Enhanced solid plasma matrix and preparation method thereof
By stretching the solid plasma matrix, a dense fibrin network is constructed, which solves the problems of fast degradation and weak barrier effects, achieving higher stability and better clinical application effects.
Patent Information
- Application Number
- CN202411680549.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-22
- Publication Date
- 2025-02-25
AI Technical Summary
The existing plasma matrix products degrade rapidly and have loose internal structures, resulting in weak barrier effects, limiting their wider clinical applications.
Strengthen the solid plasma matrix through tensile treatment, build a denser fibrin network, enhance its stability and barrier function, and prolong the degradation time.
It improves the toughness and barrier effect of solid plasma matrix, extends its service life, and enhances its regeneration effect in guiding bone/tissue regeneration.
Smart Images
Figure CN119499454A_ABST
Abstract
Description
Technical Field
[0001] The present disclosure relates to the technical field of blood products, and in particular to an enhanced solid plasma matrix and a preparation method thereof. Background Art
[0002] Plasma matrix is widely used in the dental field because it does not contain anticoagulants and xenobiotics. It provides a rich reservoir of autologous growth factors (GFs), which are essential for bone regeneration, including platelet-derived growth factor (PDGF), transforming growth factor β (TGF-β), and vascular endothelial growth factor (VEGF). Plasma matrix is widely used in oral guided bone regeneration and tissue regeneration, mainly for mixing with implanted biomaterials or as a barrier membrane covering the surface of implanted materials to promote wound healing. However, the plasma matrix products currently used degrade quickly and have a loose internal structure, resulting in a weak barrier effect, which limits their wider clinical application.
[0003] Therefore, how to provide a method to enhance the barrier effect of plasma matrix products and prolong their degradation time has become a problem that needs to be solved urgently. Summary of the invention
[0004] In order to solve the above technical problems, the present disclosure provides an enhanced solid plasma matrix and a preparation method thereof. The preparation method strengthens the toughness of the solid plasma matrix through stretching treatment, while making it have a better barrier effect and prolonging its degradation time, so that it can obtain better regeneration effect when applied to clinical guided bone / tissue regeneration.
[0005] In a first aspect, the present disclosure provides a method for preparing a solid plasma matrix, the preparation method comprising the following steps:
[0006] (1) obtaining a whole blood sample and obtaining an upper solid plasma matrix after centrifugation;
[0007] (2) The solid plasma matrix obtained in step (1) is subjected to a stretching treatment to obtain an enhanced solid plasma matrix.
[0008] The preparation method disclosed in the present invention adopts stretching technology to construct a denser and stronger fibrin network inside the plasma matrix, thereby enhancing the stability of the solid plasma matrix product, making it more durable, and maintaining its original performance and activity for a longer period of time, reducing the risk of deterioration, and extending its service life. At the same time, it also enhances its barrier effect, reduces the entry of fibroblasts and bacteria into the bone graft area and around the implant through the barrier, and has good application prospects.
[0009] The following are preferred technical solutions of the present disclosure, but are not intended to be limitations of the technical solutions provided by the present disclosure. Through the following technical solutions, the technical objectives and beneficial effects of the present disclosure can be better achieved and realized.
[0010] As a preferred technical solution of the present disclosure, the centrifugal force of the centrifugation in step (1) is 60-1000×g, for example, 60×g, 100×g, 200×g, 300×g, 500×g, 700×g or 1000×g, but is not limited to the listed values, and other values not listed within the range are also applicable.
[0011] As a preferred technical solution of the present invention, the centrifugation time in step (1) is 2-10 min, for example, 2 min, 3 min, 4 min, 5 min, 6 min, 7 min, 8 min, 9 min or 10 min, but is not limited to the listed values, and other values not listed within the range are also applicable.
[0012] As a preferred technical solution of the present disclosure, the preparation method further comprises: before performing step (2), preparing the obtained solid plasma matrix into a solid plasma matrix product.
[0013] As a preferred technical solution of the present disclosure, the solid plasma matrix product includes a solid plasma matrix membrane.
[0014] As a preferred technical solution of the present disclosure, the preparation of the solid plasma matrix membrane includes: using a solid plasma matrix pressing machine to press the obtained solid plasma matrix into a solid plasma matrix membrane.
