CN112794732B - Calcium silicate ceramic with surface modified by microbial mineralization and application thereof - Google Patents
Calcium silicate ceramic with surface modified by microbial mineralization and application thereof Download PDFInfo
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Abstract
The invention relates to a calcium silicate ceramic with a microorganism mineralization modified surface and application thereof2The mineralization time is controlled to be 48-72 hours, and the content of the initial strain is 7 multiplied by 1012～2.1×1013CFU/ml, and forming a mineralized layer with a micro-nano structure on the surface of the calcium silicate ceramic.
Description
Technical Field
The invention relates to calcium silicate ceramic with a microbial mineralization micro-nano structure formed on the surface and capable of promoting bone activity and application thereof, belonging to the field of biomedical materials.
Background
The artificially synthesized calcium silicate ceramic is a degradable material with good biological activity, but because the degradable material has high degradation speed and lacks biocompatibility, the material is not beneficial to the adhesion, proliferation and differentiation of cells on the surface of the material, and therefore, the material lacks good osteogenesis inducing activity [ biomaterials.2008; 29:4392]. Research shows that the construction of a micro-nano structure on the surface of a material can obviously improve the performances of cell adhesion, proliferation and osteogenic differentiation, and further improve the osteoinductive activity of the material [ Acta biomaterials.2012; 8:3794]. Calcium carbonate is the most widely-occurring biomineralization product in nature, is also one of the main components of human bone tissues, and has natural biocompatibility, biodegradability and osteoconductivity [ chem.commun.2010; 46:6578]. However, in a solution with a given concentration, calcium carbonate crystals grow rapidly and form a firm adhesive force with a substrate material difficultly, and the construction of a micro-nano calcium carbonate structure on the surface of a ceramic material still faces a challenge. Therefore, the development of a simple, feasible and low-cost method for preparing the calcium carbonate micro-nano structure on the surface of the calcium silicate ceramic so as to have good induced osteogenesis activity has important significance and application value.
Disclosure of Invention
The invention mainly aims to provide a calcium silicate biological ceramic with a functionalized micro-nano structure constructed on the surface through microbial mineralization.
The present inventors have conducted extensive and intensive studies in order to achieve the above object. Urease-producing bacteria (such as Staphylococcus cohnii) can slowly decompose substrate urea to produce NH3The formation of pH and CO around the bacteria3 2+An increased microenvironment. The cell wall contains a large amount of electronegative groups which can be connected with Ca in the environment2+Interactions take place to participate in the nucleation and growth of calcium carbonate [ Earth-sci. rev.2015; 148:1]. But at present, the micro-nano calcium silicate ceramic on the surface constructed by the assistance of microorganisms is lack of research. The present inventors have intensively studied and found that the above problems can be solved by the invention described below, thereby completing the present invention.
The invention relates to a calcium silicate biological ceramic with a microorganism mineralization modified surface, which is characterized in that the calcium silicate biological ceramic with the microorganism mineralization modified surface is prepared by immersing calcium silicate ceramic into a solution containing urease-producing bacteria, urea and CaCl2The mineralization time is controlled to be 48-72 hours, and the initial strain content (namely the initial content of the urease producing bacteria in the culture medium solution) is 7 multiplied by 1012～2.1×1013CFU/ml, and forming a mineralized layer with a micro-nano structure on the surface of the calcium silicate ceramic.
The invention relates to a micro-nano calcium silicate biological ceramic with a surface constructed by the assistance of microorganisms, which is prepared by immersing calcium silicate ceramic into a solution containing urease-producing bacteria, urea and CaCl2In the culture medium solution, the mineralization time and the strain content are regulated and controlled, so that the microorganisms gradually form a micro-nano calcium carbonate structure on the surface of the calcium silicate ceramic through mineralization, the micro-nano topological morphology and the calcium carbonate components effectively stimulate cell adhesion, proliferation and osteogenic differentiation, and the bionic interface with good biological activity is favorable for improving the osteogenic activity and the osseointegration performance of the implant. The calcium silicate biological ceramic with the micro-nano calcium carbonate structure formed on the surface can obviously promote the adhesion, proliferation and osteogenic differentiation of osteoblasts and mesenchymal stem cells, and can be applied to the field of hard tissue defect repair.
