CN114288472A - Platelet-rich plasma-based artificial bone repair material and preparation method thereof - Google Patents
Platelet-rich plasma-based artificial bone repair material and preparation method thereof Download PDFInfo
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Abstract
The application relates to the field of biomedical materials, in particular to an artificial bone repair material based on platelet-rich plasma and a preparation method thereof; the raw materials of the repair material comprise: platelet rich plasma, type I collagen and artificial bone material; the method comprises the following steps: separating to obtain isolated blood; centrifuging and screening the isolated blood to obtain platelet-rich plasma; respectively obtaining artificial bone material and I type collagen solution; adding the I-type collagen solution into the artificial bone material, standing, and then stirring and mixing for the first time to obtain mixed bone paste; carrying out second stirring and mixing on the mixed bone paste and the platelet-rich blood serum to obtain a repair material with mutually matched components; form the bone mud through mixing between with I type collagen and the artifical bone material, utilize containing a large amount of osteogenesis factors in the I type collagen, can increase the activity of osteogenesis factor in the mixed bone mud together with the growth factor in the platelet rich plasma to guarantee the completion of the microenvironment of osteogenesis process, effectively promote bone tissue regeneration.
Description
Technical Field
The application relates to the field of biomedical materials, in particular to an artificial bone repair material based on platelet-rich plasma and a preparation method thereof.
Background
Platelet Rich Plasma (PRP) is plasma rich in platelets at high concentration obtained by centrifuging whole blood of animals or humans, and becomes a gel after thrombin is added thereto, and thus is also called platelet rich gel or platelet rich leukocyte gel (PLG), and PRP contains a large amount of growth factors such as platelet-derived growth factor (PDGF), transforming growth factor β (TGF- β), insulin-like growth factor 1(IGF-1), and the like.
The primary mechanism by which PRP promotes bone regeneration is that it releases some growth factors for the local microenvironment of the bone defect, with the growth factors that play a major role being PDGF and TGF- β present in the α granule; when PRP is not activated, the amount of the growth factor released by alpha granules in the platelets is less, the release amount of the growth factor greatly increases after the PRP is activated, the proliferation promoting effect of the activated PRP is obviously stronger than that of an inactivated group, and the amount of the growth factor released by the PRP is increased along with the increase of the thrombin concentration.
Hydroxyapatite is the main component of natural bone inorganic salt, has good bone conductivity and biocompatibility, is considered as an ideal material for bone defect repair, and particularly, the nano-scale hydroxyapatite is similar to the inorganic component in natural bone, can be introduced into a bone repair material to enable the material to have great superiority in the aspects of mechanics and biology, and is beneficial to the growth of new bone tissues and vascular tissues; although nano-hydroxyapatite has good osteoconductivity and biocompatibility, single hydroxyapatite has large brittleness and low toughness, so that an artificial bone material to be constructed is generally compounded with a collagen component, or growth factors are simply added, or osteogenic protein is added, although partial osteogenic factors can be compensated by adding platelet-rich plasma, sufficient osteogenic factors cannot be provided, a complete microenvironment cannot be provided for an osteogenesis process, and rapid regeneration and repair of bone tissues are not facilitated.
Therefore, how to ensure the integrity of the microenvironment of the osteogenesis process of the artificial bone repair material is a technical problem which needs to be solved at present.
Disclosure of Invention
The application provides an artificial bone repair material based on platelet rich plasma and a preparation method thereof, which aim to solve the technical problem of poor suitability between the platelet rich plasma and the artificial bone material in the prior art.
In a first aspect, the present application provides an artificial bone repair material based on platelet rich plasma, which comprises the following raw materials in parts by weight: platelet rich plasma: 15-20 parts of type I collagen: 25-35 parts of artificial bone material: 45 to 55 portions.
Optionally, the artificial bone material comprises hydroxyapatite or calcium phosphate bioactive ceramics.
Optionally, the type I collagen is a type I collagen solution, and comprises type I collagen solutes and a solvent, wherein the mass concentration of the type I collagen solutes is 0.5% -5%.
Optionally, the solvent comprises acetic acid, and the molar concentration of the acetic acid is 0.2 mol/L-1.0 mol/L.
