CN107952115B - Bionic biomineralization artificial bone repair material and preparation method and application thereof - Google Patents

Bionic biomineralization artificial bone repair material and preparation method and application thereof Download PDF

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CN107952115B
CN107952115B CN201610902145.XA CN201610902145A CN107952115B CN 107952115 B CN107952115 B CN 107952115B CN 201610902145 A CN201610902145 A CN 201610902145A CN 107952115 B CN107952115 B CN 107952115B
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collagen
biomineralization
hydroxyapatite
aqueous solution
artificial bone
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CN107952115A (en
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张自强
张以河
陈飞旭
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Beijing Pashion Biotech Co ltd
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
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    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

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Abstract

The invention provides a biomimetic biomineralization artificial bone repair material and a preparation method and application thereof, wherein the material comprises a cross-linked collagen matrix and biomineralization collagen/hydroxyapatite composite powder dispersed in the matrix, and the mass ratio of the collagen matrix to the biomineralization collagen/hydroxyapatite composite powder is 9: 1-1: 9. The bionic biomineralization artificial bone repair material is collagen-based bionic biomineralization artificial bone, can meet the requirement of suturing artificial bone and tissue in GTR or GBR clinical operation, comprises hydroxyapatite and pure collagen, and the two components are main components of natural tissue of a human body, are nontoxic, do not influence the structure and the osteogenesis effect of the material, and have good biocompatibility. Due to the existence of the cross-linked collagen matrix, the problems of easy collapsibility, high degradation speed, poor mechanical property and the like of the material are solved, and the bionic construction of the nano hydroxyapatite and the collagen is realized.

Description

Bionic biomineralization artificial bone repair material and preparation method and application thereof
Technical Field
The invention relates to a bionic biomineralization artificial bone and a preparation method and application thereof, belonging to the field of biomedical materials.
Background
In clinical practice, bone defects are mainly caused by trauma, infection and tumor, and for the situation, people prepare various substitute materials for bone repair by various methods or ways and achieve certain effects.
Bone implant materials can be broadly classified into autogenous bone, allogeneic bone, xenogeneic bone, and synthetic materials. Autologous bone grafting is almost impossible due to the limited availability of autologous donors; allogeneic bone has good osteogenesis effect, but also has the risks of rejection reaction and virus infection, and is difficult to realize clinical large-scale use due to limited sources; the xenogeneic bone is rarely used due to the large rejection reaction. Therefore, the artificially synthesized bone repair material which has good biological activity and is similar to natural bone components is produced, the material can provide a microenvironment which is beneficial to the adhesion, proliferation and function exertion of osteoblasts and is similar to natural bone, the material can be directly used as a bone defect repair material and a good bone tissue engineering carrier material, and the wide prospect is developed for the development of bone tissue engineering.
Artificially synthesized bone materials are the hot spot of the current research and can be mainly divided into four categories of metal alloy materials, inorganic ceramic materials, high polymer materials and composite materials containing metal/inorganic ceramic/high polymer organic combination. In the aspect of metal materials, materials mainly used at present are stainless steel, cobalt alloy, titanium alloy, tantalum alloy, memory alloy and the like to replace gold. Because common stainless steel is easy to corrode as iron-based alloy, currently, the molybdenum (Mo) -containing bone repair stainless steel is frequently used, and the biochemical corrosion resistance of the stainless steel is stronger than that of the common stainless steel. The metal material has excellent mechanical property, can meet the requirement, but is higher than bone tissues too much to cause stress shielding. In addition, metal materials are difficult to be protected from corrosion in biochemical environments, and toxic metal ions are another big problem of metal-based biological materials. In the aspect of ceramic materials, the ceramic materials mainly comprise calcium sulfate filler, inert biological ceramic, bioactive ceramic, absorbable ceramic and other inorganic ceramic materials. Inorganic ceramic materials generally have non-toxic components, but the strength and bioactivity of the material do not appear to be compatible. Generally, the material with good biological activity has poor mechanical properties, and the material with good mechanical properties has no biological activity. In the aspect of polymer materials, polymer materials applied to orthopedic repair can be roughly divided into two types, namely natural polymer materials and synthetic polymer materials. The orthopedic polymer material generally has no toxicity, but has lower bioactivity, and the strength of the material is far different from that of bone tissues. Therefore, people develop a plurality of new processes and methods to synthesize various composite materials by utilizing the respective advantages and disadvantages of the materials, and the materials with different properties are matched and compounded in a smart method, so that the materials are expected to mutually make up for the deficiencies and meet different application conditions. New processes such as nano-compounding, gradient compounding, 3D printing and the like have gradually become the mainstream of orthopedic repair materials, such as levorotatory polylactic acid/hydroxyapatite whisker/collagen liquid crystal scaffold (wudi, preparation and characterization of levorotatory polylactic acid/hydroxyapatite whisker/collagen liquid crystal scaffold [ D ]. river south university, 2014.).
The research on bone scaffolds at home and abroad mainly focuses on hydroxyapatite and collagen composite scaffolds, and although various types of collagen/hydroxyapatite composite bone scaffolds are manufactured by CN1562387A, CN101229392A, CN101417145A and the like, due to the defects that the materials have the defects of loose structure, poor strength, very weak toughness, excessively high degradation speed and the like, and cannot be well matched with the bone growth speed, the materials are difficult to become good bone tissue engineering scaffolds. Researches show that the three-dimensional scaffold space structure is the basis of bone repair materials and osteoconductive performance, in recent years, researchers improve the performance of collagen-based nano hydroxyapatite composite materials by methods of biomimetic synthesis, cross-linking treatment, introduction of a third phase and the like, but still have the problems of high brittleness, disorientation of hydroxyapatite microcrystals, poor binding degree with collagen and the like, and the scaffolds are formed by a compaction method, do not form a porous form and are not beneficial to cell adhesion and proliferation. CN102205150A, CN103785059A and CN102240415A use polylactic acid or a copolymer thereof as a matrix to prepare a bone material with a three-dimensional space structure, although the bone material has better mechanical property, the material has poor hydrophilic property, and degradation products can continuously generate acidic substances to cause poor biocompatibility. CN105457097A and CN101554493A use anti-ulcer agent, dispersant and/or modifier, etc. to mix well to obtain nano hydroxyapatite/collagen scaffold, but the anti-ulcer agent, dispersant and/or modifier in the scaffold are components of non-natural bone structure, which affects material structure and bone formation effect.
