Platelet gel containing zinc ions and preparation method thereof

Abstract

The invention relates to the technical field of platelet gel preparation, in particular to platelet gel containing zinc ions and a preparation method thereof.The preparation method comprises the steps that the blood sampling amount of single-time blood sampling is determined according to the wound volume; blood is collected according to the blood collection amount, and the collected blood is stored in a preparation device; a zinc-containing reagent strip is prepared, and the zinc-containing reagent strip is placed in the preparation device to be assembled; centrifuging the preparation device, taking out the centrifuged preparation device, and standing; and separating a red blood cell layer generated in the centrifugation process from the platelet gel containing the zinc ions to complete the preparation. Zinc ions are added into the platelet gel, the preparation efficiency and the treatment effect of the gel are promoted and improved, the antibacterial performance of the gel is supplemented, the whole preparation process of the preparation method is conducted in a sealed mode, and the sterility degree is improved; the preparation efficiency is improved, the operation process is simplified, and consistent operation can be realized; the gel efficiency and the antibacterial performance are improved by adding zinc ions.

Description

Technical Field

[0001] The invention relates to the technical field of platelet gel preparation, and in particular to a platelet gel containing zinc ions and a preparation method thereof. Background Art

[0002] Diabetic foot ulcer (DFU) is a common and serious complication of diabetic patients. Traditional treatments such as surgical debridement and dressing coverage have limited effects, and many patients eventually face amputation. Platelet concentrates, including platelet-rich plasma (PRP), platelet-rich gel (PRG), and platelet-rich fibrin (PRF), have been shown to significantly promote wound healing because they contain a variety of growth factors and cytokines. However, these treatments have problems such as complex preparation, high risk of sample contamination, and uneven quality of clinical trials. In addition, zinc supplementation shows potential in promoting DFU healing. It not only inhibits inflammation and antibacterial properties, but also improves the preparation efficiency and stability of platelet gel.

[0003] Chinese Patent Publication No.: CN113827773A discloses an autologous platelet factor gel and its preparation method and application. The preparation method comprises the following steps: step S1, collecting autologous whole blood, first centrifuging to obtain plasma; the plasma contains platelet-poor plasma (PPP) and platelet-rich plasma (PRP); step S2, centrifuging the obtained plasma again to obtain a centrifuge, and storing the upper layer liquid PPP and the lower layer liquid PRP separately for later use; step S3, physically modifying the obtained upper layer liquid to form a gel; step S4, physically lyzing the obtained lower layer liquid to release the factors; step S5, The gel obtained in step S3 and the lower layer liquid after physical lysis in step S4 are mixed to obtain autologous platelet factor gel; it can be seen that the autologous platelet factor gel and its preparation method and application have the following problems: the gel components lack components that inhibit inflammation, and the treatment effect on DFU is poor; the existing preparation process requires the addition of chemical reagents such as anticoagulants and activators, the operation is cumbersome, the risk of sample contamination is high, and there are uncertainties in subsequent treatments and potential risks to human safety; there is no strictly designed clinical standardized treatment process and management system, and it cannot be widely promoted and implemented in clinical practice. Summary of the invention

[0004] To this end, the present invention provides a platelet gel containing zinc ions and a preparation method thereof, so as to overcome the problem that the gel components in the prior art lack components that inhibit inflammation and have poor therapeutic effects on DFU.

[0005] To achieve the above object, on the one hand, the present invention provides a method for preparing a platelet gel containing zinc ions, comprising: Step S1, detecting the wound volume of diabetic foot, and determining the blood collection volume for a single blood collection according to the wound volume; Step S2, collecting blood according to the blood collection amount, and storing the collected blood in a preparation device; Step S3, preparing a zinc-containing reagent strip, placing the zinc-containing reagent strip in the preparation device for assembly; Step S4, centrifuging the preparation device assembled with the zinc-containing reagent strip; Step S5, taking out the preparation device after centrifugation and letting it stand; Step S6, separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions to complete the preparation.

