CN115230142A - Nanofiber surface porous bone repair scaffold and preparation method thereof - Google Patents

Nanofiber surface porous bone repair scaffold and preparation method thereof Download PDF

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CN115230142A
CN115230142A CN202210853278.8A CN202210853278A CN115230142A CN 115230142 A CN115230142 A CN 115230142A CN 202210853278 A CN202210853278 A CN 202210853278A CN 115230142 A CN115230142 A CN 115230142A
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printing
bone repair
porous bone
temperature
nanofiber surface
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黄文华
吴耀彬
李梓岳
利时雨
郑蔚晗
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Southern Medical University
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Southern Medical University
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/10—Processes of additive manufacturing
    • B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/30—Auxiliary operations or equipment
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/30—Auxiliary operations or equipment
    • B29C64/307—Handling of material to be used in additive manufacturing
    • B29C64/314—Preparation
    • 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
    • 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
    • B33Y40/00—Auxiliary operations or equipment, e.g. for material handling
    • B33Y40/10—Pre-treatment
    • 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
    • B33Y40/00—Auxiliary operations or equipment, e.g. for material handling
    • B33Y40/20—Post-treatment, e.g. curing, coating or polishing
    • 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
    • B33Y70/10—Composites of different types of material, e.g. mixtures of ceramics and polymers or mixtures of metals and biomaterials
    • 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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  • Optics & Photonics (AREA)
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Abstract

A nanofiber surface porous bone repair scaffold and a preparation method thereof are disclosed, wherein the preparation method comprises the following steps: mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution; adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1) to obtain a membrane casting solution; step (3) cooling the 3D printing platform in advance, then taking the casting film liquid obtained in the step (2) as printing ink, and performing 3D printing in the 3D printing platform according to printing parameters to obtain a printing support; and (4) cooling and washing the printing support obtained in the step (3) to obtain the nanofiber surface porous bone repair support. The scaffold forms a micro-nano-grade fiber porous structure by utilizing a low-temperature thermally induced phase separation technology, and the nano-fiber surface porous bone repair scaffold has good biocompatibility, can simultaneously have a good bone differentiation promoting effect, and is beneficial to generation and growth of bone tissues.

Description

Nanofiber surface porous bone repair scaffold and preparation method thereof
Technical Field
The invention relates to the technical field of nanofiber materials, in particular to a nanofiber surface porous bone repair support and a preparation method thereof.
Background
Bone deficiency due to trauma or surgery is called bone defect. Due to the presence of bone defects, nonunion, delayed or no healing of the resulting bone, and localized dysfunction are often caused. Bone defects caused in pathological processes, such as comminuted fracture, open fracture and massive bone tissue defect caused by trauma, inflammation, bone diseases and other factors, osteonecrosis and abscission and separation caused by inflammation, and defects caused by massive osteonecrosis caused by bone infarction or osteoischemic necrosis, belong to bone defects caused by diseases. The bone defect caused by the operation is caused by artificial factors.
The artificial material transplantation is carried out in an important means for clinically treating large-area bone defects. However, the artificial materials are generally lack of biocompatibility and surface activity, the seed cells are difficult to adhere and grow, and the channels for exchanging tissue nutrients are lacked, so that the occurrence of peripheral micro-blood vessels is not facilitated.
Therefore, it is necessary to provide a nanofiber surface porous bone repair scaffold and a preparation method thereof to solve the defects of the prior art.
Disclosure of Invention
One of the purposes of the invention is to provide a preparation method of a nanofiber surface porous bone repair scaffold to avoid the defects of the prior art. The nanofiber surface porous bone repair scaffold can be prepared by the preparation method, has good biocompatibility, can simultaneously have good bone differentiation promoting effect, and is beneficial to generation and growth of bone tissues.
The above object of the present invention is achieved by the following technical measures:
provides a preparation method of a nanofiber surface porous bone repair scaffold, which comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1) to obtain a membrane casting solution;
step (3) cooling the 3D printing platform in advance, then taking the casting film liquid obtained in the step (2) as printing ink, and performing 3D printing in the 3D printing platform according to printing parameters to obtain a printing support;
and (4) cooling and washing the printing support obtained in the step (3) to obtain the nanofiber surface porous bone repair support.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 50% -75%;
sodium chloride: 5% -20%;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
beta-tricalcium phosphate: 2% -15%;
polylactic acid: 20 to 40 percent.
