CN110776315A

CN110776315A - Magnesium strontium-silicon phosphate material, preparation method thereof and structure-controllable porous bone repair composite scaffold material containing magnesium strontium-silicon phosphate material

Info

Publication number: CN110776315A
Application number: CN201911060768.7A
Authority: CN
Inventors: 张鹏; 李健; 白雪岭; 成文翔; 王新峦; 赖毓霄
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2019-11-01
Filing date: 2019-11-01
Publication date: 2020-02-11
Anticipated expiration: 2039-11-01
Also published as: CN110776315B; WO2021083369A1

Abstract

The invention provides a magnesium strontium-phosphate silicate material, a preparation method thereof and a porous bone repair composite scaffold material with a controllable structure containing the magnesium strontium-phosphate silicate material. The magnesium strontium-silicon phosphate salt material has the following structural formula. In the magnesium strontium-phosphate silicate material provided by the invention, the magnesium ions and the strontium ions have a synergistic interaction effect, and the synergistic interaction of the magnesium ions and the strontium ions can promote the proliferation and differentiation of osteoblasts, is beneficial to the functions of osteogenesis and angiogenesis in the bone repair process, and shortens the bone defect repair time.

Description

Magnesium strontium-silicon phosphate material, preparation method thereof and structure-controllable porous bone repair composite scaffold material containing magnesium strontium-silicon phosphate material

Technical Field

The invention belongs to the technical field of biomedical tissue engineering, and relates to a magnesium strontium-phosphate silicate material, a preparation method thereof, and a porous bone repair composite scaffold material with a controllable structure containing the magnesium strontium-phosphate silicate material.

Background

Various types of bone defects caused by trauma, congenital malformation, osteoporosis, bone tumor excision and the like are quite common, and the repair and functional reconstruction of large-range and large-area bone defects are one of the common problems faced by orthopedic surgery, craniofacial surgery and plastic surgery. According to the American Association of orthopedics physicians (AAOS), 630 million people in the United states have fractures, with over 50 million patients requiring bone graft, and up to $ 25 billion in 2005 costs for bone grafts (Stevens B, Yang YZ, Mohandas A, Stucker B, Nguyen KT. A review of materials, simulation to bone graft regeneration in methods, and strategies used engineered bone tissues J Biomed. Mater B2008; 85B: 573-82.). China has more than 350 ten thousand of bone-defect patients every year (Shuxing, research and development of bionic tissue engineering bone, Chinese scientific and technological achievements 2011; 12:1-2.), hundreds of thousands of new cases every year, 7.2 percent of annual growth rate of trauma hospitalization, the second place of high living quarters, and high incidence of bone defects, so that the bone graft becomes the graft with the largest demand after blood transfusion, and heavy medical burden is brought to the society. Therefore, the repair treatment of orthopedic disorders, particularly large bone defects, has become one of the most major clinical medical needs in the world.

Autologous bone grafting has been used as the gold standard for the treatment of bone defects, but still has problems such as pain in the bone-removed area, bleeding, limited sources and the like, and is greatly limited in application. The replacement of autologous bone transplantation with allogeneic bone involves risks of infection, immune rejection, disease transmission and the like. Therefore, the research and development of the biological composite scaffold material with osteoinductive activity has important clinical research significance. With the continuous and deep research of bone tissue engineering in recent years, a new hope is brought to the repair of large-section bone defects, the traditional mode of the traditional wound repair is changed, a large-range tissue defects can be repaired by a small amount of tissue cells, an ideal shape can be achieved by shaping according to needs, and a theory and a method are provided for realizing wound repair and perfect biological reconstruction.

