Modified plasma matrix as well as preparation method and application thereof
By combining the plasma matrix with calcium phosphate modifier and using freeze-drying technology, the problems of insufficient mechanical strength and short degradation time of plasma matrix are solved, forming a composite of high mechanical strength and long degradation time for orthopedics and plastic surgery for orthopedics and plastic surgery. Materials meet a wider clinical application needs.
Patent Information
- Application Number
- CN202411486079.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-23
- Publication Date
- 2024-12-27
AI Technical Summary
The existing plasma matrix has problems of insufficient mechanical strength and short degradation time, which limits its application in orthopedics and plastic surgery; although calcium phosphate biomaterials avoid opening up the second surgical area, they have low biodegradability and osteogenesis induction. A problem of weak sexuality.
By combining the plasma matrix with calcium phosphate modifier and using freeze-drying technology, the mechanical strength of the plasma matrix is enhanced and its degradation time is extended, forming a kind of high mechanical strength, long degradation time, good biocompatibility, and osteogenesis induction Strong composite material.
It has achieved a wider range of clinical application scenarios, provided better therapeutic effects and better bone regeneration support, and solved the problem of weak osteogenic induction of calcium phosphate biological materials.
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Figure CN119185661A_ABST
Abstract
Description
Technical Field
[0001] The present disclosure relates to the technical field of blood products, and in particular to a modified plasma matrix and a preparation method and application thereof. Background Art
[0002] Plasma matrix is a concentrated extract derived from human autologous blood. It has attracted much attention as an emerging bioregenerative material. It also has good biocompatibility and biosafety, and will not cause autoimmune rejection. The main component of plasma matrix is platelets, which can play a powerful procoagulant role after being successfully activated. More importantly, plasma matrix is rich in growth factors, including transforming growth factor (TGF-β), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PGDF), etc., and plasma matrix is simple to prepare and has no rejection reaction. It has developed rapidly in recent years and has been widely used in various clinical scenarios, including soft and hard tissue regeneration, refractory wound healing, plastic surgery and other medical fields. However, it has disadvantages such as insufficient strength and short degradation time, so its scope of application is limited.
[0003] Severe periodontal bone resorption and difficult-to-heal fractures are one of the major challenges in orthopedics, dentistry and plastic surgery. The "gold standard" for bone transplantation treatment is autologous bone tissue transplantation. However, autologous bone transplantation often requires the opening of a second surgical area, which increases the harm to the patient. At the same time, autologous bone is also prone to problems such as insufficient absorption. Calcium phosphate (CaP) biomaterials are considered to be a suitable alternative to autologous bone tissue transplantation, avoiding the need to open a second surgical area. At the same time, calcium phosphate biomaterials in the form of injectable pastes can better adapt to complex bone defect morphology and can minimize additional damage to patients through minimally invasive injection. However, calcium phosphate biomaterials have the characteristics of low biodegradability and low microporosity, and lack bioactive agents for recruiting progenitor cells and promoting the ingrowth of new tissue, which hinders complete bone regeneration.
[0004] Therefore, in view of the defects and advantages of the above two materials, how to provide a material with high mechanical strength, long degradation time, good biocompatibility and strong osteogenic induction to meet a wider range of clinical application scenarios 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 modified plasma matrix and a preparation method and application thereof. The preparation method combines the plasma matrix with a calcium phosphate modifier, which makes up for the shortcomings of the two and gives full play to the advantages of the two, thereby obtaining a composite material with high mechanical strength, long degradation time, good biocompatibility and strong osteogenic inductivity, which has good application prospects.
[0006] In a first aspect, the present disclosure provides a method for preparing a modified plasma matrix, the preparation method comprising the following steps:
[0007] (1) obtaining a whole blood sample and obtaining a solid plasma matrix or a liquid plasma matrix after centrifugation;
[0008] (2) If a solid plasma matrix is obtained:
[0009] The solid plasma matrix obtained in step (1) is mixed with a calcium phosphate modifier, and freeze-dried to obtain a modified plasma matrix;
[0010] Alternatively, the solid plasma matrix obtained in step (1) is first freeze-dried and crushed in sequence to obtain freeze-dried plasma matrix particles; the obtained freeze-dried plasma matrix particles are then mixed with a calcium phosphate modifier to obtain a modified plasma matrix;
[0011] If liquid plasma matrix is available:
[0012] The liquid plasma matrix obtained in step (1) is mixed with a calcium phosphate modifier, and then freeze-dried to obtain a modified plasma matrix.
