Vitamin C-containing plasma matrix as well as preparation method and application thereof

Abstract

The invention relates to a plasma matrix containing vitamin C as well as a preparation method and application thereof. The preparation method of the plasma matrix containing the vitamin C comprises the following steps: preparing the plasma matrix containing the vitamin C through the interaction of the vitamin C and the plasma matrix; wherein the plasma matrix comprises at least one of a liquid plasma matrix and a solid plasma matrix. The vitamin C-containing plasma matrix obtained by the preparation method disclosed by the embodiment of the invention has relatively high capabilities of resisting inflammation, promoting wound healing and tissue regeneration and promoting collagen regeneration.

Description

Technical Field

[0001] The present invention relates to the technical field of blood products, and in particular to a plasma matrix containing vitamin C, a preparation method and an application thereof. Background Art

[0002] Autologous blood products are rich in cells, growth factors, fibrin scaffolds and other materials in autologous blood, and have the advantages of simple preparation and no foreign body reaction. Autologous blood products are widely used in the fields of soft and hard tissue regeneration, refractory wound healing, facial beauty, osteoarthritis treatment, etc. In the face of most clinical application scenarios, especially facial beauty, dealing with inflammation and promoting collagen regeneration are often the two major problems that need to be solved in clinical applications. Summary of the invention

[0003] In order to solve at least one of the above technical problems, the present invention provides a plasma matrix containing vitamin C and a preparation method and application thereof. The plasma matrix containing vitamin C in the embodiment of the present invention has high anti-inflammatory, wound healing and tissue regeneration promoting, and collagen regeneration promoting capabilities.

[0004] As one aspect, the present invention provides a method for preparing a plasma matrix containing vitamin C, and the preparation method comprises any one of the following methods I to III: Method I: spraying or injecting vitamin C into a centrifuge tube to obtain a centrifuge tube modified with vitamin C; adding blood to the centrifuge tube modified with vitamin C, centrifuging, and obtaining a plasma matrix containing vitamin C; or, Method II: placing blood in a centrifuge tube, centrifuging, and obtaining a solid plasma matrix; mixing the solid plasma matrix and vitamin C uniformly to obtain a plasma matrix containing vitamin C; or, Method III: placing blood in a centrifuge tube, centrifuging, and obtaining a liquid plasma matrix; mixing the liquid plasma matrix and vitamin C uniformly to obtain a plasma matrix containing vitamin C.

[0005] In some embodiments of the present invention, the centrifugal acceleration is 20-950g, for example, it can be 149g, 378g, 563g, 719g, 836g, 928g, etc.; the centrifugal time is 2-8min, for example, it can be 2.3min, 3.7min, 4.3min, 5.8min, 6.9min, 7.8min, etc.

[0006] In some embodiments of the present invention, in method I, the ratio of the mass of vitamin C to the volume of blood is in the range of 5 to 50 μg / mL, for example, it can be 9 μg / mL, 13 μg / mL, 28 μg / mL, 37 μg / mL, 42 μg / mL, 49 μg / mL, etc.

[0007] In some embodiments of the present invention, in method II, the method for uniformly mixing the solid plasma matrix and vitamin C is selected from any one of the following: soaking the solid plasma matrix in a vitamin C solution; or, cutting the solid plasma matrix into pieces to obtain solid plasma matrix fragments, and uniformly mixing the solid plasma matrix fragments and vitamin C.

[0008] In some embodiments of the present invention, in method II, the concentration range of the vitamin C solution is 5 to 50 μg / mL, for example, it can be 8 μg / mL, 12 μg / mL, 27 μg / mL, 36 μg / mL, 43 μg / mL, 48 μg / mL, etc.; and / or, the mass ratio of the solid plasma matrix to the volume ratio of the vitamin C solution is in the range of 5 to 50 μg / mL, for example, it can be 7 μg / mL, 14 μg / mL, 26 μg / mL, 35 μg / mL, 41 μg / mL, 47 μg / mL, etc.

[0009] In some embodiments of the present invention, in method II, the mass ratio of solid plasma matrix fragments and vitamin C ranges from (10 to 1):1, for example, it can be 1.6:1, 2.3:1, 3.9:1, 4.8:1, 6.3:1, 8.3:1, 9.7:1, etc.

[0010] In some embodiments of the present invention, in method III, the ratio of the mass of liquid plasma matrix to the mass of vitamin C is in the range of (10-1):1, for example, it can be 1.8:1, 2.4:1, 3.6:1, 4.7:1, 6.2:1, 8.1:1, 9.8:1, etc.

