CN108355166B - Mesoporous bioactive glass/metal organic framework support material and preparation method thereof - Google Patents
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A—HUMAN NECESSITIES
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y50/00—Data acquisition or data processing for additive manufacturing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y70/00—Materials specially adapted for additive manufacturing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y80/00—Products made by additive manufacturing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Abstract
The invention discloses a mesoporous bioactive glass/metal organic framework composite scaffold material and a preparation method thereof. The preparation method comprises the steps of preparing the mesoporous bioactive glass/metal organic framework composite printing ink; designing the appearance and the internal structure of the bracket material by using CAD/CAM computer aided software; preparing the mesoporous bioactive glass/metal organic framework composite scaffold material by a low-temperature 3D printing technology. The mesoporous bioactive glass/metal organic framework composite scaffold material prepared by the invention has a three-dimensional communicated and controllable macroporous structure, high mechanical strength and good biocompatibility, can promote the proliferation, differentiation and osteogenesis of mesenchymal stem cells of human body, and is expected to provide a new strategy for the clinical repair and treatment of large-section bone defects.
Description
Technical Field
The invention belongs to the field of materials, relates to a bone tissue engineering scaffold material, and particularly relates to a mesoporous bioactive glass/metal organic framework composite scaffold material.
Background
The repair treatment of large bone defects caused by bone tumors, traffic accidents, etc. is a great clinical challenge. At present, the repair treatment of bone defects by using bone tissue engineering scaffold materials is considered to be one of effective methods for solving the related problems. In general, bone tissue engineering scaffold materials need to have basic requirements such as three-dimensional communicated porous structures, good mechanical support properties, biodegradability, bioactivity and the like. Therefore, designing and preparing bone tissue engineering scaffold materials meeting the basic requirements is a key problem for repairing and treating bone defects.
The Mesoporous Bioactive Glass (MBG) has a large specific surface area and highly ordered mesoporous channels, has higher bioactivity than common bioglass, and simultaneously has a mesoporous structure which endows the medicament with efficient loading and slow release characteristics and an in-situ medicament slow release function. Research shows that MBG can promote the proliferation and differentiation of osteoblasts and induce new bone regeneration, but the degradation products induce the increase of the alkalinity of the surrounding environment and are not beneficial to the response of osteocytes. The Metal-Organic Framework (MOF) is a porous crystal material formed by self-assembly of Metal ions or clusters and Organic ligands through coordination, wherein part of the iron-based, zinc-based and zirconium-based MOF materials have biocompatibility and biodegradability and show bright application prospects in the field of biomedicine; meanwhile, organic ligands in the MOF material degradation products are weakly acidic, and the degraded metal ions can promote bone cell response in a certain range. Therefore, the mesoporous bioglass and the metal organic framework are combined to prepare the porous composite scaffold material which has structural mechanics, bioactivity, pH microenvironment and bone cell response, and the porous composite scaffold material is expected to be applied to repair and treatment of large-section bone defects.
The low-temperature 3D printing technology is used as an emerging rapid forming technical means, and the external shape, the internal structure and the hole connectivity of the stent material can be accurately controlled. Compared with other 3D printing technologies, the low-temperature 3D printing operation is convenient, high temperature is not needed, the damage to the material is small, and the preparation method can be effectively used for preparing the drug-loaded composite stent material.
Under the background of the prior art, the mesoporous bioglass/metal organic framework composite scaffold material is prepared by using a low-temperature 3D printing technology, and is expected to provide a new strategy for the clinical repair and treatment of large-section bone defects.
Disclosure of Invention
The invention provides a mesoporous bioactive glass/metal organic framework composite scaffold material and a preparation method thereof, aiming at the preparation problem of a scaffold material for bone tissue engineering, and the preparation method of the composite scaffold material solves the problems that the scaffold material for bone tissue engineering prepared in the prior art cannot accurately regulate and control the internal structure of the scaffold material, and the mechanical property, pH microenvironment regulation, bioactivity and the like are not ideal.
