WO2023069027A1

WO2023069027A1 - A tube for advanced method of obtaining platelet rich fibrin

Info

Publication number: WO2023069027A1
Application number: PCT/TR2021/051070
Authority: WO
Inventors: Salih ÖZOVALI; Timur Veysel DOĞRUOK
Original assignee: T-Bi̇yoteknoloji̇ Laboratuvar Esteti̇k Medi̇kal Kozmeti̇k San. Ve Ti̇c. Ltd. Şti̇.
Priority date: 2021-10-19
Filing date: 2021-10-19
Publication date: 2023-04-27
Anticipated expiration: 2024-04-19
Also published as: EP4419254A1

Abstract

The invention comprises the following; longitudinally arranged tube body (11) with a mouth (113), a tube cap (12) connected with the tubular body (11), needle inlet (121) at the top of the tube cap (12) body, sealing element (122) on the body of the tube cap (12), at least one destruction element (13) on the inner surface of the tube body (11) and extending at least partially towards the center of the tube in order to shorten the coagulation process by breaking down the platelets in the blood during centrifugation, a longitudinal pipe body (111) in the tube body (11), body base (112) at the bottom of said pipe body (111) preferably provided in a curved structure, tubular body (111) and body base (112) provided in single piece.

Description

A TUBE FOR ADVANCED METHOD OF OBTAINING PLATELET RICH FIBRIN

FIELD OF THE INVENTION

The present invention relates to a method for obtaining platelet rich fibrin from autologous blood by centrifugation process, and to a centrifuge tube for use in the related method.

PRIOR ART

Platelet Rich Fibrin (PRF) means a polymerized structure due to activation of platelets to form a gel-lookalike structure formed of blood during centrifugation without any presence of any anticoagulants or other substances within the protocol, however there may have some other supportive protocols to be applied along with the designated tube.

PRF is a form of activated platelets and used to enhance various treatments due the growth factors within the fibrin form inside the recipient tissue. Since plenty of growth factors are presented with their own mechanism of actions; a fibrin structure as its solid form is used in various aspects for regenerative medicine. Most particularly, widely used in facial plastic surgery, oral and maxillofacial surgery, dentistry and dental surgery, regenerative orthopedics, regenerative aesthetics. The basis of the treatment is based on the preparation of Platelet Rich Fibrin from one's own processed blood to be applied onto the wounded/injured/impaired area in order to stimulate the regenerative potential due to growth factor secretion to utilize the healing cascade.

It consists of three parts in the tube as a consequence of the centrifugation due to density of the different layers affected by the centrifugational force. After the designated protocol including the centrifugation, the bottom layer is formed of Red Blood Cells due to higher molecular weight and the total number of the cells compared to other cell types within the blood. The middle section is formed of fibrin structure, mainly formed of platelets due to their activation and 3D formation of a scaffold. Platelets are non-nucleated cells with an outer membrane. Due to activation of platelets in-vitro; the membrane of the platelets are disrupted and protruding pseudopods formation occurs to form a 3-dimensional clot or fibrin structure as a scaffold. On the top layer, serum or liquid remnant formed of plasma is presented.

PRF is made suitable for use as a result of the centrifugation process, the section consisting of fibrins in the middle of the tube was taken from the three sections formed in the tube, and dewatering process is optionally carried out by removing the liquid inside with certain plates or equipment designed as a compression or crushing system.

Most platelet-rich tissue obtaining methods, there is a need for faster clotting in PRF production because fibrin tissue is obtained quickly and its use increases the effectiveness of the treatment steps and provides saving time. For these reasons, centrifugal tubes containing coagulation chemicals are generally used in advanced platelet-rich fibrin extraction methods.

One of the tubes generally used in the technical field, metered vacuum (6 ml volume) and gel Coagulation Activator tubes with added silica particles are used. It makes the sample ready for centrifugation by preventing the blood cells from adhering to the inner surface of the tube and providing a rapid coagulation in a short time such as 5-10 minutes by the additive (silica gel) it contains, and the gel contained therein easily enters between the shaped elements of the blood (erythrocyte, leukocyte, thrombocyte) and the rest of the blood during the centrifugation process, separating the two in a healthy way. This is the general working principle of silica gel tubes. However, particles of that chemical remain in the PRF produced as a result of the use of these tubes and this has negative consequences in terms of health. Glass tubes have been used so as to prevent this situation however, the yellowing of the glass color of the glass tubes in long use reveals a bad and dirty appearance and it is not preferred for us since it frequently breaks during use.