In view of the above, there is a need in the art for a bone repair scaffold material with a three-dimensional structure and a good biomimetic effect, which has not been effectively satisfied for a long time, and therefore, there is a need to develop a new bone repair scaffold material to meet the practical needs.
Disclosure of Invention
The invention mainly aims to provide a biomimetic biomineralization artificial bone repair material so as to have good biomimetic effect.
The invention also aims to provide a preparation method of the biomimetic biomineralization artificial bone repair material.
The invention also aims to provide application of the bionic biomineralization artificial bone repair material.
Therefore, on one hand, the invention provides a biomimetic biomineralization artificial bone repair material which is a three-dimensional porous scaffold material and comprises a cross-linked collagen matrix and biomineralization collagen/hydroxyapatite composite powder wrapped in the matrix, wherein the mass ratio of the collagen matrix to the biomineralization collagen/hydroxyapatite composite powder is 9: 1-1: 9, preferably 1-5: 5-9; more preferably 3 to 5:5 to 7.
The mass ratio of the collagen matrix to the composite powder is controlled within the range of 3-5: 5-7, so that the mechanical property, flexibility and porosity of the material are well balanced, and the material can meet the requirements of practical application.
The bionic biomineralization artificial bone repair material is a collagen-based bionic biomineralization artificial bone, solves the technical problem that the three-dimensional bone repair scaffold material which directly takes collagen and hydroxyapatite as main components is provided for a long time in the field but is not realized, can meet the requirement of artificial bone and tissue suture in GTR or GBR clinical operation, comprises hydroxyapatite and pure collagen, and the two components are main components of human body natural tissues, are nontoxic, do not influence the material structure and the osteogenesis effect, and have good biocompatibility. Due to the existence of the cross-linked collagen matrix, the problems of easy collapsibility, high degradation speed, poor mechanical property and the like of the material are solved, and the bionic construction of the nano hydroxyapatite and the collagen is realized.
The term "comprising" means that some additional components which are not specifically defined in the present invention can be added to the material of the present invention without affecting the contribution of the present invention to the prior art, and particularly, the term "comprising" means that the material consists of the listed raw materials.
The biomimetic biomineralization artificial bone has a good three-dimensional structure, mature collagen fibers of triple helix are arranged in order, biomineralization collagen/hydroxyapatite composite powder is tightly attached to the collagen fibers and fully filled among the collagen fibers, and has a collagen-hydroxyapatite-collagen three-level composite structure which is a combination of multiple dimensions, so that the biomineralization collagen/hydroxyapatite composite powder can be stabilized in a three-dimensional porous scaffold structure and is not easy to fall off, the stability of the three-dimensional scaffold structure is increased, the mechanical property of the material is improved, and the material is not easy to damage.
The biomineralization collagen/hydroxyapatite composite powder is prepared in a biomineralization process, through interaction of organic macromolecules of collagen and inorganic ions at an interface, crystallization and growth of inorganic mineral phases of hydroxyapatite are controlled from a molecular level, so that the collagen/hydroxyapatite composite powder has a special hierarchical structure and an assembly mode, can be prepared according to the existing method, for example, the collagen/hydroxyapatite composite powder is prepared through steps (1) to (4) in CN1106861C, and is equivalent to calcium-phosphate dry powder in the case, and a person skilled in the art knows that the addition sequence of phosphate and calcium ions has no substantial influence on biomineralization.
The collagen/biomineralization hydroxyapatite composite powder is prepared by biomineralization of collagen, the hydroxyapatite takes the collagen as a template and grows on a molecular structure of the collagen in an ordered crystallization way, the collagen and the hydroxyapatite are in molecular orientation, and the molecules are changed into an ordered structure again through a biomimetic biomineralization process, so that the collagen/biomineralization hydroxyapatite composite powder has more consistent and wider collagen fiber orientation, and the artificial bone has higher mechanical strength.
According to the specific implementation mode of the invention, the diameter of the hole in the biomimetic biomineralization artificial bone repair material is 10-500 μm, and the porosity of the hole is 50-97%.
According to the specific implementation mode of the invention, in the biomineralization artificial bone repair material, the mass fraction of hydroxyapatite in the biomineralization collagen/hydroxyapatite composite powder is 50-99 wt%, preferably 80-99 wt%. The mass fraction of the hydroxyapatite in the composite powder is controlled within the range of 80-99 wt%, so that the particle size of the composite powder becomes smaller, and the composite powder is dispersed in a collagen matrix more uniformly.
According to the specific embodiment of the invention, the XRD pattern of the biomimetic biomineralization artificial bone repair material is basically as shown in col-HA in figure 4 or any one of the bionic bone 19, the bionic bone 37 or the bionic bone 55 in figure 4. From fig. 3 or fig. 4, it is obvious that the biomimetic biomineralization artificial bone repair material has a high degree of similarity with natural bone, in particular to the bionic bone 37 or the bionic bone 55.
According to a specific embodiment of the invention, in the biomimetic biomineralization artificial bone repair material, collagen in the collagen matrix or the biomineralization collagen/hydroxyapatite composite powder is natural type I collagen without telopeptide and/or recombinant human type I collagen without telopeptide. Preferably, the native type I collagen is atelocollagen extracted from the skin and/or achilles tendon of an animal (e.g., pig, cow, sheep, horse, etc.) by an enzymatic process (which may further include dialysis purification, etc.).
