Enhanced solid plasma matrix and preparation method thereof
Through heat treatment, the tensile strength and degradation time of solid plasma matrix are enhanced, and the problems of insufficient mechanical strength and short degradation time of plasma matrix products are solved, achieving wider clinical applications and better therapeutic effects.
Patent Information
- Application Number
- CN202411483482.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-23
- Publication Date
- 2024-12-17
AI Technical Summary
The existing plasma matrix products have insufficient mechanical strength and short degradation time, making it difficult to meet the needs of a wider range of clinical application scenarios.
The tensile strength and ductility of solid plasma matrix are enhanced by heat treatment and prolonged their degradation time, forming a denser fibrin network, improving mechanical strength and retention time.
It has achieved the improvement of mechanical strength of plasma matrix and the extension of degradation time, which is suitable for a wider range of clinical application scenarios and provides better therapeutic effects.
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Figure CN119139349A_ABST
Abstract
Description
Technical Field
[0001] The present disclosure relates to the technical field of blood products, and in particular to a reinforced solid plasma matrix and a preparation method thereof. Background Art
[0002] With the continuous development of biomedicine and materials science, great changes have taken place in the preparation methods, product characteristics and biological performance of plasma matrix. Autologous plasma matrix products have developed rapidly in recent years because they are rich in blood cells, growth factors and three-dimensional fibrin scaffolds, and are simple to prepare and have no rejection reaction. They are widely used in the fields of soft and hard tissue regeneration, refractory wound healing and osteoarthritis treatment.
[0003] Fibrin in the plasma matrix is the activated mode of fibrinogen in the blood. This soluble fibrin molecule exists in large quantities in plasma and α particles. When the coagulation cascade reaction begins, it can be converted into insoluble fibrin to surround cells, and growth factors form a three-dimensional network structure. The microporous structure formed by fibrin in the plasma matrix can serve as a scaffold for cell migration, proliferation and differentiation, and can also serve as a medium for the release of growth factors. In recent years, the use of plasma matrix products as barrier membranes, defect filling materials, and other forms of research and clinical applications have gradually increased.
[0004] However, at present, plasma matrix products still have shortcomings such as insufficient strength and short degradation time. Therefore, how to provide a method to improve the mechanical strength of plasma matrix products and prolong the degradation time to meet a wider range of clinical application scenarios and obtain better therapeutic effects has become an urgent problem to be solved. Summary of the invention
[0005] In order to solve the above technical problems, the present disclosure provides a reinforced solid plasma matrix and a preparation method. The present disclosure enhances the tensile strength and ductility of the solid plasma matrix through heat treatment, and at the same time prolongs its degradation time, so that it has a more ideal application effect in application scenarios such as clinical GBR.
[0006] In a first aspect, the present disclosure provides a method for preparing a fortified solid plasma matrix, the method comprising the following steps:
[0007] (1) obtaining a whole blood sample and obtaining a solid plasma matrix after centrifugation;
[0008] (2) heat-treating the solid plasma matrix obtained in step (1) to achieve strengthening.
[0009] The preparation method disclosed in the present invention does not require the addition of any additional reagents, and only promotes protein denaturation through heat treatment, thereby making the fibrin network inside the plasma matrix more dense, increasing the mechanical strength of the plasma matrix product, and prolonging the degradation time to meet a wider range of clinical application scenarios and obtain better therapeutic effects.
[0010] 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.
[0011] As a preferred technical solution of the present invention, the centrifugal force of the centrifugation in step (1) is 20-950×g, for example, 20×g, 50×g, 100×g, 200×g, 300×g, 400×g, 500×g, 600×g, 700×g, 800×g, 900×g or 950×g, 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 invention, the centrifugation time in step (1) is 2-8 min, for example, 2 min, 3 min, 4 min, 5 min, 6 min, 7 min or 8 min, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0013] As a preferred technical solution of the present disclosure, step (2) also includes preparing the obtained solid plasma matrix into a solid plasma matrix product.