[0011] In some embodiments of the present invention, the surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

[0012] As another aspect, the present invention provides a plasma matrix containing vitamin C, and the plasma matrix containing vitamin C is obtained using the above-mentioned preparation method.

[0013] As yet another aspect, the present invention provides a use of the above-mentioned plasma matrix containing vitamin C in soft tissue regeneration materials.

[0014] The technical solution of the embodiment of the present invention has the following advantages:

[0015] The plasma matrix containing vitamin C and the preparation method and application thereof of the embodiments of the present invention prepare the plasma matrix containing vitamin C through the interaction between vitamin C and the plasma matrix.

[0016] Plasma matrix containing vitamin C improves the anti-inflammatory and antioxidant effects of the plasma matrix, and can promote wound healing and tissue regeneration. Specifically, by adding vitamin C to the plasma matrix, while improving the anti-inflammatory and antioxidant effects of the plasma matrix, it also improves the beneficial effects of the plasma matrix on fibroblasts and the promoting effect of the plasma matrix on soft tissue healing and volume increase. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0018] 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, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0019] Figure 1 This is a comparison diagram of the secretion of type I collagen by fibroblasts in the experimental group and the control group of Example 1 of the present invention;

[0020] Figure 2 This is a comparison chart of the proliferation of fibroblasts in the experimental group and the control group of Example 2 of the present invention. DETAILED DESCRIPTION

[0021] In order to more clearly understand the above-mentioned objectives, features and advantages of the present invention, the scheme of the present invention will be further described below. It should be noted that the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

[0022] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein; it is obvious that the embodiments in the specification are only part of the embodiments of the present invention, rather than all of the embodiments.

[0023] The white blood cells, platelets and corresponding growth factors contained in the plasma matrix products have the effects of promoting tissue regeneration, reducing inflammation and antibacterial effects.

[0024] Vitamin C is an effective scavenger of reactive oxygen species, especially superoxide radicals. During wound healing, ROS can activate the pathways for the production of pro-inflammatory cytokines, leading to delayed wound healing. Therefore, vitamin C helps maintain the balance of pro-oxidant and anti-oxidant activity within cells. In addition, vitamin C can promote collagen formation in fibroblasts, which helps postoperative wound healing and soft tissue regeneration.

[0025] According to an embodiment of the present invention, a method for preparing a plasma matrix containing vitamin C is provided, and the preparation method comprises any one of the following methods I to III: Method I: spraying or injecting vitamin C into a centrifuge tube to obtain a centrifuge tube modified with vitamin C; adding blood to the centrifuge tube modified with vitamin C, centrifuging, and obtaining a plasma matrix containing vitamin C; or, Method II: placing blood in a centrifuge tube, centrifuging, and obtaining a solid plasma matrix; mixing the solid plasma matrix and vitamin C uniformly to obtain a plasma matrix containing vitamin C; or, Method III: placing blood in a centrifuge tube, centrifuging, and obtaining a liquid plasma matrix; mixing the liquid plasma matrix and vitamin C uniformly to obtain a plasma matrix containing vitamin C.

[0026] In some embodiments of the present invention, the plasma matrix containing vitamin C includes but is not limited to at least one of the following forms: a liquid plasma matrix containing vitamin C, a solid plasma matrix containing vitamin C, and a solid plasma matrix membrane containing vitamin C.

[0027] In some embodiments of the present invention, a method for preparing a plasma matrix containing vitamin C comprises the following steps:

[0028] (1) Spraying vitamin C in a centrifuge tube to obtain a centrifuge tube modified with vitamin C;

[0029] Preferably, the surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

[0030] (2) collecting blood into the centrifuge tube in step (1);

[0031] Preferably, the ratio of the mass of vitamin C in step (1) to the volume of blood in step (2) is in the range of 5 to 50 μg / mL.

[0032] (3) The centrifuge tube containing blood obtained in step (2) is mixed evenly, and then centrifuged at a centrifugal acceleration of 20 to 950 g for 2 to 8 minutes to obtain a centrifuge product; wherein the upper yellow portion of the centrifuge product is the plasma matrix containing vitamin C.

[0033] In some embodiments of the present invention, a method for preparing a plasma matrix containing vitamin C comprises the following steps:

[0034] (1) Injecting vitamin C into a centrifuge tube to obtain a centrifuge tube modified with vitamin C;

[0035] Preferably, the surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

[0036] (2) collecting blood into the centrifuge tube in step (1);

[0037] Preferably, the ratio of the mass of vitamin C in step (1) to the first volume of blood in step (2) is in the range of 5 to 50 μg / mL.