On the other hand, the invention provides a preparation method of the biomimetic biomineralization artificial bone repair material, wherein the method comprises the following steps:
(a) providing biomineralization collagen/hydroxyapatite composite powder and a collagen aqueous solution, preferably, the mass fraction of collagen in the collagen aqueous solution is 0.8-5%;
(b) adding the biomineralization collagen/hydroxyapatite composite powder into the collagen aqueous solution to form a mixed system, wherein the mass ratio of the biomineralization collagen/hydroxyapatite composite powder to the collagen in the collagen aqueous solution is 9-1: 1-9, preferably 1-5: 5-9, more preferably 3-5: 5-7;
(c) adjusting the pH value of the mixed system obtained in the step (b) to 7-9, and then stirring to form mineralized collagen composite sol; preferably, the pH value of the obtained mixed system is adjusted to 7-9 by NaOH, KOH or ammonia water;
(d) freeze-drying the mineralized collagen composite sol to obtain a mineralized collagen composite porous material;
optionally, pre-freezing the mineralized collagen composite sol obtained in the step (c) to obtain mineralized collagen gel, and then performing freeze drying;
(e) and crosslinking the obtained mineralized collagen composite porous material to obtain the bionic biomineralization artificial bone repair material.
The bionic biomineralization artificial bone repair material can be cut into a required shape and is packaged in an aseptic mode.
According to a particular embodiment of the invention, in the method according to the invention, the crosslinking in step (e) comprises physical crosslinking and/or chemical crosslinking.
Preferably, the physical crosslinking comprises one or more of uv irradiation, thermal dehydrogenation and radiation sterilization crosslinking.
Preferably, the chemical crosslinking comprises crosslinking using one or more of carbodiimide, diamine, epoxy compound, hydroxysuccinimide, diphenyl phosphate (DPPA), glutaraldehyde, formaldehyde, glyoxylic acid, and genipin, and after the chemical crosslinking agent is used, the residual crosslinking agent is removed by an elution procedure.
According to the specific embodiment of the invention, preferably, the biomineralization collagen/hydroxyapatite composite powder is prepared by the following method:
(i) placing collagen in an acid to form an acidic aqueous solution of collagen; preferably, the mass fraction of the collagen in the acidic aqueous solution is 0.01-2%, and the pH value is 2.5-5.5;
(ii) slowly dripping an aqueous solution containing calcium ions into the acidic aqueous solution of the collagen; preferably, the addition amount of calcium ions is 0.01-0.1 mol per gram of collagen in the acidic aqueous solution of collagen;
(iii) adding a phosphoric acid solution into the solution obtained in the step (ii), and stirring and uniformly mixing to form a mixed system; preferably, the adding amount of the phosphoric acid solution is such that the molar ratio of calcium ions to phosphate ions in the system is 1.5-2;
(iv) adjusting the pH value of the system obtained in the step (iii) to 7-10 to obtain a white suspension;
(v) (iv) standing the white suspension obtained in the step (iii), preferably standing at 15-25 ℃ for 22-26 hours, filtering, freeze-drying the filter cake, preferably fully freeze-drying at-50 to-20 ℃ for 24-48 hours, and then grinding to obtain dry powder, namely the biomineralization collagen/hydroxyapatite composite powder;
preferably, the acid in step (i) comprises one or more of sulphuric acid, hydrochloric acid, nitric acid, acetic acid, propionic acid and citric acid;
preferably, the calcium ion-containing aqueous solution in step (ii) is CaCl2、Ca(NO3)2And CaCO3One or more of (a).
The method for preparing the biomineralization collagen/hydroxyapatite can effectively control the precipitation effect of the powder, and is more beneficial to obtaining the composite powder with smaller particle size compared with the prior art, so that the obtained composite powder is more uniformly dispersed in a collagen matrix.
In the step (iv), when the pH is adjusted, the mixed system starts to precipitate when the pH is 5 to 6, and when the pH is 7 to 10, a white suspension appears in the mixed system.
According to the specific implementation mode of the invention, the preparation method of the biomimetic biomineralization artificial bone repair material comprises the following steps:
(S1) preparation of biomineralization collagen/hydroxyapatite composite powder, which comprises:
(S1-1) dissolving collagen in an acid solution to prepare an acidic aqueous solution of collagen; preferably, the mass fraction of the collagen in the acidic aqueous solution is 0.01-2%, and the pH value is 2.5-5.5;
(S1-2) continuously stirring the solution obtained in the step (S1-1), and slowly dropping an aqueous solution containing calcium ions; preferably, the adding amount of calcium ions is 0.01-0.1 mol per gram of collagen in the acidic aqueous solution of the collagen;
(S1-3) continuously stirring the solution obtained in the step (S1-2), adding a phosphoric acid solution, and uniformly stirring to form a mixed system; preferably, the adding amount of the phosphoric acid solution is determined by the adding amount of the calcium ions in the step (S1-2), so that the molar ratio of the calcium ions to the phosphate ions is 1.5-2;
(S1-4) continuously stirring the solution obtained in the step (S1-3), and adjusting the pH of the mixed system to 7-10 to obtain a white suspension;
(S1-5) standing the mixed system obtained in the step (S1-4), preferably standing at 15-25 ℃ for 22-26 hours, filtering, freeze-drying a filter cake, preferably fully freeze-drying at-50 to-20 ℃ for 24-48 hours, and then grinding to obtain dry powder, namely the biomineralization collagen/hydroxyapatite composite powder;
(S2) complexing of collagen and biomineralized collagen/hydroxyapatite composite powder, comprising:
(S2-1) dissolving collagen in purified water to prepare an aqueous solution of collagen; preferably, the mass fraction of the collagen in the collagen aqueous solution is 0.8-5%;
(S2-2) continuously stirring the solution obtained in the step (S2-1), adding the biomineralization collagen/hydroxyapatite composite powder prepared in the step (S1-5), and uniformly stirring to form a mixed system, wherein the mass ratio of the added biomineralization collagen/hydroxyapatite composite powder to the collagen in the collagen aqueous solution obtained in the step (S2-1) is 9: 1-1: 9, preferably 1-5: 5-9, and more preferably 3-5: 5-7;
(S2-3) continuously stirring the solution obtained in the step (S2-2), and adjusting the pH of the mixed system to 7-9;
(S2-4) standing and stirring the mixed system obtained in the step (S2-3) to form a mineralized collagen composite solution; preferably, standing for 10-48 hours at 15-25 ℃, and stirring for 5-30 minutes to fully and uniformly mix the components to form a mineralized collagen composite solution;
(S3) preparation of a biomimetic biomineralization artificial bone repair material, which comprises:
(S3-1) pre-freezing the mineralized collagen composite solution obtained in the step (S2-4), preferably at the temperature of 1-5 ℃ for 3-24 hours, so as to obtain mineralized collagen gel;
(S3-2) freeze-drying the mineralized collagen gel obtained in the step (S3-1), preferably fully freeze-drying at-50 to-20 ℃ for 22 to 26 hours to obtain a mineralized collagen composite porous material;
(S3-3) crosslinking the mineralized collagen composite porous material by using a physical and/or chemical method to obtain the biomimetic biomineralization artificial bone repair material.