Preferably, the urease-producing bacteria are Staphylococcus cohnii (Staphylococcus cohnii), and the urease activity of the bacteria is moderate, and calcium carbonate particles with micro-nano scale can be induced.
The thickness of the mineralized layer may be 1-5 μm.
The mineralized layer is provided with a protruding part, the width of the protruding part can be 50-100 nm, and the height of the protruding part can be 50-100 nm.
The mineralized calcium silicate ceramic can be cleaned, sterilized at high temperature and high pressure and dried in the air to obtain the microbial mineralized layer. After the calcium silicate ceramic is mineralized and modified by microorganisms, a micro-nano calcium carbonate structure is formed on the surface of the calcium silicate ceramic, and a good morphology structure is still maintained after high-temperature and high-pressure sterilization.
The calcium silicate in the calcium silicate ceramic can be alpha-CaSiO3。
The calcium silicate ceramic may be a three-dimensional printed ceramic scaffold.
The calcium silicate ceramic can be prepared by the following method:
reacting beta-CaSiO3Mixing the powder with 3D printing resin, preparing photocuring printing slurry, and performing photocuring printing to obtain a biscuit body; placing the biscuit inAnd preserving the heat for 2-5 hours at the temperature of 1200-1300 ℃ to obtain the calcium silicate biological ceramic.
The calcium silicate ceramic can be prepared by the following method:
reacting beta-CaSiO3Mixing the powder with a binder, and performing dry pressing to obtain a biscuit;
and (3) preserving the heat of the biscuit at the temperature of 1200-1300 ℃ for 2-5 hours to obtain the calcium silicate biological ceramic.
The calcium silicate biological ceramic with the surface modified by the microbial mineralization can be applied to the preparation of materials for repairing hard tissue defects.
Compared with the traditional calcium silicate ceramic material with a flat plate structure, the calcium silicate biological ceramic with the surface micro-nano structure has the advantages that the biological effects of spreading, proliferation, osteogenic differentiation and the like of mesenchymal stem cells of bone marrow are better promoted, and the in-vivo osteogenic activity is promoted, so that the calcium silicate biological ceramic is more suitable for repairing large bone defects. In addition, the preparation process for constructing the surface micro-nano calcium silicate ceramic with the assistance of the microorganisms is simple and easy to implement, low in cost and convenient to popularize, can be completed in a common environment, and does not need expensive equipment.
Drawings
FIG. 1 is a SEM scan of the surface morphology of samples from different mineralizing solutions of example 1 (CS:2D α -CS ceramic soaked in 2% urea, 15mM CaCl2The mineralized liquid is added for 72 hours; soaking the M-CS:2D alpha-CS ceramic in 2% urea and 15mM CaCl2And the sample after 72 hours in the mineralized liquid of the culture medium; soaking CBM-CS 2D alpha-CS ceramic in 7X 1012CFU/ml Staphylococcus cohnii, 2% urea, 15mM CaCl2Samples 72h after being mixed with the mineralized liquid of the culture medium);
FIG. 2 is an XRD pattern of α -CS ceramics obtained in different mineralizing solutions of example 1;
FIG. 3 is a SEM scan of the morphology of cells on the surface of three 2D α -CS ceramic plates of example 1;
FIG. 4 is a graph showing the proliferation effect of bone marrow stromal stem cells on three 2D α -CS ceramic sheets of example 1, respectively;
FIG. 5 shows the differentiation of bone marrow stromal stem cells on CS, M-CS and CBM-CS of example 1;
FIG. 6 shows different mineralizing solutions of example 1 (CS: 3D. alpha. -CS ceramic soaked in 2% Urea, 15mM CaCl)2The mineralized liquid is added for 72 hours; soaking the M-CS:3D alpha-CS ceramic in 2% urea and 15mM CaCl2And the sample after 72 hours in the mineralized liquid of the culture medium; soaking CBM-CS 3D alpha-CS ceramic in 1.4X 1013CFU/ml Staphylococcus cohnii, 2% urea, 15mM CaCl2Sample 72h after in the mineralized liquid with the culture medium) optical, SEM pictures of the prepared 3D scaffold;
