KR101087653B1 - Preparing for prp - Google Patents

Abstract

The present invention provides a method for producing multi-platelet plasma, comprising: separating the collected blood into a blood cell layer / PRP layer / PPP layer, separating only the PRP, and then separating only the blood cell layer; Accommodating the blood cell layer in a container having a predetermined space in front of the first piston and shielding the upper part of the blood cell layer with a second piston, and then centrifuging the blood cell layer; And applying positive pressure to the centrifuged blood cell layer to collect PRP in the front of the first piston, or applying negative pressure to recover PRP in another container.

Description

 Production method of multi-platelet plasma {PREPARING FOR PRP}

The present invention relates to a method for producing multiplatelet plasma, and more particularly, to a method for producing multiplatelet plasma which can improve platelet recovery.

PRP (Patelet Rich Plasma) refers to the centrifugation of blood after special treatment, which refers to the plasma components that make platelets richer than normal blood. Originally, platelets contain several growth factors, which play an important role in wound healing and skin regeneration. In addition, it is reported that such platelets are contained in high concentrations in PRP, and various growth factors stimulate the proliferation of surrounding cells and stimulate abundant synthesis of collagen and other components. Accordingly, PRP has been used in various fields such as pain treatment such as low back pain, hair loss treatment, skin disease such as skin regeneration or burn treatment.

The method of forming PRP has been proposed in terms of physical and chemical methods, and the conventional method is a physical method, which centrifuges collected blood to form blood components and platelet-poor plasma (PPP). The method of separating and extracting only PRP using a filter is used. However, the conventional method has a problem that the blood may be exposed to the atmosphere in the process of moving the blood to the centrifuge vessel, the transfer to the separation vessel including the filter after the blood collection. Exposure to the atmosphere may cause side effects due to secondary infections or contamination due to safety issues for the human body, such as a risk of microbial contamination. That is, the conventional method cannot use the syringe for blood collection, so it is necessary to move the container, and it is difficult to directly use it at the procedure site because it must be moved from the blood collection equipment to the separation equipment and the syringe for injection again. There is a risk of causing sepsis.

In addition, platelets have a significant difference in specific gravity from red blood cells, so that a considerable amount of platelets are included in a layer in which red blood cells are stacked, but the conventional physical method has a disadvantage in that platelets included in the red blood cell layer cannot be separated. In the use of PRP, the exclusion of red blood cells is the most important. Therefore, the removal of red blood cells by the conventional method was used to utilize the high concentration of platelets in the remaining plasma. Although there are more than 10 times the number of platelets, no method of removing them has been suggested.

On the other hand, as a chemical method, International Publication No. WO2004 / 012750 proposes a method for selectively promoting aggregation and sedimentation of red blood cells by adding sodium poly-L-glutamate to blood. However, the chemical method cannot be a preferable means in that the same external exposure as the conventional physical method coexists to separate only PRP.

For these reasons, the conventional PRP recovery rate is reported to be about 78% even in the case of AutoloGel ^™ by Cytomedix. Therefore, about 20% or more platelets are discarded in a state where they cannot be recovered.

Platelet recovery

An object of the present invention to solve the above problems is to provide a method for producing multi-platelet plasma which can recover the platelet components remaining in the plasma layer to be discarded.

Another object of the present invention is to provide a method for producing multi-platelet plasma having improved stability by blocking the possibility of air pollution while minimizing PRP damage.

Still another object of the present invention is to provide a method for obtaining PRP at a procedure site in a short time.

In order to achieve the above object, the present invention provides a method for producing multi-platelet plasma, comprising: preparing a blood cell layer; Accommodating the blood cell layer in a container having a predetermined space in front of the first piston and shielding the upper part of the blood cell layer with a second piston, and then centrifuging the blood cell layer; And applying positive pressure to the centrifuged blood cell layer to collect PRP in the front of the first piston, or applying negative pressure to recover PRP in another container.

In another aspect, the present invention provides a method for producing multi-platelet plasma, comprising the steps of preparing a blood cell layer; The blood cell layer is accommodated in a container having a predetermined space in front of the first piston, the upper part of the blood cell layer is shielded with a second piston, and then PPP or isotonic solution to be discarded is added to the blood cell layer, followed by a saking process. Floating platelets and centrifuging them; And applying positive pressure to the centrifuged blood cell layer to collect PRP in the front of the first piston, or applying negative pressure to recover PRP in another container.