The method has the advantages of simple production process, easy large-scale production and low production cost, and is very favorable for reducing the medical cost of patients.
The preferred technical means of the present invention may be combined with each other to achieve further effects.
In another aspect, the invention provides the use of the biomimetic biomineralization artificial bone repair material or the biomimetic biomineralization artificial bone repair material prepared by the method in the preparation of an implant for supporting a bone defect position of a human being or an animal.
When the bone repair material of the present invention is placed as an implant at a bone defect site, it is used as a three-dimensional scaffold for bone formation, bone regeneration, bone repair or bone replacement, or for repair and regeneration of bone defects such as bone, extraction sockets and the like.
The invention has the following beneficial effects:
(1) the biomimetic biomineralization artificial bone has a collagen-hydroxyapatite-collagen three-level composite structure, mature collagen fibers of triple helices are arranged in order, biomineralization collagen/hydroxyapatite composite powder is tightly attached to the collagen fibers and fully filled between the collagen fibers, and the biomineralization collagen/hydroxyapatite composite bone has the collagen-hydroxyapatite-collagen three-level composite structure and is the combination of multiple dimensions, so that the biomineralization collagen/hydroxyapatite composite powder can be stabilized in a three-dimensional porous scaffold structure, hydroxyapatite crystals are not easy to fall off, and the stability of hydroxyapatite in the three-dimensional scaffold structure is improved.
(2) The bionic biomineralization artificial bone is characterized in that a porous three-dimensional communication structure with orderly arranged collagen and hydroxyapatite is adopted, and an internal connection pore structure is favorable for the communication of blood vessels growing into the deep part of the material so as to ensure the nutrient supply of the deep tissue of the growing material; besides the combination of the material and the implanted bed, the porous structure creates conditions for the growth of the bone tissue of the organism, the formation of mechanical internal lock and the combination of the enhanced implanted material, and the retention; the porous surface is favorable for the growth of blood vessels and soft tissues, the mutual communication of the pore diameters is a prerequisite condition for the inward growth of bones, the mutual communication is favorable for the flow of cells and body fluid in vivo and the metabolism of tissues, the micropores for the growth of bone and a large amount of external callus jointly form firm biological fixation, and the micropores form a huge surface area to provide a good matrix for bone deposition.
(3) The bionic biomineralization artificial bone is composed of hydroxyapatite and pure collagen, does not use polylactic acid, chitosan, chitin, an anti-collapse agent, a template agent, a dispersing agent, a modifier and the like, does not influence the structure of a material and the osteogenesis effect, is nontoxic and has good biocompatibility; the collagen is prepared by atelocollagen, and has no immunogenicity; realizes the simultaneous bionics of components and structures.
(4) The majority of the biomineralization collagen/hydroxyapatite composite powder is in nanometer level; the nanometer material has unique surface effect, small size effect, quantum effect and other performance, and the nanometer level hydroxyapatite particle has relatively high bioactivity compared with common hydroxyapatite.
(5) Through physical or chemical crosslinking, the mechanical property of the material is greatly improved, the material has strong degradation resistance and is not easy to collapse, and the bionic construction of the nano hydroxyapatite and the collagen is realized.
(6) The production process is simple, the large-scale production is easy, the production cost is low, and the medical cost of patients is reduced.
Drawings
FIG. 1 is a flowchart of examples 1 to 7;
FIG. 2 is an electron micrograph of the product obtained in example 1;
FIGS. 3 and 4 are XRD contrast patterns in example 9;
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description of the embodiments of the present invention taken in conjunction with the accompanying drawings, which are included to illustrate and not to limit the scope of the present invention. In the examples, each raw reagent material is commercially available, and the experimental method not specifying the specific conditions is a conventional method and a conventional condition well known in the art, or a condition recommended by an instrument manufacturer.
The reagents and equipment in the following examples and comparative examples are as follows:
hydroxyapatite (Beijing surge Biotechnology, Inc.), type I collagen (Beijing surge Biotechnology, Inc.), NaOH (analytically pure), and the like.
FT/IR-6800 type Fourier transform infrared spectrometer (Jasco, Japan), D8 type FocusX-ray diffractometer (Bruker, Germany), HP-500 type digital display push-pull dynamometer (Edinburg instruments, Leqing), GLZ-2 type vacuum freeze-drying machine (Shanghai Pudong freeze-drying equipment, Inc.), DZF-6050AF type vacuum drying cabinet (Tianjin Eng laboratory instruments, Inc.), 2XZ-2 type rotary vane vacuum pump (Shanghai double goose refrigeration equipment, Inc.), etc.
Example 1
In this example, the acid solution was hydrochloric acid and the calcium ion solution was CaCl2Aqueous solution, alkaline solution adopts NaOH aqueous solution, and ultraviolet radiation physical crosslinking method is used. FIG. 1 is a flow chart of the preparation method of the biomimetic biomineralization artificial bone of the present invention.