In a second aspect, the present application provides a method for preparing an artificial bone repair material based on platelet rich plasma, the method comprising:
separating to obtain isolated blood;
centrifuging and screening the isolated blood to obtain platelet-rich plasma;
respectively obtaining artificial bone material and I type collagen solution;
adding the type I collagen solution into the artificial bone material, standing, and then carrying out first stirring and mixing to obtain mixed bone paste;
and carrying out second stirring and mixing on the mixed bone paste and the platelet-rich blood slurry to obtain the repair material with mutually matched components.
Optionally, the rotation speed of the first stirring and mixing is 35rpm to 40rpm, and the time of the first stirring and mixing is 2min to 5 min.
Optionally, the rotation speed of the second stirring and mixing is 60rpm to 80rpm, and the time of the second stirring and mixing is 6min to 10 min.
Optionally, the standing time is 0.5 min-5 min.
Optionally, the mean platelet number of the platelet rich plasma is 310 × 109/L to 860 × 109And (2) per liter.
In a third aspect, the present application provides a use of an artificial bone repair material based on platelet rich plasma, said use comprising the use of the repair material according to the first aspect in the repair of a bone defect site.
Compared with the prior art, the technical scheme provided by the embodiment of the application has the following advantages:
the embodiment of the application provides a pair of artificial bone repair material based on platelet rich plasma, through the collocation between I type collagen and the artificial bone material, form the bone paste, utilize I type collagen to act as calcium salt deposit support and the mineralized promoter and the mineralized template of bone matrix mineralize mineralization of calcium-containing artificial bone material, I type collagen can cooperate with the growth factor of platelet rich plasma simultaneously, contain a large amount of osteogenesis factors in the reason I type collagen, can be together with the growth factor of platelet rich plasma, thereby balanced platelet rich plasma and artificial bone material, increase the activity of osteogenesis factor in the mixed bone paste, thereby guarantee the complete of the microenvironment of the osteogenesis process of artificial bone repair material, effectively promote bone tissue regeneration.
Drawings
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.
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive exercise.
Fig. 1 is a schematic flow chart of a method provided in an embodiment of the present application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are some embodiments of the present application, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
The inventive thinking of the application is that: because the inorganic material is not easy to be absorbed, especially the inorganic material burnt by high temperature exists with the interface of the surrounding tissue for a long time after being implanted; although the induced osteogenesis performance of the organic material is better, the organic material lacks enough mechanical strength in the early stage of implantation and has less extraction amount; therefore, the research trend of the artificial bone is towards the development of composite materials, even if the materials contain organic and inorganic components, the materials have the advantages of both the organic and inorganic components; the prior composite material technology mainly comprises the following steps:
(1) calcium phosphate composite artificial bone: mainly comprises TCP, HA, collagen, bone growth factor and other composite artificial bones. The basic process of Collagen Hydroxyapatite (CHA) induced osteogenesis is observed by transmission electron microscope and scanning electron microscope, and the observation shows that collagen HAs chemotactic effect and differentiation promoting effect on interstitial cells, and HA plays a core role and participates in matrix calcification to promote new bone formation. Wangdan and the like report the osteogenesis inducing capability of degradable porous beta-TCP/rhBMP-2 artificial bones, and the results prove that an experimental group has a large amount of new cartilage and bone formation, and a control group has no cartilage and bone formation. The beta-TCP/rhBMP-2 is considered to have good bone induction effect and is an ideal bone grafting substitute material.
(2) Polymer composite artificial bone: biodegradable polymers are a hot spot in the field of biomaterial research in recent years, and various polymers with various structural forms and certain biodegradation characteristics can be synthesized through technical processing. However, they have no osteoinductive activity and need to be combined with other osteoinductive factors to obtain good effect. Isobe et al repaired 5mm large rat femur defect with PLGA capsule containing 3ug/rhBMP-2, samples were taken for X-ray examination and histological examination at 4 weeks and 8 weeks after surgery, and the results showed: the PLGA-BMP group has formed bone healing, while the control group has no bone connection, rhBMP-2/PLGA capsule is considered to be a promising bone regeneration and release system, Hollinger et al uses the composite material of antigen-free autodigestion bone (AA bone) and PLA/PGA to repair skull defect of monkey with the diameter of 24mm, and new bones formed by the composite material group are mutually fused at 6 weeks after operation, and inner and outer bone plates and a middle bone marrow cavity with good development appear.
