CN114848898A - Artificial bone scaffold manufactured based on 3D printing process and method - Google Patents
Artificial bone scaffold manufactured based on 3D printing process and method Download PDFInfo
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- 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
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Abstract
The invention provides an artificial bone scaffold manufactured based on a 3D printing process and a method, wherein the method comprises the following steps: uniformly mixing beta-TCP, HA whiskers and medical glycerin to obtain a mixture, wherein the mass ratio of the beta-TCP to the medical glycerin is (2-3) to (1-2), the HA whiskers account for 6% -11.25% of the mass of the mixture, and then defoaming to obtain a printing paste; printing the printing paste body by a 3D printing process according to parameters required by design to obtain an initially formed artificial bone scaffold; the preliminary formed artificial bone scaffold is sequentially subjected to refrigeration, freeze drying and sterilization, so that the operation is simple, the component cost caused by the binder can be saved, the mechanical strength of the artificial bone scaffold is enhanced through the HA whiskers, and the artificial bone scaffold with degradability and osteoconductivity is obtained.
Description
Technical Field
The invention belongs to the technical field of medical treatment, and particularly relates to an artificial bone scaffold manufactured based on a 3D printing process and a method.
Background
In recent years, artificial bone repair has become one of the major trends in the field of medical technology. Through research and study of documents, synthetic materials such as polyvinyl alcohol (PVA), Polycaprolactone (PCL) and polylactic acid (PLA) are the most widely used adhesives for conventional artificial bone scaffolds. These materials are not only expensive, but also add to the cost; and the time is consumed for preparation in the production process of the artificial bone scaffold, so that the working hours are increased. When the formed artificial bone scaffold is implanted into a human body, the process of the substance in the human body for a long time is not clear.
The raw material of the artificial bone scaffold is one of the most important factors, and HA (hydroxyl), β -TCP (β -tricalcium phosphate) and bioactive glass are being noted. Through research, the beta-TCP is nontoxic, has good biocompatibility, degradability, bone inductivity, bone transferability and the like, and has excellent repairing effect on bone defects after being implanted into a human body. The strength of human cancellous bone is 2-20 MPa, but the strength of an artificial bone scaffold formed by the traditional beta-TCP is less than 1MPa, so that the initial supporting effect cannot be met.
Disclosure of Invention
The invention provides an artificial bone scaffold manufactured based on a 3D printing process and a method thereof, aiming at the problems in the prior art, the artificial bone scaffold is mainly prepared by taking beta-TCP and HA whiskers as raw materials and medical glycerin as a binder by using the 3D printing method, the operation is simple, the component cost caused by the binder can be saved, the mechanical strength of the artificial bone scaffold is enhanced by the HA whiskers, the cancellous bone strength of a human body is satisfied by 2-20 MPa, and the artificial bone scaffold HAs degradability and bone conductivity.
The invention is realized by the following technical scheme:
a method for manufacturing an artificial bone scaffold based on a 3D printing process comprises the following steps:
uniformly mixing beta-TCP, HA whiskers and medical glycerin to obtain a mixture, wherein the mass ratio of the beta-TCP to the medical glycerin is (2-3) to (1-2), the HA whiskers account for 6-11.25% of the mass of the mixture, and defoaming to obtain a printing paste; printing the printing paste body by a 3D printing process according to parameters required by design to obtain an initially formed artificial bone scaffold; and sequentially refrigerating, freeze-drying and sterilizing the preliminarily formed artificial bone scaffold to obtain the artificial bone scaffold.
Preferably, the medical glycerin solution with the mass fraction of 10% -15% is obtained by adding sterilized water for injection into the medical glycerin, and then the beta-TCP, the HA whiskers and the medical glycerin solution are uniformly mixed.
Preferably, the beta-TCP and the medical glycerin are mixed and stirred for 3-5 times at 1000-3000 r/min, and each time lasts for 1-3 min.
Preferably, the printing paste is obtained by uniformly mixing the β -TCP, the HA whisker and the medical glycerin and then defoaming the mixture in a defoaming machine.
Preferably, the primarily formed artificial bone scaffold is refrigerated at the temperature of-25 to-20 ℃ for 2 to 4 hours and then freeze-dried.