According to the steps shown in figure 1, the preparation method of the biomimetic biomineralization artificial bone comprises the following steps:
step S1 (preparation of biomineralization collagen/hydroxyapatite composite powder) specifically includes:
step S1-1 (collagen solubilization): dissolving 2g of collagen in 1000mL of hydrochloric acid solution with the concentration of 0.001mol/L to prepare acid solution of collagen, wherein the concentration of the collagen is 0.2%, and the pH value of the solution is 3;
step S1-2 (addition of calcium ions): the solution obtained in step S1-1 was continuously stirred, and 200mL of 0.2mol/L CaCl was slowly added dropwise2An aqueous solution;
step S1-3 (addition of phosphoric acid): continuously stirring the solution obtained in the step S1-2, adding 400mL of phosphoric acid solution with the concentration of 0.06mol/L, and uniformly stirring to form a mixed system, wherein the adding amount of the phosphoric acid solution depends on the adding amount of the calcium ions in the step S1-2, so that the molar ratio of the calcium ions to the phosphate ions reaches 1.67;
step S1-4 (pH adjustment): continuously stirring the solution obtained in the step S1-3, slowly dropwise adding a NaOH solution with the concentration of 0.03mol/L until the pH value of the mixed system is 7-10, wherein when the pH value is 5-6, the mixed system begins to precipitate, and when the pH value is 7-10, the mixed system appears as a white suspension;
step S1-5 (filtering, freeze drying, grinding to obtain composite powder): and (4) standing the mixed system obtained in the step S1-4 at 15-25 ℃ for 24 hours, filtering, fully freezing and drying the filter cake at-40 ℃ for 24 hours, and then grinding to obtain dry powder, namely the biomineralization collagen/hydroxyapatite composite powder.
Step S2 (compounding of collagen and biomineralized collagen/hydroxyapatite composite powder) specifically includes:
step S2-1 (collagen solubilization): dissolving 3g collagen in 97mL purified water to obtain collagen solution with collagen concentration of 3%
Step S2-2 (blend): continuously stirring the solution obtained in the step S2-1, adding 3g of the biomineralization collagen/hydroxyapatite composite powder prepared in the step S1-5, and uniformly stirring to form a mixed system;
step S2-3 (pH adjustment): continuously stirring the solution obtained in the step S2-2, and slowly dropwise adding a 0.01mol/L NaOH aqueous solution until the pH value of the mixed system is 9;
step S2-4 (standing, stirring): standing the mixed system obtained in the step S2-3 at 25 ℃ for 24 hours, pouring the mixed system into a stirrer, and stirring for 15 minutes to fully and uniformly mix the mixed system, wherein the mixed system is changed into white emulsion to form mineralized collagen composite solution;
step S3 (preparation of mineralized collagen composite porous material) specifically includes:
step S3-1 (mold filling and pre-freezing): injecting the mineralized collagen composite solution obtained in the step S2-4 into a rectangular mould, paving, and pre-freezing for 3 hours at 4 ℃ to obtain mineralized collagen gel;
step S3-2 (freeze-drying): fully freezing and drying the mineralized collagen gel after the mold filling in the step S3-1 at the temperature of minus 50 ℃ for 24 hours to obtain the mineralized collagen composite porous material;
step S3-3 (crosslinking): crosslinking the mineralized collagen composite porous material by using an ultraviolet irradiation method, wherein the ultraviolet irradiation condition is as follows: an ultraviolet lamp with the wavelength of 254nm irradiates for 24 hours at 10W;
step S3-4 (cutting, aseptic packaging, sterilization): cutting the cross-linked material into desired shape, aseptically packaging, and sterilizing.
The bionic biomineralization artificial bone (numbered as bionic bone 55) has a collagen-hydroxyapatite-collagen three-layer composite structure, takes block materials with regular shapes, respectively measures length, width, height and weight, and has a calculated apparent density of 0.13g/cm3The porosity was 65%, and the electron micrograph thereof is shown in FIG. 2.
Example 2
In this example, the acid solution was hydrochloric acid and the calcium ion solution was CaCl2Aqueous solution, alkaline solution adopts NaOH aqueous solution, and glutaraldehyde is used for chemical crosslinking.
Preparing a mineralized collagen composite porous material according to steps S1-1 to S3-2 in example 1, and then chemically crosslinking the mineralized collagen composite porous material by using glutaraldehyde, specifically, soaking the mineralized collagen composite porous material obtained in step S3-2 in an absolute ethanol solution of 0.05 wt% of glutaraldehyde for 48 hours to crosslink; then taking out the crosslinked mineralized collagen composite porous material, placing the material in a chromatographic column, and washing the material for 48 hours by using flowing pure water to remove the residual crosslinking agent; and (3) carrying out vacuum drying on the crosslinked mineralized collagen composite porous material at 110 ℃ for 48 hours to obtain the bionic biomineralization artificial bone. The test shows that the apparent density of the bionic biomineralization artificial bone of the embodiment is 0.24g/cm3The porosity was 85%.
Example 3
In this example, the acid solution was aqueous nitric acid solution, and the calcium ion solution was Ca (NO)3)2The aqueous solution and the alkaline solution adopt KOH aqueous solution, and the physical crosslinking is carried out by using a thermal dehydrogenation method.
A biomimetic biomineralization artificial bone was prepared by following substantially the same procedure as in example 1 except that the acid solution in step S1-1 was replaced with 1000mL of a 0.001mol/L nitric acid aqueous solution and the calcium ion solution in step S1-2 was replaced with 20mL of 0.1mol/L Ca (NO)3)2Replacing CaCl with aqueous solution2Aqueous solution, the alkaline aqueous solution in the step S1-4 and the step S2-3 replaces the NaOH aqueous solution with KOH aqueous solution with the concentration of 0.01mol/L, and after the step S3-2, the mineralized collagen composite porous material is physically crosslinked by a thermal dehydrogenation method, wherein the parameters of the thermal dehydrogenation method are 130 ℃, the vacuum degree is-0.1 MPa, and the rest steps are the same as the example 1. The apparent density of the bionic biomineralization artificial bone prepared by the embodiment is tested to be 0.15g/cm3The porosity was 70%.