In another aspect, the present invention comprises the steps of adding the PPP or isotonic solution to be discarded in the blood cell layer and floating the platelets through a saking process and centrifuging them; And applying positive pressure to the centrifuged blood cell layer to collect PRP in the front part of the first piston, or applying negative pressure to recover PRP in another container.

In addition, the present invention includes a filter of 4 to 10㎛, more preferably 3 to 5㎛ on the first piston when the positive pressure is applied, or to 10㎛, more preferably 3 to the second piston when the negative pressure is applied It provides a method for producing multi-platelet plasma containing a filter of 5㎛.

In addition, the present invention comprises the steps of collecting blood from the human or animal to separate only the blood cell layer; Centrifuging the blood contained in the container to separate the blood cell layer / PRP layer / PPP layer; Centrifuging the separated blood again, separating the blood cells / PRP layer / PPP layer between the filters by using a piston including filters of different sizes mounted in a container; It provides a method for producing multi-platelet plasma comprising the step of removing the exposed PPP component and transferring only the PRP component to another container through a Luerac adapter.

In addition, the present invention is the piston is a piston body without a plunger, a packing for maintaining airtightness with the cylinder inner wall of the container is coupled to the outer surface of the piston body, the outlet port is connected to the front and rear of the piston body, It includes an opening and closing means such as a bolt of the discharge port, and a filter formed in the discharge passage of the blood, a cap is formed at the front end of the body, the cap is formed with a plurality of outlet holes, the front end is attached to the tip of the container It provides a method for producing a multi-platelet plasma formed with a stopper.

As described above, the method for preparing multi-platelet plasma according to an embodiment of the present invention has an advantage of obtaining stability of PRP by blocking the opportunity of blood contacting the outside in obtaining PRP as an active ingredient. have.

In addition, the method according to the embodiment of the present invention can be obtained as a simple tool in the separation process can be obtained immediately PRP, the acquisition of the method is easy to reduce the cost as well as the degree of training of the operator There is an advantage.

The PRP obtained according to the present invention can directly use the blood collected in the procedure space, and thus, it is possible to secure the stability of the immune system that can be obtained by using autologous blood.

The terms "about "," substantially ", etc. used to the extent that they are used herein are intended to be taken to mean an approximation of, or approximation to, the numerical values of manufacturing and material tolerances inherent in the meanings mentioned, Accurate or absolute numbers are used to help prevent unauthorized exploitation by unauthorized intruders of the referenced disclosure.

In the blood collection according to the present invention, the blood may be blood of human or animal, and may be collected through a syringe. The syringe which can be used at this time includes all types capable of collecting blood, and as a non-limiting example, Korean Patent Application Nos. 2003-5029, 2005-61134, 2005-34848, It may be a syringe disclosed in 2006-64946, Korean Utility Model Application No. 2003-21484, 2004-10685, 2006-26454. A characteristic of the syringe is that at the same time as the blood is drawn, the collected blood is blocked from the atmosphere.

On the other hand, the piston that can be used in the syringe may be those shown in Figures 7 and 8 as a non-limiting example. The piston 200 has a piston body 210 without a plunger, a packing 220 coupled to the outer surface of the piston body to maintain airtightness with the cylinder inner wall of the container, and the front and rear of the piston body. It is a structure including a discharge port 230 is connected to, the opening and closing means 240, such as the bolt of the discharge port, and the filter 250 is formed in the discharge passage of blood, the weight of the piston body separately in the rear ( 260 may be formed in a coupled structure. In addition, a cap 270 is formed at the front end of the main body 210, and the cap 270 is formed with a plurality of outlet holes 271, and a stopper 273 at the front part so as to be attached to the front end of the container. ) Is formed. The piston may be used more than one in one syringe in carrying out the method according to the invention. When one or more pistons are used, the filter of the first piston is made of a mesh structure of 4 to 10㎛, and the second piston may be a filter made of a mesh structure of 0.1 to 0.3㎛.

Since the piston according to the present invention does not have a plunger, a luer lock adapter 300 or a plunger driver 400 or 500 as shown in FIGS. 4 to 6 may be used. The plunger drive may be disclosed in Utility Model Registration No. 323838 previously proposed by the present inventors. The detailed configuration and use method will be described later.