The silicate biological ceramic material is very similar to natural bone mineral components in physical and chemical properties, has good biocompatibility and osteoconductivity, can promote direct chemical bonding with bone, and is widely concerned by researchers in the field of bone tissue engineering. Because the silicate ceramic can adjust the chemical composition, ions with the activity of promoting bone formation and blood vessels are introduced into the ceramic components in different contents, so that the mechanical strength and the degradation speed of the material can be adjusted and controlled, the biological effect of the ceramic material on the aspect of bone repair can be improved, and the bone tissue regeneration capacity can be improved. Strontium-phosphosilicate (SPS, Sr) ₅(PO ₄) ₂SiO ₄) In the chemical composition of bioceramics, Sr performs a function similar to that of Ca in bone, and the pharmacological effects of Sr have been shown to have beneficial effects in improving bone mass, bone quality and bone resistance in osteoporosis models and patients (Bonnelye E, channel A, Saltel F, Judic P. Dual effect of bone mineral: stimulation of bone differentiation and inhibition of bone differentiation and restriction in bone 2008; 42:129-38.), (Thormann U, Ray S, Sommer U, Elkhassawna T, Rehling T, Hundgeburth M, et al. bone formation induced gradient modified calcium phosphate center in crystalline-size metallic defect defects in enriched rates. biomaterials 2013; 34:8589-98.). However, the research on SPS materials in bone tissue engineering is very limited, and most of the early researchers focused on SPS and its modified physical opticsBehavior (Huang YL, Gan JH, Seo HJ. luminescence exploration of Eu) ^-Activated Sr ₅(PO ₄) ₂SiO ₄Phosphor by Combustion Synthesis.J Am Ceram Soc 2011；94:1143-8)、(Gan JH,Huang YL,Shi L,Qiao XB,Seo HJ.Luminescence properties ofEu ²⁺-activated Sr ₅(PO ₄) ₂SiO ₄for green-emitting phosphor. mater Lett 2009; 63:2160-2.). Magnesium (Mg) ²⁺) Is an essential element of the human body, and the content of the element is the element arranged at the 10 th position in the human body, and about 65 percent of magnesium exists in bones and teeth. Studies have shown magnesium (Mg) ²⁺) Not only can induce vascular endothelial cells to generate nitric oxide and enhance the migration and proliferation of the endothelial cells; and can mobilize Endothelial Progenitor Cells (EPCs), increase VEGF production, promote the formation of new blood vessels, and exert important physiological functions in maintaining vascular function (Katakawa M, Fukuda N, Tsunemi A, Mori M, Maruyama T, Matsumoto T, et al. tau and magnesium fertilization enhancement processing of end gene cells through antibiotic oxidation in biochemical and specific biological high tension rates. Hypertension Res 2016; 39:848-56.), (Cooke JP, Losordo DW. nucleic acid oxide and angiogenesis. circulation 2002; 105: 2133-5.). Further, magnesium (Mg) ²⁺) Can also regulate osteoblast proliferation, differentiation and morphogenesis, promote bone formation (Zhang Y, Xu J, Ruan YC, Yu MK, O' Laughlin M, Wise H, et al, Implant-derived magnesium induced local production of CGRP to animal bone-derived healing induction, Nature media 2016; 22: 1160-9.); however, there is no report on the application of both Sr and Mg to bone repair materials.

The prior biological ceramic scaffold materials have the characteristics of weak mechanical strength, fracture toughness, high brittleness and the like, so that the clinical application of the materials is limited. Therefore, the multifunctional composite scaffold material begins to be widely applied to the basic research and clinical application of tissue repair. Especially, the polymer-bioactive ceramic composite material has the advantages of all components, so that the composite scaffold material has good clinical application prospects in the aspects of tissue compatibility, mechanical strength, bone conduction and bone induction, and becomes a hotspot of bone tissue engineering research and a new clinical treatment means.

Therefore, a new bone repair composite scaffold material needs to be developed to meet the application requirements.

Disclosure of Invention

The composite scaffold material provided by the invention has excellent biocompatibility and degradability, strontium (Sr) ions and magnesium (Mg) ions released by being transplanted into a body have the activity of promoting osteogenesis and vascularization, and degradation products of the composite scaffold material can also promote the proliferation, differentiation and mineralization of osteoblasts, inhibit the activity of osteoclasts and have the function of resisting osteoporosis, so the composite scaffold material provided by the invention has great application potential in the aspect of preparing bone defect repair scaffolds, particularly osteoporosis bone defect repair scaffolds.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides a magnesium strontium-phosphate silicate material having the following structural formula:

Sr _5-xMg _x(PO ₄) ₂SiO ₄，0<x<5。

the 0< x <5 can be 0.01, 0.05, 0.1, 0.2, 0.4, 0.5, 0.8, 1, 1.2, 1.4, 1.5, 1.8, 2.0, 2.2, 2.5, 2.8, 3.0, 3.2, 3.4, 3.5, 3.8, 4.0, 4.2, 4.5, 4.8, 4.9, and the like.