Example 4
In the embodiment, the bionic biomineralization artificial bone is prepared by adopting CaCO as calcium ion aqueous solution3Physical crosslinking is performed using a thermal dehydrogenation process.
A biomimetic biomineralization artificial bone was prepared by following substantially the same procedure as in example 1, except that the calcium ion solution in step S1-2 was CaCO3And (3) after the particle step S3-2, carrying out material crosslinking on the mineralized collagen composite porous material by using a thermal dehydrogenation method, wherein the parameters of the thermal dehydrogenation method are 130 ℃, and the vacuum degree is-0.1 MPa, and the rest steps are the same as those in the example 1. The test shows that the apparent density of the bionic biomineralization artificial bone of the embodiment is 0.14g/cm3The porosity was 72%.
Example 5
In this example, the acid solution was acetic acid aqueous solution, and the calcium ion aqueous solution was CaCl2Aqueous solution, alkaline aqueous solution adopts Na2CO3Aqueous solution, chemical crosslinking using formaldehyde.
A biomimetic biomineralization artificial bone was prepared by following substantially the same procedure as in example 1 except that the acid solution in step S1-1 was replaced with 1000mL of an aqueous acetic acid solution having a concentration of 0.01mol/L in place of the hydrochloric acid solution, and the alkaline solution in steps S1-4 and S2-3 was replaced with Na having a concentration of 0.5mol/L in2CO3Mixing aqueous solution instead of NaOH aqueous solutionThe procedure of example 1 was followed except that the mineralized collagen composite porous material was chemically crosslinked using 0.025 wt% formaldehyde-absolute ethanol solution after the pH adjustment step S3-2. The test shows that the apparent density of the bionic biomineralization artificial bone of the embodiment is 0.28g/cm3The porosity was 67%.
Example 6
In this example, the acid solution was acetic acid aqueous solution, calcium ion aqueous solution was CaCl2The alkaline aqueous solution is formed by using KOH, and chemical crosslinking is carried out by using formaldehyde.
A biomimetic biomineralization artificial bone was prepared according to substantially the same procedure as in example 1, except that the acid solution in step S1-1 was replaced with 1000mL of acetic acid having a concentration of 0.01mol/L instead of the hydrochloric acid solution, the alkaline solution in steps S1-4 and S2-3 was replaced with KOH having a concentration of 0.02mol/L instead of the NaOH aqueous solution, and after step S3-2, the mineralized collagen composite porous material was chemically cross-linked using a 0.025 wt% formaldehyde absolute ethanol solution, and the remaining steps were the same as in example 1. The test shows that the apparent density of the bionic biomineralization artificial bone of the embodiment is 0.31g/cm3The porosity was 66%.
Example 7
In the preparation of the biomimetic biomineralization artificial bone, the acid solution is citric acid aqueous solution, and the calcium ion aqueous solution is CaSO4Aqueous solution and alkaline solution adopt ammonia water, and genipin is used for chemical crosslinking.
The biomimetic biomineralization artificial bone was prepared according to substantially the same procedure as in example 1 except that the acid solution in step S1-1 was replaced with 1000mL of citric acid having a concentration of 0.01mol/L instead of the hydrochloric acid solution, the alkaline solution in steps S1-4 and S2-3 was replaced with aqueous ammonia solution instead of the aqueous NaOH solution, and after step S3-2, the mineralized collagen composite porous material was chemically cross-linked using 0.5 wt% genipin aqueous solution. The test shows that the apparent density of the bionic biomineralization artificial bone of the embodiment is 0.25g/cm3The porosity was 74%.
Example 8
This example is the same as example 1 except that step S2-1 and step S2-2 are different from example 1; the steps S2-1 and S2-2 specifically comprise the following steps:
step S2-1: dissolving 1g of collagen in 99mL of purified water to prepare a collagen solution, wherein the concentration of the collagen is 1%;
step S2-2 (blend): continuously stirring the solution obtained in the step S2-1, adding 9g of the biomineralization collagen/hydroxyapatite composite powder prepared in the step S1-5, and uniformly stirring to form a mixed system;
the biomimetic biomineralization artificial bone prepared in this example is numbered as biomimetic bone 19.
Example 9
This example is the same as example 1 except that step S2-1 and step S2-2 are different from example 1; the steps S2-1 and S2-2 specifically comprise the following steps:
step S2-1: dissolving 3g of collagen in 97mL of purified water to prepare a collagen solution, wherein the concentration of the collagen is 3%;
step S2-2 (blend): continuously stirring the solution obtained in the step S2-1, adding 7g of the biomineralization collagen/hydroxyapatite composite powder prepared in the step S1-5, and uniformly stirring to form a mixed system;
the biomimetic biomineralization artificial bone prepared in this example is numbered as biomimetic bone 37.
The crystal phase structure and the crystallinity of the bionic biomineralization artificial bone are researched by adopting an X-ray diffractometer (XRD) of Focus.
FIG. 3 is an XRD pattern of natural bone (pig bone), Hydroxyapatite (HA) and the sample (Col-HA) prepared in example 1. Comparing the two spectrums of HA and Col-HA in figure 3, the diffraction peak of the biomimetic biomineralization artificial bone is found to be consistent with the diffraction peak spectrum of the standard hydroxyapatite crystal, which indicates that the inorganic phase in the biomimetic biomineralization artificial bone is mainly hydroxyapatite. Comparing the Col-HA and pig bone in FIG. 3, the positions and the number of diffraction peaks of the biomimetic biomineralization artificial bone and the natural bone are consistent, which shows that the inorganic phase in the composite material is similar to the inorganic phase in the natural bone.