On the other hand, as a non-limiting example of the method for producing multi-platelet plasma according to the present invention can be prepared by the following method.

First, blood is collected from a human or animal with a syringe made of the piston. The tip of the syringe where blood collection is completed is shielded by means such as a lid.

The opening and closing means of the piston in the container containing the collected blood forms a sealed state (meaning bolts in the case of bolts) and through the centrifugation as shown in Figure 1 from the bottom blood cell layer 110 The PRP layer 130 and the PPP layer 150 are separated. Centrifugation can proceed for about 15 minutes at 4,500 rpm.

A first piston having a filter having a diameter of about 5 μm and a second piston having an opening and closing means of the piston 200 and equipped with a filter of about 0.2 μm to remove residual components except PRP from the centrifuged blood. Mount on the top of the piston. At this time, the opening and closing means of the second piston forms an open state.

After the centrifugation for 5 minutes at about 3,500rpm again, as shown in FIG. 1, the PPP layer 150 is formed on the second piston 300 and the first piston 200 as shown in FIG. 1. In between, the PRP layer 130 is formed in a state where the blood cell layer 110 is separated from each other under the first piston 200. It cannot pass through the second piston because the size of platelets is about 2 to 3 μm and red blood cells cannot pass through the first piston because it is about 6 to 9 μm. Using this principle, only the effective ingredient PRP is separated.

Among the components separated between the pistons, the PPP layer is removed because it is in an open state even without a separate device, and the active ingredient PRP is removed by using a Luerac adapter in a state in which the opening / closing means is removed. Fork syringe) can be moved. The transfer method can be transferred to another vessel pressurizing the Luerac adapter. The luerac adapter 300 is the end portion 310 is formed of a shape 311 corresponding to the piston and the end of the end portion is formed with a screw thread 313, the end portion 330 is a syringe It is formed to correspond to the inlet. Body tube 350 is in the form of a hollow cylinder.

PRP component formed as described above is characterized in that the opportunity to be exposed to the atmosphere in the process of moving from the blood collection step to the injection syringe finally.

On the other hand, when explaining the method for producing PRP as another embodiment, the blood collection step and centrifugation step is as described above. However, in the above example, the component moved to another syringe was the PPP component, but in this embodiment, both PPP and PRP are transferred to another container. In another container, a piston equipped with a 0.2 μm filter is mounted, and when the piston is opened and opened, the centrifugal separation is performed at about 3,500 rpm for 5 minutes. Are separated. The PPP component formed on the top of the piston is in an open state and can be removed (see FIG. 2).

Meanwhile, as another embodiment, a method of manufacturing PRP is proposed. First, the blood collection step is as described above. However, in this embodiment, there is a difference in that the first piston including the 5 μm filter and the second piston including the 0.2 μm filter are simultaneously mounted in the syringe containing the collected blood. The opening and closing means of the first and second pistons are kept open and centrifuged for 15 minutes at about 4,500 rpm. The centrifuged blood is formed from the bottom of the blood cell layer / PRP layer / PPP layer, the active ingredient PRP can be extracted in the same manner as described above (see Figure 3).

Hereinafter, a method of further recovering PRP to increase the recovery rate will be described.

Meanwhile, FIG. 9 illustrates a blood cell layer 100 which is discarded in the related art, and the blood cell layer includes red blood cells 111 and platelets 113. Therefore, the recovery rate can be improved by further recovering the platelets of the discarded blood cell layer. At the same time, this recovery rate should be improved, along with the prevention of contamination that may be caused by exposure to the atmosphere during a series of processes.

As a method of recovering PRP from the blood cell layer, the blood cell layer 110 is accommodated in a container having a predetermined space in front of the first piston 610, and centrifuged to thereby centrifuge PRP having a smaller specific gravity than red blood cells. Gather at the points. In this case, a second piston 630 may be further formed on the blood cell layer for sealing. The first piston and the second piston may include a filter of 4 to 10㎛, more preferably 3 to 5㎛ to separate platelets and red blood cells.

The method of separating only PRP may apply positive pressure to the vessel so that the PRP collects at the front of the first piston (in this case, the filter may be included in the first piston), or the negative pressure may be used to recover the PRP to another vessel. (In this case, the filter may be included in the second piston) (hereinafter referred to as 'first recovery method', see FIG. 10).