Further, in the mixed solution, the weight ratio is:
tetrahydrofuran (tetrahydrofuran): 60% -70%;
sodium chloride: 8 to 12 percent;
n, N-dimethylformamide: and the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 4% -7%;
polylactic acid: 25 to 32 percent.
Furthermore, in the mixed solution, the weight ratio is as follows:
tetrahydrofuran (tetrahydrofuran): 67.5 percent;
sodium chloride: 22.5 percent;
n, N-dimethylformamide: the balance;
in the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 5 percent;
polylactic acid: 30 percent.
Preferably, in the step (2), the beta-tricalcium phosphate and the polylactic acid are added into the mixed solution obtained in the step (1), the stirring speed is 450-550 rpm, the temperature is 35-45 ℃, and the stirring time is 18-24 hours, so as to obtain the membrane casting solution.
Preferably, the step (3) is specifically to reduce the temperature of the 3D printing platform to-20-4 ℃ in advance, and then perform 3D printing on the casting solution obtained in the step (2) as printing ink according to printing parameters to obtain a printing support;
the printing parameters comprise that the temperature of the storage bin and the printing nozzle is 30-60 ℃, the moving speed of the printing nozzle is 2-3 mm/s, and the extrusion pressure of the printing nozzle is 4.3-6 bar.
Preferably, the step (4) includes:
step 4.1, cooling the printing support obtained in the step 3 to below-20 ℃, standing for 6-24 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is more than or equal to 4 and less than or equal to 10, is an integer, and t is more than or equal to 2 ℃ and less than or equal to 5 ℃, so as to obtain the nanofiber surface porous bone repair support.
Preferably, the step 4.2 is to soak the printing support in water with the temperature t, and replace the original soaking water with deionized water with the temperature t every other alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2 nd hour period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein t is more than or equal to 2 ℃ and less than or equal to 5 ℃, alpha is 10min, beta is 20min, and gamma is 30min.
Preferably, the pore diameter of the nanofiber surface porous bone repair scaffold is 390.63 μm +/-35.42 μm.
Preferably, the tensile strength of the nanofiber surface porous bone repair scaffold is 2.0MPa-2.1MPa.
Another object of the present invention is to provide a nanofiber surface porous bone repair scaffold which avoids the disadvantages of the prior art. The nanofiber surface porous bone repair scaffold has good biocompatibility, can simultaneously have good effect of promoting bone differentiation, and is beneficial to generation and growth of bone tissues.
The above object of the present invention is achieved by the following technical measures:
the nanofiber surface porous bone repair scaffold is prepared by the preparation method of the nanofiber surface porous bone repair scaffold.
The invention relates to a nanofiber surface porous bone repair scaffold and a preparation method thereof, wherein the preparation method comprises the following steps: mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution; adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1) to obtain a membrane casting solution; step (3) cooling the 3D printing platform in advance, then taking the casting film liquid obtained in the step (2) as printing ink, and performing 3D printing in the 3D printing platform according to printing parameters to obtain a printing support; and (4) cooling and washing the printing support obtained in the step (3) to obtain the nanofiber surface porous bone repair support. The nanofiber surface porous bone repair scaffold forms a micro-nano-grade fiber porous structure by using a low-temperature thermally induced phase separation technology, has good biocompatibility, can simultaneously have a good bone differentiation promoting effect, and is beneficial to generation and growth of bone tissues.
Drawings
The invention is further illustrated by means of the attached drawings, the content of which is not in any way limiting.
FIG. 1 is a scanning electron microscope image of the nanofiber surface porous bone repair scaffold of example 3.
Fig. 2 is a spectrum diagram of the nanofiber surface porous bone repair scaffold of example 3.
Fig. 3 is a scanning electron microscope image of the nanofiber surface porous bone repair scaffold of example 8.
FIG. 4 is a scanning electron micrograph of a stent of comparative example 1.