Fig. 4 is an XRD spectrum of collagen, hydroxyapatite, bionic bone 19, bionic bone 37, bionic bone 55 and natural rabbit bone, and it can be seen from fig. 4 that the ratio of biomineralization collagen/hydroxyapatite composite powder is more similar to the crystal phase structure and crystallinity of natural bone at 3:7 and 5:5, and the biomimetic effect is more obvious.
The method inspects the stability of the hydroxyapatite in the prepared bionic bones 19, 37 and 55 by observing the turbidity of the solution after the soaking and shaking of the physiological saline, and comprises the following specific steps:
putting 1g of bionic bone into a test tube, adding 10ml of normal saline, standing at room temperature of 15-25 ℃ for 24 hours, shaking for 1 minute by using a vortex mixer, standing for 5 minutes, and observing whether white precipitate exists at the bottom of the test tube, wherein the white precipitate is hydroxyapatite falling. The experimental result shows that no white precipitate is generated in the bionic bone 55 and the free bone 37, no hydroxyapatite is dropped, the free bone 19 has a slight white precipitate, and the dropping rate of the hydroxyapatite is less than 10% through drying calculation.
Finally, the description is as follows: although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that: modifications and equivalents may be made thereto without departing from the spirit and scope of the invention and it is intended to cover any modifications or equivalents as may fall within the scope of the invention.

Claims (18)

1. A bionic biomineralization artificial bone repair material is a three-dimensional porous scaffold material, which consists of a crosslinked collagen matrix and biomineralization collagen/hydroxyapatite composite powder wrapped in the matrix, and has a collagen-hydroxyapatite-collagen three-level composite structure, collagen fibers of triple helix are orderly arranged, and the biomineralization collagen/hydroxyapatite composite powder is closely attached to the collagen fibers and is fully filled among the collagen fibers;
the mass ratio of the collagen matrix to the biomineralization collagen/hydroxyapatite composite powder is 3-5: 5-7; the mass fraction of hydroxyapatite in the biomineralization collagen/hydroxyapatite composite powder is 80-99 wt%;
the bionic biomineralization artificial bone repair material is prepared by the following steps:
(a) providing a collagen aqueous solution and biomineralization collagen/hydroxyapatite composite powder;
(b) adding the biomineralization collagen/hydroxyapatite composite powder into the collagen aqueous solution to form a mixed system;
(c) adjusting the pH value of the mixed system obtained in the step (b) to 7-9, and then stirring to form mineralized collagen composite sol;
(d) pre-freezing the mineralized collagen composite sol obtained in the step (c) to obtain mineralized collagen gel, and then performing freeze drying to obtain a mineralized collagen composite porous material;
(e) crosslinking the obtained mineralized collagen composite porous material to obtain the bionic biomineralization artificial bone repair material; the diameter of a hole in the bionic biomineralization artificial bone repair material is 10-500 mu m, and the porosity of the hole is 50-97%;
the collagen in the collagen matrix or the biomineralization collagen/hydroxyapatite composite powder is natural type I collagen without telopeptide and/or recombinant human type I collagen without telopeptide.
2. The biomimetic biomineralized artificial bone repair material of claim 1, wherein the atelocytic type I collagen is atelocytic collagen extracted from skin and/or achilles tendon of an animal and prepared by an enzymatic hydrolysis process.
3. A method of preparing a biomimetic biomineralised artificial bone repair material according to claim 1 or 2, the method comprising the steps of:
(a) providing a collagen aqueous solution and biomineralization collagen/hydroxyapatite composite powder; the biomineralization collagen/hydroxyapatite composite powder is prepared by the following method:
(i) placing collagen in an acid to form an acidic aqueous solution of collagen; the mass fraction of the collagen in the acidic aqueous solution is 0.01-2%, and the pH value is 2.5-5.5;
(ii) slowly dripping an aqueous solution containing calcium ions into the acidic aqueous solution of the collagen;
(iii) adding a phosphoric acid solution into the solution obtained in the step (ii), and stirring and uniformly mixing to form a mixed system;
(iv) adjusting the pH value of the system obtained in the step (iii) to 7-10 to obtain a white suspension;
(v) (iv) standing the white suspension obtained in the step (iv), filtering, freeze-drying a filter cake, and then grinding to obtain dry powder, namely the biomineralization collagen/hydroxyapatite composite powder;
(b) adding the biomineralization collagen/hydroxyapatite composite powder into the collagen aqueous solution to form a mixed system;
(c) adjusting the pH value of the mixed system obtained in the step (b) to 7-9, and then stirring to form mineralized collagen composite sol;
(d) pre-freezing the mineralized collagen composite sol obtained in the step (c) to obtain mineralized collagen gel, and then performing freeze drying to obtain a mineralized collagen composite porous material; freeze-drying for 22-26 hours at-50 to-20 ℃;
(e) crosslinking the obtained mineralized collagen composite porous material to obtain the bionic biomineralization artificial bone repair material;
the crosslinking comprises physical crosslinking and/or chemical crosslinking;
the physical crosslinking comprises one or more of ultraviolet irradiation, thermal dehydrogenation method and irradiation sterilization method crosslinking;
the chemical crosslinking comprises crosslinking by using one or more of carbodiimide, diamine, epoxy compound, hydroxysuccinimide, diphenyl phosphate, glutaraldehyde, formaldehyde, glyoxylic acid and genipin, and removing residual crosslinking agent by an elution procedure after using the chemical crosslinking agent.
4. The method according to claim 3, wherein the pH of the mixed system obtained in step (c) is adjusted to 7-9 by NaOH, KOH or ammonia water.
5. A process according to claim 3, wherein the acid in step (i) comprises one or more of sulphuric acid, hydrochloric acid, nitric acid, acetic acid, propionic acid and citric acid.
6. The method according to claim 3, wherein in step (ii), the amount of calcium ions added is 0.01-0.1 mol per gram of collagen in the acidic aqueous solution of collagen.