The invention provides a preparation method of a mesoporous bioactive glass/metal organic framework composite scaffold material, which comprises the following steps:
1) preparing mesoporous bioactive glass/metal organic framework composite printing ink, uniformly mixing mesoporous bioactive glass and metal organic framework powder according to a certain mass ratio, sieving with a 200-600-mesh sieve, adding the mixed powder with a certain mass into a trichloromethane solution of polycaprolactone,
stirring uniformly to obtain printing ink, and then sealing and storing;
2) designing the appearance and the internal structure of the mesoporous bioactive glass/metal organic framework bracket material by using CAD/CAM computer aided software, wherein the size of the bracket material model is 15 multiplied by 15
The ink has any size and shape within cm, the inner aperture is 50-1500 mu m, and the ink strike included angle of two adjacent layers is 0-180 degrees;
3) preparing a mesoporous bioactive glass/metal organic framework support material by a low-temperature 3D printing technology, namely filling prepared printing ink into a charging barrel of a 3D printer, then installing a needle head with the diameter of 100-1000 mu m, and precooling for 5-15 minutes at 4-8 ℃; operating a 3D printing program, adjusting the air pressure of a printer to be 1.2-5.0 bar, and adjusting the printing speed to be 2.0-8.0 mm/s; printing the ink line on a glass culture dish of a loading platform, accurately forming according to a layer-by-layer stacking mode to obtain the needed mesoporous bioactive glass/metal organic framework composite support material, and drying at 37 ℃ for 12-24 hours for later use.
Furthermore, the mass ratio of the mesoporous bioactive glass to the metal organic framework powder is in any ratio of 99:1 to 30: 70.
Further, the mass-volume ratio between the polycaprolactone and the trichloromethane is 0.1g/mL to 0.15g/mL, and the mass-volume ratio between the mesoporous bioactive glass/metal organic framework mixed powder and the polycaprolactone solution is 0.1g/mL to 1 g/mL.
Further, the metal organic framework powder material comprises any one of iron-based MOF materials MIL-100(Fe), MIL-88(Fe), zinc-based MOF materials ZIF-8(Zn), ZIF-90(Zn), zirconium-based MOF materials Uio-66(Zr) and PCN-223 (Zr).
The invention also provides the mesoporous bioactive glass/metal organic framework composite scaffold material prepared by the method.
The mesoporous bioactive glass/metal organic framework support material disclosed by the invention is prepared by combining the excellent bioactivity of mesoporous bioactive glass and the degradability of a metal organic framework by adopting a low-temperature 3D printing technology. The detection of the physicochemical property and the biological property of the scaffold material shows that the scaffold material has a three-dimensional communicated and controllable macroporous structure, high mechanical strength and good biocompatibility, and can promote the proliferation, differentiation and osteogenesis of human bone marrow mesenchymal stem cells.
Compared with the prior art, the invention has remarkable technical progress. The invention provides a 3D printing preparation method of a mesoporous bioactive glass/metal organic framework support material. The mesoporous bioactive glass/metal organic framework scaffold material obtained by the method is expected to provide a new strategy for repairing and treating large bone defects clinically.
Drawings
FIG. 1 is an optical photograph of the mesoporous bioactive glass/metal organic framework scaffold material prepared in example 1.
FIG. 2 is a Scanning Electron Microscope (SEM) image of the mesoporous bioactive glass/metal organic framework scaffold material prepared in example 1.
FIG. 3 is a Scanning Electron Microscope (SEM) image of human mesenchymal stem cells after being seeded on the mesoporous bioactive glass/metal organic framework scaffold material prepared in example 1 for 3 days.
Detailed Description
In order to make the technical solutions of the present invention better understood by researchers in the related fields, the present invention is further described below with reference to specific examples.
Example 1
A preparation method of a mesoporous bioactive glass/metal organic framework composite scaffold material comprises the following steps:
preparing mesoporous bioactive glass/metal organic framework (MBG/MOF) composite printing ink. The MBG powder and the MOF powder are uniformly mixed according to the mass ratio of 90:10, the mixture is sieved by a 400-mesh sieve, then 0.7 g of the mixed powder is added into 2.0 mL of Polycaprolactone (PCL)/trichloromethane solution (0.15g/mL), the mixture is uniformly stirred, and then the mixture is sealed and stored.
Step two, the appearance and the internal structure of the MBG/MOF composite scaffold material are designed by using CAD/CAM computer aided software, and the scaffold material model is a cylinder (
h is 10mm), the inner aperture is 400 μm, and the ink strike angle of two adjacent layers is 60 °.
And step three, preparing the MBG/MOF composite scaffold material by using a low-temperature 3D printing technology. Filling the prepared printing ink into a charging barrel of a 3D printer, then installing a needle head with the diameter of 400 microns, and precooling for 10 minutes at 4 ℃; operating a 3D printing program, adjusting the air pressure of a printer to be 1.8-2.4 bar, and adjusting the printing speed to be 4.6-6.2 mm/s; and printing the ink line on a glass culture dish of a carrying platform, accurately forming the required MBG/MOF composite scaffold material in a layer-by-layer stacking mode, and drying the MBG/MOF composite scaffold material at 37 ℃ for 12 hours for later use.