[0006] Furthermore, in step S1, the step of determining the blood sampling volume for a single blood sampling according to the wound volume includes: Detection of diabetic foot wound volume; Determining parameters corresponding to the wound volume and the blood volume; Determining the blood collection volume of the single blood collection according to the wound volume and the volume corresponding parameter; The amount of blood collected in each single blood draw corresponds one to one to the volume of a single wound.

[0007] Furthermore, in step S2, the step of collecting blood according to the blood collection amount includes: Determining the required blood sampling scale corresponding to the blood sampling process of the preparation device according to the blood sampling amount of the single blood sampling; Placing a blood collection needle in a rubber stopper at the blood collection end of the preparation device to collect blood; When the blood volume in the preparation device reaches the required blood collection scale, blood collection is completed.

[0008] Furthermore, in step S3, the step of preparing the zinc-containing reagent strip includes: Using ultrasonic spraying, spraying the zinc-containing solution onto the prefabricated reagent strip to obtain an initial reagent strip; Performing a quality test on the initial reagent strip, and judging whether the spraying is qualified according to the test result; If the test result meets the requirements, the spraying is judged to be qualified, and the initial reagent strip is the zinc-containing reagent strip.

[0009] Furthermore, the spraying parameters of the ultrasonic spraying include a nozzle moving speed of 25 mm / s and a nozzle shaft speed of 800 r / min.

[0010] Furthermore, if the test result does not meet the requirements, the spraying is judged to be unqualified, the ultrasonic spraying process is repeated on the initial reagent strip, and the quality test is performed again after the spraying is completed, and it is determined whether the zinc-containing reagent strip is obtained based on the test result.

[0011] Furthermore, in step S4, the step of centrifuging the preparation device to obtain initial platelet-rich fibrin comprises: The preparation device is placed in a centrifuge and centrifuged under preset centrifugal conditions.

[0012] Furthermore, in the step S5, the preparation device is left to stand for 10 minutes.

[0013] Furthermore, in step S6, the step of separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions includes: Extracting the separated red blood cell layer from the tail end of the preparation device; Open the tube cap of the preparation device to obtain the platelet gel containing zinc ions.

[0014] On the other hand, the present invention also provides a platelet gel containing zinc ions, which is prepared according to the preparation method, and the platelet gel containing zinc ions consists of a pure blood centrifugation product and zinc ions.

[0015] Compared with the prior art, the invention has the beneficial effect that the invention improves the preparation efficiency, simplifies the operation process, and can achieve consistent operation. The entire preparation process is sealed to ensure the sterility of the platelet gel; in addition, the addition of zinc ions to the platelet gel promotes and improves the preparation efficiency and treatment of the gel on the one hand, and supplements the antibacterial properties of the gel on the other hand. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a flow chart of a platelet gel containing zinc ions and a preparation method thereof according to an embodiment of the present invention; Figure 2 It is a structural schematic diagram of a preparation device according to an embodiment of the present invention; Figure 3 A comparison chart of fresh PRF cytokine levels in the preparation results of the embodiments of the present invention; Figure 4 This is a comparison chart of PRF cytokine levels after standing for 72 hours in the preparation results of the embodiment of the present invention; Figure 5 A flow chart of centrifuging a preparation device according to an embodiment of the present invention; Figure 6 This is a wound change diagram of Example 1 of the present invention; Figure 7 This is a wound change diagram of Example 2 of the present invention; In the figure: 1, blood collection port; 2, rubber plug at the blood collection end; 3, tube cap; 4, tube body; 5, wound volume scale; 6, blood collection volume scale; 7, blood drawing port; 8, tube lumen; 9, rubber plug at the blood drawing end; 10, groove on the inner wall of the tube; 11, reagent strip. DETAILED DESCRIPTION

[0017] In order to make the objects and advantages of the present invention more clearly understood, the present invention is further described below in conjunction with embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] The preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are only used to explain the technical principles of the present invention and are not intended to limit the protection scope of the present invention.