[0013] In the present disclosure, by combining the calcium phosphate modifier with the plasma matrix, the mechanical strength of the plasma matrix can be effectively enhanced and its degradation time can be prolonged; at the same time, the present disclosure utilizes the good biocompatibility and strong growth factor release capacity of the plasma matrix to solve the disadvantage of the weak osteoinductivity of the calcium phosphate biomaterial. In addition, the present disclosure also utilizes freeze-drying technology to concentrate the plasma matrix, so that the fibrin network inside the plasma matrix is formed more densely, further increasing the mechanical strength of the plasma matrix product and prolonging the degradation time; and the freeze-drying technology makes the mixture of the calcium phosphate modifier and the plasma matrix into a product that is easy to store and use, so that it can meet a wider range of clinical application scenarios and obtain better therapeutic effects.
[0014] 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.
[0015] As a preferred technical solution of the present invention, the centrifugal force of the centrifugation in step (1) is 50 to 950×g, for example, 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.
[0016] Preferably, the centrifugation time in step (1) is 5 to 10 min, for example, 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.
[0017] As a preferred technical solution of the present disclosure, the calcium phosphate modifier in step (2) includes one or more of calcium phosphate bone cement, calcium phosphate ceramics or calcium phosphate hyaluronic acid microspheres, such as a combination of calcium phosphate bone cement and calcium phosphate ceramics, a combination of calcium phosphate ceramics and calcium phosphate hyaluronic acid microspheres, etc.
[0018] As a preferred technical solution of the present disclosure, the mass ratio of the solid plasma matrix or the liquid plasma matrix to the calcium phosphate modifier in step (2) is 10:(0.01-0.2), for example, 10:0.01, 10:0.05, 10:0.1 or 10:0.2, etc., but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0019] As a preferred technical solution of the present disclosure, the mixing temperature in step (2) is independently 20-37°C, for example, 20°C, 25°C, 30°C, 35°C or 37°C, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0020] Preferably, the mixing time in step (2) is independently 5 to 10 minutes, for example 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes or 10 minutes, but is not limited to the listed values, and other values not listed within the range are also applicable.
[0021] As a preferred technical solution of the present invention, the freeze-drying temperature in step (2) is independently -40 to -10°C, such as -40°C, -30°C, -20°C or -10°C, but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0022] Preferably, the freeze-drying time in step (2) is independently 60 to 120 min, for example 60 min, 70 min, 80 min, 90 min, 100 min, 110 min or 120 min, etc., but is not limited to the listed values, and other unlisted values within the range are also applicable.
[0023] Preferably, the freeze-drying method in step (2) includes but is not limited to air-cooled freeze-drying or water-cooled freeze-drying.
[0024] 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 plasma matrix membrane, and then crushing it.
[0025] Preferably, the modified plasma matrix prepared using the solid plasma matrix needs to be further crushed to form a granular product.
[0026] Preferably, when the solid plasma matrix is mixed with the calcium phosphate modifier, in order to ensure sufficient mixing, the solid plasma matrix fragments are generally first slowly added to the calcium phosphate modifier at a low speed (2 to 10 rpm) to avoid the generation of bubbles, and then stirred at a high speed (20 to 40 rpm).
[0027] As a preferred technical solution of the present disclosure, if a liquid plasma matrix is obtained, the calcium phosphate modifier is in block form;
[0028] The mixing method of the liquid plasma matrix and the block calcium phosphate modifier includes soaking or injection, that is, soaking the block calcium phosphate modifier in the liquid plasma matrix, or injecting the liquid plasma matrix into the block calcium phosphate modifier.
[0029] In a second aspect, the present disclosure provides a modified plasma matrix prepared by the preparation method described in the first aspect.
[0030] In a third aspect, the present disclosure provides a use of the modified plasma matrix as described in the second aspect, wherein the modified plasma matrix is used as a soft and hard tissue augmentation material.
[0031] In the present disclosure, the modified plasma matrix can be mixed with physiological saline or PBS buffer to prepare the desired plasma matrix product for use; the modified plasma matrix can also be used directly.
[0032] The modified plasma matrix provided by the present disclosure is applied to various soft and hard tissue augmentation.
[0033] Compared with the prior art, the technical solution provided by the embodiments of the present disclosure has the following advantages:
[0034] On the one hand, the preparation method described in the present disclosure can effectively enhance the mechanical strength of the plasma matrix and prolong its degradation time by combining the calcium phosphate modifier with the plasma matrix; at the same time, the present disclosure utilizes the good biocompatibility and strong growth factor release ability of the plasma matrix to solve the shortcoming of the weak osteogenic inductivity of calcium phosphate biomaterials.