[0015] Preferably, the pressing pressure is 1-10N, such as 1N, 2N, 3N, 4N, 5N, 6N, 7N, 8N, 9N or 10N, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0016] Preferably, the pressing time is 30-120 s, such as 30 s, 60 s, 90 s or 120 s, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0017] As a preferred technical solution of the present disclosure, the stretching force of the stretching treatment in step (2) is 0.5-2N, such as 0.5N, 1N, 1.5N or 2N, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0018] As a preferred technical solution of the present invention, the time of the stretching treatment in step (2) is 10-180s, for example, 10s, 20s, 30s, 40s, 50s, 60s, 70s, 80s, 100s, 120s, 140s, 160s or 180s, etc., but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0019] In the present disclosure, selecting appropriate stretching force and stretching time helps to further enhance the performance of the solid plasma matrix. If the stretching force is too large or the stretching time is too long, the plasma matrix membrane will be broken.
[0020] In a second aspect, the present disclosure provides an enhanced solid plasma matrix, which is prepared by the preparation method described in the first aspect.
[0021] In a third aspect, the present disclosure provides a use of the enhanced solid plasma matrix as described in the second aspect, wherein the enhanced solid plasma matrix is used for regeneration of soft and hard tissues, and is particularly suitable for regeneration of soft and hard tissues in the mouth.
[0022] Furthermore, the enhanced solid plasma matrix can be used alone or in combination with other biomaterials to form a composite material for covering or filling soft and hard tissue defects.
[0023] In the present disclosure, the problem of loose structure and easy degradation of the currently prepared solid plasma matrix is solved by stretching. Under the action of the stretching force, the fibrin structure in the solid plasma matrix becomes tighter, which can enhance the toughness of the plasma matrix and prolong the degradation time. When used in guided bone / tissue regeneration, it can more effectively block the entry of fibroblasts, provide long-term growth factors and stable space for osteogenesis, and has great application prospects in clinical practice.
[0024] Compared with the prior art, the technical solution provided by the embodiments of the present disclosure has the following advantages:
[0025] (1) The preparation method disclosed in the present invention uses stretching technology to construct a denser and stronger fibrin network inside the plasma matrix, thereby enhancing the stability of the plasma matrix product, making it more durable, and maintaining its original performance and activity for a longer period of time, reducing the risk of deterioration, and extending the service life. At the same time, it also enhances its barrier effect and reduces the entry of fibroblasts and bacteria into the bone graft area and around the implant through the barrier;
[0026] (2) The preparation method disclosed in the present invention is simple to operate and does not require the addition of any additional substances, which is conducive to industrial production and application. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present disclosure and, together with the description, serve to explain the principles of the present disclosure.
[0028] 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.
[0029] Figure 1 This is a SEM image of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure;
[0030] Figure 2 This is a SEM image of the enhanced solid plasma matrix membrane described in Example 1 of the present disclosure;
[0031] Figure 3 This is a graph showing the gingival fibroblast penetration performance of the solid plasma matrix membrane obtained in Example 1 and Comparative Example 1 of the present disclosure;
[0032] Figure 4 This is a graph showing the Staphylococcus aureus penetration performance of the solid plasma matrix membrane obtained in Example 1 and Comparative Example 1 of the present disclosure;
[0033] Figure 5 This is a degradation test diagram of the solid plasma matrix membrane obtained in Examples 1-2 and Comparative Example 1 of the present disclosure. DETAILED DESCRIPTION
[0034] In order to more clearly understand the above-mentioned objectives, features and advantages of the present disclosure, the scheme of the present disclosure will be further described below. It should be noted that the embodiments of the present disclosure and the features in the embodiments can be combined with each other without conflict.
[0035] In the following description, many specific details are set forth to facilitate a full understanding of the present disclosure, but the present disclosure 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 disclosure, rather than all of the embodiments.
[0036] Example 1
[0037] This embodiment provides a method for preparing an enhanced solid plasma matrix, the preparation method comprising the following steps:
[0038] (1) 10 mL of blood was collected from the volunteer's elbow using a hydrophilic blood collection tube, and then quickly placed in a horizontal centrifuge and centrifuged at 700 × g for 8 min to obtain the upper solid plasma matrix;
[0039] (2) pressing the solid plasma matrix obtained in step (1) into a solid plasma matrix membrane using a plasma matrix membrane pressing device kit at a pressure of 5 N for 90 seconds;
[0040] The obtained solid plasma matrix membrane was stretched for 60 seconds using a tensile force of 1 N to obtain a reinforced plasma matrix.
[0041] Example 2
[0042] This example provides a method for preparing an enhanced solid plasma matrix. The preparation method is similar to that of Example 1, except that the stretching force in step (2) is 0.5N.
[0043] Example 3
[0044] This example provides a method for preparing an enhanced solid plasma matrix. The preparation method is similar to that of Example 1, except that the stretching force in step (2) is 1 N and the stretching time is 120 s.