FIG. 5 is an energy spectrum of the stent of comparative example 1.
FIG. 6 is a cell seeding map of the nanofiber surface porous bone repair scaffold of example 3.
Fig. 7 is a staining pattern of living cells of the nanofiber surface porous bone repair scaffold of example 3 after cell seeding.
FIG. 8 is a diagram of scaffold seeded cells of comparative example 1.
FIG. 9 is a graph showing staining of viable cells after cell seeding in comparative example 1.
FIG. 10 is a graph showing the growth of the nanofiber surface porous bone repair scaffold of example 3 after being seeded with cells, and the growth of the conventional two-dimensional cultured cells, compared to the scaffold of comparative example 1.
FIGS. 11, 12 and 13 are comparative graphs showing the detection of osteogenic differentiation-related genes after induction culture of cells of the scaffolds of comparative example 1 and the scaffolds for nanofiber surface porous bone repair of example 3, respectively.
Detailed Description
The technical solution of the present invention is further illustrated by the following examples.
The experimental procedures in the following examples are conventional unless otherwise specified. The raw materials, reagent materials and the like used in the following examples can be purchased from conventional biochemical reagent stores or pharmaceutical operation companies unless otherwise specified. Wherein, the beta-tricalcium phosphate and the polylactic acid are respectively purchased from the Jinan Dai handle biotech GmbH; the cells of the present invention were 3T3 cells, and 3T3 cells were purchased from Yuan-Jing Biotechnology Ltd
Example 1.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps: mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1) to obtain a membrane casting solution;
step (3) cooling the 3D printing platform in advance, then taking the casting film liquid obtained in the step (2) as printing ink, and performing 3D printing in the 3D printing platform according to printing parameters to obtain a printing support;
and (4) cooling and washing the printing support obtained in the step (3) to obtain the nanofiber surface porous bone repair support.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 50% -75%;
sodium chloride: 5% -20%;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 2% -15%;
polylactic acid: 20 to 40 percent.
It should be noted that, because the 3D printing ink is stored in the printing bin and extruded through the nozzle, β -tricalcium phosphate and polylactic acid in the casting solution may gradually solidify with volatilization of tetrahydrofuran, causing nozzle blockage; and the printing ink with the too strong fluidity of the casting solution is difficult to be rapidly molded on a printing platform after being extruded, and the mechanical strength of the molded bracket is not enough when the molded bracket is used for implantation.
The concentration of tetrahydrofuran and N, N-dimethylformamide in the mixed solution is in the range, and the weight ratio of beta-tricalcium phosphate to polylactic acid in the casting solution is in the range, so that the spray head can be prevented from being blocked by volatilization of tetrahydrofuran, and the smoothness of the extrusion process is ensured; meanwhile, the composite material can be quickly molded after being extruded. The sodium chloride is added to improve the cohesive force, so that the mechanical strength of the printing support is improved, and the printing support is convenient to implant into a loaded bone defect part.
Specifically, the step (2) of the invention is to add beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), wherein the stirring speed is 450-550 rpm, the temperature is 35-45 ℃, and the stirring time is 18-24 h, so as to obtain the membrane casting solution.
It should be noted that, the temperature in the preparation process of the casting solution needs to be controlled between 35 ℃ and 45 ℃, because too high temperature can cause volatilization of a mixture system in the heating process, so that the yield of the casting solution is reduced, and the obtained casting solution is difficult to form when the fluidity of the casting solution is too strong for printing; if the temperature is too low, the beta-tricalcium phosphate and the polylactic acid are difficult to dissolve in the mixed liquid, and printing ink cannot be formed. In addition, the mixture splashes due to the fact that the stirring speed is too high in the process, and the dissolving efficiency is influenced by the fact that the stirring speed is too low.
Specifically, the step (3) is that the temperature of the 3D printing platform is reduced to-20-4 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters, and the printing support is obtained.
The printing parameters of the invention include that the temperature of the storage bin and the printing nozzle is 30-60 ℃, the moving speed of the printing nozzle is 2-3 mm/s, and the extrusion pressure of the printing nozzle is 4.3-6 bar.