FIG. 7 is an XRD pattern of the 3D α -CS ceramic obtained in different mineralizing solutions of example 1;
FIG. 8 is a SEM scan of the morphology of cells on the surface of three 3D α -CS ceramic plates in example 1;
FIG. 9 shows the proliferation effect of bone marrow stromal stem cells on three 3D α -CS ceramic sheets of example 1, respectively;
FIG. 10 is an optical photograph and a CT scan three-dimensional photograph of different animal experiments of example 1 in a rabbit femur experiment;
FIG. 11 is a statistical analysis of quantitative data for osteogenesis for the different components of example 1 in a rabbit femoral experiment;
FIG. 12 is a SEM scan of the surface morphology of the sample obtained in example 2;
fig. 13 is a SEM scan of the morphology of the sample surface obtained in example 3.
Detailed Description
The present invention is further illustrated by the following examples, which are to be understood as merely illustrative and not restrictive.
The invention relates to a micro-nano calcium silicate ceramic with a surface constructed by microorganism assistance, and a preparation method and application thereof, belonging to the field of biomedical materials. Immersing calcium silicate ceramic in the solution containing urease-producing bacteria, urea and CaCl2In the culture medium solution, the pH of a microenvironment and the concentration of carbonate are slowly improved by the ureolysis of microorganisms (Staphylococcus cohnii) to mineralize and synthesize the micro-nano biological calcium carbonate coating on the surface of the calcium silicate ceramic scaffold.
According to the surface micro-nano calcium silicate ceramic disclosed by the invention, a micro-nano calcium carbonate structure formed by microbial mineralization can obviously promote the adhesion, proliferation and osteogenic differentiation of osteoblasts and bone marrow mesenchymal stem cells, so that the osseointegration performance of a calcium silicate ceramic implant is improved. The invention has the characteristics of simple and easy process and convenient popularization. The preparation method of the calcium silicate bioceramic with the microorganism mineralization modified surface provided by the invention is exemplarily illustrated below.
Preparing calcium silicate ceramic. The calcium silicate ceramic may be formed in various shapes such as a two-dimensional ceramic sheet, a three-dimensional ceramic support, and the like. The calcium silicate in the calcium silicate ceramic can be alpha-CaSiO3(α -CS). The alpha-CS ceramic can be slowly degraded in a mineralized solution, and Si and Ca ions are released to form a calcium-enriched surface, so that the crystallization of calcium carbonate is facilitated. The disclosure may employ beta-CaSiO3(beta-CS) powder is used as raw material to prepare two-dimensional and three-dimensional alpha-CaSiO3(alpha-CS) ceramic as a base material. The preparation method comprises the following steps: reacting beta-CaSiO3Mixing the (beta-CS) powder with a binder, and performing dry pressing to obtain a biscuit; and (3) preserving the temperature of the biscuit at 1200-1300 ℃ for 2-5 hours to obtain the calcium silicate biological ceramic. The binder may be PVA (polyvinyl alcohol). In some embodiments, the polyvinyl alcohol aqueous solution with a concentration of 5 to 10 wt.% is added, and the addition amount may be 5 to 10 wt.% of the β -CS powder. The order of mixing is not particularly limited, and may be, for example, in the form of beta-CaSiO3And (beta-CS) powder is added with a binder and stirred uniformly. The ceramic biscuit can be obtained by dry-pressing the mixed powder with a certain mass under the pressure of 2-10 MPa. CaSiO when sintering at 1200-1300 DEG C3The crystal phase is formed by beta-CaSiO3To alpha-CaSiO3. In some embodiments, two-dimensional (2D) α -CS ceramic sheets are prepared using a biscuit made of β -CS powder and a binder.