[0045] Example 4
[0046] This example provides a method for preparing an enhanced solid plasma matrix. The preparation method is similar to that of Example 1, except that the stretching force in step (2) is 2 N and the stretching time is 10 s.
[0047] Example 5
[0048] This example provides a method for preparing an enhanced solid plasma matrix. The preparation method is similar to that of Example 1, except that the stretching force in step (2) is 0.5 N and the stretching time is 180 s.
[0049] Comparative Example 1
[0050] This comparative example provides a solid plasma matrix membrane, which is obtained by the same method as in Example 1, but without any stretching treatment.
[0051] Performance Test:
[0052] (i) The solid plasma matrix membrane of comparative example 1 without stretching treatment and the enhanced solid plasma matrix membrane obtained in example 1 were placed in a glutaraldehyde solution with a concentration of 2.5 wt % for 4 hours, and then freeze-dried at -51°C and observed by scanning electron microscopy. The results were as follows: Figure 1 and Figure 2 shown.
[0053] from Figure 1 and Figure 2 It can be seen that compared with Comparative Example 1, the solid plasma matrix membrane fibrin network after stretching in Example 1 is more dense.
[0054] (ii) Staphylococcus aureus and gingival fibroblasts were used to measure the amount of bacteria and cells on both sides of the solid plasma matrix membrane of comparative example 1 without stretching treatment and the enhanced solid plasma matrix membrane obtained in example 1. The specific test method includes:
[0055] Obtain cell suspension and bacterial suspension of a certain concentration:
[0056] The solid plasma matrix membrane to be tested is placed in the middle, and an equal volume of cell suspension or bacterial suspension is added above the solid plasma matrix membrane to be tested, and an equal volume of ordinary culture medium is placed below; then it is placed in an incubator, and the cell concentration or bacterial concentration in the lower layer is measured after 48 hours, and the penetration rate = lower suspension concentration / upper suspension concentration×100%.
[0057] Among them, the penetration performance of gingival fibroblasts is shown in Figure 2. Figure 3 As shown in Figure 2, the penetration performance of Staphylococcus aureus is shown in Figure 2. Figure 4 shown.
[0058] from Figure 3 and Figure 4 It can be seen that the solid plasma matrix membrane after stretching treatment in Example 1 can better resist the penetration of bacteria and fibroblasts.
[0059] (III) The solid plasma matrix membrane of comparative example 1 without stretching treatment and the enhanced solid plasma matrix membrane obtained in Example 1-2 were placed in a culture medium without serum, cultured at 37° C. and 5 vol.% CO2, and then weighed and tested every week for 8 weeks. The measurement results are as follows: Figure 5 shown.
[0060] from Figure 5 It can be seen that the enhanced solid plasma matrix membrane obtained in Example 1-2 degrades slowly, can be maintained for a longer time, and has a broader application prospect in clinical practice.
[0061] 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.
[0062] The above description is only a specific embodiment of the present disclosure, so that those skilled in the art can understand or implement the present disclosure. 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 disclosure. Therefore, the present disclosure will not be limited to the embodiments described herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A method for preparing an enhanced solid plasma matrix, characterized in that: The preparation method comprises the following steps: (1) obtaining a whole blood sample and obtaining an upper solid plasma matrix after centrifugation; (2) The solid plasma matrix obtained in step (1) is subjected to a stretching treatment to obtain an enhanced solid plasma matrix.
2. The preparation method according to claim 1, characterized in that: The centrifugal force of the centrifugation in step (1) is 60-1000×g.
3. The preparation method according to claim 1 or 2, characterized in that: The centrifugation time in step (1) is 2-10 min.
4. The preparation method according to any one of claims 1 to 3, characterized in that The preparation method further comprises: before carrying out step (2), preparing the obtained solid plasma matrix into a solid plasma matrix product.
5. The preparation method according to claim 4, characterized in that: The solid plasma matrix product comprises a solid plasma matrix membrane.
6. The preparation method according to claim 5, characterized in that: The preparation of the solid plasma matrix membrane comprises: using a solid plasma matrix pressing machine to press the obtained solid plasma matrix into a solid plasma matrix membrane; Preferably, the pressing pressure is 1-10N; Preferably, the pressing time is 30-120s.
7. The preparation method according to any one of claims 1 to 6, characterized in that: The stretching force of the stretching treatment in step (2) is 0.5-2N.
8. The preparation method according to any one of claims 1 to 7, characterized in that: The stretching treatment time in step (2) is 10-180s.
9. An enhanced solid plasma matrix, characterized in that The enhanced solid plasma matrix is prepared by the preparation method according to any one of claims 1 to 8.