Wherein, step (4) includes:
step 4.1, cooling the printing support obtained in the step 3 to below-20 ℃, standing for 6-24 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is more than or equal to 4 and less than or equal to 10, is an integer, and t is more than or equal to 2 ℃ and less than or equal to 5 ℃, so as to obtain the nanofiber surface porous bone repair support.
Specifically, the step 4.2 is to soak the printing support in water with the temperature t, and replace the original soaking water with deionized water with the temperature t every other alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within a time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein t is more than or equal to 2 ℃ and less than or equal to 5 ℃, alpha is 10min, beta is 20min, and gamma is 30min.
It should be noted that, the printing support is soaked in water, tetrahydrofuran inside the printing support is gradually replaced with water, and in the experimental process, it is found that the replacement speed of tetrahydrofuran is fast in the front and slow in the back, so that the soaking water needs to be quickly replaced in the front in a period from the starting point to delta hours, and the printing support is prevented from being soaked in the soaking water containing replaced tetrahydrofuran for a long time, and the structure of the printing support is damaged due to tetrahydrofuran. After replacing tetrahydrofuran, a fiber sample micro-morphology of about 1 micron is formed on the surface of the printing support.
The pore diameter of the nanofiber surface porous bone repair scaffold is 390.63 microns +/-35.42 microns. The tensile strength of the porous bone repair bracket on the surface of the nano fiber is 2.0MPa-2.1MPa.
Polylactic acid (PLA), also called polylactide, is a polymer polymerized from lactic acid as a main raw material, and has excellent biodegradability and biocompatibility. Beta-tricalcium phosphate (beta-TCP) also has excellent biodegradability, and calcium and phosphorus ions degraded in the implanted body can enter a body circulation system to stimulate the osteogenesis process. Polylactic acid and beta-tricalcium phosphate are dissolved and then used as printing ink, a high-precision complex bracket structure is obtained through 3D printing, complex porous materials with micro-nano scale are formed by combining a low-temperature thermally induced phase separation technology, and the polylactic acid and beta-tricalcium phosphate composite material has good biocompatibility and good bone promoting effect.
In addition, the low-temperature thermally induced phase separation technique means that the temperature of the casting solution is lower than the melting points of the beta-tricalcium phosphate and the polylactic acid. In the step (2), the temperature of the mixed solution is lower than the melting points of the beta-tricalcium phosphate and the polylactic acid and is higher than the cloud point temperature of the casting solution, and the temperature of the 3D printing platform is obviously lower than the cloud point temperature of the casting solution. Thus, when the casting solution enters the 3D printing platform, a thermally-induced phase separation mechanism and a non-solvent-induced phase separation mechanism occur simultaneously, thereby forming a porous structure.
The preparation method of the nanofiber surface porous bone repair scaffold has the following beneficial effects:
(1) Through computer aided design and direct molding by 3D printing, the molding structure is consistent with the design structure, the structure and the mechanical property of the bracket can be ensured to be matched with the height of a defect part, and no pollution is caused by cutting;
(2) The micro-nano level fiber porous structure is formed by a low-temperature thermally induced phase separation technology, the surface of the bracket has high porosity and strong adsorbability, and the pores are communicated with each other, so that the adhesive growth of seed cells, the growth of blood vessels and the exchange of nutrient substances after implantation are facilitated;
(3) The biocompatibility is good, and meanwhile, the bone formation promoting effect is good;
(4) The preparation method is simple and convenient, the preparation process is stable, and the preparation method is non-toxic and pollution-free.
Example 2.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), wherein the stirring speed is 480-500 rpm, the temperature is 38-42 ℃, and the stirring time is 20-22 h, so as to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-15-0 ℃ in advance, then the casting solution obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise the temperature of a storage bin and a printing nozzle of 40-50 ℃, the moving speed of the printing nozzle of 2.3-2.7 mm/s, and the extrusion pressure of the printing nozzle of 5-5.8 bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-21 ℃, standing for 10 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to the time within delta hours, wherein delta is 6 and t is 3 ℃ to obtain the nanofiber surface porous bone repair support.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every other alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta, and obtaining the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 60% -70%;
sodium chloride: 8 to 12 percent;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 4 to 7 percent;
polylactic acid: 25 to 32 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore diameter, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 3.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 490rpm and the temperature of 45 ℃ for 20 hours to obtain the membrane casting solution.