(3) Red bone marrow composite artificial bone: bone Marrow (BM) is composed of two parts, the hematopoietic and stromal systems. Stromal cells of healthy red bone marrow of humans and animals contain both committed osteoprogenitor cells (DOPC) and inducible osteoprogenitor cells (IOPC). DOPC has the ability to differentiate directionally into bone tissue, and IOPC can differentiate into bone under the action of an inducing factor (such as HBMP). Grundel et al, adopting TCP (40%) and HA (60%) to synthesize biphasic calcium phosphate ceramic, and implanting the biphasic calcium phosphate ceramic and BM after compounding to treat bone defects, and finding out 24 weeks after surgery that in 6 cases of bone marrow and block ceramic compound groups, 3 cases show bone connection, and 3 cases have fiber connection; in 6 cases of the bone marrow and granular ceramic composite group, 5 cases obtain bony connection, 1 case obtains fibrous connection, and 5 cases of single bone marrow implantation all obtain bony connection; blank control 3 cases had no 1 connection. Dongzhongchuan binds bone marrow cells to HA and separately adds bFGF and/or osteopontin-1 (OP-1), and the biological activities of them were determined by measuring the amount of thymidine incorporated into DNA, ALP activity and the formation of new bone. The results showed that bFGF stimulates the proliferation of bone marrow cells, OP-1 increases the activity of ALP and stimulates the formation of new bone and promotes the differentiation of bone marrow cells.
(4) Other kinds of composite artificial bones: mainly comprises an artificial bone (such as a composite artificial bone of ceramics, collagen and growth factors or related cells) consisting of more than two materials, an artificial bone compounded with a plurality of growth factors, and the like. Experimental studies of composite bone prefabricated iliac valve were reported by mozzanine et al. The bear building method is characterized in that rhBMP-2, collagen and coral composite bone with a certain shape is implanted into the ilium area of a dog, and the situation that bone tissue is converted and the ilium shape is changed after 3 months of composite bone operation and new bone is transformed into mature bone 4 months later is shown. The rhBMP-2, collagen and coral composite bone is considered to be suitable for prefabricating a bone tissue flap with a certain shape and structure in vivo.
The above-mentioned methods are all that the artificial bone material is compounded with collagen component, or only growth factor is added, or osteogenic protein is added, so that it is lack of sufficient osteogenic factor.
The type I collagen is a main structural protein of spine animals, is extracellular matrix secreted by osteoblasts in the osteogenesis process, is a promoter of a calcium salt deposition bracket and a bone matrix double layer, is also a double-template double layer template for manufacturing periosteum by adopting a double-template co-assembly technology, can promote cell migration, adsorption and differentiation, can regulate cell growth, has the advantages of low immunogenicity, no toxic or side effect on in-vivo degradation products and the like, but has poor mechanical property and high degradation rate.
If the mineralization process is adopted, hydroxyapatite can be guaranteed to be dispersed in an integral system in a nanoscale state to form mineralized collagen, and meanwhile, the mineralized collagen and the I-type collagen are compounded for use, so that the problem of insufficient performance of a single material can be solved, the advantage complementation of various materials is realized, the bone repair is promoted, and although the I-type collagen can provide partial osteogenesis factors, enough osteogenesis factors are still lacked, and the development of artificial bones is promoted.
The preparation method of PRP has not formed a unified standard, and the preparation method mainly comprises 2 methods of a density gradient centrifugation method and a plasma separation and replacement method, wherein the density gradient centrifugation method is to separate and extract PRP from whole blood according to different sedimentation coefficients of components in the blood, a secondary centrifugation method is generally adopted at present, however, the platelet concentration and the activity of PRP prepared by different centrifugal forces and centrifugal time are different, but the collection process of PRP adopts anticoagulant to prevent blood coagulation, and because the artificial bone material contains calcium salt, the activity of the anticoagulant is reduced by the dissolution of the calcium salt, so that the coagulation reaction of PRP is promoted, the viscosity of bone paste is further improved, and the osteogenesis factor of the artificial bone repair material is completely supplemented.
In one embodiment of the present application, there is provided a platelet rich plasma-based artificial bone repair material, which comprises the following raw materials in parts by weight: platelet rich plasma: 15-20 parts of type I collagen: 25-35 parts of artificial bone material: 45 to 55 portions.