The preparation method can also comprise the following steps: reacting beta-CaSiO3Mixing the powder with 3D printing resin, preparing photocuring printing slurry, and performing photocuring printing to obtain a biscuit body; and (3) preserving the heat of the biscuit at the temperature of 1200-1300 ℃ for 2-5 hours to obtain the calcium silicate biological ceramic. The addition amount of the 3D printing resin may be 100 wt.% of the β -CS powder. Can be prepared by mixing resin and beta-CaSiO3Photocuring printing prepared by mixing powder and solventAnd (3) slurry. The solids content of the slurry may be 40% to 70 wt.%. The order of mixing is not particularly limited. The 3D structure of the scaffold can be designed using 3D Max (Autodex) to obtain STL files, and the slurry is polymerized and cross-linked by a 3D printer at a setting of 50-150 μm thick per piece, exposure time 3-5s, light source wavelength 405 nm. CaSiO when sintering at 1200-1300 DEG C3The crystal phase is formed by beta-CaSiO3To alpha-CaSiO3. In some embodiments, a three-dimensional (3D) α -CS scaffold is prepared by photocuring printing using a β -CS paste incorporating a resin. In addition, commercially available α -CaSiO may be used3A ceramic.
And preparing a microbial mineralized surface micro-nano structure. Immersing calcium silicate ceramic (substrate material) in a solution containing urease-producing bacteria, urea and CaCl2The culture medium solution (microorganism mineralization solution) is added, mineralization is controlled for 48 to 72 hours, and the content of initial strains is 7 multiplied by 1012～2.1×1013CFU/ml, and forming a mineralized layer with a micro-nano structure on the surface of the calcium silicate ceramic. The urease producing bacteria can be Staphylococcus cohnii (Staphylococcus cohnii), and the bacteria have low ureolysis speed, so that the regulation and control of calcium carbonate in a micro-nano scale are facilitated. The mineralized solution of microorganisms may contain 7 × 1012～2.1×1013CFU/ml urease-producing bacteria, 1-5 wt.% urea and 10-50 mM CaCl21-5g/L peptone, 1-5g/L beef extract, 5-10g/L NaCl and initial pH of 7-8. Can be mineralized at normal temperature. In the mineralization process, the reaction time and the strain concentration can obviously influence the micro-nano morphology of the calcium carbonate on the surface of the material. The higher the concentration of the strain is, the more easily calcium carbonate crystals formed on the surface of the material are aggregated into large-size particles. Preferably, the initial content of the strains in the mineralized solution of the microorganisms is 7 multiplied by 1012～1.4×1013CFU/ml, thereby further forming a uniform micro-nano calcium carbonate structure. The mineralized calcium silicate ceramic can be cleaned, sterilized at high temperature and high pressure (134 ℃ for 20min) and dried to obtain the microbial mineralized layer. After the calcium silicate ceramic is mineralized and modified by microorganisms, a micro-nano calcium carbonate structure is formed on the surface of the calcium silicate ceramic, and a good morphology structure is still maintained after high-temperature and high-pressure sterilization.
In the embodiment, a uniform calcium carbonate micro-nano structure grows on the surface of the calcium silicate ceramic through the biomineralization of the urease-producing bacteria Staphylococcus cohnii. The calcium carbonate micro-nano structure is formed into a mineralized layer with a micro-nano structure. The mineralized layer has a uniform concave-convex structure. The mineralized layer is calcite type calcium carbonate. The thickness of the mineralized layer may be 1 to 5 μm, the average width of the protrusions is 50 to 100nm, and the average height is 50 to 100 nm. During the degradation process of calcium silicate in solution, a large amount of Si-OH is formed on the surface, which is beneficial to the nodule of calcium carbonate crystals.