Further, the freeze drying is carried out according to the following processes:
starting from-40 ℃, the temperature is raised to-30 ℃ at a constant rate within 4min, then raised from-30 ℃ to-20 ℃ at a constant rate within 8min, then raised from-20 ℃ to 0 ℃ at a constant rate within 5min, kept at 0 ℃ for 3min, then raised to 20 ℃ at a constant rate within 3min starting from 0 ℃, then raised to 25 ℃ at a constant rate for 3min, and finally raised from 25 ℃ to 37 ℃ at a constant rate for 3 min.
Preferably, the artificial bone scaffold formed preliminarily is subjected to refrigeration, freeze drying and gamma ray sterilization to obtain the artificial bone scaffold.
Preferably, the 3D printing process uses an extrusion biological 3D printer.
An artificial bone scaffold obtained by the method for manufacturing the artificial bone scaffold based on the 3D printing process.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention relates to a method for manufacturing an artificial bone scaffold based on a 3D printing process, which mainly takes beta-TCP as a raw material, has the components similar to bone mineral composition, good biocompatibility and obvious degradation behavior under the biological environment of a human body, and achieves the aim of repairing bone defects while generating a large amount of new bone tissues. The beta-TCP and the medical glycerin are uniformly mixed, the medical glycerin is used as a binder, the material cost can be reduced, the operation is simple, the working hours for preparing a solution are not needed, the manpower, material resources and financial resources are reduced, and then the printing paste can be obtained through deaeration; according to parameters required by design, the printing paste can be printed by a 3D printing process to obtain an initially formed artificial bone scaffold; the artificial bone scaffold formed preliminarily is subjected to cold storage, freeze drying and sterilization in sequence to obtain the artificial bone scaffold, HA whiskers can enhance the mechanical strength of the artificial bone scaffold, and the mechanical strength of the artificial bone can reach 6.34-8.71 MPa.
Drawings
Fig. 1 is a flow chart of the preparation of the 3D printed artificial bone scaffold according to the present invention.
FIG. 2 is a graph of the freeze-drying temperature of the present invention as a function of time.
Fig. 3a is a diagram of an artificial bone scaffold obtained in example 1 of the present invention.
Fig. 3b is a top view of fig. 3 a.
Fig. 4 is a diagram of an artificial bone scaffold obtained in example 2 of the present invention.
Fig. 5 is a diagram of an artificial bone scaffold obtained in example 3 of the present invention.
Detailed Description
The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.
The 3D printing process is also one of preparation methods which are more concerned in recent years, has microstructure regulation and control capability, can regulate and control the size, the shape, the internal connectivity, the porosity and other characteristics of pores in the bracket from the microstructure, and can be customized individually. The artificial bone scaffold not only needs to be simple to operate, but also needs to control the cost, and the performance can meet the requirement.
Therefore, the invention discloses a method for manufacturing an artificial bone scaffold based on a 3D printing process, wherein raw materials comprise beta-TCP, HA whiskers and medical glycerin. beta-TCP is used as a raw material, the components of the beta-TCP are similar to those of bone minerals, the biocompatibility is good, the existing related experiments show that the beta-TCP has very obvious degradation behavior under the human body biological environment, the aim of repairing bone defects is achieved while a large amount of new bone tissues are generated, and the beta-TCP is widely used for repairing bone defects. The medical glycerin is a colorless, transparent and viscous liquid, is usually extracted from grease, has high purity, is non-toxic and safe, is an important raw and auxiliary material for medicines, and is widely applied to the aspects of infusion, injection and the like in the pharmaceutical industry. A large number of widely applied examples show that the bone cement is nontoxic, safe, convenient to obtain and low in price, and is suitable for serving as an artificial bone scaffold adhesive. The medical glycerin is used as the binder, so that the material cost caused by using PVA, PCL and the like as the binder can be reduced, the operation is simple, and the labor and time for preparing the solution are not required.