[0019] It should be noted that, in the description of the present invention, terms such as "up", "down", "left", "right", "inside" and "outside" indicating directions or positional relationships are based on the directions or positional relationships shown in the drawings. This is merely for the convenience of description and does not indicate or imply that the device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it cannot be understood as a limitation on the present invention.

[0020] See also Figure 1 and Figure 2 As shown, Figure 1 The figure is a flow chart of a platelet gel containing zinc ions and a preparation method thereof according to an embodiment of the present invention. Figure 2 The present invention is a schematic diagram of the structure of a preparation device according to an embodiment of the present invention; the present invention provides a method for preparing a platelet gel containing zinc ions, comprising: Step S1, detecting the wound volume of diabetic foot, and determining the blood collection volume for a single blood collection according to the wound volume; Step S2, collecting blood according to the blood collection amount, and storing the collected blood in a preparation device; Step S3, preparing a zinc-containing reagent strip, placing the zinc-containing reagent strip in the preparation device for assembly; Step S4, centrifuging the preparation device assembled with the zinc-containing reagent strip; Step S5, taking out the preparation device after centrifugation and letting it stand; Step S6, separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions to complete the preparation.

[0021] The main function of platelets is coagulation and hemostasis. In addition, platelet degranulation growth factors and cytokines, including platelet-derived growth factor (PDGF), transforming growth factor (TGF-β), fibroblast growth factor (FGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF), can exert a variety of biological effects including tissue regeneration, stimulation of collagen synthesis, and promotion of capillary proliferation in the injured area [1].

[0022] Platelet concentrates (PC) and various platelet-derived growth factors such as EGF, PDGF and FGF have been widely used in the treatment of wounds and ulcers and have achieved good clinical efficacy. PC includes platelet-rich plasma (PRP), platelet-rich plasma gel (PRG, or autologous platelet-rich plasma gel, APG), and the recently reported platelet-rich fibrin (PRF).

[0023] It is understood that during the formation of PRF, blood samples are centrifuged and platelets and fibrin form a gel-like network structure. Therefore, PRF can be said to be a gel. This gel-like substance has good biocompatibility and bioactivity, which helps promote tissue repair and regeneration.

[0024] Zinc can inhibit the growth of different types of bacteria. One of its molecular mechanisms of action is that zinc participates in the regulation of pro-inflammatory cytokines and exhibits free radical scavenger activity against reactive oxygen species (ROS). When zinc is applied directly to wounds and damaged periwound skin, the clearance of inactivated tissue is promoted, the risk of superinfection is reduced, inflammation is alleviated, and epithelialization is activated. Multiple studies have shown its debridement effect on pressure injuries, diabetic foot, and burns. Zinc gluconate is highly water soluble and is the best choice for topical application to wounds.

[0025] At the same time, zinc also helps platelet activation and gel formation stability, improving the preparation efficiency of the second-generation gel PRF.

[0026] A blood sampling port 1 is provided at the center of the top end of the tube cap 3 of the preparation device, which is closed by a rubber stopper 2 at the blood sampling end. A wound volume scale 5 and a blood sampling volume scale 6 are provided on the outside of the tube body 4. A blood drawing port 7 is provided at the bottom of the tube body, which is closed by a rubber stopper 9 at the blood drawing end. A groove 10 on the inner wall of the tube is located in the tube cavity 8 and is arranged on the inner side of the tube body. A reagent strip 11 is installed in the groove on the inner wall of the tube. The tube body is a glass tube, and the interior of the preparation device is in a vacuum state.

[0027] See also Figure 3 and Figure 4 As shown, Figure 3 This is a comparison chart of fresh PRF cytokine levels in the preparation results of the embodiment of the present invention, Figure 4 This is a comparison chart of the PRF cytokine levels after standing for 72 hours in the preparation results of the embodiment of the present invention. When preparing the second-generation gel PRF, adding zinc can not only inhibit inflammation and antibacterial, but also increase the preparation efficiency of PRF. ELISA was used to detect the cytokines of fresh PRF and PRF standing at room temperature for 72 hours. It was found that compared with the traditional preparation method, the preparation method of the present invention can significantly improve the level of cytokines in PRF. After adding zinc ions, the cytokine level can be further improved. After standing at room temperature, it was detected again, and it was found that the cytokines in PRF obtained by the preparation device were still higher than those of the traditional preparation method.