According to the structure of the invention, a micro-nano calcium carbonate structure is constructed on the surface of calcium silicate ceramic through the mineralization of microorganisms, so that a bionic interface with good bioactivity is formed to improve the bone integration performance of the implant. The prepared calcium silicate ceramic with the surface micro-nano calcium carbonate structure can remarkably promote the adhesion, proliferation and osteogenic differentiation of osteoblasts and mesenchymal stem cells, and can be applied to the field of hard tissue defect repair and the like. The calcium silicate ceramic with the microbial mineralization micro-nano structure on the surface prepared by the method has the advantages of simple and feasible preparation process, low cost, convenience in popularization, capability of being finished in a common environment and no need of expensive equipment.
The present invention will be described in detail by way of examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art may be made in light of the above teachings. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values of the following examples;
in the following examples, reagents, materials and instruments used are all conventional reagents, conventional materials and conventional instruments, which are commercially available, if not specifically mentioned, and the reagents involved therein can also be synthesized by conventional synthesis methods.
Carrying out the process steps
2D α -CS ceramic preparation example:
weighing 0.1g of beta-CaSiO3The powder is prepared by uniformly stirring a PVA aqueous solution with the concentration of 10 wt.% of 6% (mass percentage) as a binder. And carrying out dry pressing molding on the mixed powder under the pressure of 10MPa to obtain the two-dimensional ceramic biscuit. Then the biscuit is insulated for 3 hours at the temperature of 1250 ℃ to obtain alpha-CaSiO3And (5) ceramic plates.
Preparation example of 3D α -CS ceramic:
weighing 50g of beta-CaSiO3The powder is ball-milled with 50g of 3D printing resin (WANHAO company) to obtain slurry, and a three-dimensional support biscuit is prepared by a photocuring printer of ten-dimensional science and technology limited company. Then the biscuit is preserved for 3 hours at the temperature of 1250 ℃ to obtain alpha-CaSiO3A ceramic;
immersing the sintered 2D and 3D alpha-CS ceramics into mineralized solution containing microorganisms (7 x 10)12-2.1×1013CFU/ml) and culturing for 48-72 h at normal temperature to obtain the alpha-CS ceramic with the micro-nano structure induced by the microorganisms on the surface. In addition, alpha-CS ceramics prepared from two mineralization solutions without bacteria were used as two control samples, urea and CaCl only2And containing urea and CaCl2And the mineralized liquid of the culture medium.
Evaluation of Performance
(1) Microbe-induced micro-nano structure evaluation
Scanning Electron Microscopy (SEM) is adopted to observe the surface appearance of the 2D and 3D alpha-CS ceramics with the surfaces having the micro-nano structures induced by microorganisms. Analyzing the mineral structure induced by the microorganism by using an X-ray diffraction pattern;
(2) biological evaluation:
research on the adhesion, proliferation and differentiation of a micro-nano structure induced by microorganisms of the alpha-CS ceramic to bone marrow mesenchymal stem cells from rabbits;
(3) in vivo evaluation:
the surface of the alpha-CS bracket with a micro-nano structure induced by microorganisms has osteogenic activity at the femoral part of a rabbit.
Example 1:
micro-nano induced by microorganismRice calcium carbonate modified 2D alpha-CS ceramics: preparing a microbial mineralization solution of 7 x 1012CFU/ml Staphylococcus cohnii, 2 wt.% urea, 15mM CaCl23g/L peptone, 1g/L beef extract, 5g/L NaCl, pH 7.2. And immersing the fired alpha-CS ceramic plate into the solution, and mineralizing for 72h at 37 ℃. Taking out the ceramic wafer, leaching, sterilizing at high temperature and high pressure, and naturally drying to obtain the microbial mineralized coating (figure 1). The surface was determined to be a uniform calcite relief (fig. 2), with an average width of the projections of about 50nm, an average height of about 100nm and a mineralized layer thickness of 1.5 μm.
Compared with the control group, the product has better cell expansion state and larger cytoskeleton (figure 3), the cell number is obviously increased after the cells are cultured for 1 day, 3 days and 7 days (figure 4), and the expression of genes such as Alpha v, COL 1, OPN, ALP and the like related to osteogenesis is increased (figure 5). Compared with a control group, the micro-nano structure induced by the microorganism has better biological characteristics of promoting cell adhesion, proliferation and osteogenic differentiation.