In the magnesium strontium-phosphate silicate material provided by the invention, the magnesium ions and the strontium ions have a synergistic interaction effect, and the synergistic interaction of the magnesium ions and the strontium ions can promote the proliferation and differentiation of osteoblasts, is beneficial to the functions of osteogenesis and angiogenesis in the bone repair process, and shortens the bone defect repair time.

And, compared to the use of Sr alone ₅(PO ₄) ₂SiO ₄And Mg ₅(PO ₄) ₂SiO ₄Magnesium ion and strontium ion Sr as provided herein _5-xMg _x(PO ₄) ₂SiO ₄The bone repair liquid has better effect of promoting the proliferation and differentiation of osteoblasts for bone repair.

In a second aspect, the present invention provides a method for preparing a magnesium strontium-silicon phosphate salt material according to the first aspect, the method comprising the steps of:

(1) mixing a mixed solution of an Sr source and an Mg source with a mixed solution of a P source and an Si source, aging, reacting, and then carrying out polycondensation reaction to obtain gel;

(2) and drying the gel to obtain the magnesium strontium-silicon phosphate salt material.

Preferably, the molar ratio of the Sr source and Mg source is (5-x): x, 0< x <5, e.g. 4.99:0.01, 4.5:0.5, 4:1, 3:2, 2:3, 1:4, 0.5:4.5, 0.01: 4.99.

Preferably, the solvent of the mixed solution of the Sr source and the Mg source is double distilled water.

Preferably, the Sr source is selected from strontium nitrate; the Mg source is selected from magnesium nitrate hexahydrate.

Preferably, the molar ratio of the P source to the Si source is 2: 1.

Preferably, the source of P is selected from triethyl phosphate and the source of Si is selected from ethyl orthosilicate.

Preferably, the mixing time is 1-3h, such as 1.5h, 1.8h, 2.0h, 2.5h, etc.

Preferably, the aging is at room temperature for a period of 24-30 hours, e.g., 25 hours, 28 hours, 29 hours, etc.

Preferably, the reaction is carried out at a temperature of 80-90 deg.C, e.g., 82 deg.C, 85 deg.C, 88 deg.C, etc., for a period of 16-20 hours, e.g., 17 hours, 18 hours, 19 hours, etc.

Preferably, the polycondensation is carried out at a temperature of 75 to 85 ℃, e.g., 78 ℃, 80 ℃, 82 ℃, etc., for a period of 5 to 10 hours, e.g., 6 hours, 7 hours, 8 hours, 9 hours, etc.

Preferably, the drying is performed by first drying at 100-120 deg.C (e.g., 105 deg.C, 110 deg.C, 115 deg.C) for 5-8h (e.g., 6h, 7h, etc.), and then drying at 850-900 deg.C (e.g., 860 deg.C, 870 deg.C, 880 deg.C, 890 deg.C, etc.) for 2-3h (e.g., 2.2h, 2.5h, 2.8h, etc.).

The invention synthesizes the biological ceramic material containing magnesium ions and strontium ions by using a sol-gel method.

In a third aspect, the invention provides an application of the magnesium strontium-phosphate silicate material according to the first aspect in preparing a bone repair composite scaffold material.

In a fourth aspect, the invention provides a structure-controllable porous bone repair composite scaffold material, which comprises polyhydroxyalkanoate and the magnesium strontium-phosphate silicate material of the first aspect.

The polyhydroxyalkanoate used in the invention has good biocompatibility and degradability, the degradation product 3-hydroxybutyric acid (3HB) is one of the main components of ketone bodies in mammals, has no toxic effect on organisms, can promote the proliferation, differentiation and mineralization of osteoblasts, can inhibit the activity of osteoclasts, and has the effect of resisting osteoporosis.

According to the invention, the polyhydroxyalkanoate and the magnesium strontium-phosphate silicate material are compounded, so that the advantages of the biological ceramic scaffold material are ensured, and meanwhile, the mechanical property of the biological ceramic scaffold material is overcome by the polyhydroxyalkanoate, so that the composite scaffold material provided by the invention has good biocompatibility, can promote the activity of osteogenesis and angiogenesis, has good mechanical strength, and can meet the application requirements.