[0038] (3) The centrifuge tube containing blood obtained in step (2) is mixed evenly, and then centrifuged at a centrifugal acceleration of 20 to 950 g for 2 to 8 minutes to obtain a centrifuge product; wherein the upper yellow portion of the centrifuge product is the plasma matrix containing vitamin C.

[0039] In some embodiments of the present invention, a method for preparing a plasma matrix containing vitamin C comprises the following steps:

[0040] (1) Collect blood and place it in a centrifuge tube;

[0041] Preferably, the surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

[0042] (2) Centrifuging the centrifuge tube containing blood obtained in step (1) at a centrifugal acceleration of 20 to 950 g for 2 to 8 minutes to obtain a centrifuge product; wherein the upper yellow portion of the centrifuge product is the solid plasma matrix.

[0043] (3) immersing the solid plasma matrix obtained in step (2) in a vitamin C solution to obtain a plasma matrix containing vitamin C;

[0044] Preferably, the concentration of the vitamin C solution is in the range of 5 to 50 μg / mL.

[0045] In some embodiments of the present invention, a method for preparing a plasma matrix containing vitamin C comprises the following steps:

[0046] (1) Collect blood and place it in a centrifuge tube;

[0047] Preferably, the surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

[0048] (2) Centrifuging the centrifuge tube containing blood obtained in step (1) at a centrifugal acceleration of 20 to 950 g for 2 to 8 minutes to obtain a centrifuge product; wherein the upper yellow portion of the centrifuge product is the solid plasma matrix.

[0049] (3) The solid plasma matrix obtained in step (2) is collected and pressed into a film using a pressing device box to prepare a solid plasma matrix membrane.

[0050] (4) Cutting the solid plasma matrix membrane obtained in step (3) into pieces to obtain solid plasma matrix fragments.

[0051] (5) uniformly mixing the solid plasma matrix fragments obtained in step (4) and vitamin C to obtain a plasma matrix containing vitamin C;

[0052] Preferably, the mass ratio of the solid plasma matrix fragments to vitamin C is in the range of (10-1):1.

[0053] In some embodiments of the present invention, a method for preparing a vitamin C-modified plasma matrix comprises the following steps:

[0054] (1) collecting blood and placing it in a centrifuge tube, centrifuging it to obtain a liquid plasma matrix;

[0055] Preferably, the centrifuge tube is made of a material selected from plastic or glass.

[0056] (2) uniformly mixing the liquid plasma matrix obtained in step (1) and vitamin C to prepare a plasma matrix containing vitamin C;

[0057] Preferably, the ratio of the mass of the liquid plasma matrix to the mass of vitamin C is in the range of (10-1):1.

[0058] In order to further illustrate the effect of the present invention, specific embodiments are used for illustration below.

[0059] Example 1

[0060] This embodiment provides a method for preparing a plasma matrix bone block containing vitamin C, comprising the following steps:

[0061] (1) Collect 10 mL of blood from the volunteer’s elbow using a hydrophobic centrifuge tube;

[0062] (2) centrifuging the centrifuge tube containing blood obtained in step (1) at a centrifugal acceleration of 700 g for 8 min to obtain a centrifugal product;

[0063] (3) separating and collecting the upper yellow portion of the centrifugal product obtained in step (2) to obtain a liquid plasma matrix;

[0064] (4) mixing 3 g of the liquid plasma matrix obtained in step (3) and 15 to 150 μg of vitamin C to obtain a plasma matrix containing vitamin C;

[0065] (5) Cultivating fibroblasts in vitro:

[0066] Culture medium for control group: conventional culture medium;

[0067] The culture medium of the experimental group: the plasma matrix containing vitamin C obtained in step (4) was added to the conventional culture medium;

[0068] After 48 hours of in vitro fibroblast culture, the morphology of fibroblasts and the secretion of type I collagen in each group were observed by immunofluorescence. The test results of the experimental group and the control group are as follows Figure 1 As shown; among them, Figure 1 The “vitamin C treatment group” in the figure corresponds to the “experimental group”.

[0069] according to Figure 1 The data analysis in the results showed that the plasma matrix containing vitamin C can promote the proliferation of fibroblasts in vitro and also promote the production of type I collagen.

[0070] Example 2

[0071] This embodiment provides a method for preparing a plasma matrix containing vitamin C, comprising the following steps:

[0072] (1) Collect 10 mL of blood from the volunteer’s elbow using a hydrophilic centrifuge tube;

[0073] (2) The centrifuge tube containing blood obtained in step (1) is mixed evenly, and then centrifuged at a centrifugal acceleration of 700 g for 8 minutes to obtain a centrifuge product; wherein the upper yellow portion of the centrifuge product is the solid plasma matrix.