The artificial bone scaffold plays a supporting role in the initial stage of being implanted into a human body. As shown in fig. 1, the specific method comprises the following steps:
step (1), preparing a binder:
adding water for sterilization injection into purchased medical glycerin to ensure that the mass fraction of the medical glycerin in the obtained solution is 10-15%;
step (2), weighing:
weighing beta-TCP and HA whiskers and the binder obtained in the step (1) by using an electronic balance, enabling the mass ratio of the beta-TCP to the binder obtained in the step (1) to be (2-3) to (1-2), enabling the HA whiskers to account for 6% -11.25% of the total mass, and mixing the beta-TCP and the HA whiskers with the binder obtained in the step (1);
step (3), preparing slurry:
stirring the mixture obtained in the step (2) in a homogenizer at 1000-3000 r/min for 3-5 times, wherein each time lasts for 1-3 min, so as to obtain a mixed system, then filling the mixed system into a 30mL needle cylinder, and defoaming the mixed system in a defoaming machine, so as to obtain a printing paste;
step (4), constructing a printing model:
designing an artificial bone scaffold which is customized individually through three-dimensional design software, wherein the artificial bone scaffold comprises pore diameter, porosity and size;
step (5), 3D printing:
printing the paste in the step (3) into the artificial bone scaffold constructed in the step (4) by a biological 3D printer;
step (6), freeze drying:
refrigerating the artificial bone scaffold printed in the step (5) for 2-4 hours at-25 to-20 ℃, then putting the artificial bone scaffold into a freeze dryer, and freeze-drying the artificial bone scaffold at-40 to 37 ℃ for 28.5-29.5 hours, wherein the freezing, sublimation and drying are performed and the artificial bone scaffold is changed in a gradient manner, and the specific process is shown in figure 2;
and (7) sterilizing:
and (4) sterilizing the artificial bone scaffold obtained in the step (6) by gamma rays to obtain the finally required sterile artificial bone scaffold.
All the operations are operated in a sterile and clean environment.
The artificial bone is simple in material, low in price, simple and convenient to operate in the manufacturing method, and has degradability and certain biomechanical strength in performance.
Example 1
The invention relates to a method for manufacturing an artificial bone scaffold based on a 3D printing process, which comprises the following steps:
step (1), preparing a binder:
3g of medical glycerol with the purity of 95 percent and 25g of sterilized water for injection are weighed respectively and poured into a blue-cap bottle, and the medical glycerol is diluted into a binder with the mass fraction of 10 percent.
Step (2), weighing:
weighing 27g of beta-TCP, 3g of HA whiskers and 20g of binder by using an electronic balance, wherein the mass fraction of the binder is 10%, and the beta-TCP, the HA whiskers and the binder are mixed to obtain a mixture.
Step (3), preparing the paste:
stirring the obtained mixture in a homogenizer at 2000r/min for 4 times, each time for 1min, taking out, filling into a syringe, and defoaming in a defoaming machine to prepare the printing paste.
Step (4), constructing a printing model:
an artificial bone scaffold model with the pore diameter of 700 mu m, the porosity of 70 percent, the pore size of 10X 10mm and the quadrilateral shape is designed through design software, and the STL format is derived.
Step (5), 3D printing:
and printing the artificial bone scaffold by using an extrusion type biological 3D printer. Firstly, loading the printing paste in the step (3) into a printing bin, then loading the three-dimensional model STL format file of the artificial bone scaffold designed in the step (4) into an extrusion type biological 3D printer, and setting the printing parameters as follows: the printing speed is 10mm/min, the printing layer thickness is 0.3mm, the printing paste is uniformly extruded at a constant speed through a spiral propeller, the workbench performs synthetic motion along an x-y axis, the printing head moves along a z axis, the printing is sequentially performed layer by layer, the printing of the biological ceramic artificial bone support is finally completed, and the filling rate and the extrusion width need to be continuously adjusted in the printing process according to actual conditions and are not fixed values.
Step (6), freeze drying:
placing the printed artificial bone scaffold into a low-temperature storage box, contacting the bottom surface of the artificial bone scaffold with the low-temperature storage box, refrigerating for 4h at-20 ℃, then placing the artificial bone scaffold into a freeze dryer to carry out freeze drying according to a curve shown in figure 2, namely, heating to-30 ℃ at a constant speed within 4min from-40 ℃, then heating to-20 ℃ at a constant speed within 8min, then heating to 0 ℃ at a constant speed within 5min, keeping the temperature at 0 ℃ for 3min, then heating to 20 ℃ at a constant speed within 3min from 0 ℃, then heating to 25 ℃ at a constant speed for 3min, and finally heating to 37 ℃ at a constant speed for 3 min.