Preferably, the mass ratio of the polyhydroxyalkanoate to the magnesium strontium-silicon phosphate salt material is (0.5-5):1, such as 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, etc., preferably (1-3): 1.

Preferably, the polyhydroxyalkanoate is selected from any one of poly-3-hydroxybutyrate, poly-3-hydroxyvalerate, poly-3-hydroxyhexanoate, 3-hydroxybutyrate-3-hydroxyvalerate or 3-hydroxybutyrate-3-hydroxyhexanoate copolyester or a combination of at least two thereof.

The selected polyhydroxyalkanoate has good mechanical strength, and when the polyhydroxyalkanoate is applied to a bone repair composite scaffold material, the polyhydroxyalkanoate has good mechanical strength and good application effect.

Preferably, the porosity of the structurally controllable porous bone repair composite scaffold material is 60-80%, such as 62%, 64%, 66%, 68%, 70%, 72%, 75%, 78%, etc.

Preferably, the pore connectivity of the structurally controllable porous bone repair composite scaffold material is 60-90%, such as 62%, 64%, 66%, 68%, 70%, 72%, 75%, 78%, 80%, 82%, 85%, 88%, and the like.

Preferably, the pore diameter of the porous bone repair composite scaffold material with controllable structure is 200-400 μm, such as 220 μm, 250 μm, 280 μm, 300 μm, 320 μm, 350 μm, 380 μm and the like.

In a fifth aspect, the invention provides a method for preparing the porous bone repair composite scaffold material with the controllable structure according to the fourth aspect, wherein the method for preparing the porous bone repair composite scaffold material comprises the following steps:

(1) mixing the polyhydroxyalkanoate solution with the magnesium strontium-silicon phosphate salt material to obtain a solution to be printed;

(2) printing by using a low-temperature 3D printer, and then carrying out freeze drying to obtain the porous bone repair composite scaffold material with the controllable structure.

Preferably, the solvent of the polyhydroxyalkanoate solution is 1, 4-dioxane and/or chloroform.

Preferably, the concentration of the polyhydroxyalkanoate in the polyhydroxyalkanoate solution is 1-2g/mL, such as 1.1g/mL, 1.2g/mL, 1.4g/mL, 1.6g/mL, 1.8g/mL, and the like.

Preferably, the parameters of the printing are: nozzle diameter 50 μm, nozzle moving speed 10mm/s, spinning speed 25mm ³(s) print size 20X 30 mm.

The structure of the porous bone repair composite scaffold material can be controlled by controlling the printing parameters of the low-temperature printer.

Because the macrostructure of the bone repair composite scaffold can be controlled by 3D printing, the "structurally controllable" of the present invention means that its porosity, pore size, and pore connectivity are controllable.

Preferably, prior to said freeze-drying, it is first frozen at a temperature lower than-20 ℃ (which may be-25 ℃, -30 ℃, -40 ℃, -50 ℃ and the like) for at least 6h (which may be 6.5h, 7h, 8h, 10h and the like).

In a sixth aspect, the invention provides the application of the porous bone repair composite scaffold material with a controllable structure according to the fourth aspect in preparing a bone defect filling material or a bone defect repair material.

Preferably, the bone defect is an osteoporotic bone defect.

Compared with the prior art, the invention has the following beneficial effects:

(1) in the magnesium strontium-phosphate silicate material provided by the invention, the magnesium ions and the strontium ions have a synergistic interaction effect, and the synergistic interaction of the magnesium ions and the strontium ions can promote the proliferation and differentiation of osteoblasts, is beneficial to the functions of osteogenesis and angiogenesis in the bone repair process, and shortens the bone defect repair time.

(2) According to the invention, the polyhydroxyalkanoate and the magnesium strontium-phosphate silicate material are compounded, so that the advantages of the biological ceramic scaffold material are ensured, and meanwhile, the mechanical property of the biological ceramic scaffold material is overcome by the polyhydroxyalkanoate, so that the composite scaffold material provided by the invention has good biocompatibility, can promote the activity of osteogenesis and angiogenesis, has good mechanical strength, and can meet the application requirements.

Drawings

FIG. 1 is an XRD pattern of the magnesium strontium-silicon phosphate salt materials provided in examples 1-3.