[0014] As a preferred technical solution of the present disclosure, the solid plasma matrix product includes any one of a solid plasma matrix membrane, a solid plasma matrix plug, a solid plasma matrix fragment or a solid plasma matrix composite material.
[0015] The solid plasma matrix composite material is composed of a solid plasma matrix and other biological materials, and the other biological materials include but are not limited to bone materials.
[0016] As a preferred technical solution of the present disclosure, the temperature of the heat treatment in step (2) is 40-120°C, for example, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 110°C or 120°C, but is not limited to the listed values. Other values not listed in the range are also applicable, preferably 50-90°C, more preferably 80-90°C,
[0017] As a preferred technical solution of the present invention, the time of the heat treatment in step (2) is 8-15s, for example 8s, 9s, 10s, 11s, 12s, 13s, 14s or 15s, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0018] In the present disclosure, selecting an appropriate heat treatment temperature and heat treatment time helps to further enhance the performance of the solid plasma matrix. If the heat treatment temperature is too high or the heat treatment time is too long, it will cause the fibrin structure inside the plasma matrix to denature, water loss, cell activity to decrease, and mechanical strength and tensile properties to decrease, which is not conducive to being used as a barrier material or bone replacement material.
[0019] As a preferred technical solution of the present disclosure, the heat treatment method in step (2) includes direct heating or indirect heating.
[0020] Preferably, the indirect heating comprises water bath heating or oil bath heating.
[0021] As a preferred technical solution of the present disclosure, the form of heat treatment in step (2) includes local heating or overall heating.
[0022] Furthermore, for example, the solid plasma matrix membrane can be heated locally, heated on one side, heated on both sides, etc.
[0023] The present disclosure can locally heat or completely heat the solid plasma matrix to selectively strengthen specific parts of the solid plasma matrix to meet different needs.
[0024] In a second aspect, the present disclosure provides a reinforced solid plasma matrix, which is prepared by the preparation method described in the first aspect.
[0025] In a third aspect, the present disclosure provides an application of the reinforced solid plasma matrix described in the second aspect, wherein the reinforced solid plasma matrix is used in the fields of soft and hard tissue regeneration, facial beauty, hair regeneration, refractory wound healing, and osteoarthritis treatment.
[0026] Compared with the prior art, the technical solution provided by the embodiments of the present disclosure has the following advantages:
[0027] In view of the shortcomings of the plasma matrix in the existing technology, such as insufficient strength and short degradation time, heat treatment is used to densify the fibrin structure inside the plasma matrix through protein denaturation without adding other substances, thereby improving the mechanical strength of the plasma matrix and prolonging the degradation time to meet a wider range of clinical application scenarios and obtain better therapeutic effects. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] 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.
[0029] 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.
[0030] Figure 1 This is a SEM image of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure.
[0031] Figure 2 This is a SEM image of the reinforced solid plasma matrix membrane described in Example 1 of the present disclosure.
[0032] Figure 3 This is a SEM image of the reinforced solid plasma matrix membrane described in Example 2 of the present disclosure.
[0033] Figure 4 This is a SEM image of the reinforced solid plasma matrix membrane described in Example 3 of the present disclosure.
[0034] Figure 5 This is a SEM image of the reinforced solid plasma matrix membrane described in Example 4 of the present disclosure.
[0035] Figure 6 It is a graph showing strain results of tensile tests on the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membrane described in Examples 1-4.
[0036] Figure 7 It is a stress result diagram of the tensile test of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membrane described in Examples 1-4.
[0037] Figure 8 This is a graph showing the degradation test results of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membrane described in Examples 1-4.
[0038] Fig. 9 These are SEM images of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membranes described in Examples 3 and 6.
[0039] Fig.10 It is a graph showing strain results of tensile tests on the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membranes described in Examples 3 and 6.
[0040] Fig.11 This is a stress result diagram of the tensile test of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membrane described in Examples 3 and 6.