Step (3), the temperature of a 3D printing platform is reduced to-20 ℃ in advance, then the casting solution obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperatures of a storage bin and a printing nozzle are 41 ℃, the moving speed of the printing nozzle is 2.4mm/s, and the extrusion pressure of the printing nozzle is 5.2bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-20 ℃, standing for 5 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to the time within delta hours, wherein delta is 5 and t is 4 ℃ to obtain the porous bone repair support on the surface of the nanofiber.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every other alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 67.5 percent;
sodium chloride: 22.5 percent;
n, N-dimethylformamide: and the balance.
In the casting solution, the mass volume ratio is as follows:
beta-tricalcium phosphate: 5 percent;
polylactic acid: 30 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore size, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 4.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 450rpm and the temperature of 35 ℃ for 18 hours to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-20 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise the temperatures of a storage bin and a printing nozzle of 40 ℃, the moving speed of the printing nozzle of 2mm/s, and the extrusion pressure of the printing nozzle of 4.3bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-25 ℃, standing for 6 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and is 4; and t is 2 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature t every gamma minute within the 3-hour time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta, and obtaining the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 50 percent;
sodium chloride: 5 percent;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 15 percent;
polylactic acid: 20 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore diameter, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 5.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 550rpm at the temperature of 45 ℃ for 24 hours to obtain a membrane casting solution.
Step (3), reducing the temperature of the 3D printing platform to 4 ℃ in advance, then using the casting film liquid obtained in the step (2) as printing ink and carrying out 3D printing according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperatures of a storage bin and a printing nozzle are 60 ℃, the moving speed of the printing nozzle is 3mm/s, and the extrusion pressure of the printing nozzle is 6bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-21 ℃, standing for 24 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and delta is 10; t is 1 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every other alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2 nd hour period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 75 percent;
sodium chloride: 20 percent;
n, N-dimethylformamide: and the balance.
In the casting solution, the mass volume ratio is as follows:
β -tricalcium phosphate: 2 percent;
polylactic acid: 40 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore diameter, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 6.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 500rpm and the temperature of 40 ℃ for 20 hours to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-10 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperature of a storage bin and a printing nozzle is 50 ℃, the moving speed of the printing nozzle is 2.5mm/s, and the extrusion pressure of the printing nozzle is 5.5bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-22 ℃, standing for 8 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and is 7; and t is 4 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2 nd hour period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta, and obtaining the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 60 percent;
sodium chloride: 12 percent;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 4 percent;
polylactic acid: 25 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore size, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 7.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 500rpm and the temperature of 40 ℃ for 20 hours to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-10 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperature of a storage bin and a printing nozzle is 50 ℃, the moving speed of the printing nozzle is 2.5mm/s, and the extrusion pressure of the printing nozzle is 5.5bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-22 ℃, standing for 8 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and is 7; and t is 4 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta, and obtaining the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 70 percent;
sodium chloride: 8 percent;
n, N-dimethylformamide: and the balance.
In the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 7 percent;
polylactic acid: 32 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore diameter, tensile strength, biocompatibility and bone differentiation promoting effect.
Example 8.
A preparation method of a nanofiber surface porous bone repair scaffold comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 490rpm and the temperature of 45 ℃ for 20 hours to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to 4 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperatures of a storage bin and a printing nozzle are 41 ℃, the moving speed of the printing nozzle is 2.4mm/s, and the extrusion pressure of the printing nozzle is 5.2bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-20 ℃, standing for 5 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to the time within delta hours, wherein delta is 5 and t is 4 ℃ to obtain the porous bone repair support on the surface of the nanofiber.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature t every gamma minute within the 3-hour time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 67.5 percent;
sodium chloride: 22.5 percent;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
beta-tricalcium phosphate: 5 percent;
polylactic acid: 30 percent.
Compared with the embodiment 1, the nanofiber surface porous bone repair scaffold obtained in the embodiment has better pore diameter, tensile strength, biocompatibility and bone differentiation promoting effect.