Fig. 2A is an SEM image of the magnesium strontium-silicon phosphate salt material provided in example 1.

Fig. 2B is an SEM image of the magnesium strontium-silicon phosphate salt material provided in example 2.

Fig. 2C is an SEM image of the magnesium strontium-silicon phosphate salt material provided in example 3.

Figure 3A is an EDS plot of the magnesium strontium-silicon phosphate salt material provided in example 1.

Figure 3B is an EDS plot of the magnesium strontium-silicon phosphate salt material provided in example 2.

Figure 3C is an EDS plot of the magnesium strontium-silicon phosphate salt material provided in example 3.

FIG. 4 is an apparent topography of the porous bone repair composite scaffold provided in example 1

FIG. 5A is a graph showing the results of the samples provided in examples 1 to 3 and comparative example 1 for enhancing the osteogenic differentiation capacity of mouse BMSCs (ALP expression staining activity).

FIG. 5B is a graph showing the results of the samples provided in examples 1-3 and comparative example 1 for enhancing osteogenic differentiation of mouse BMSCs (extracellular calcium nodule alizarin red staining).

FIG. 6 is a graph of the ability of the samples provided in examples 1-3 and comparative example 1 to induce HUVEC 6h microvascular formation in vitro.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

Preparation example 1

A magnesium strontium-silicon phosphate salt material has the following structural formula: sr _4.99Mg _0.01(PO ₄) ₂SiO ₄(x ═ 0.01), the preparation method was as follows:

(1) sr source (Sr (NO) ₃) ₂) With a source of Mg (NO) ₃) ₂·6H ₂O) is evenly mixed and dissolved in ddH according to the proportion of 4.99:0.01 ₂O is in;

(2) p source (C) ₆H ₁₅O ₄P) and Si source (TEOS, Si (OC) ₂H ₅) ₄) Uniformly dissolving in ddH according to the ratio of 2:1 ₂O is in;

(3) slowly adding the mixed solution of the Sr/Mg source into the stirred P/Si source solution, continuously stirring for 1h, and aging for 24h at room temperature;

(4) refluxing the aged solution at 85 ℃ for 16h to promote ionic groups to be bonded with each other to form sol, evaporating the sol at 80 ℃ for 8h, and performing polycondensation to form gel;

(5) and (3) drying the gel in an oven at 100 ℃ for 8h, and heating at 900 ℃ for 2h to obtain the magnesium strontium-silicon phosphate salt material.

Preparation examples 2 to 5

The difference from preparation example 1 is only that the molar ratio of the Sr source and the Mg source is controlled so that x of the finally obtained magnesium strontium-phosphosilicate material is 0.03 (preparation example 2), 0.05 (preparation example 3), 2.5 (preparation example 4), and 4.99 (preparation example 5).

Comparative preparation example 1

A strontium-silicon phosphate salt material having the formula:

Sr ₅(PO ₄) ₂SiO ₄

the preparation method comprises the following steps:

(1) adding 0.5M Sr source (Sr (NO) ₃) ₂) 0.2M P source (C) ₆H ₁₅O ₄P) and 0.1M Si source (Si (OC) ₂H ₅) ₄) Respectively dissolved in ddH ₂O; 0.2M of cetyltrimethylammonium bromide (CTAB), a cationic surfactant, was dissolved in ddH ₂In O, treating for 10min by ultrasonic wave to form a uniform solution;

(2) in the preparation process, a 0.2M P source prepared initially is added into a CTAB solution of 0.2M, and the mixture is magnetically stirred for 30 min; then slowly dripping 0.1M Si source solution into the solution; 0.5M Sr source solution is dripped into the P and Si-CTAB solution, and the mixture is continuously stirred to form viscous gel solution; then, ammonia water is dripped into the gel solution, the pH value of the solution is adjusted to 10, and the solution is stirred for 4 hours at room temperature;

(3) placing the white gel precipitate at room temperature for 24h, and then placing the white gel precipitate in an oven to bake at 110 ℃ for 12 h;

(4) the above dry mixture was reused in ddH ₂Washing with water, removing residual ammonia water, and drying at 100 deg.C for 2 hr; and drying the dry powder at 900 ℃ and 1000 ℃ for 3h and 5h respectively to obtain pure SPS powder.