7. The method of claim 3, wherein the calcium ion-containing aqueous solution of step (ii) is CaCl2And/or Ca (NO)3)2And (4) forming.
8. The method according to claim 3, wherein the phosphoric acid solution in step (iii) is added in an amount such that the molar ratio of calcium ions to phosphate ions in the system is 1.5 to 2.
9. The method according to claim 3, wherein the white suspension in the step (v) is allowed to stand at 15 to 25 ℃ for 22 to 26 hours.
10. A method according to claim 3, wherein the method comprises the steps of:
(S1) preparation of biomineralization collagen/hydroxyapatite composite powder, which comprises:
(S1-1) dissolving collagen in an acid solution to prepare an acidic aqueous solution of collagen;
(S1-2) continuously stirring the solution obtained in the step (S1-1), and slowly dropping an aqueous solution containing calcium ions;
(S1-3) continuously stirring the solution obtained in the step (S1-2), adding a phosphoric acid solution, and uniformly stirring to form a mixed system;
(S1-4) continuously stirring the solution obtained in the step (S1-3), and adjusting the pH of the mixed system to 7-10 to obtain a white suspension;
(S1-5) standing the white suspension obtained in the step (S1-4), filtering, freeze-drying a filter cake, and grinding to obtain dry powder, namely the biomineralization collagen/hydroxyapatite composite powder;
(S2) complexing of collagen and biomineralized collagen/hydroxyapatite composite powder, comprising:
(S2-1) dissolving collagen in purified water to prepare an aqueous solution of collagen;
(S2-2) continuously stirring the solution obtained in the step (S2-1), adding the biomineralization collagen/hydroxyapatite composite powder prepared in the step (S1-5), uniformly stirring to form a mixed system, wherein the mass ratio of the added biomineralization collagen/hydroxyapatite composite powder to the collagen in the collagen aqueous solution obtained in the step (S2-1) is 5-7: 3-5,
(S2-3) continuously stirring the mixed system obtained in the step (S2-2), and adjusting the pH of the mixed system to 7-9;
(S2-4) standing and stirring the mixed system obtained in the step (S2-3) to form mineralized collagen composite sol;
(S3) preparation of a biomimetic biomineralization artificial bone repair material, which comprises:
(S3-1) pre-freezing the mineralized collagen composite sol obtained in the step (S2-4) to obtain mineralized collagen gel;
(S3-2) freeze-drying the mineralized collagen gel obtained in the step (S3-1) to obtain a mineralized collagen composite porous material;
(S3-3) crosslinking the mineralized collagen composite porous material by using a physical and/or chemical method to obtain the biomimetic biomineralization artificial bone repair material.
11. The method according to claim 10, wherein in the step (S1-2), calcium ion is added in an amount of 0.01 to 0.1mol per gram of collagen in the acidic aqueous solution of collagen.
12. The method according to claim 10, wherein the phosphoric acid solution is added in an amount such that the molar ratio of calcium ions to phosphate ions in the system is 1.5 to 2 in step (S1-3).
13. The method according to claim 10, wherein the white suspension is left to stand at 15 to 25 ℃ for 22 to 26 hours in the step (S1-5).
14. The method according to claim 10, wherein the freeze-drying in the step (S1-5) is carried out at-50 to-20 ℃ for 24 to 48 hours.
15. The method according to claim 10, wherein the collagen aqueous solution in the step (S2-1) has a collagen mass fraction of 0.8% to 5%.
16. The method according to claim 10, wherein the mixed system is left to stand at 15 to 25 ℃ for 10 to 48 hours in the step (S2-4), and is stirred for 5 to 30 minutes to be fully and uniformly mixed to form the mineralized collagen composite sol.
17. The method according to claim 10, wherein the mineralized collagen composite sol is pre-frozen at 1-5 ℃ for 3-24 hours in step (S3-1) to obtain mineralized collagen gel.
18. Use of a biomimetic biomineralization artificial bone repair material according to claim 1 or 2 or a biomimetic biomineralization artificial bone repair material prepared by the method according to any of claims 3-17 for preparing an implant for supporting a bone defect site of a human being or an animal.
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Families Citing this family (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109381745B (en) * 2018-09-21 2021-03-19 北京市意华健科贸有限责任公司 Collagen coral hydroxyapatite composite bone filling scaffold material and preparation method thereof
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CN112773938B (en) * 2019-11-06 2023-03-24 湘潭大学 Strychnine-based 3D printing artificial bone composition and method for preparing strychnine-based 3D printing artificial bone
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1775306A (en) * 2005-10-10 2006-05-24 胡庆柳 Method for preparing porous collagen composite nano hydroox apatite artificial bone
CN103830775A (en) * 2012-11-27 2014-06-04 北京奥精医药科技有限公司 High-strength collagen base artificial bone repair material

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6300315B1 (en) * 1999-08-28 2001-10-09 Ceramedical, Inc. Mineralized collagen membrane and method of making same
US7718616B2 (en) * 2006-12-21 2010-05-18 Zimmer Orthobiologics, Inc. Bone growth particles and osteoinductive composition thereof
CN101417145B (en) * 2008-12-04 2012-11-07 山东大学 Bracket material for bone tissue engineer and preparation method thereof
CN101791438B (en) * 2010-03-16 2013-02-27 浙江大学 Method for preparing bioactive poly(lactic-co-glycolic acid)/collagen/hydroxyapatite composite fiber membrane for bone repair
CN104096268B (en) * 2014-06-19 2016-11-23 北京奥精医药科技有限公司 A kind of mineralized collagen artificial periosteum and preparation method thereof
CN105457097A (en) * 2015-11-12 2016-04-06 谢宝钢 Injectable mineralized collagen bone repair material and preparing method

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1775306A (en) * 2005-10-10 2006-05-24 胡庆柳 Method for preparing porous collagen composite nano hydroox apatite artificial bone
CN103830775A (en) * 2012-11-27 2014-06-04 北京奥精医药科技有限公司 High-strength collagen base artificial bone repair material

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