[0074] (3) in a membrane pressing device box, pressing the solid plasma matrix obtained in step (2) into a membrane to obtain a solid plasma matrix membrane;

[0075] (4) cutting the solid plasma matrix membrane obtained in step (3) into pieces to obtain solid plasma matrix fragments;

[0076] (5) 0.2 g of the solid plasma matrix fragments obtained in step (4) and 20 μg of vitamin C are mixed evenly to obtain a plasma matrix containing vitamin C.

[0077] (6) Extracting primary gingival fibroblasts from the gingival tissue of volunteers and isolating and culturing them;

[0078] Culture medium for control group: conventional culture medium;

[0079] The culture medium of the experimental group: the plasma matrix containing vitamin C obtained in step (5) was added to the conventional culture medium;

[0080] The proliferation of fibroblasts in the experimental group and the control group was tested after 1 day, 3 days, and 5 days of culture. Figure 2 As shown; among them, Figure 2 The “vitamin C treatment group” in the figure corresponds to the “experimental group”.

[0081] according to Figure 2 The data analysis in the results showed that the plasma matrix containing vitamin C can promote the proliferation of fibroblasts in vitro.

[0082] In summary, the plasma matrix containing vitamin C and the preparation method and application thereof provided in the embodiments of the present invention are prepared by adding vitamin C to the plasma matrix. In the process of wound healing, free radicals or reactive oxygen species (ROS) in the extracellular environment can activate the pathway of pro-inflammatory cytokine production, leading to delayed wound healing. Vitamin C is a powerful scavenger of reactive oxygen species, especially superoxide radicals, and therefore helps to maintain the balance of pro-oxidation and anti-oxidation in cells; at the same time, vitamin C can also be used as a surface decolorizing agent.

[0083] The plasma matrix containing vitamin C in the embodiment of the present invention is applied to the field of soft tissue regeneration in beauty or oral soft tissue, and can achieve the effects of facial rejuvenation, removal of pigmentation, and soft tissue increment.

[0084] It should be noted that, in this article, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0085] The above description is only a specific embodiment of the present invention, so that those skilled in the art can understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments described herein, but should conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a plasma matrix containing vitamin C, characterized in that: The preparation method includes any one of the following methods I to III: Method I: Spraying or injecting vitamin C into the centrifuge tube to obtain a centrifuge tube modified with vitamin C; Adding blood into the centrifuge tube modified with vitamin C, centrifuging, and obtaining a plasma matrix containing vitamin C; Or, Method II: The blood is placed in a centrifuge tube and centrifuged to obtain a solid plasma matrix; Mixing the solid plasma matrix and vitamin C evenly to obtain a plasma matrix containing vitamin C; Or, Method III: The blood is placed in a centrifuge tube and centrifuged to obtain a liquid plasma matrix; The liquid plasma matrix and vitamin C are mixed evenly to obtain a plasma matrix containing vitamin C.

2. The preparation method according to claim 1, characterized in that: The centrifugal acceleration of the centrifugation is 20-950g, and the centrifugal time is 2-8min.

3. The preparation method according to claim 1 or 2, characterized in that: In the method I, The ratio of the mass of the vitamin C to the volume of the blood is in the range of 5 to 50 μg / mL.

4. The preparation method according to any one of claims 1 to 3, characterized in that: In the method II, the method for uniformly mixing the solid plasma matrix and vitamin C is selected from any one of the following: soaking the solid plasma matrix in a vitamin C solution; or, The solid plasma matrix is ​​cut into pieces to obtain solid plasma matrix fragments, and the solid plasma matrix fragments and vitamin C are mixed evenly.

5. The preparation method according to claim 4, characterized in that: In the method II, The concentration range of the vitamin C solution is 5 to 50 μg / mL; And / or, the mass ratio of the solid plasma matrix to the volume of the vitamin C solution is in the range of 5 to 50 μg / mL.

6. The preparation method according to claim 4 or 5, characterized in that: In the method II, The mass ratio of the solid plasma matrix fragments to vitamin C is in the range of (10-1):

1.

7. The preparation method according to any one of claims 1 to 6, characterized in that: In the method III, The ratio of the mass of the liquid plasma matrix to the mass of vitamin C is in the range of (10-1):

1.

8. The preparation method according to any one of claims 1 to 7, characterized in that: The surface of the centrifuge tube is selected from a hydrophilic surface or a hydrophobic surface.

9. A plasma matrix containing vitamin C, characterized in that The vitamin C-containing plasma matrix is ​​obtained by the preparation method according to any one of claims 1 to 8.

10. Use of the plasma matrix containing vitamin C according to claim 9 in soft tissue regeneration materials.

Images