Example 2
A preparation method of a mesoporous bioactive glass/metal organic framework composite scaffold material comprises the following steps:
step one, preparing MBG/MOF composite printing ink. The MBG powder and the MOF powder are uniformly mixed according to the mass ratio of 70:30, the mixture is sieved by a 400-mesh sieve, then 0.7 g of the mixture is added into 3.0 mL of Polycaprolactone (PCL)/trichloromethane solution (0.1g/mL), the mixture is uniformly stirred, and then the mixture is sealed and stored.
Step two, the appearance and the internal structure of the MBG/MOF composite scaffold material are designed by using computer aided software such as CAD/CAM and the like, and the scaffold material model is a cylinder (
h is 10mm), the inner aperture is 300 mu m, and the running angle of the two adjacent layers of ink is 60 degrees.
And step three, preparing the MBG/MOF composite scaffold material by using a low-temperature 3D printing technology. Filling the prepared printing ink into a charging barrel of a 3D printer, then installing a needle head with the diameter of 300 microns, and precooling for 15 minutes at 8 ℃; operating a 3D printing program, adjusting the air pressure of a printer to be 1.8-2.0 bar, and adjusting the printing speed to be 4.0-5.6 mm/s; and printing the ink line on a glass culture dish of a carrying platform, accurately forming the required MBG/MOF composite scaffold material in a layer-by-layer stacking mode, and drying the MBG/MOF composite scaffold material at 37 ℃ for 12 hours for later use.
Example 3
A preparation method of a mesoporous bioactive glass/metal organic framework composite scaffold material comprises the following steps:
step one, preparing MBG/MOF composite printing ink. The MBG powder and the MOF powder are uniformly mixed according to the mass ratio of 50:50, the mixture is sieved by a 300-mesh sieve, then 0.7 g of the mixture is added into 3.0 ml of Polycaprolactone (PCL)/trichloromethane solution (0.1g/ml), the mixture is uniformly stirred, and then the mixture is sealed and stored.
Step two, the appearance and the internal structure of the MBG/MOF composite scaffold material are designed by using CAD/CAM computer aided software, and the scaffold material model is a cylinder (
h is 10mm), the inner aperture is 200 μm, and the ink strike angle of two adjacent layers is 90 °.
And step three, preparing the MBG/MOF composite scaffold material by using a low-temperature 3D printing technology. Filling the prepared printing ink into a charging barrel of a 3D printer, then installing a needle head with the diameter of 200 microns, and precooling for 15 minutes at 4 ℃; operating a 3D printing program, adjusting the air pressure of a printer to be 1.9-2.2 bar, and adjusting the printing speed to be 4.4-5.6 mm/s; and printing the ink line on a glass culture dish of a carrying platform, accurately forming the required MBG/MOF composite scaffold material in a layer-by-layer stacking mode, and drying the MBG/MOF composite scaffold material at 37 ℃ for 12 hours for later use.
Example 4 cell adhesion test example
The study on the adhesion condition of human mesenchymal stem cells on the mesoporous bioactive glass/metal organic framework composite scaffold material prepared in example 1 comprises the following steps:
step one, human mesenchymal stem cells are extracted according to the literature reports [ Matsubara T, subarta K, Ishii M, et al. Alvecor Bone marrow as a cell source for regenerating medium cells: differences between interstitial cells and interstitial Bone marrow cells. journal of Bone and Mineral Research,2005,20, 399-.
Step two, irradiating the MBG/MOF composite scaffold material prepared in the example 1 under an ultraviolet lamp for 24 hours for sterilization, then putting the MBG/MOF composite scaffold material into a 24-hole culture plate, adding l ml of DMEM cell culture medium containing 10% fetal calf serum, pre-wetting for 24 hours, and then removing the culture medium. Then, the mixture will contain 1 × 105100 microliters of culture solution of human mesenchymal stem cells was dropped on each scaffold material, and 1 ml of DMEM cell culture medium containing 10% fetal bovine serum was added after the cells were pre-attached for 4 hours, and 5% CO was added at 37 deg.C2Culturing in an atmosphere incubator.
Step three, after the cells were cultured on the scaffold material for 3 days, the cells were washed three times with PBS buffer at 37 ℃, followed by fixation with PBS containing 2.5% glutaraldehyde for 1 hour, the fixative was removed by PBS containing 4% sucrose, and then fixed by PBS buffer containing 1% osmium tetroxide, followed by dehydration treatment by gradient ethanol solution (50%, 70%, 90%, 95%, 100%) and hexamethyldisilazane solution (HMDS).
And step four, observing the adhesion condition of the cells on the surface of the support material by using a scanning electron microscope (FEI Quanta 450), adhering the support material sample on a metal base by using a conductive adhesive, and carrying out gold spraying treatment for 60 seconds before observation.