Comparative example 1.
Mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
and (2) adding polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 500rpm and the temperature of 45 ℃ for 20 hours to obtain a casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-20 ℃ in advance, then the casting solution obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperature of a storage bin and a printing nozzle is 50 ℃, the moving speed of the printing nozzle is 2.5mm/s, and the extrusion pressure of the printing nozzle is 5.5bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-22 ℃, standing for 5 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and is 7; and t is 4 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature of t every gamma minute within the 3 h time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the stent of the comparative example 1, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 67.5 percent;
sodium chloride: 10 percent;
n, N-dimethylformamide: and (4) the balance.
In the casting solution, the mass-to-volume ratio is as follows:
polylactic acid: 30 percent.
In comparison with example 3, comparative example 1 did not add β -tricalcium phosphate and the other reaction conditions were the same.
Comparative example 2.
Step (1), mixing tetrahydrofuran and N, N-dimethylformamide to obtain a mixed solution;
and (2) adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 500rpm and the temperature of 45 ℃ for 20 hours to obtain the membrane casting solution.
Step (3), the temperature of the 3D printing platform is reduced to-20 ℃ in advance, then the casting film liquid obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support, wherein the printing parameters comprise that the temperature of a storage bin and a printing nozzle is 50 ℃, the moving speed of the printing nozzle is 2.5mm/s, and the extrusion pressure of the printing nozzle is 5.5bar;
the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to-22 ℃, standing for 5 hours, and entering the step 4.2;
step 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is an integer and is 7; and t is 4 ℃, and the porous bone repair scaffold with the nanofiber surface is obtained.
Specifically, step 4.2, soaking the printing support in water with the temperature t, and replacing the original soaking water with deionized water with the temperature t every alpha minutes within the 1 st hour period; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2 nd hour period; replacing the original soaking water with deionized water at the temperature t every gamma minute within the 3-hour time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within the time period from delta-3 hours to delta to obtain the stent of the comparative example 2, wherein alpha is 10min, beta is 20min, and gamma is 30min.
In the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 75 percent;
n, N-dimethylformamide: the balance;
in the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 5 percent;
polylactic acid: 30 percent.
In comparison with example 3, in comparative example 2, the sample of comparative example 2 was obtained without adding sodium chloride, and the ratio of tetrahydrofuran to N, N-dimethylformamide was different, and the other reaction conditions were the same.
Verification and conclusion
1. Micro-nanofiber surface characterization
The nanofiber surface porous bone repair scaffold prepared in the examples 3 to 8, the scaffold in the comparative example 1 and the scaffold in the comparative example 2 are characterized under a scanning electron microscope,
wherein the nanofiber surface porous bone repair scaffold prepared in the embodiments 3 to 8 respectively have the fiber scaffold pore diameter of 390.63 +/-35.42 mu m and uniform fiber distribution.
FIG. 1 is a scanning electron microscope image of the nanofiber surface porous bone repair scaffold of example 3, and the scale bar is 4 μm.
Fig. 2 is a spectrum diagram of the nanofiber surface porous bone repair scaffold of example 3.
FIG. 3 is a scanning electron microscope image of the scaffold 1 for the nanofiber surface porous bone repair of example 8, which is scaled to 2 μm.
FIG. 4 is a scanning electron micrograph of a stent of comparative example 1, which is scaled to 4 μm.
FIG. 5 is an energy spectrum of the stent of comparative example 1. From the comparison of fig. 2 and 5, a calcium ion peak appears in the energy spectrum of fig. 2, but a calcium ion peak does not appear in fig. 5, thus demonstrating that beta-calcium phosphate is loaded into the nanofiber surface porous bone repair scaffold of the present invention.
Meanwhile, comparing fig. 3 with fig. 1, the difference of the surface structures of the two is not great, thereby showing that the precooling temperature of the printing platform at-20 ℃ and-4 ℃ has no obvious influence on the forming of the microstructure of the printing support.
2. Biocompatibility
The nanofiber surface porous bone repair scaffold prepared in examples 3 to 8, the scaffold of comparative example 1 and the scaffold of comparative example 2 were immersed in 95% alcohol and soaked for 12 hours, respectively, and then irradiated under an ultraviolet lamp for 12 hours, and finally cells were inoculated and culture induced.