[0028] The present invention improves the preparation efficiency, simplifies the operation process, and can achieve consistent operation. The entire preparation process is sealed to ensure the sterility of the platelet gel; in addition, zinc ions are added to the platelet gel, which, on the one hand, promotes and improves the preparation efficiency and treatment of the gel; on the other hand, it supplements the antibacterial properties of the gel.

[0029] Specifically, in step S1, the step of determining the blood sampling volume for a single blood sampling according to the wound volume includes: Detection of diabetic foot wound volume; Determining parameters corresponding to the wound volume and the blood volume; Determining the blood collection volume of the single blood collection according to the wound volume and the volume corresponding parameter; The amount of blood collected in each single blood draw corresponds one to one to the volume of a single wound.

[0030] In practice, 1cm 3 The wound volume requires 0.6 mL of venous blood; the wound volume of diabetic foot is measured by a ruler and probe; The preparation device is provided with wound volume scales and blood sampling volume scales, and the blood sampling volume is determined based on the wound volume detected before blood sampling and the wound volume scales.

[0031] Specifically, the step of collecting blood according to the blood collection amount includes: Determining the required blood sampling scale corresponding to the blood sampling process of the preparation device according to the blood sampling amount of the single blood sampling; Placing a blood collection needle in a rubber stopper at the blood collection end of the preparation device to collect blood; When the blood volume in the preparation device reaches the required blood collection scale, blood collection is completed.

[0032] During implementation, the blood collection needle is inserted into the lumen through the rubber stopper at the blood collection end to collect blood under negative pressure.

[0033] Specifically, in step S3, the steps of preparing the zinc-containing reagent strip include: Using ultrasonic spraying, spraying the zinc-containing solution onto the prefabricated reagent strip to obtain an initial reagent strip; Performing a quality test on the initial reagent strip, and judging whether the spraying is qualified according to the test result; If the test result meets the requirements, the spraying is judged to be qualified, and the initial reagent strip is the zinc-containing reagent strip.

[0034] In implementation, the zinc-containing solution is preferably a zinc gluconate solution; and the prefabricated reagent strip is preferably an alginate dressing reagent strip.

[0035] 1mL blood volume corresponds to 3mg zinc gluconate (C 12 H 22 O 14 Zn), determine the standard mass based on the mass of zinc gluconate corresponding to the blood volume and the mass of the prefabricated reagent strip; when the mass of the zinc-containing reagent strip is greater than or equal to the standard mass, the test result meets the requirements.

[0036] See also Figure 5 As shown, it is a flow chart of centrifuging the preparation device according to an embodiment of the present invention, and the spraying parameters of the ultrasonic spraying include a nozzle moving speed of 25 mm / s and a nozzle shaft speed of 800 r / min.

[0037] In practice, during the spraying process of ultrasonic spraying, both sides of the prefabricated reagent strip are sprayed twice to ensure uniform spraying.

[0038] The zinc content of the zinc alginate test strip is 0.1% to 1%, with the optimal value being 0.5%.

[0039] Specifically, if the test result does not meet the requirements, the spraying is judged to be unqualified, the ultrasonic spraying process is repeated on the initial reagent strip, and the quality test is performed again after the spraying is completed, and it is determined whether the zinc-containing reagent strip is obtained based on the test result.

[0040] In practice, if the mass of the initial reagent strip after the second spraying is greater than the standard mass, the test result meets the requirements and it is determined that a zinc-containing reagent strip is obtained.

[0041] The zinc-containing reagent strip is assembled with the preparation device to install the zinc-containing reagent strip into the embedded groove on the inner wall of the tube of the preparation device.