[0041] Fig.12 This is a graph showing the degradation test results of the solid plasma matrix membrane described in Comparative Example 1 of the present disclosure and the reinforced solid plasma matrix membranes described in Examples 3 and 6. DETAILED DESCRIPTION
[0042] 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.
[0043] 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.
[0044] Example 1
[0045] This embodiment provides a method for preparing a reinforced solid plasma matrix, the preparation method comprising the following steps:
[0046] (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 a solid plasma matrix;
[0047] (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 6 N for 60 seconds;
[0048] The obtained solid plasma matrix membrane was heat-treated (double-sided heating) at 50° C. for 10 seconds using a metal heater (model: H2O3-H, purchased from: Kayoudi Biotechnology Yixing Co., Ltd.) to strengthen the solid plasma matrix membrane.
[0049] Example 2
[0050] This example provides a method for preparing a reinforced solid plasma matrix. The preparation method is similar to that of Example 1, except that the heat treatment temperature in step (2) is 75°C.
[0051] Example 3
[0052] This example provides a method for preparing a reinforced solid plasma matrix. The preparation method is similar to that of Example 1, except that the heat treatment temperature in step (2) is 90°C.
[0053] Example 4
[0054] This example provides a method for preparing a reinforced solid plasma matrix. The preparation method is similar to that of Example 1, except that the heat treatment temperature in step (2) is 105°C.
[0055] Example 5
[0056] This embodiment provides a method for preparing a reinforced solid plasma matrix. The preparation method is similar to that of Embodiment 3, except that the heat treatment in step (2) is performed by heating only one side.
[0057] Comparative Example 1
[0058] This comparative example provides a solid plasma matrix membrane, which is obtained by the same method as in Example 1, but without any heat treatment.
[0059] Performance Testing:
[0060] Investigate the effect of different heating temperatures on the performance of solid plasma matrix membrane under double-sided heating:
[0061] (i) The solid plasma matrix membrane without heat treatment in Comparative Example 1 and the reinforced solid plasma matrix membrane obtained in Examples 1-4 were respectively placed in a glutaraldehyde solution with a concentration of 2.5 wt % for 4 hours and then observed by scanning electron microscopy. The results were as follows: Figure 1-Figure 5 shown.
[0062] from Figure 1-Figure 5 It can be seen that, compared with Comparative Example 1, within a suitable temperature range, as the temperature increases, the solid plasma matrix membrane fibrin network gradually becomes denser.
[0063] (ii) The unheat-treated solid plasma matrix membrane of Comparative Example 1 and the reinforced solid plasma matrix membrane obtained in Examples 1-4 were respectively measured for their tensile properties using a tensile tester (model: ZQ-990, purchased from Dongguan Zhiqu Precision Instrument Co., Ltd.). The tensile tester was set to have a maximum load capacity of 5N when the standard tensile rate was 1mm / min. Before the tensile test, the original length between the two clamps was set to 5mm. After the tensile test, the strain results were as follows: Figure 6 The stress results are shown in Figure 7 shown.
[0064] Depend on Figure 6 and Figure 7 It can be seen that the solid plasma matrix membrane after heat treatment enhances the tensile strain and tensile stress, enabling it to better resist the stretching and tearing of the membrane by external mechanical stress, and has broader application prospects in clinical practice.
[0065] (III) The solid plasma matrix membrane without heat treatment in Comparative Example 1 and the reinforced solid plasma matrix membrane obtained in Examples 1-4 were placed in serum DMEM with a concentration of 10 wt% for degradation test, and photographed and recorded on the 1st day, 3rd day, 5th day, 7th day and 14th day to observe the degradation. The change of the mass of each solid plasma matrix membrane over time is shown in the figure Figure 8 shown.
[0066] from Figure 8 It can be seen that the thermally treated Examples 1-4 degraded more slowly than Comparative Example 1. This means that the solid plasma matrix membrane degradation time was prolonged after thermal treatment, which was more conducive to retention in the body and provided a longer barrier support effect for the healing process such as bone regeneration.