Meanwhile, another method of recovering PRP is to add PPP or isotonic solution to be discarded to the blood cell layer 110, float the platelets through a saking process, centrifuge it, and further recover PRP by separating only the PRP described above. (Hereinafter referred to as a 'second recovery method') The second recovery method may be executed independently, or may be performed on the remaining blood cell layer after the first recovery method.

Meanwhile, FIG. 11 shows a state in which a piston is mounted in a syringe in a reverse direction, which shows an example, and a mounting direction of the piston may be forward or reverse.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the inventions. It will be clear to those who have knowledge of.

1 is a conceptual diagram of a PRP manufacturing process according to an embodiment of the present invention.

2 is a conceptual diagram of a PRP manufacturing process according to another preferred embodiment of the present invention.

3 is a conceptual diagram of a PRP manufacturing process according to another preferred embodiment of the present invention.

Figure 4 is a front view of the Luer lock adapter according to a preferred embodiment of the present invention.

5 and 6 are front views of the flanger drive according to an embodiment of the present invention.

7 and 8 are an exploded perspective view and a cross-sectional view of the piston of the present invention.

9 is a conceptual diagram of a blood cell layer.

10 is a conceptual diagram of a PRP recovery according to an embodiment of the present invention.

11 is a conceptual view in which the piston is mounted in the reverse direction.

Claims (6)

In the method of producing multi-platelet plasma, Preparing a blood cell layer; Accommodating the blood cell layer in a container having a predetermined space in front of the first piston and shielding the upper part of the blood cell layer with a second piston, and then centrifuging the blood cell layer; And Positive pressure is applied to the centrifuged blood cell layer so that PRP (multiple platelet plasma) is collected at the front of the first piston, or negative pressure is applied to the recovery of PRP (multiple platelet plasma) to another container. Manufacturing method. In the method of producing multi-platelet plasma, Preparing a blood cell layer; The blood cell layer is accommodated in a container having a predetermined space in front of the first piston, and the upper part of the blood cell layer is shielded with a second piston, and then PPP (anemia platelet plasma) or isotonic fluid to be discarded is added to the blood cell layer. Floating the platelets through a shaking process and centrifuging them; And Positive pressure is applied to the centrifuged blood cell layer so that PRP (multiple platelet plasma) is collected at the front of the first piston, or negative pressure is applied to the recovery of PRP (multiple platelet plasma) to another container. Manufacturing method. The method of claim 1, Adding PPP (anemia platelet plasma) or isotonic solution to be discarded to the blood cell layer, floating the platelets through a saking process, and centrifuging the platelets; And Applying positive pressure to the centrifuged blood cell layer to collect PRP (multiple platelet plasma) in the front part of the first piston, or by applying negative pressure to multi-platelet plasma further comprising the step of recovering PRP (multiple platelet plasma) in another container Manufacturing method. The method according to any one of claims 1 to 3, When the positive pressure is applied to the first piston comprises a filter of 4 to 10㎛, or when the negative pressure is applied to the second piston is a method for producing multi-platelet plasma comprising a filter of 4 to 10㎛. The method according to claim 1 or 2, To prepare the blood cell layer Taking blood from a human or animal; Centrifuging the blood contained in the container to separate into blood cell layer / PRP (polyplatelet plasma) layer / PPP (anemia platelet) layer; Centrifuging the separated blood again, using a piston comprising a filter of different sizes mounted in the container, each of the blood cell layer / PRP (polyplatelet plasma) layer / PPP (anemia platelet) layer between the filters Separating to locate; Removing the exposed PPP (anemia platelet plasma) component and transferring only the PRP (multiplatelet plasma) component to another container through a Luerac adapter to separate only the blood cell layer. The method according to claim 1 or 2, The piston includes a piston body without a plunger, a packing coupled to an outer surface of the piston body for maintaining airtightness with the cylinder inner wall of the container, an outlet port connected to the front and rear of the piston body, and bolts of the outlet port; The same opening and closing means, and including a filter formed in the discharge passage of the blood, Cap is formed at the front end of the main body, the cap is formed with a plurality of outlet holes, the front portion is a method for producing multi-platelet plasma formed with a stopper to be attached to the front end of the container.

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