[0035] Secondly, the present invention also utilizes freeze-drying technology to concentrate the plasma matrix, making the fibrin network inside the plasma matrix more dense, further increasing the mechanical strength of the plasma matrix product, and prolonging the degradation time; at the same time, freeze-drying technology converts the mixture of calcium phosphate modifier and plasma matrix into a product that is easy to store and use, so that it can meet a wider range of clinical application scenarios and obtain better therapeutic effects.
[0036] Thirdly, compared with the traditional method for preparing plasma matrix products, the preparation method disclosed in the present invention can prepare more plasma matrix products with excellent performance under the same blood collection volume, and has a good application prospect. BRIEF DESCRIPTION OF THE DRAWINGS
[0037] 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.
[0038] 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.
[0039] Figure 1 This is a SEM image of the plasma matrix membrane described in Comparative Example 1 of the present disclosure.
[0040] Figure 2 This is a SEM image of a modified plasma matrix membrane made using the modified plasma matrix described in Example 1 of the present disclosure.
[0041] Figure 3 This is a graph showing the tensile strain results of the plasma matrix membrane described in Comparative Example 1 of the present disclosure and the modified plasma matrix membrane made using the modified plasma matrix described in Example 1.
[0042] Figure 4 This is a graph showing the tensile stress results of the plasma matrix membrane described in Comparative Example 1 of the present disclosure and the modified plasma matrix membrane made using the modified plasma matrix described in Example 1.
[0043] Figure 5 It is a graph showing the degradation test results of the plasma matrix membrane described in Comparative Example 1 of the present disclosure and the modified plasma matrix membrane made using the modified plasma matrix described in Example 1. DETAILED DESCRIPTION
[0044] 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.
[0045] 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.
[0046] Example 1
[0047] This embodiment provides a method for preparing a modified plasma matrix, the preparation method comprising the following steps:
[0048] (1) Collect 10 ml of fresh blood from the volunteer's elbow using a hydrophilic blood collection tube and immediately centrifuge it at 700 × g for 8 min to obtain a solid plasma matrix;
[0049] (2) compressing the solid plasma matrix obtained in step (1) for 90 seconds using a solid plasma matrix pressing device to obtain a plasma matrix membrane;
[0050] The obtained plasma matrix membrane was cut into pieces, and then mixed with calcium phosphate bone cement at a mass ratio of 10:0.05 at 25°C, and stirred for 8 minutes to obtain a mixture; the obtained mixture was freeze-dried at -20°C for 60 minutes, and then crushed to obtain a modified plasma matrix.
[0051] Example 2
[0052] This embodiment provides a method for preparing a modified plasma matrix, the preparation method comprising the following steps:
[0053] (1) Collect 10 ml of fresh blood from the volunteer's elbow using a hydrophilic blood collection tube and immediately centrifuge it at 700 × g for 8 min to obtain a solid plasma matrix;
[0054] (2) compressing the solid plasma matrix obtained in step (1) for 90 seconds using a solid plasma matrix pressing device to obtain a plasma matrix membrane;
[0055] The obtained plasma matrix membrane was freeze-dried at -10°C for 90 minutes and then crushed to obtain plasma matrix freeze-dried particles; the obtained plasma matrix freeze-dried particles were then mixed with calcium phosphate bone cement at a mass ratio of 10:0.2 at 30°C to obtain a modified plasma matrix.
[0056] Example 3
[0057] This embodiment provides a method for preparing a modified plasma matrix, the preparation method comprising the following steps:
[0058] (1) Collect 10 ml of fresh blood from the volunteer's elbow using a hydrophilic blood collection tube and immediately centrifuge it at 700 × g for 8 min to obtain liquid plasma matrix;
[0059] (2) at 27° C., injecting the liquid plasma matrix obtained in step (1) into the block calcium phosphate bone cement to ensure that the mass ratio of the liquid plasma matrix to the block calcium phosphate bone cement is 10:0.1, standing for 7 minutes, and then freeze-drying at -30° C. for 120 minutes to obtain a modified plasma matrix.
[0060] Comparative Example 1
[0061] This comparative example provides a plasma matrix membrane, which is obtained by referring to the preparation method of Example 1, but is not doped with any modifier and is not freeze-dried.
[0062] Performance Test:
[0063] Test preparation:
[0064] The modified plasma matrix described in Example 1 was mixed with 0.5 ml of physiological saline and re-compressed to obtain a modified plasma matrix membrane.
[0065] (1) The plasma matrix membrane described in Comparative Example 1 and the modified plasma matrix membrane obtained in Example 1 were characterized using a scanning electron microscope. The results are as follows: Figure 1 and Figure 2 shown.