Centrifuge is basically a separation method. Particles in the sample are separated according to their shape, size and density using the centrifugal force from the rotational motion.

The radial acceleration causes denser particles in the sample tubes to settle at the bottom, while lower density substances remain at the top.

The blood to be separated is first placed in the centrifuge tubes in the centrifugation process. These tubes are specific tubes that are suitable for the centrifuge device. The tubes are placed in the centrifuge device. There is a point to be taken into consideration while placing the same. Tubes should be placed symmetrically so as to ensure a healthy centrifugal balance. RCF (Relative Centrifugal Force), are determined and the process is initiated.

Centrifuge tubes are conical or cylindrical tubes that are made of pressure-resistant glass or plastic, in which the material to be centrifuged is placed. The product to be worked on and the amount of product are effective in tube selection.

When classifying the fields of use and types of centrifuge tubes, they are generally classified according to the cap colors. Examples of these tubes are as follows:

Red capped tubes are used to coagulate blood and obtain serum by centrifugation, separate the serum from the blood with its Special Gel formulation and yellow capped tubes are for obtaining serum, blue capped tubes are used for coagulation tests (prothrombin time), the tube containing thrombin, orange capped tubes that allow the blood to clot quickly.

Some fibrin structures are obtained by separating them to be used in medical fields or different areas as a result of the centrifugation process. PRF is widely used, especially in medical fields.

A different or improved centrifuge tube is required for the production of PRF obtained in the centrifugation process specifically in medical fields.

The following documents are found during the preliminary patent research made.

Use of titanium or titanium alloy tubes to ensure faster coagulation of the blood to be centrifuged is proposed in the document with publication number WO2013062495A2. Although effective results are obtained, titanium tubes are not preferred since they are produced at very high costs.

Tubes and devices that are used in tissue regeneration, collection of whole blood in the system in a tube containing thixotropic gel, and centrifugation of this tube until thrombin serum remains, used for the separation of thrombin and platelets are disclosed in the PCT document with publication number KR1020180127538A. It causes health problems as the gel particles remain in the PRF at the end of the centrifugation process due to the gel it contains. In PCT document with publication number WO2014168409A1 , a blood separation vessel designed for platelet self-removal is disclosed. This vessel has a high cost and a more complex structure since it consists of various layers and elements.

PCT document with publication number KR101279652B1 relates to a plasma separation apparatus designed to shorten the coagulation time of blood plasmas by shortening the coagulation time of blood plasmas and to increase its efficiency, it has a higher cost and a more complex structure due to the fact that the separation apparatus consists of various layers and elements.

As a result, all abovementioned problems have made it necessary to make an improvement in the relevant technical field.

AIM OF THE INVENTION

The present invention aims to eliminate the abovementioned problems and to make a development in the relevant technical field.

The main aim of the invention is to provide an improved method of obtaining platelet rich fibrin, which does not require the use of coagulation accelerator chemicals and/or substances.

Another aim of the invention is to produce the PRF tissue obtained as a result of the centrifugation process more efficiently in terms of vision, dimension and weight.

Another aim of the invention is to provide a centrifuge tube structure that can provide faster coagulation without the use of chemicals and/or substances and is cost-effective.

BRIEF DESCRIPTION OF INVENTION

The present invention method above and which can be construed from the detailed description below is a centrifugal tube for use in a centrifugation process so as to obtain platelet rich fibrin. Accordingly, the present invention comprises, a longitudinally arranged tubular body with a mouth, a tube cap connected with the tubular body, at least one destruction element extended on the inner surface of the tubular body and at least partially extended towards the tube center to shorten the coagulation process by breaking down the platelets in the blood during centrifugation. In a preferred embodiment of the invention, said destruction element is arranged to extend at least partially along the tubular body.

A preferred embodiment of the invention, said destruction element is arranged in such a way that it extends linearly from the mouth to the base of the body.

A preferred embodiment of the invention comprises a plurality of destruction elements.

A preferred embodiment of the invention, said destruction element is provided angularly spaced on the inner surface of the tubular body.

A preferred embodiment of the invention, said destruction element is not provided on at least a part of the inner surface of said tubular body.

A preferred embodiment of the invention, said destruction element is provided helically on the inner surface of the tubular body.

A preferred embodiment of the invention, the centrifugal tube is made of plastic, preferably polyethylene terephthalate.