Microorganism-induced micro-nano calcium carbonate modified 3D alpha-CS ceramic: preparing the microbial mineralization solution into 1.4 multiplied by 1013CFU/ml Staphylococcus cohnii, 2% Urea, 15mM CaCl2Culture medium (3g/L peptone, 1g/L beef extract, 5g/L NaCl), pH 7.2. Immersing the fired alpha-CS ceramic bracket into the solution, and mineralizing for 72h at 37 ℃. And taking out the ceramic support, leaching, sterilizing at high temperature and high pressure, and naturally airing to obtain the microbial mineralized coating (figure 6). The surface was determined to be a uniform calcite relief (fig. 7), with an average width of the projections of about 100nm, an average height of about 100nm and a mineralized layer thickness of 1 μm.
The product cultured cells in a more expanded state and a larger cytoskeleton than the control group (FIG. 8). After 1 day, 3 days and 7 days of cell culture, the number of cells was significantly increased compared to the control group (fig. 9).
Compared with the control group, the alpha-CS bracket with the structure of the micro-nano calcium carbonate induced by the microorganism has better osteogenesis performance in the animal body (figure 10, figure 11).
FIG. 1 shows different mineralizing solutions of example 1 (CS: 2D. alpha. -CS ceramic soaked in 2% Urea, 15mM CaCl)2The mineralized liquid is added for 72 hours; soaking the M-CS:2D alpha-CS ceramic in 2% urea and 15mM CaCl2And the sample after 72 hours in the mineralized liquid of the culture medium; soaking CBM-CS 2D alpha-CS ceramic in 7X 1012CFU/ml Staphylococcus cohnii, 2% urea, 15mM CaCl2And 72h in the mineralized medium). As can be seen from the figure, the prepared microorganism-induced surface micro-nano structure has an obvious concave-convex structure;
figure 2 is an XRD pattern of the alpha-CS ceramic obtained in different mineralising solutions of example 1. As can be seen from the figure, the micro-nano structure formed on the surface of the alpha-CS ceramic by the prepared microorganism is calcite, while a control group does not form a new crystal;
FIG. 3 is a SEM scan of the morphology of cells on the surface of three 2D α -CS ceramic plates of example 1;
FIG. 4 is a graph showing the proliferation effect of bone marrow stromal stem cells on three 2D α -CS ceramic sheets of example 1, respectively;
FIG. 5 shows differentiation results of bone marrow stromal cells on CS, M-CS and CBM-CS of example 1, which indicates that calcium carbonate micro-nanostructure induced by microorganism improves osteogenic related gene expression of bone marrow stromal cells;
FIG. 6 shows different mineralizing solutions of example 1 (CS: 3D. alpha. -CS ceramic soaked in 2% Urea, 15mM CaCl)2The mineralized liquid is added for 72 hours; soaking the M-CS:3D alpha-CS ceramic in 2% urea and 15mM CaCl2And the sample after 72 hours in the mineralized liquid of the culture medium; soaking CBM-CS 3D alpha-CS ceramic in 1.4X 1013CFU/ml Staphylococcus cohnii, 2% urea, 15mM CaCl2And a sample after 72h in the mineralized solution in the medium) of the culture medium. As can be seen from the figure, the prepared microorganism-induced surface micro-nano structure has an obvious concave-convex structure;
figure 7 is an XRD pattern of the 3D α -CS ceramic obtained in different mineralising solutions of example 1. As can be seen from the figure, the micro-nano structure formed on the surface of the 3D alpha-CS ceramic by the prepared microorganism is calcite, while a control group does not form a new crystal;
FIG. 8 is a SEM scan of the morphology of cells on the surface of three 3D α -CS ceramic plates in example 1;
FIG. 9 shows the proliferation effect of bone marrow stromal stem cells on three 3D α -CS ceramic sheets of example 1, respectively;
FIG. 10 is an optical photograph and a CT scan three-dimensional photograph of different animal experiments of example 1 in a rabbit femur experiment;
fig. 11 is a statistical analysis of the quantitative data of osteogenesis for the different components of example 1 in a rabbit femoral experiment.