Wherein, FIG. 6 is a cell seeding map of the nanofiber surface porous bone repair scaffold of example 3, and the scale bar is 50 μm.
FIG. 7 is a staining pattern of viable cells of the nanofiber surface porous bone repair scaffold of example 3 after cell seeding, and the scale bar is 500 μm.
Fig. 8 is a graph of the scaffold seeded cells of comparative example 1, with scale bar =50 μm.
Fig. 9 is a graph of staining of viable cells after seeding the cells of comparative example 1, with scale bar =500 μm.
FIG. 10 is a graph showing the growth of the nanofiber surface porous bone repair scaffold of example 3 after being seeded with cells, and the growth of the conventional two-dimensional cultured cells, compared with the scaffold of comparative example 1.
FIGS. 11, 12 and 13 are comparison graphs of osteogenic differentiation-related gene assays after induction culture of cells of the scaffold of comparative example 1 and the nanofiber surface porous bone repair scaffold of example 3, respectively.
As can be seen from fig. 6, 7, 8 and 9, in comparative example 1, no β -tricalcium phosphate was added, the other reaction conditions were the same, and after no β -tricalcium phosphate was added in comparative example 1, the surface of the extruded scaffold still had a significant nanofiber microporous structure, and the biocompatibility was good.
It can be seen from fig. 10 that the cell growth rate of both example 3 and comparative example 1 is much higher than that of the conventional two-dimensional cultured cells, and that the cell growth rate of example 3 with β -tricalcium phosphate is higher than that of comparative example 1 without addition of β -tricalcium phosphate after 7 days of growth.
Runx2, COL-I and OCN of FIGS. 11 to 13 are all common bone cell-specific transcription factors. The two-dimensional culture is a traditional cell culture mode, the non-two-dimensional culture (PLA and PLA/beta-TCP) is cell culture on a scaffold, wherein the PLA/beta-TCP is the nanofiber surface porous bone repair scaffold in example 3, the PLA is the scaffold in comparative example 1, the non-induction of the two-dimensional culture refers to induction culture by using an osteogenesis induction medium under the traditional culture condition, the induction of the two-dimensional culture refers to the use of a normal medium, and the PLA/beta-TCP in figures 11 to 13 are all high-expression, which shows that the bone strengthening is realized, namely, the nanofiber surface porous bone repair scaffold is more beneficial to osteogenesis.
The results show that after the scaffold prepared in examples 3 to 8 is inoculated with cells, the biocompatibility is good, the cell proliferation speed is high compared with that of the traditional two-dimensional culture, and the effect of promoting bone differentiation is obvious.
3. Tensile strength
The nanofiber surface porous bone repair scaffold prepared in examples 3 to 8 and the scaffold of comparative example 2 were subjected to mechanical property correlation tests, respectively.
The nanofiber surface porous bone repair scaffold prepared in examples 3 to 8 measured tensile strength of 2.0Mpa to 2.1Mpa.
The stent of comparative example 2, to which no sodium chloride was added, had a tensile strength of less than 2.0Mpa, and the mechanical strength of the stent after extrusion was insufficient.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (10)

1. A preparation method of a nanofiber surface porous bone repair scaffold is characterized by comprising the following steps: the method comprises the following steps:
mixing tetrahydrofuran, N-dimethylformamide and sodium chloride to obtain a mixed solution;
adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1) to obtain a membrane casting solution;
step (3) cooling the 3D printing platform in advance, then taking the casting film liquid obtained in the step (2) as printing ink, and performing 3D printing in the 3D printing platform according to printing parameters to obtain a printing support;
and (4) cooling and washing the printing support obtained in the step (3) to obtain the nanofiber surface porous bone repair support.
2. The method for preparing the nanofiber surface porous bone repair scaffold according to claim 1, wherein in the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 50% -75%;
sodium chloride: 5% -20%;
n, N-dimethylformamide: the balance;
in the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 2% -15%;
polylactic acid: 20 to 40 percent.