A preferred embodiment of the invention, it comprises a needle inlet that is positioned in the center of the tube cap so as to prevent microorganisms from entering the tubular body with blood by opening the tube cover during the process of discharging blood into the tubular body, allows the syringe needle to enter easily with its elastic structure and the needle hole to be closed automatically after exiting, preventing the entry of microorganisms.

The present invention method above and which can be construed from the detailed description below is a method for obtaining platelet rich fibrin. Accordingly, the present method comprises the following steps;

- Transferring the blood drawn from the donor to the centrifugal tube according to any of the preceding claims,

- Centrifugation of the centrifuge tube in such a way that the platelets in the blood are destroyed by the destruction element and that the blood is separated into three layers, at least a cell-free plasma, red blood cells and platelet rich fibrin and

- Separating said platelet-rich fibrin from the remaining layers.

A preferred embodiment of the invention, dewatering obtained platelet-rich fibrin. A preferred embodiment of the invention, said centrifugation process is carried out between 300-4000 RCF and between 5-60 minutes.

A preferred embodiment of the invention, said centrifugation process is repeated twice at intervals.

A preferred embodiment of the invention, the first step of said centrifugation process is carried out between 1 -20 minutes, between 100-2000 RCF, second step is carried out between 300-4000 RCF and between 5-60 minutes.

A preferred embodiment of the invention, said centrifugation process is repeated three times at intervals.

A preferred embodiment of the invention, the first step of said centrifugation process is carried out between 100-2000 RCF and between 1 -20 minutes, second step is carried out between 300-4000 RCF and between 5-60 minutes, third step is carried out between 100-2000 RCF and between 1-20 minutes.

BRIEF DESCRIPTION OF DRAWINGS

In Figure 1 , an isometric view of an embodiment of the inventive centrifugal tube is given.

A cross-sectional view of Figure 1 is given in Figure 2.

Figure 3 shows the top view of Figure 1 without the tube cap.

Figure 4, is a top view of an embodiment of the inventive centrifugal tube.

DESCRIPTION OF THE REFERENCES IN FIGURES

10. Centrifugal tube

11. Tubular body

111. Pipe body

112. Body base

113. Mouth

12. Tube cap

121. Needle inlet

122. Sealing element 13. Destruction element

DETAILED DESCRIPTION OF THE INVENTION

In this detailed description, the inventive advanced method of obtaining platelet rich fibrin and a centrifugal tube suitable for this method (10) is described by means of examples only for clarifying the subject matter such that no limiting effect is created.

The subject of the invention relates to a centrifuge tube (10) designed to obtain the improved platelet rich fibrins in the blood in a healthy, easy and low cost manner by centrifugation process.

The present invention relates to a centrifuge tube (10) designed to obtain the improved platelet rich fibrins in the blood in a healthy, easy and low cost manner by centrifugation process, characterized in that; it comprises the following;

• A longitudinally arranged tubular body (11 ) with a mouth (113),

• One tube cap (12) connected with the tubular body (11 ),

• At least one destruction element (13) extended on the inner surface of the tubular body (11 ) and at least partially extended towards the tube center to shorten the coagulation process by breaking down the platelets in the blood during centrifugation

The inventive centrifugal tube (10) comprises a tubular body (11 ) with reference to Figures 1 and 2. If said tubular body (11 ) is provided with a longitudinal tubular body (111 ) and is at the base of said tubular body (111 ), preferably a body base (112) is provided in a curved structure, tubular body (111 ) and body base (112) are provided in single-piece. The tubular body (11 ) further comprises an opening (113) to be closed with a tube cap (12).

Said centrifugal tube is preferably made of plastic, in particular polyethylene terephthalate, known as PET. Here, different materials similar to PET can be used, but PET is an advantageous material in terms of strength and cost. Here, the main criterion in the selection of plastic-based materials is that the material can be shaped in the injection molding method. The centrifuge tubes (10) are vacuumed before closing its mouth (113). Another advantage of PET is capability of maintain vacuum. Other materials like PP causes loss of the vacuum effect. The tube cap (12) can be used so as to close the mouth part (113). Said tube cap (12) may simply be a stopper, preferably it has structure that allows the needle to be entered for the transfer of blood without being opened and then provides sealing when the needle is removed.

A preferred tube cap (12) is shown in Figure 2. Said tube cap (12) has a cylindrical body and there is an opening at the top of the body as a needle inlet (121 ). A sealing element (122) that can be reached through said opening is provided within the body of the tube cap (12). The sealing element (122) is made of an elastic material. Here, after the needle inlet (121 ), the sealing element (122) recovers its form in a manner such that it minimizes the opened hole as much as possible.