Example 2:
microorganism-induced micro-nano calcium carbonate modified 3D alpha-CS ceramic: preparing the microbial mineralization solution into 2.1 × 1013CFU/ml Staphylococcus cohnii, 2% Urea, 15mM CaCl23g/L peptone, 1gL beef extract, 5g/L NaCl, pH 7.2. And immersing the fired alpha-CS ceramic plate into the solution, and mineralizing for 72h at 37 ℃. And taking out the ceramic wafer, leaching, sterilizing at high temperature and high pressure, and naturally drying to obtain the microbial mineralized coating. The surface of the ground layer was determined to have poor morphological uniformity of the mineralized layer, about 100nm calcium carbonate particles, however, the formation of large calcium carbonate particles (about 10 μm) occurred (fig. 12).
Example 3:
microorganism-induced micro-nano calcium carbonate modified 2D alpha-CS ceramics: preparing a microbial mineralization solution of 7 x 1012CFU/ml Staphylococcus cohnii, 2% Urea, 15mM CaCl23g/L peptone, 1g/L beef extract, 5g/L NaCl, pH 7.2. And immersing the fired alpha-CS ceramic plate into the solution, and mineralizing for 48 hours at 37 ℃. And taking out the ceramic wafer, leaching, sterilizing at high temperature and high pressure, and naturally drying to obtain the microbial mineralized coating. The surface was determined to be a uniform mineralized layer with an average width of about 50nm and an average height of about 50nm (FIG. 13).
Claims (8)
1. The calcium silicate biological ceramic with the microbial mineralization modified surface is characterized in that the calcium silicate biological ceramic with the microbial mineralization modified surface is prepared by immersing calcium silicate ceramic into a solution containing urease-producing bacteria, 1-5 wt.% of urea and 10-50 mM CaCl2The initial pH value of the culture medium is 7-8, the mineralization time is controlled to be 48-72 hours, and the initial strain content is 7 multiplied by 1012～2.1×1013CFU/ml, and forming a calcium carbonate mineralized layer with a micro-nano structure on the surface of the calcium silicate ceramic.
2. The calcium silicate bioceramic of claim 1, wherein the urease producing bacteria are staphylococcus cohnii.
3. The calcium silicate bioceramic according to claim 1, wherein the calcium carbonate mineralized layer has a thickness of 1-5 μm.
4. The calcium silicate bioceramic according to claim 1, wherein the calcium carbonate mineralized layer has micro-nano particle protrusions, wherein the protrusions have a width of 50-100 nm and a height of 50-100 nm.
5. The calcium silicate bioceramic according to claim 1, wherein the calcium silicate ceramic is a three-dimensional printed ceramic scaffold.
6. The calcium silicate bioceramic according to any one of claims 1 to 5, wherein the calcium silicate ceramic is prepared by a method comprising:
reacting beta-CaSiO3Mixing the powder with 3D printing resin, preparing photocuring printing slurry, and performing photocuring printing to obtain a biscuit body;
and (3) preserving the heat of the biscuit at the temperature of 1200-1300 ℃ for 2-5 hours to obtain the calcium silicate biological ceramic.
7. The calcium silicate bioceramic according to any one of claims 1 to 4, wherein the calcium silicate ceramic is prepared by a method comprising:
reacting beta-CaSiO3Mixing the powder with a binder, and performing dry pressing to obtain a biscuit;
and (3) preserving the heat of the biscuit at the temperature of 1200-1300 ℃ for 2-5 hours to obtain the calcium silicate biological ceramic.
8. Use of the calcium silicate bioceramic with a microbial mineralization-modified surface according to any one of claims 1-7 in preparation of a material for repairing defects of hard tissues.
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