3. The method for preparing the nanofiber surface porous bone repair scaffold according to claim 2, wherein in the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 60% -70%;
sodium chloride: 8 to 12 percent;
n, N-dimethylformamide: the balance;
in the casting solution, the mass-to-volume ratio is as follows:
β -tricalcium phosphate: 4% -7%;
polylactic acid: 25 to 32 percent.
4. The method for preparing the nanofiber surface porous bone repair scaffold according to claim 3, wherein in the mixed solution, the weight ratio is as follows:
tetrahydrofuran: 67.5 percent;
sodium chloride: 22.5 percent;
n, N-dimethylformamide: the balance;
in the casting solution, the mass volume ratio is as follows:
beta-tricalcium phosphate: 5 percent;
polylactic acid: 30 percent.
5. The preparation method of the nanofiber surface porous bone repair scaffold according to any one of claims 1 to 4, wherein: and (2) specifically, adding beta-tricalcium phosphate and polylactic acid into the mixed solution obtained in the step (1), stirring at the speed of 450-550 rpm at the temperature of 35-45 ℃, and stirring for 18-24 h to obtain the membrane casting solution.
6. The preparation method of the nanofiber surface porous bone repair scaffold according to any one of claims 1 to 4, wherein: the step (3) is specifically that the temperature of the 3D printing platform is reduced to-20-4 ℃ in advance, then the casting solution obtained in the step (2) is used as printing ink, and 3D printing is carried out according to printing parameters to obtain a printing support;
the printing parameters comprise the temperature of the stock bin and the printing nozzle is 30-60 ℃, the moving speed of the printing nozzle is 2-3 mm/s, and the extrusion pressure of the printing nozzle is 4.3-6 bar.
7. The preparation method of the nanofiber surface porous bone repair scaffold according to any one of claims 1 to 4, wherein: the step (4) comprises the following steps:
step 4.1, cooling the printing support obtained in the step 3 to below minus 20 ℃, standing for 6-24 hours, and entering the step 4.2;
and 4.2, soaking the printing support in water with the temperature t, starting timing, and replacing the soaking water for multiple times from the starting point to a time period within delta hours, wherein delta is more than or equal to 4 and less than or equal to 10, is an integer, and t is more than or equal to 2 ℃ and less than or equal to 5 ℃, so as to obtain the nanofiber surface porous bone repair support.
8. The preparation method of the nanofiber surface porous bone repair scaffold according to claim 7, characterized in that: the step 4.2 is to soak the printing support in water with the temperature t, and in the 1 st hour period, replace the original soaking water with deionized water with the temperature t every other alpha minutes; replacing the original soaking water with deionized water at the temperature of t every other beta minutes within the 2h time period; replacing the original soaking water with deionized water at the temperature t every gamma minute within the 3-hour time period; and replacing the original soaking water with deionized water at the temperature t every 1 hour within a time period from delta-3 hours to delta to obtain the nanofiber surface porous bone repair scaffold, wherein t is more than or equal to 2 ℃ and less than or equal to 5 ℃, alpha is 10min, beta is 20min, and gamma is 30min.
9. The preparation method of the nanofiber surface porous bone repair scaffold according to any one of claims 1 to 4, wherein: the pore diameter of the porous bone repair scaffold on the surface of the nanofiber is 390.63 microns +/-35.42 microns;
the tensile strength of the porous bone repair scaffold with the nanofiber surface is 2.0-2.1 MPa.
10. The nanofiber surface porous bone repair scaffold is characterized in that: the nanofiber surface porous bone repair scaffold is prepared by the preparation method of any one of claims 1 to 9.
CN202210853278.8A 2022-07-20 2022-07-20 Nanofiber surface porous bone repair scaffold and preparation method thereof Pending CN115230142A (en)

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CN113370533A (en) * 2021-04-25 2021-09-10 中国人民解放军空军军医大学 Preparation method of 3D printing shapeable guide bone regeneration membrane
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WO2018078130A1 (en) * 2016-10-28 2018-05-03 Paul Gatenholm Preparation and applications of 3d bioprinting bioinks for repair of bone defects, based on cellulose nanofibrils hydrogels with natural or synthetic calcium phosphate particles
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