[0067] To explore the effects of different heating methods on the performance of solid plasma matrix membranes when the heat treatment temperature is constant (all 90°C):
[0068] (IV) The solid plasma matrix membrane without heat treatment in Comparative Example 1 and the reinforced solid plasma matrix membranes obtained in Examples 3 and 6 were respectively placed in a glutaraldehyde solution with a concentration of 2.5 wt % for 4 hours and then observed by scanning electron microscopy. The results are as follows: Fig. 9 shown.
[0069] from Fig. 9 It can be seen that the solid plasma matrix membrane fibrin network structure subjected to double-sided heating in Example 3 is more compact; while the solid plasma matrix membrane fibrin network structure subjected to only single-sided heating in Example 6 has a compact heated surface and a loose non-heated surface.
[0070] (V) The unheat-treated solid plasma matrix membrane of Comparative Example 1 and the reinforced solid plasma matrix membranes obtained in Examples 3 and 6 were respectively measured for their tensile properties using a tensile tester (ZQ-990, ZhiQu). The tensile tester was set to have a maximum load capacity of 5N when the standard tensile rate was 1mm / min. Before the tensile test, the original length between the two clamps was set to 5mm. After the tensile test, the strain performance results are as follows: Fig.10 The stress performance results are shown in Fig.11 shown.
[0071] Depend on Fig.10 and Fig.11 It can be seen that the tensile properties of the solid plasma matrix membrane heated on both sides in Example 3 are better than those of the solid plasma matrix membrane heated on only one side in Example 6.
[0072] (VI) The solid plasma matrix membrane without heat treatment in Comparative Example 1 and the reinforced solid plasma matrix membranes obtained in Examples 3 and 6 were placed in serum DMEM with a concentration of 10 wt% for degradation test, and photographed and recorded on the 1st day, 3rd day, 5th day, 7th day and 14th day to observe the degradation. The change of the mass of each solid plasma matrix membrane over time is shown in the figure below: Fig.12 shown.
[0073] from Fig.12 It can be seen that the solid plasma matrix membrane without heat treatment in Comparative Example 1 degrades faster, while the solid plasma matrix membrane after heat treatment in Examples 3 and 6 degrades slower, which is more conducive to retention in the body, and can provide a longer barrier support effect for healing processes such as bone regeneration; further, the solid plasma matrix membrane heated on both sides in Example 3 degrades more slowly than the solid plasma matrix membrane heated on only one side in Example 6.
[0074] 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.
[0075] 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 a reinforced solid plasma matrix, characterized in that: The preparation method comprises the following steps: (1) obtaining a whole blood sample and obtaining a solid plasma matrix after centrifugation; (2) heat-treating the solid plasma matrix obtained in step (1) to achieve strengthening.
2. The preparation method according to claim 1, characterized in that: The centrifugal force of the centrifugation in step (1) is 20-950×g.
3. The preparation method according to claim 1 or 2, characterized in that: The centrifugation time in step (1) is 2-8 min.
4. The preparation method according to any one of claims 1 to 3, characterized in that Step (2) also includes 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 includes any one of a solid plasma matrix membrane, a solid plasma matrix plug, a solid plasma matrix fragment or a solid plasma matrix composite material.
6. The preparation method according to any one of claims 1 to 5, characterized in that: The temperature of the heat treatment in step (2) is 40-120°C, preferably 50-90°C, and more preferably 80-90°C.
7. The preparation method according to any one of claims 1 to 6, characterized in that: The heat treatment time in step (2) is 8-15s.
8. The preparation method according to any one of claims 1 to 7, characterized in that: The heat treatment in step (2) includes direct heating or indirect heating; Preferably, the indirect heating comprises water bath heating or oil bath heating.
9. The preparation method according to any one of claims 1 to 8, characterized in that: The heat treatment in step (2) may be in the form of local heating or overall heating.
10. A fortified solid plasma matrix, characterized in that The reinforced solid plasma matrix is prepared by the preparation method according to any one of claims 1 to 9.