Said tube cap (121 ) preferably closes the mouth (113) section with screw threads provided on its inner part and on the outer surface of the tubular body (11 ) or in a click- fit manner.

At least one, preferably multiple drive elements (13) are formed on the inner surface of the inventive centrifugal tube (10) especially on the inner surface of the tubular body (111 ). The drive elements (13) are mainly configured to break down the platelets of blood from the donor and thus it extends at least partially relative to the center of the centrifugal tube (10). The membranes of the platelets that hit these protrusion/extension formed destruction elements (13) become activated by being damaged and initiate coagulation and accordingly, a faster and more efficient coagulation process is provided compared to normal tubes.

Said destruction elements (13) can be provided as linearly extending structures as can be seen in Figure 2. Here, linear destruction elements extend especially from the mouth (113) to the body base (112). Alternatively, linear destruction elements (13) can be arranged horizontally and angularly, the production of the centrifugal tube with such destruction elements (13), especially by injection molding, is more difficult and costly than the structure with the destruction element (13) extending from the mouth (113) to the body base (112).

In another alternative, said destruction elements (13) can be provided in a helical form, similarly, the production of such a destruction element (13) is more difficult than linear structures. Referring to Figure 3; said destruction elements (13) are angularly distributed on the inner surface of the tubular body (111 ) and preferably the intervals between each of them are equal. Besides, the efficiency of the centrifuge tube (10) can be increased by forming the maximum number of destruction elements (13) by providing said destruction elements (13) in contact with each other in such a way that there is no angular distance between them.

As in Figure 4, there may be a longer angular distance between at least two of the destruction elements (13) than the others. In this case, it is seen more comfortably inside the centrifugal tube (10) in said wide range and, as will be explained later, the formation of fibrin tissue can be more easily controlled by the user.

The destruction elements (13) shown in Figures 3 and 4 are curved. Here, it is also possible to arrange destruction elements (13) with sharper lines, eg triangular crosssections, besides the curve.

The centrifuge tube (10) mentioned above and in the claims is configured for use in a method so as to obtain platelet rich fibrin.

In order to obtain said platelet rich fibrin, first of all, the blood drawn from a donor with a needle (with phlebotomy set or syringe) and transferred to the tube body (11 ) with the help of the needle from the needle inlet (121 ) located in the middle of the tube cap (12) which is on the upper part of the centrifugal tube (10) and has an elastic structure. Here, the centrifugal tube (10) does not contain any coagulation accelerator chemicals. After the transfer process is completed, the centrifuge tube (10) is placed in the centrifuge machine for centrifugation.

Here, centrifuge machines with angled slots or at least capable of keeping the centrifugal tube (10) at an angle to the floor during centrifugation are advantageous, because the centrifugal force will push the blood directly towards the destruction elements (13) in an angled centrifuge and a faster and more effective platelet activation process will be ensured. The coagulation process is accelerated without the need to use any chemicals; the blood in the tubular body (11 ) collides with the destruction elements (13) on the inner surface of the tubular body (11 ) during the centrifugation process. Three sections are formed in the centrifugal tube (10) after the centrifugation process is finished and middle section, which contain platelet rich fibrin, is separated. In some embodiments, the platelet rich fibrin is cut after separation.

Before using the platelet rich fibrin, optionally separated middle section is dewatered by apply pressure on it or crushing.

Three patients were randomly selected to compare the test results with a normal centrifuge tube with a commercially used BD brand of Coagulation Activator, and 10 ml of blood was drawn from each patient in order to obtain an advanced method of obtaining platelet rich fibrin and a centrifugal tube (10) and PRF suitable for this method.

Blood drawn from the same patient is to be used for each protocol; the following protocols were applied to 2 different tubes, the centrifugal tube (10) and a normal centrifuge tube with BD® brand Coagulation Activator;

1. Protocol

Centrifugal tube (10) and a normal centrifuge tube with BD brand Coagulation Activator; A single centrifugation process is applied in this protocol, centrifugation is performed at 1600 RCF for 25 minutes.

2. Protocol

Centrifugal tube (10) and a normal centrifuge tube with BD brand Coagulation Activator; Two centrifugation processes are applied intermittently in this protocol, centrifugation is carried out at 400 RCF for 2 minutes in the first centrifuge, then, centrifugation is carried out for 20 minutes at 1600 RCF in the second centrifuge.