[0042] Specifically, in step S4, the step of centrifuging the preparation device includes: The preparation device is placed in a centrifuge and centrifuged under preset centrifugal conditions.

[0043] In implementation, the preset centrifugation conditions include a centrifugal force of 400 g and a centrifugation time of 12 minutes.

[0044] Specifically, in step S5, the preparation device is left to stand for 10 minutes.

[0045] Specifically, in step S6, the step of separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions includes: Extracting the separated red blood cell layer from the tail end of the preparation device; Open the tube cap of the preparation device to obtain the platelet gel containing zinc ions.

[0046] During implementation, the red blood cell layer (red part) at the tail end of the preparation device is extracted from the rubber stopper at the blood drawing end, and the platelet gel part containing zinc ions (yellow part and orange part) is retained. The complete high-quality PRF can be obtained by opening the tube cap.

[0047] A platelet gel containing zinc ions is prepared by a method for preparing a platelet gel containing zinc ions. The platelet gel containing zinc ions consists of a pure blood centrifugation product and zinc ions.

[0048] Example 1 Patient Zhang X, female, 45 years old, diagnosed with type 2 diabetes, diabetic foot ulcer (left little toe), the wound type was a flat wound, the depth of the flat wound was set at 1cm, and she received four PRF treatments in 13 days.

[0049] Before PRF treatment, the wound area was 1.5cm×1.2cm, 1.8cm 2 ; The first PRF treatment, wound area 1.1cm×1.1cm, 1.21cm 2 ; The second PRF treatment, wound area 0.8cm×0.7cm, 0.56cm 2 ; The third PRF treatment, wound area 0.8cm×0.4cm, 0.32cm2 ; The fourth PRF treatment, wound area 0.2cm×0.2cm, 0.04cm 2 .

[0050] Example 2 Mr. Liu, 54 years old, male, diagnosed with type 2 diabetes and diabetic foot ulcer (left big toe), received 4 PRF treatments in 13 days.

[0051] Before PRF treatment, the volume of the left wound was 1cm×0.9cm×2.5cm, 2.25cm 3 The right wound volume is 0.9cm×0.9cm×2.4cm, 1.94cm 3 ; The first PRF treatment, the left wound volume 0.9cm×0.8cm×1.4cm, 1.01cm 3 , right wound volume 0.8cm×0.7cm×1.2cm, 0.67cm 3 ; The second PRF treatment, the left wound volume 1cm×0.9cm×1.2cm, 1.08cm 3 , right wound volume 0.4cm×0.4cm×0.8cm, 0.13cm 3 ; The third PRF treatment, the left wound volume 0.5cm×0.5cm×0.5cm, 0.125cm 3 , right wound volume 0.3cm×0.4cm×0.7cm, 0.084cm 3 ; The fourth PRF treatment, the left wound volume 0.5cm×0.5cm×0.2cm, 0.05cm 3 .

[0052] See also Figure 6 and Figure 7 As shown, Figure 6 This is a wound change diagram of Example 1 of the present invention, Figure 7 The wound change diagram of Example 2 of the present invention is shown in Figure 2. According to the treatment results of each embodiment of the present invention, two diabetic patients received platelet-rich fibrin (PRF) treatment to promote the healing of their diabetic foot ulcers. During the 13-day treatment cycle, patients Zhang X and Liu received four PRF treatments respectively. The treatment results showed that the wound area of ​​both patients was significantly reduced, indicating that PRF treatment has a significant effect on diabetic foot ulcers.

[0053] Specifically, Zhang X's wound was reduced from the initial 1.8 square centimeters to 0.04 square centimeters, while Liu's left foot wound volume was reduced from 2.25 cubic centimeters to 0.05 cubic centimeters. These data show that PRF treatment can not only reduce the surface area of ​​the wound, but also reduce its depth, thereby promoting more comprehensive healing. Through this innovative treatment method, the patient's diabetic foot ulcer was effectively controlled, reducing the risk of further complications.