Comparative preparation example 2

A magnesium-silicon phosphate salt material having the formula:

Mg ₅(PO ₄) ₂SiO ₄

the preparation method comprises the following steps:

(1) mixing 0.5M Mg source (Mg (NO) ₃) ₂) 0.2M P source (C) ₆H ₁₅O ₄P) and 0.1M Si source (Si (OC) ₂H ₅) ₄) Respectively dissolved in ddH ₂O; 0.2M of cetyltrimethylammonium bromide (CTAB), a cationic surfactant, was dissolved in ddH ₂In O, treating for 10min by ultrasonic wave to form a uniform solution;

(2) in the preparation process, a 0.2M P source prepared initially is added into a CTAB solution of 0.2M, and the mixture is magnetically stirred for 30 min; then slowly dripping 0.1M Si source solution into the solution; dripping 0.5M Mg source solution into the P and Si-CTAB solution, and continuously stirring to form viscous gel solution; then, ammonia water is dripped into the gel solution, the pH value of the solution is adjusted to 10, and the solution is stirred for 4 hours at room temperature;

(4) the above dry mixture was reused in ddH ₂Washing with water, removing residual ammonia water, and drying at 100 deg.C for 2 hr; drying the above dry powder at 900 deg.C and 1000 deg.C for 3 hr and 5 hr respectively to obtain pure Mg ₅(PO ₄) ₂SiO ₄And (3) powder.

Performance test 1

The samples provided in preparations 1 to 5 and comparative preparations 1 to 2 were subjected to the performance test as follows:

(1) XRD analysis: analyzing the phase composition of the synthesized material by using an X-ray diffractometer to determine whether the elemental composition of the material is correct;

FIG. 1 is an XRD pattern of the MgSr-SiPO 4 salt materials provided in examples 1-3, which shows that the MgSr-SiPO 4 salt materials are successfully prepared by the present invention.

(2) Microscopic morphology: analyzing the microscopic morphology of the sample by using a Scanning Electron Microscope (SEM);

FIGS. 2A-2C are SEM images of the magnesium strontium-silicon phosphate salt materials provided in examples 1-3, showing that magnesium strontium-silicon phosphate salt materials with different Mg contents have different micro-morphologies.

(3) The elements comprise: the elemental composition of the samples was examined using EDS.

FIG. 3 is an EDS chart of the MgSr-SiPO 4 materials provided in examples 1-3, which shows that the samples provided by the present invention have both Mg and Sr.

Examples 1 to 5

This example provides a porous bone repair composite scaffold material comprising poly-3-hydroxybutyrate and the magnesium strontium phosphate silicate material (Sr) provided in preparation examples 1-5 _5-xMg _x(PO ₄) ₂SiO ₄) And (4) forming.

The preparation method comprises the following steps:

(1) magnetically stirring and refluxing the poly-3-hydroxybutyrate and 1, 4-dioxane at 55 ℃ for 4h for dissolution to form a uniform and transparent solution; wherein the mass-to-volume (w/v) ratio of the poly-3-hydroxybutyrate to the 1, 4-dioxahexaane is 1: 6;

(2) adding magnesium strontium-phosphosilicate material powder into the solution formed in the step (1), and magnetically stirring for 2 hours at 37 ℃ to form a uniform solution; wherein the mass ratio of the poly-3-hydroxybutyrate to the magnesium strontium-phosphorus silicate material is 1: 1;

(3) adding the mixed solution obtained in the step (2) into 3D printing equipment, and printing the three-dimensional support; wherein the printing parameters are nozzle diameter of 50 μm, nozzle moving speed of 10mm/s, and spinning speed of 25mm ³(s) print size 20X 30 mm;

(4) and (3) putting the 3D printed composite scaffold into a low-temperature refrigerator at the temperature of-40 ℃ for 8h, and carrying out vacuum drying for a week to obtain the porous bone repair composite scaffold material.

Examples 6 to 10

The difference from example 1 is that in this example, the mass ratio of poly-3-hydroxybutyrate to magnesium strontium-phosphosilicate material was 0.5:1 (example 6), 5:1 (example 7), 3:1 (example 8), 0.3:1 (example 9), 7:1 (example 10).

Examples 11 to 13

The difference from example 1 is that in this example, poly-3-hydroxybutyrate was replaced with poly-3-hydroxyvalerate (example 11), 3-hydroxybutyrate-3-hydroxyhexanoate copolyester (example 12), polylactic acid (example 13).