The platelet-rich plasma 15-20 parts by weight has the positive effects that within the range of the parts by weight, the platelet-rich plasma can provide sufficient osteogenic factors and simultaneously provide sufficient growth factors, such as PDGF, TGF-beta 1, TGF-beta 2, IGF, EGF, VEGF and the like, wherein the growth factors can promote the formation of new blood vessels, provide sufficient nutrition for cell regeneration, prevent the apoptosis of femoral head cells and promote the bone regeneration; the growth factor can promote the proliferation and differentiation of bone marrow mesenchymal cells (BMSC), can also induce the BMSC to differentiate to chondrocytes and promote cartilage regeneration, can also locally inhibit apoptosis of cells in osteonecrosis and promote the generation of bone defect parts, and when the platelet rich plasma is mixed with artificial aggregate, the muddy artificial aggregate flows to the defect parts before coagulation reaction according to the time difference of the coagulation reaction because the platelet rich plasma has coagulation reaction, and the artificial aggregate is fixed at the defect parts after coagulation reaction, thereby preventing bone mud from migrating or entering a body circulation system to generate complicated inflammation; when the value of the weight part is larger than the maximum value of the end point of the range, the adverse effect is that the content of the bone formation factor can be further increased by the overhigh platelet rich plasma, but the overhigh platelet rich plasma can cause the migration of the muddy artificial aggregate before coagulation reaction agglutination, if the platelet rich plasma enters the internal circulation system of the human body, inflammation is complicated, and when the value of the weight part is smaller than the minimum value of the end point of the range, the adverse effect is that the content of the bone formation factor is reduced by the overlow platelet rich plasma, and meanwhile, the muddy artificial aggregate before coagulation reaction agglutination cannot flow to a proper position, and the fixation of the artificial aggregate after final coagulation reaction agglutination is influenced.
The weight portion of the I-type collagen is 25-35 parts, and the positive effect is that in the weight portion range, the I-type collagen can provide a part of osteogenic factors, is also used as a bracket of the calcium salt-containing artificial aggregate and a bone matrix mineralization promoter, and is also used as a calcification template, and can promote the repair of the bone defect part after the artificial aggregate is filled; when the value of the weight portion is larger than the maximum value of the end point of the range, the adverse effect is that the content of the I-type collagen is too much, the calcium salt in the artificial aggregate is too much deposited, the mobility of the artificial aggregate is influenced, and the defect part cannot be effectively filled, and when the value of the weight portion is smaller than the minimum value of the end point of the range, the adverse effect is that the content of the I-type collagen is too little, the mineralization speed of the bone matrix of the artificial aggregate is too slow, and the recovery time after the artificial aggregate is agglutinated and fixed is too long.
The artificial bone material has the advantages that the artificial bone material can serve as a framework of a repair material and form bone paste with good fluidity with I type collagen within the range of the weight portion, and the artificial bone material can rapidly generate agglutination reaction under the action of platelet-rich plasma and the I type collagen so as to be filled and fixed at a defect part; when the value of the weight part is larger than the maximum value of the end point of the range, the adverse effect is that excessive artificial bone materials cannot generate coagulation reaction under the action of platelet-rich plasma and type I collagen to agglutinate, so that bone mud migrates or enters the internal circulation system to generate complicated inflammation, and when the value of the weight part is smaller than the minimum value of the end point of the range, the adverse effect is that the excessive artificial bone materials can cause bone mud migrating, so that the complicated inflammation is generated.
In some embodiments, the artificial bone material comprises a hydroxyapatite or a calcium phosphate bioactive ceramic.
In some embodiments, the type I collagen is a type I collagen solution comprising a solute of type I collagen and a solvent, wherein the solute of type I collagen is present at a concentration of 0.5% to 5% by mass.
In the application, the positive effect that the mass concentration of the solute of the type I collagen is 0.5-5% is that in the concentration range, the type I collagen can play a role of a promoter, and meanwhile, the solute of the type I collagen can form bone mud with better fluidity with an artificial bone material, so that the subsequent filling of a bone defect part is facilitated; when the value of concentration is greater than the endpoint maximum value of this scope, the adverse effect that will lead to is that the concentration of type I collagen is too big, though can also form the better bone mud of mobility, but because type I collagen's mechanical properties is relatively poor, too much type I collagen forms the mechanical properties of bone mud with artificial bone material relatively poor, influence subsequent artificial bone's hardness, when the value of weight part is less than the endpoint minimum value of this scope, the adverse effect that will lead to is that the concentration of type I collagen is not enough, the mobility of bone mud is relatively poor, can't effectual promotion artificial bone material shaping simultaneously.