And (7) sterilizing:
and (4) packaging the artificial bone scaffold obtained in the step (6) at 37 ℃, and sterilizing by gamma rays to obtain a finished product.
Fig. 3a is a diagram of an object of the artificial bone scaffold obtained in embodiment 1 of the present invention, and fig. 3b is a top view of fig. 3 a. The holes in 3a cannot be seen due to the problems of angle and resolution, but fig. 3b shows that the holes are uniformly distributed in each section of the artificial bone scaffold, and are through up and down, and the holes on the side are also through, so the holes are uniformly distributed in the artificial bone scaffold. The strength of the support is subjected to a compression test by using a universal mechanical testing machine, the loading speed is 1mm/min, the strength of the compressed support when the displacement changes by 2 percent is measured, and the mechanical strength is 6.34 MPa.
Example 2
The invention relates to a method for manufacturing an artificial bone scaffold based on a 3D printing process, which comprises the following steps:
step (1), preparing a binder:
5g of medical glycerol with the purity of 95 percent and 34.6g of sterilized water for injection are weighed respectively and poured into a blue-cap bottle, and the medical glycerol is diluted into a binder with the mass fraction of 12 percent.
Step (2), weighing:
beta-TCP, HA whiskers and a binder with the mass fraction of 12% were weighed by an electronic balance, wherein the mass of the beta-TCP was 26.4g, the mass of the HA whiskers was 3.6g, and the mass of the binder was 20g, to obtain a mixture.
Step (3), preparing the paste:
stirring the obtained mixture in a homogenizer at 2000r/min for 4 times, each time for 1min, taking out, filling into a syringe, and defoaming in a defoaming machine to prepare the printing paste.
Step (4), constructing a printing model:
an artificial bone scaffold model with the pore diameter of 500 microns, the porosity of 80 percent, the pore size of 50 x 20 x 3mm and a quadrilateral shape is designed through design software, and an STL format is derived.
Step (5), 3D printing:
and printing the artificial bone scaffold by using an extrusion type biological 3D printer. Firstly, loading the printing paste in the step (3) into a printing stock bin, then loading the STL format file of the artificial bone scaffold three-dimensional model designed in the step (4) into an extrusion type biological 3D printer, and setting the printing parameters as follows: the printing speed is 10mm/min, the printing layer thickness is 0.25mm, the printing paste is uniformly extruded at a constant speed through a spiral propeller, the workbench performs synthetic motion along an x-y axis, the printing head moves along a z axis, the printing is sequentially performed layer by layer, the printing of the biological ceramic type artificial bone support is finally completed, and the filling rate and the extrusion width need to be continuously adjusted in the printing process according to actual conditions and are not fixed values.
Step (6), freeze drying:
placing the printed artificial bone scaffold into a low-temperature storage box, contacting the bottom surface of the low-temperature storage box, refrigerating at-20 deg.C for 4 hr, and freeze-drying in a freeze-drying machine according to the curve shown in FIG. 2.
And (7) sterilizing:
and (4) packaging the artificial bone scaffold obtained in the step (6) at 37 ℃, and sterilizing by gamma rays to obtain a finished product.
Fig. 4 is a schematic view of the artificial bone scaffold obtained in example 2 of the present invention, in which holes cannot be seen due to problems of angle and resolution, but actually, the holes are uniformly distributed in each cross section of the artificial bone scaffold, and the holes are vertically and laterally penetrated, so that the holes are uniformly distributed in the artificial bone scaffold. The mechanical strength of the compression test of the universal mechanical testing machine is 7.43 MPa.
Example 3
The invention relates to a method for manufacturing an artificial bone scaffold based on a 3D printing process, which comprises the following steps:
step (1), preparing a binder:
6g of medical glycerol with the purity of 95% and 32g of sterilized water for injection are weighed respectively and poured into a blue-cap bottle, and the medical glycerol is diluted into a binder with the mass fraction of 15%.
And (2) weighing beta-TCP, HA whiskers and 15% of binder by mass by using an electronic balance, wherein the mass of the beta-TCP is 30.6g, the mass of the HA whiskers is 5.4g, and the mass of the binder is 12g to obtain a mixture.