Comparative example 1

The difference from example 1 is that in this comparative example the magnesium strontium-phosphosilicate material was replaced by the strontium-phosphosilicate material provided in comparative preparation example 1.

Comparative example 2

The difference from example 1 is that in this comparative example, the magnesium strontium-phosphosilicate material was replaced with the magnesium phosphosilicate material provided in comparative preparation example 2.

Comparative example 3

The difference from example 1 is that in this comparative example the magnesium strontium-phosphorus silicate material was replaced by a combination of a strontium-phosphorus silicate material and a magnesium-phosphorus silicate material, wherein the molar ratio of the two was 4.99: 0.01.

Comparative example 4

The difference from example 1 is that in this comparative example, no magnesium strontium-phosphosilicate material is added.

Comparative example 5

The difference from example 1 is that in this comparative example, no poly-3-hydroxybutyrate ester was added.

Performance test 2

The samples provided in examples 1-13 and comparative examples 1-5 were tested for performance by the following method:

(1) apparent morphology: and observing the apparent appearance.

Fig. 4 is an apparent topography of the porous bone repair composite scaffold provided in example 1, which shows that the composite scaffold provided by the present invention is porous.

(2) Osteogenic differentiation capacity:

a: soaking the sterilized 3D composite scaffold in Simulated Body Fluid (SBF) according to the mass-volume ratio of 1:10, sealing, standing in an incubator at 37 ℃, and collecting leaching liquor after soaking for 4 weeks;

b: uniformly mixing the leaching liquor and osteogenic induction liquid according to the volume ratio of 1: 3;

c: dropping the above inducing liquid to a container containing 1X 10 per well ⁵Changing liquid once every three days in bone marrow mesenchymal stem cells (BMSCs), and carrying out induced culture for 14 days;

d: removing the inducing solution, rinsing with PBS three times, and fixing with 4% paraformaldehyde for 15 min;

e: removing the paraformaldehyde, rinsing with PBS for three times, and adding 1mL alizarin red solution for dyeing for 15 min;

f: removing the alizarin red solution, rinsing with PBS for three times, and adding 0.5mL of 10% cetyl pyridinium chloride solution into the alizarin red solution for extraction;

g: measuring the absorbance of the solution by enzyme-labeling instrument at 562nm wavelength to quantify osteogenic differentiation capacity

Fig. 5A and 5B are graphs showing the results of the samples provided in examples 1 to 3 and comparative example 1 on the enhancement of the osteogenic differentiation capacity of mouse BMSCs, wherein fig. 5A shows the ALP expression staining activity, and fig. 5B shows the extracellular calcium nodule alizarin red staining, and it can be seen from the graphs that the osteogenic capacity is enhanced with the increase of the Mg content.

(3) Microvascular formation ability:

b: uniformly mixing the leaching liquor and the angiogenesis medium according to the volume ratio of 1: 3;

c: the above medium was added dropwise to a mixture containing 1X 10 of the medium per well ⁴Culturing Human Umbilical Vein Endothelial Cells (HUVEC) at 37 deg.C for 8 hr;

d: the amount of microvessels formed was observed and photographed with an optical microscope (magnification 10).

FIG. 6 is a graph of the analysis of HUVEC 6h microvascular formation induced in vitro by the samples provided in examples 1-3 and comparative example 1, and it can be seen that the vascularization ability is enhanced with the increase of Mg content.

(4) Modulus of elasticity: the center of the stent material was compressed at a rate of 10mm/min using a mechanical tester, and the elastic modulus value was recorded at a load of 5mm diameter area.

The results of the tests on the examples and comparative examples are shown in Table 1:

TABLE 1

The embodiment and performance tests show that the porous bone repair composite scaffold material provided by the invention has good biocompatibility, can promote the activity of osteogenic angiogenesis, has good mechanical strength and can meet the application requirements.