[0066] from Figure 1 and Figure 2 It can be seen that compared with Comparative Example 1, the fibrin network inside the modified plasma matrix obtained in Example 1 is more densely formed.
[0067] (2) The tensile properties of the plasma matrix membrane obtained in Comparative Example 1 and the modified plasma matrix membrane obtained in Example 1 were measured using a tensile measuring instrument (model: ZQ-990, purchased from Dongguan Zhiqu Precision Instrument Co., Ltd.), wherein the tensile measuring instrument was set to have a maximum load capacity of 5 N when the standard tensile rate was 1 mm / min. Before the tensile test, the original length between the two clamps was set to 5 mm. The tensile strain diagram is shown in FIG. Figure 3 As shown, the tensile stress diagram is Figure 4 shown.
[0068] Depend on Figure 3 and Figure 4It can be seen that the tensile properties of the modified plasma matrix obtained in Example 1 are better than those of the plasma matrix membrane described in Comparative Example 1. The modified plasma matrix membrane obtained in the present disclosure can better resist the stretching and tearing of external mechanical stress, better maintain the integrity of the membrane, expand the clinical application scenarios, provide stronger mechanical protection for the bone augmentation area, and fit various forms of bone defect surfaces.
[0069] (3) The plasma matrix membrane obtained in Comparative Example 1 and the modified plasma matrix membrane obtained in Example 1 were placed in a 0.25% trypsin culture medium and incubated at 37°C. The mass of the plasma matrix membrane remaining in the culture medium at 0 days, 1 day, 3 days, 5 days, 7 days, 14 days, 21 days and 28 days was recorded to evaluate its degradation and record the results as follows: Figure 5 shown.
[0070] from Figure 5 It can be seen that the degradation rate of the modified plasma matrix membrane obtained in Example 1 is significantly lower than that of the plasma matrix membrane in Comparative Example 1. This indicates that the modified plasma matrix obtained by the preparation method disclosed herein can prolong the degradation time of the plasma matrix product, is more conducive to the retention of the plasma matrix product in the body, and provides a longer barrier support effect for guiding the healing process such as bone tissue regeneration.
[0071] 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.
[0072] 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 modified plasma matrix, characterized in that: The preparation method comprises the following steps: (1) obtaining a whole blood sample and obtaining a solid plasma matrix or a liquid plasma matrix after centrifugation; (2) If a solid plasma matrix is obtained: The solid plasma matrix obtained in step (1) is mixed with a calcium phosphate modifier, and freeze-dried to obtain a modified plasma matrix; Alternatively, the solid plasma matrix obtained in step (1) is first freeze-dried and crushed in sequence to obtain freeze-dried plasma matrix particles; the obtained freeze-dried plasma matrix particles are then mixed with a calcium phosphate modifier to obtain a modified plasma matrix; If liquid plasma matrix is available: The liquid plasma matrix obtained in step (1) is mixed with a calcium phosphate modifier, and then freeze-dried to obtain a modified plasma matrix.
2. The preparation method according to claim 1, characterized in that: The centrifugal force of the centrifugation in step (1) is 50 to 950×g; Preferably, the centrifugation time in step (1) is 5 to 10 minutes.
3. The preparation method according to claim 1 or 2, characterized in that: The calcium phosphate modifier in step (2) includes one or more of calcium phosphate bone cement, calcium phosphate ceramics or calcium phosphate hyaluronic acid microspheres.
4. The preparation method according to any one of claims 1 to 3, characterized in that In step (2), the mass ratio of the solid plasma matrix or the liquid plasma matrix to the calcium phosphate modifier is 10:(0.01-0.2).
5. The preparation method according to any one of claims 1 to 4, characterized in that: The mixing temperature in step (2) is independently 20 to 37° C. Preferably, the mixing time in step (2) is independently 5 to 10 minutes.
6. The preparation method according to any one of claims 1 to 5, characterized in that: The freeze-drying temperature in step (2) is independently -40 to -10°C.
7. The preparation method according to any one of claims 1 to 6, characterized in that: The freeze-drying time in step (2) is independently 60 to 120 minutes.
8. The preparation method according to any one of claims 1 to 7, characterized in that: The preparation method further comprises: before carrying out step (2), preparing the obtained solid plasma matrix into a plasma matrix membrane, and then breaking it.
9. A modified plasma matrix prepared by the preparation method according to any one of claims 1 to 8.
10. A use of the modified plasma matrix according to claim 9, characterized in that: The modified plasma matrix is used as a soft and hard tissue augmentation material.