In some embodiments, the type I collagen solvent comprises acetic acid at a molar concentration of 0.2mol/L to 1.0 mol/L.
In the application, the active effect that the molar concentration of the acetic acid is 0.2 mol/L-1.0 mol/L is that the I-type collagen can be completely dissolved in the range of the concentration, and meanwhile, the I-type collagen taking the acetic acid as a solvent can further promote the repair of the bone defect part because the acetic acid has affinity to the human body; when the value of concentration is greater than the endpoint maximum value of this scope, the adverse effect that will lead to is that too high concentration is though can be quick dissolve type I collagen, but because the too high pH that will lead to the solution of acetic acid content changes, cause the dissolution of type I collagen to receive the influence, when the value of concentration is less than the endpoint minimum value of this scope, the adverse effect that will lead to is that too low acetic acid concentration will not make type I collagen dissolve fully, lead to the content of type I collagen not enough, thereby lead to the osteogenesis factor content not enough, can't guarantee the completeness of the microenvironment of osteogenesis process, thereby can't effectively promote the regeneration of bone tissue.
In one embodiment of the present application, as shown in fig. 1, there is provided a method for preparing a platelet rich plasma-based artificial bone repair material, the method comprising:
s1, separating to obtain isolated blood;
s2, centrifuging and screening the in vitro blood to obtain platelet-rich plasma;
s2, respectively obtaining an artificial bone material and an I-type collagen solution;
s4, adding the I-type collagen solution into the artificial bone material, standing, and then carrying out first stirring and mixing to obtain mixed bone paste;
s5, carrying out second stirring and mixing on the mixed bone paste and the platelet-rich blood serum to obtain a repair material with mutually matched components,
wherein the isolated blood is autologous blood of a patient with bone defect to be repaired.
In some embodiments, the first agitation and mixing is performed at a speed of 35rpm to 40rpm for 2min to 5 min.
In the application, the positive effect that the first stirring and mixing rotating speed is 35 rpm-40 rpm is that in the rotating speed range, the artificial bone material and the I-type collagen solution can be fully mixed, so that bone cement with proper fluidity is formed; when the value of the rotating speed is larger than the maximum value of the end point of the range, the adverse effect to be caused is that the I type collagen solution is spilled due to the excessively fast rotating speed, the defect of the raw material is caused, and when the value of the rotating speed is smaller than the minimum value of the end point of the range, the adverse effect to be caused is that the mixing between the artificial bone material and the I type collagen solution is insufficient due to the excessively slow rotating speed, so that the subsequent bone mud cannot effectively flow to the bone defect, and the repairing effect is influenced.
The first stirring and mixing time is 2-5 min, and the positive effect is that in the time range, the artificial bone material and the I-type collagen solution can be fully mixed, so that bone cement with proper fluidity is formed; when the time value is larger than the maximum value of the end point of the range, the adverse effect is that the overall time of the process is too long due to too long time, the time for repairing is influenced, and when the time value is smaller than the minimum value of the end point of the range, the adverse effect is that the artificial bone material and the type I collagen solution cannot be fully mixed due to too short time, and the subsequent repairing effect is influenced.
In some embodiments, the second agitation and mixing is performed at a rotation speed of 60rpm to 80rpm, and the second agitation and mixing is performed for 6min to 10 min.
In the application, the second stirring and mixing rotation speed of 60rpm to 80rpm has the positive effect that bone cement and platelet-rich plasma can be fully mixed in the rotation speed range, so that a repair material with proper fluidity is formed; when the value of the rotating speed is larger than the maximum value of the end point of the range, the adverse effect is that the bone mud and the platelet-rich plasma are spilled out due to the excessively high rotating speed, so that the raw materials are damaged, and when the value of the rotating speed is smaller than the minimum value of the end point of the range, the adverse effect is that the bone mud and the platelet-rich plasma are not sufficiently mixed due to the excessively low rotating speed, so that the formed repairing materials cannot be effectively distributed, and the repairing effect is influenced.