3. Protocol

Centrifugal tube (10) and a normal centrifuge tube with BD brand Coagulation Activator; Three intermittent centrifugation processes are applied in this protocol, centrifugation is carried out at 400 RCF for 2 minutes in the first centrifuge, then, centrifugation is performed for 10 minutes at 1600 RCF in the second centrifuge, the centrifugation at 400 RCF for 2 minutes is performed.

Protocols have been implemented. It is checked whether there is rupture and fragmentation in the PRF tissue obtained at the end of the protocols, the size and weight of the tissue were measured and continued in the absence of any rupture or fragmentation

In Table 1 , the size and weight results of the PRF tissue obtained in accordance with the protocols applied for 2 different tubes are given.

As a result of the test, 3 different centrifugation protocols were applied to the tube (10) and the BD ® brand Coagulation Activator tube in 3 different patients. As the obtained PRF tissues can be seen in Table 1 , it is observed that the centrifugal tube (10) gives better results than the BD brand Coagulation Activator tube in terms of size and weight.

The protection scope of the invention is specified in the claims and cannot be limited to the description made for illustrative purposes in this detailed description. It is clear that a person skilled in the art can present similar embodiments in the light of the above descriptions without departing from the main theme of the invention.

Claims

CLAIMS A centrifugal tube (10) for use in a centrifugation process so as to obtain platelet rich fibrin, characterized in that; it comprises the following;

• A longitudinally arranged tubular body (11 ) with a mouth (113),

• A tube cap (12) connected with the tubular body (11 ),

• At least one destruction element (13) extended on the inner surface of the tubular body (11 ) and at least partially extended towards the tube center to shorten the coagulation process by breaking down the platelets in the blood during centrifugation. A centrifugal tube (10) according to claim 1 , characterized in that; said destruction element (13) is arranged to extend at least partially along the tubular body (10). A centrifugal tube (10) according to claim 1 or 2, characterized in that; said destruction element (13) is arranged in such a way that it extends linearly from the mouth to the base of the body. A centrifugal tube (10) according to any of the preceding claims, characterized in that; it comprises a plurality of destruction elements (13). A centrifugal tube (10) according to claim 4, characterized in that; said destruction element (13) is provided angularly spaced on the inner surface of the tubular body (11 ). A centrifugal tube (10) according to claim 4 or 5, characterized in that; said destruction element (13) is not provided on at least a part of the inner surface of said tubular body (11 ). A centrifugal tube (10) according to claim 1 , characterized in that; said destruction element (13) is provided helically on the inner surface of the tubular body (11 ).

8. A centrifugal tube (10) according to any of the preceding claims, characterized in that; the centrifugal tube (10) is made of plastic, preferably polyethylene terephthalate.

9. A centrifugal tube (10) according to claim 1 , characterized in that; it comprises a needle inlet (121 ) that is positioned in the center of the tube cap (12) so as to prevent microorganisms from entering the tubular body (11 ) with blood by opening the tube cover (12) during the process of discharging blood into the tubular body (11 ), allows the syringe needle to enter easily with its elastic structure and the needle hole to be closed automatically after exiting, preventing the entry of microorganisms.

10. A method for obtaining platelet rich fibrin, characterized in that; it comprises the following steps;

- Transferring the blood drawn from the donor to the centrifugal tube (10) according to any of the preceding claims,

- Centrifugation of the centrifuge tube in such a way that the platelets in the blood are destroyed by the destruction element (13) and that the blood is separated into three layers, at least a cell-free plasma, red blood cells and platelet rich fibrin and

- Separating said platelet-rich fibrin from the remaining layers.

11. A method according to claim 10, characterized in that; dewatering obtained platelet-rich fibrin.

12. A method according to claim 10, characterized in that; said centrifugation process is carried out between 300-4000 RCF and between 5-60 minutes.

13. A method according to claim 10, characterized in that; said centrifugation process is repeated twice at intervals.

14. A method according to claim 13, characterized in that; the first step of said centrifugation process is carried out between 1-20 minutes, between 100-2000 RCF, second step is carried out between 300-4000 RCF and between 5-60 minutes. A method according to claim 10, characterized in that; said centrifugation process is repeated three times at intervals. A method according to claim 15, characterized in that; the first step of said centrifugation process is carried out between 100-2000 RCF and between 1-20 minutes, second step is carried out between 300-4000 RCF and between 5-60 minutes, third step is carried out between 100-2000 RCF and between 1-20 minutes.

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