As can be seen from the comparison between example 1 and comparative examples 1-2, magnesium ions and strontium ions have synergistic interaction, and the synergistic interaction of the magnesium ions and the strontium ions can promote the proliferation and differentiation of osteoblasts, is beneficial to the functions of osteogenesis and angiogenesis in the bone repair process, shortens the bone defect repair time, and only promotes bone or angiogenesis and cannot form a coupling process if magnesium ions or strontium ions are used alone; from the comparison of example 1 and comparative example 3, only magnesium ions and strontium ions are represented by Sr _5-xMg _x(PO ₄) ₂SiO ₄The form of the compound is more beneficial to the proliferation and differentiation of osteoblasts, the osteogenic and angiogenisis functions in the bone repair process, and the bone defect repair time is shortened; it is clear from the comparison of example 1 with comparative examples 4 to 5 that the absence of magnesium strontium phosphosilicate material and polyhydroxyalkanoates leads to a decrease in osteogenic differentiation capacity on the one hand and a lower elastic modulus and poorer mechanical properties on the other hand.

The applicant states that the magnesium strontium phosphate silicate material and the preparation method thereof, and the structure-controllable porous bone repair composite scaffold material containing the same are illustrated by the above examples, but the present invention is not limited to the above process steps, i.e. the present invention does not mean that the present invention must rely on the above process steps to be implemented. It will be apparent to those skilled in the art that any modification of the present invention, equivalent substitutions of selected materials and additions of auxiliary components, selection of specific modes and the like, which are within the scope and disclosure of the present invention, are contemplated by the present invention.

Claims

1. The magnesium strontium-silicon phosphate salt material is characterized by having the following structural formula:

Sr _5-xMg _x(PO ₄) ₂SiO ₄，0<x<5。

2. the method for preparing a magnesium strontium-silicon phosphate salt material according to claim 1, comprising the steps of:

3. The method according to claim 2, wherein the molar ratio of the Sr source to the Mg source is (5-x) x, 0< x < 5;

preferably, the solvent of the mixed solution of the Sr source and the Mg source is double distilled water;

preferably, the Sr source is selected from strontium nitrate and the Mg source is selected from magnesium nitrate hexahydrate;

preferably, the molar ratio of the P source to the Si source is 2: 1;

preferably, the source of P is selected from triethyl phosphate and the source of Si is selected from ethyl orthosilicate;

preferably, the mixing time is 1-3 h;

preferably, the aging temperature is room temperature, and the aging time is 24-30 h;

preferably, the reaction temperature is 80-90 ℃ and the reaction time is 16-20 h;

preferably, the temperature of the polycondensation is 75-85 ℃ and the time is 5-10 h;

preferably, the drying is first drying at 100-120 ℃ for 5-8h, and then drying at 850-900 ℃ for 2-3 h.

4. Use of the magnesium strontium-phosphate silicate material according to claim 1 in the preparation of a bone repair composite scaffold material.

5. A structurally controlled porous bone repair composite scaffold material comprising a polyhydroxyalkanoate and the magnesium strontium-silicon phosphate salt material of claim 1.

6. The structurally controlled porous bone repair composite scaffold material according to claim 5, wherein the mass ratio of polyhydroxyalkanoate to the magnesium strontium phosphate silicate material is (0.5-5):1, preferably (1-3): 1;

7. The structurally controllable porous bone repair composite scaffold material according to claim 5 or 6, wherein the porosity of the structurally controllable porous bone repair composite scaffold material is 60-80%;

preferably, the pore communication rate of the porous bone repair composite scaffold material with the controllable structure is 60-90%;

preferably, the pore diameter of the porous bone repair composite scaffold material with the controllable structure is 200-400 μm.

8. The method for preparing a structurally controllable porous bone repair composite scaffold material according to any one of claims 5 to 7, comprising the steps of:

9. The method according to claim 8, wherein the solvent of the polyhydroxyalkanoate solution is 1, 4-dioxahexaalkane and/or trichloromethane;

preferably, the concentration of the polyhydroxyalkanoate in the polyhydroxyalkanoate solution is 1-2 g/mL;

preferably, the parameters of the printing are: nozzle diameter 50 μm, nozzle moving speed 10mm/s, spinning speed 25mm ³(s) print size 20X 30 mm;

preferably, prior to said freeze-drying, it is first frozen at a temperature below-20 ℃ for at least 6 h.

10. Use of a structurally controllable porous bone repair composite scaffold material according to any one of claims 5-7 for the preparation of a bone defect filling material or a bone defect repair material;

preferably, the bone defect is an osteoporotic bone defect.