The second stirring and mixing time is 6-10 min, and the positive effect is that in the time range, the bone paste and the platelet-rich plasma can be fully mixed, so that a repair material with proper fluidity is formed; when the time value is greater than the maximum value of the end point of the range, the adverse effect is that the overall time of the process is too long due to too long time, the time for repairing is affected, and when the time value is less than the minimum value of the end point of the range, the adverse effect is that the bone mud and the platelet-rich plasma cannot be fully mixed due to too short time, and the subsequent repairing effect is affected.
In some embodiments, the time of standing is 0.5min to 5 min.
In the application, the standing time is 0.5-5 min, so that the artificial bone material can be fully soaked by the type I collagen solution within the time range, the artificial bone material can form a preliminary bracket, the subsequent formation of bone mud is facilitated, and the fluidity of the bone mud is ensured; when the value of the time is larger than the maximum value of the end point of the range, the adverse effect is that the overall time of the process is too long due to too long time, the time for repairing is influenced, and when the value of the time is smaller than the minimum value of the end point of the range, the adverse effect is that the artificial bone material cannot be fully soaked by the type I collagen solution due to too short time, so that the mobility of the bone cement is poor, and the subsequent repairing effect is influenced.
In some embodiments, the platelet rich plasma has an average platelet count of 310 x 109each/L to 860 x 109And (2) per liter.
In the present application, the mean platelet number of the platelet-rich plasma was 310X 109each/L to 860 x 109The positive effect of the quantity/L is that the platelet-rich plasma can provide enough growth factors within the quantity range, so that the content and the variety of the osteogenic factors of the repair material are improved, and the sufficient number of the platelets can be beneficial to the proceeding of the blood coagulation reaction; when the value of the amount is larger than the maximum value of the end point of the range, the adverse effect is that the coagulation reaction is too violent due to the excessive number of the blood platelets, the flowability of the repair material is poor, and the repair effect is affected.
In one embodiment of the present application, there is provided a use of an artificial bone repair material based on platelet rich plasma, the use comprising the use of the repair material according to the first aspect in the repair of a bone defect site.
Example 1
An artificial bone repair material based on platelet rich plasma comprises the following raw materials in parts by weight: platelet rich plasma: 17 parts, type I collagen: 30 parts of artificial bone material: 53 parts.
The artificial bone material is hydroxyapatite.
The type I collagen is a type I collagen solution and comprises type I collagen solutes and a solvent, wherein the mass concentration of the type I collagen solutes is 2%.
The type I collagen solvent comprises acetic acid, and the molar concentration of the acetic acid is 0.2 mol/L-1.0 mol/L.
As shown in fig. 1, a preparation method of an artificial bone repair material based on platelet rich plasma comprises the following steps:
s1, separating to obtain isolated blood;
s2, centrifuging and screening the blood to obtain platelet-rich plasma;
s3, respectively obtaining an artificial bone material and an I-type collagen solution;
s4, adding the I-type collagen solution into the artificial bone material, standing, and then carrying out first stirring and mixing to obtain mixed bone paste;
and S5, carrying out second stirring and mixing on the mixed bone paste and the platelet-rich blood serum to obtain the repair material with mutually matched components.
The rotation speed of the first stirring and mixing was 37rpm, and the time of the first stirring and mixing was 4 min.
The rotation speed of the second stirring and mixing is 70rpm, and the time of the second stirring and mixing is 8 min.
Standing for 3 min.
The mean platelet number of the platelet-rich plasma was 860X 109And (2) per liter.
Example 2
Comparing example 2 with example 1, example 2 differs from example 1 in that:
the repair material comprises the following raw materials in parts by weight: platelet rich plasma: 15 parts, type I collagen: 25 parts of artificial bone material: 45 parts of the raw materials.
The artificial bone material is calcium phosphate bioactive ceramic.
The mass concentration of the solute of the type I collagen is 0.5 percent.
The molar concentration of acetic acid is 0.2 mol/L.
The rotation speed of the first stirring and mixing was 35rpm, and the time of the first stirring and mixing was 2 min.
The rotation speed of the second stirring and mixing was 60rpm, and the time of the second stirring and mixing was 6 min.
The standing time was 0.5 min.
The mean platelet count of the platelet rich plasma was 310X 109And (2) per liter.