The results are shown in FIG. 3, the human mesenchymal stem cells adhere well and spread out completely on the MBG/MOF composite scaffold, which indicates that the MBG/MOF composite scaffold is favorable for cell adhesion and has good biocompatibility.
In the preparation process, the MBG/MOF composite scaffold material with different component contents and different internal structures can be prepared by changing the mass ratio of the MBG powder to the MOF powder, the type of the organic solvent, adjusting the printing parameters and designing different scaffold material models.
The analysis of the physicochemical property and the biological property of the composite scaffold material shows that the mesoporous bioactive glass/metal organic framework scaffold material prepared by the invention has a three-dimensional communicated and controllable macroporous structure, the mechanical strength meets the strength of cancellous bone (2-12MPa), the biocompatibility is good, and the proliferation, differentiation and osteogenesis of human mesenchymal stem cells can be promoted. Therefore, the mesoporous bioactive glass/metal organic framework stent material prepared by the invention is expected to bring a new technology for treating large-segment bone defects clinically.
The foregoing is a more detailed description of the invention with reference to specific embodiments, which are not to be construed as limiting the invention. Accordingly, modifications and substitutions may be made thereto without departing from the general inventive concept and, therefore, are to be considered within the scope of the appended claims.
Claims (4)
1. A preparation method of a mesoporous bioactive glass/metal organic framework composite scaffold material is characterized by comprising the following steps:
1) preparing mesoporous bioactive glass/metal organic framework composite printing ink, namely uniformly mixing mesoporous bioactive glass and metal organic framework powder according to a certain mass ratio, sieving the mixture with a 200-600-mesh sieve, adding the mixed powder with a certain mass into a trichloromethane solution of polycaprolactone, uniformly stirring to obtain printing ink, and then sealing and storing;
2) designing the appearance and the internal structure of the mesoporous bioactive glass/metal organic framework support material by using CAD/CAM computer aided software, wherein the size of a support material model is 15 multiplied by 15cm and any size and shape within the range, the internal pore diameter is 50-1500 mu m, and the running included angle of two adjacent layers of ink is 0-180 degrees;
3) preparing a mesoporous bioactive glass/metal organic framework support material by a low-temperature 3D printing technology, namely filling prepared printing ink into a charging barrel of a 3D printer, then installing a needle head with the diameter of 100-1000 mu m, and precooling for 5-15 minutes at 4-8 ℃; operating a 3D printing program, adjusting the air pressure of a printer to be 1.2-5.0 bar, and adjusting the printing speed to be 2.0-8.0 mm/s; printing the ink line into a glass culture dish of a loading platform, accurately forming according to a layer-by-layer stacking mode to obtain the needed mesoporous bioactive glass/metal organic framework composite support material, and drying at 37 ℃ for 12-24 hours for later use;
in the step 1), the mass ratio of the mesoporous bioactive glass to the metal organic framework powder is in any proportion of 99:1 to 30:70, and the mass-volume ratio of the mesoporous bioactive glass/metal organic framework mixed powder to the polycaprolactone solution is 0.1g/mL to 1 g/mL.
2. The preparation method of the mesoporous bioactive glass/metal organic framework composite scaffold material according to claim 1, wherein the preparation method comprises the following steps: the mass-volume ratio of the polycaprolactone to the trichloromethane is 0.1g/mL-0.15 g/mL.
3. The preparation method of the mesoporous bioactive glass/metal organic framework composite scaffold material according to claim 1, wherein the preparation method comprises the following steps: the metal organic framework powder material comprises any one of iron-based MOF materials MIL-100(Fe), MIL-88(Fe), zinc-based MOF materials ZIF-8(Zn), ZIF-90(Zn), zirconium-based MOF materials Uio-66(Zr) and PCN-223 (Zr).
4. A mesoporous bioactive glass/metal organic framework composite scaffold material is characterized in that: prepared using the method of any one of claims 1 to 3.
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| CN105056895A (en) * | 2015-08-17 | 2015-11-18 | 中国科学院上海高等研究院 | Preparation method and application of metal organic frameworks-mesoporous silica composite material |
| CN106178132A (en) * | 2016-07-20 | 2016-12-07 | 上海理工大学 | A kind of preparation method of composite bone cement support |
| CN106421784A (en) * | 2016-08-12 | 2017-02-22 | 上海理工大学 | Nano drug carrier having photothermal effect and preparation method and application thereof |
| CN107385419A (en) * | 2017-06-28 | 2017-11-24 | 河南工业大学 | Medical magnesium alloy surface is corrosion-resistant and the coating of hydrophilicity and preparation method thereof for a kind of raising |
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