from the white blood cell layer; when the knob cap 63 is slowly turned so that the screw 62 drives the plug 61 to move toward the second cavity 13, when the boundary line between the red blood cell layer and the white blood cell layer rises significantly, the base 5 is slowly rotated to drive the piston 4 to move downward, so that the boundary line between the red blood cell layer and the white blood cell layer is always located at the bottom of the second cavity 13.

[0057] S6, flipping the extraction device 180°, starting the centrifuge, controlling the centrifugal force and time of the centrifuge for a second centrifugation, so that the platelets and leukocytes are concentrated in the bottom layer; preferably, the step of flipping the extraction device 180°, starting the centrifuge, controlling the centrifugal force and time of the centrifuge for a second centrifugation, so that the platelets and leukocytes are concentrated in the bottom layer specifically includes:

[0058] The centrifugal force of the centrifuge is controlled to be 950-1050 g, and the centrifugal time is 5-15 minutes for a second centrifugation to concentrate the platelets and leukocytes in the bottom layer.

[0059] S7, insert the syringe A from the bottom of the tube cap 2 to collect the concentrated platelets, leukocytes and plasma.

[0060] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device, characterized in that: The extraction device comprises: a centrifugal tube body (1) and a tube cap (2) which are interlocked, wherein the centrifugal tube body (1) comprises, from top to bottom, a first cavity (11), a second cavity (13) and a third cavity (12) which are sequentially connected, and the inner diameter of the second cavity (13) is smaller than the inner diameter of the third cavity (12); It also includes a piston (4) and a base (5), one end of the piston (4) is fixedly connected to the base (5); the other end is embedded in the third cavity (12), the base (5) is connected to the opening of the third cavity (12) through a threaded cover, the volume of the collected blood sample is defined as Lml, the minimum volume of the third cavity (12) at the top limit position is (0.30-0.35)*L, and the volume of the second cavity (13) is (0.2-0.3)*L; A separation device (6), comprising a screw (62) which sequentially passes through the base (5) and the piston (4) and extends to the third cavity (12), and a plug (61) and a knob cap (63) connected to both ends of the screw (62); the plug (61) is used to plug the second cavity (13); the plug (61) can move along the length direction of the second cavity (13) driven by the screw (62) to connect or close the second cavity (13) and the third cavity (12); The extraction method comprises the following steps: S1, the extraction device is arranged vertically in the forward direction, the base (5) is rotated toward the third cavity (12) to the top limit position, and then the plug (61) is moved away from the second cavity (13), so that the first cavity (11), the second cavity (13) and the third cavity (12) are connected; S2, injecting the collected blood sample into the centrifuge tube (1); S3, starting the centrifuge, controlling the centrifugal force and time of the centrifuge to perform the first centrifugation, so that the blood sample is separated into a red blood cell layer, a white blood cell layer and a plasma layer from bottom to top; S4, slowly rotating the base (5) to drive the piston (4) to move downward, so that the red blood cell layer in the second cavity (13) slowly flows into the third cavity (12), so that the boundary between the red blood cell layer and the white blood cell layer is located at the bottom of the second cavity (13); S5, slowly turning the knob cap (63) so that the screw rod (62) drives the plug (61) to move toward the second cavity (13), so that the plug (61) is blocked in the second cavity (13), thereby separating the red blood cell layer from the white blood cell layer; S6, turning the extraction device 180 degrees, starting the centrifuge, and controlling the centrifugal force and time of the centrifuge to perform a second centrifugation to concentrate the platelets and leukocytes in the bottom layer; S7, insert the syringe (A) from the bottom of the tube cap (2) to collect the concentrated platelets, leukocytes and plasma.

2. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S3, the step of controlling the centrifugal force and time of the centrifuge to perform the first centrifugation so that the blood sample is separated from bottom to top into a red blood cell layer, a white blood cell layer and a plasma layer specifically includes: The centrifugal force of the centrifuge is controlled to be 380-420 g, and the centrifugal time is 3-10 minutes for the first centrifugation, so that the blood sample is separated into a red blood cell layer, a white blood cell layer and a plasma layer from bottom to top.

3. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S6, the extraction device is turned over 180 degrees, the centrifuge is started, and the centrifugal force and time of the centrifuge are controlled to perform a second centrifugation to concentrate the platelets and leukocytes in the bottom layer. Specifically, the steps include: The centrifugal force of the centrifuge is controlled to be 950-1050 g, and the centrifugal time is 5-15 minutes for a second centrifugation to concentrate the platelets and leukocytes in the bottom layer.

4. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S2, the step of injecting the collected blood sample into the centrifuge tube (1) specifically includes: Insert the syringe (A) from the top of the tube cap (2) and inject the collected blood sample into the centrifuge tube body (1).

5. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S3, when the boundary between the red blood cell layer and the white blood cell layer is located in the third cavity (12), a preliminary blood problem is determined and the process ends.

6. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: L is 5 to 100 ml, and the minimum volume of the third cavity (12) when it reaches the top limit position is (0.33 to 0.34)*L.

7. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 6, characterized in that: The inner diameter of the second cavity (13) is 1 to 1.8 cm.

8. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S5, when the knob cap (63) is slowly rotated to cause the screw rod (62) to drive the plug (61) to move toward the second cavity (13), when the boundary line between the red blood cell layer and the white blood cell layer rises significantly, the base (5) is slowly rotated to drive the piston (4) to move downward, so that the boundary line between the red blood cell layer and the white blood cell layer is always located at the bottom of the second cavity (13).

9. A method for extracting leukocyte-rich PRP based on a leukocyte-rich PRP extraction device as claimed in claim 1, characterized in that: In step S3, when the boundary between the red blood cell layer and the white blood cell layer is located in the first cavity (11), a preliminary blood problem is determined and the process ends.

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