Example 3
Comparing example 3 with example 1, example 3 differs from example 1 in that:
the repair material comprises the following raw materials in parts by weight: platelet rich plasma: 20 parts, type I collagen: 35 parts of artificial bone material: and 55 parts.
The artificial bone material is hydroxyapatite.
The mass concentration of the solute of the type I collagen is 5 percent.
The molar concentration of acetic acid is 1.0 mol/L.
The rotation speed of the first stirring and mixing was 40rpm, and the time of the first stirring and mixing was 5 min.
The rotation speed of the second stirring and mixing is 80rpm, and the time of the second stirring and mixing is 10 min.
The standing time is 5 min.
The mean platelet number of the platelet-rich plasma was 860X 109And (2) per liter.
Example 4
Comparing example 4 with example 1, example 4 differs from example 1 in that:
the rotation speed of the first stirring and mixing was 35rpm, and the time of the first stirring and mixing was 2 min.
The rotation speed of the second stirring and mixing was 60rpm, and the time of the second stirring and mixing was 6 min.
The standing time was 0.5 min.
Example 5
Comparing example 5 with example 1, example 5 differs from example 1 in that:
the rotation speed of the first stirring and mixing was 40rpm, and the time of the first stirring and mixing was 5 min.
The rotation speed of the second stirring and mixing is 80rpm, and the time of the second stirring and mixing is 10 min.
The standing time is 5 min.
Comparative example 1
Comparative example 1 and example 1 were compared, and comparative example 1 and example 1 were distinguished in that:
no type I collagen was added.
Comparative example 2
Comparative example 2 is compared with example 1, and comparative example 2 differs from example 1 in that:
platelet rich plasma was not added.
Comparative example 3
Comparative example 3 is compared with example 1, and comparative example 3 differs from example 1 in that:
the rotation speed of the first stirring and mixing was 30rpm, and the time of the first stirring and mixing was 1 min.
The rotation speed of the second stirring and mixing was 50rpm, and the time of the second stirring and mixing was 5 min.
The standing time was 0.2 min.
Comparative example 4
Comparative example 4 is compared with example 1, and comparative example 4 differs from example 1 in that:
the rotation speed of the first stirring and mixing was 45pm, and the time of the first stirring and mixing was 10 min.
The rotation speed of the second stirring and mixing is 90rpm, and the time of the second stirring and mixing is 15 min.
The standing time was 8 min.
Related experiments:
the artificial bone repair materials obtained in comparative examples 1 to 4 of examples 1 to 5 were collected, respectively, and the viscosity thereof was measured, with the results shown in table 1.
Detection method of related experiment:
viscosity: the measurement was carried out according to GB/T23101.4-2008.
TABLE 1
| Categories | Viscosity (Pa. S) |
| Example 1 | 1500 |
| Example 2 | 1450 |
| Example 3 | 1380 |
| Example 4 | 1465 |
| Example 5 | 1360 |
| Comparative example 1 | 1088 |
| Comparative example 2 | 890 |
| Comparative example 3 | 1210 |
| Comparative example 4 | 1200 |
Specific analysis of table 1:
the viscosity refers to the strong and weak fluidity of the artificial bone repair material obtained after the platelet-rich plasma is added into the bone paste, and when the viscosity is moderate, the proper fluidity of the artificial bone repair material is indicated;
from the data of examples 1-5, it can be seen that:
if I-type collagen is added into platelet-rich plasma and the artificial bone repair material, the viscosity of the repair material can be improved, so that migration of the repair material is prevented, postoperative complications are avoided, meanwhile, the obtained repair material can make up the condition that growth factors of the traditional artificial bone material are insufficient, and the bone formation speed is accelerated.
From the data of comparative examples 1-4, it can be seen that:
if the platelet-rich plasma or the type I collagen is not added, the viscosity of the repair material is influenced, and the influence of the platelet-rich plasma on the viscosity of the repair material is large.
One or more technical solutions in the embodiments of the present application at least have the following technical effects or advantages:
(1) the artificial bone repairing material provided by the embodiment of the application makes up the shortage of osteogenic factors of the traditional artificial bone material by introducing the I-type collagen and the platelet-rich plasma, can provide various growth factors and osteogenic factors, and accelerates the bone healing speed.