[0054] So far, the technical solutions of the present invention have been described in conjunction with the preferred embodiments shown in the accompanying drawings. However, it is easy for those skilled in the art to understand that the protection scope of the present invention is obviously not limited to these specific embodiments. Without departing from the principle of the present invention, those skilled in the art can make equivalent changes or substitutions to the relevant technical features, and the technical solutions after these changes or substitutions will fall within the protection scope of the present invention.

[0055] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a platelet gel containing zinc ions, characterized in that: include: Step S1, detecting the wound volume of diabetic foot, and determining the blood collection volume for a single blood collection according to the wound volume; Step S2, collecting blood according to the blood collection amount, and storing the collected blood in a preparation device; Step S3, preparing a zinc-containing reagent strip, placing the zinc-containing reagent strip in the preparation device for assembly; Step S4, centrifuging the preparation device assembled with the zinc-containing reagent strip; Step S5, taking out the preparation device after centrifugation and letting it stand; Step S6, separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions to complete the preparation.

2. The method for preparing the platelet gel containing zinc ions according to claim 1, characterized in that: In step S1, the step of determining the blood sampling volume for a single blood sampling according to the wound volume includes: Detection of diabetic foot wound volume; Determining parameters corresponding to the wound volume and the blood volume; Determining the blood collection volume of the single blood collection according to the wound volume and the volume corresponding parameter; The amount of blood collected in each single blood draw corresponds one to one to the volume of a single wound.

3. The method for preparing the platelet gel containing zinc ions according to claim 2, characterized in that: In step S2, the step of collecting blood according to the blood collection amount includes: Determining the required blood sampling scale corresponding to the blood sampling process of the preparation device according to the blood sampling amount of the single blood sampling; Placing a blood collection needle in a rubber stopper at the blood collection end of the preparation device to collect blood; When the blood volume in the preparation device reaches the required blood collection scale, blood collection is completed.

4. The method for preparing the platelet gel containing zinc ions according to claim 3, characterized in that: In step S3, the steps of preparing the zinc-containing reagent strip include: Using ultrasonic spraying, spraying the zinc-containing solution onto the prefabricated reagent strip to obtain an initial reagent strip; Performing a quality test on the initial reagent strip, and judging whether the spraying is qualified according to the test result; If the test result meets the requirements, the spraying is judged to be qualified, and the initial reagent strip is the zinc-containing reagent strip.

5. The method for preparing the platelet gel containing zinc ions according to claim 4, characterized in that: The spraying parameters of the ultrasonic spraying include a nozzle moving speed of 25 mm / s and a nozzle shaft speed of 800 r / min.

6. The method for preparing the platelet gel containing zinc ions according to claim 5, characterized in that: If the test result does not meet the requirements, the spraying is judged to be unqualified, and the ultrasonic spraying process is repeated on the initial reagent strip. After the spraying is completed, the quality test is performed again, and it is determined whether the zinc-containing reagent strip is obtained based on the test result.

7. The method for preparing the platelet gel containing zinc ions according to claim 6, characterized in that: In step S4, the step of centrifuging the preparation device includes: The preparation device is placed in a centrifuge and centrifuged under preset centrifugal conditions.

8. The method for preparing the platelet gel containing zinc ions according to claim 7, characterized in that: In the step S5, the preparation device is left to stand for 10 minutes.

9. The method for preparing the platelet gel containing zinc ions according to claim 8, characterized in that: In step S6, the step of separating the red blood cell layer produced during the centrifugation process from the platelet gel containing zinc ions comprises: Extracting the separated red blood cell layer from the tail end of the preparation device; Open the tube cap of the preparation device to obtain the platelet gel containing zinc ions.

10. A platelet gel containing zinc ions, characterized in that: The platelet gel containing zinc ions is prepared by the method for preparing the platelet gel containing zinc ions according to any one of claims 1 to 9, wherein the platelet gel containing zinc ions is composed of a pure blood centrifugation product and zinc ions.

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