(2) The artificial bone repair material provided by the embodiment of the invention can extract platelet-rich plasma from a patient body to form a self-derived material, wherein the proportion, concentration and the like of factors related to an osteogenesis process are completely consistent with the self-derived material, and the problems of rejection and the like are avoided.
(3) The artificial bone repair material provided by the embodiment of the application provides good biocompatibility and bone induction performance through platelet-rich plasma and type I collagen, and further carries out bone tissue repair through a structure similar to human bone and a calcium-phosphorus ratio characteristic close to the human bone by mineralizing collagen, so that a good bone healing promotion effect is achieved.
(4) According to the method provided by the embodiment of the application, the I-type collagen and the artificial bone material are mixed to form the bone cement, the platelet-rich plasma is added, platelets in the platelet-rich plasma and calcium ions in the artificial bone material are utilized, the anticoagulant in the platelet-rich plasma is disabled under the action of the I-type collagen, the calcium ions in the artificial bone material enable the platelet-rich plasma to generate coagulation reaction, the artificial bone material is enabled to be agglomerated and fixed, a certain time is required in the process, therefore, the platelet-rich plasma can increase the fluidity of the bone cement in the early stage of the coagulation reaction through the synergistic reaction of the platelets in the platelet-rich plasma and the artificial bone material, the fluidity of the bone cement is rapidly reduced due to the coagulation of the platelet-rich plasma after the coagulation reaction, the bone cement is effectively prevented from migrating or entering an in-vivo circulatory system to generate complications, and the life safety of a patient is maintained.
(5) The method provided by the embodiment of the application is convenient to operate, the collected platelet-rich plasma is mixed with the bone paste formed by the artificial bone material, the effect of mixing components and procoagulant can be achieved simultaneously by one-time mixing, and meanwhile, various growth factors in the platelet-rich plasma are matched with the artificial bone paste to provide perfect bone forming matrix, so that the bone healing can be effectively accelerated, the course of disease is shortened, and the whole treatment cost of a patient is reduced.
(6) According to the method provided by the embodiment of the application, the obtained repair material can be injected into the bone defect part through an injector or other injection equipment to repair, and various growth factors in platelet-rich plasma are utilized to repair, so that the probability of bone nonunion can be effectively reduced.
It is noted that, in this document, relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The foregoing are merely exemplary embodiments of the present invention, which enable those skilled in the art to understand or practice the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. An artificial bone repair material based on platelet rich plasma is characterized in that the repair material comprises the following raw materials in parts by weight: platelet rich plasma: 15-20 parts of type I collagen: 25-35 parts of artificial bone material: 45 to 55 portions.
2. Prosthetic material according to claim 1, characterized in that said artificial bone material comprises hydroxyapatite or calcium phosphate bioactive ceramics.
3. The repair material according to claim 1, wherein the type I collagen is a type I collagen solution comprising a type I collagen solute and a solvent, and the mass concentration of the type I collagen solute is 0.5-5%.
4. The repair material according to claim 3, wherein the type I collagen solvent comprises acetic acid, and the molar concentration of the acetic acid is 0.2mol/L to 1.0 mol/L.
5. A method of preparing the repair material of any one of claims 1 to 4, comprising:
separating to obtain isolated blood;
centrifuging and screening the isolated blood to obtain platelet-rich plasma;
respectively obtaining artificial bone material and I type collagen solution;
adding the type I collagen solution into the artificial bone material, standing, and then carrying out first stirring and mixing to obtain mixed bone paste;
and carrying out second stirring and mixing on the mixed bone paste and the platelet-rich blood slurry to obtain the repair material with mutually matched components.
6. The method according to claim 5, wherein the rotation speed of the first stirring and mixing is 35rpm to 40rpm, and the time of the first stirring and mixing is 2min to 5 min.
7. The method according to claim 5, wherein the rotation speed of the second stirring and mixing is 60rpm to 80rpm, and the time of the second stirring and mixing is 6min to 10 min.
8. The method according to claim 5, wherein the standing time is 0.5min to 5 min.
9. The method of claim 5, wherein the platelet rich plasma has an average platelet count of 310 x 109each/L to 860 x 109And (2) per liter.
10. Use of an artificial bone repair material based on platelet rich plasma, characterized in that it comprises the use of a repair material according to any one of claims 1 to 4 in the repair of a bone defect site.
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