Step (3), preparing the paste:
stirring the obtained mixture in a homogenizer at 2000r/min for 4 times, each time for 1min, taking out, filling into a syringe, and defoaming in a defoaming machine to prepare the printing paste.
Step (4), constructing a printing model:
a cylindrical artificial bone scaffold model with the aperture of 600 mu m, the porosity of 75 percent, quadrangular pores and the size of phi 10 (diameter) 20mm (height) is designed through design software, and an STL format is derived.
Step (5), 3D printing:
and printing the artificial bone scaffold by using an extrusion type biological 3D printer. Firstly, loading the printing paste in the step (3) into a printing stock bin, then loading the STL format file of the artificial bone scaffold three-dimensional model designed in the step (4) into an extrusion type biological 3D printer, and setting the printing parameters as follows: the printing speed is 10mm/min, the printing layer thickness is 0.25mm, the printing paste is uniformly extruded at a constant speed through a spiral propeller, the workbench performs synthetic motion along an x-y axis, the printing head moves along a z axis, the printing is sequentially performed layer by layer, the printing of the biological ceramic type artificial bone support is finally completed, and the filling rate and the extrusion width need to be continuously adjusted in the printing process according to actual conditions and are not fixed values.
Step (6), freeze drying:
placing the printed artificial bone scaffold into a low-temperature storage box, contacting the bottom surface of the low-temperature storage box, refrigerating at-20 deg.C for 4 hr, and freeze-drying in a freeze-drying machine according to the curve shown in FIG. 2.
And (7) sterilizing:
and (4) packaging the artificial bone scaffold obtained in the step (6) at 37 ℃, and sterilizing by gamma rays to obtain a finished product.
Fig. 5 is a real image of the artificial bone scaffold obtained in example 3 of the present invention, in which holes are uniformly distributed in each cross section of the artificial bone scaffold, and are vertically and laterally perforated, so that the holes are uniformly distributed in the artificial bone scaffold. The compression test of the universal mechanical testing machine shows that the mechanical strength is 8.71 Mpa.
Claims (9)
1. A method for manufacturing an artificial bone scaffold based on a 3D printing process is characterized by comprising the following steps:
uniformly mixing beta-TCP, HA whiskers and medical glycerin to obtain a mixture, wherein the mass ratio of the beta-TCP to the medical glycerin is (2-3) to (1-2), the HA whiskers account for 6% -11.25% of the mass of the mixture, and then defoaming to obtain a printing paste; printing the printing paste body by a 3D printing process according to parameters required by design to obtain an initially formed artificial bone scaffold; and sequentially refrigerating, freeze-drying and sterilizing the preliminarily formed artificial bone scaffold to obtain the artificial bone scaffold.
2. The method for manufacturing the artificial bone scaffold based on the 3D printing process as claimed in claim 1, wherein sterile water for injection is added into medical glycerin to obtain a medical glycerin solution with the mass fraction of 10% -15%, and then the beta-TCP, the HA whiskers and the medical glycerin solution are uniformly mixed.
3. The method for manufacturing the artificial bone scaffold based on the 3D printing process as claimed in claim 1, wherein the beta-TCP and the medical glycerin are mixed and stirred for 3-5 times at 1000-3000 r/min, each time for 1-3 min.
4. The method for manufacturing the artificial bone scaffold based on the 3D printing process as claimed in claim 1, wherein the printed paste is obtained by uniformly mixing the beta-TCP, the HA whiskers and the medical glycerin and then defoaming the mixture in a defoaming machine.
5. The method for manufacturing the artificial bone scaffold based on the 3D printing process according to claim 1, wherein the preliminarily formed artificial bone scaffold is frozen and dried after being refrigerated at-25 to-20 ℃ for 2 to 4 hours.
6. The method for manufacturing an artificial bone scaffold based on 3D printing process according to claim 5, wherein the freeze-drying is performed as follows:
starting from-40 ℃, the temperature is raised to-30 ℃ at a constant rate within 4min, then raised from-30 ℃ to-20 ℃ at a constant rate within 8min, then raised from-20 ℃ to 0 ℃ at a constant rate within 5min, kept at 0 ℃ for 3min, then raised to 20 ℃ at a constant rate within 3min starting from 0 ℃, then raised to 25 ℃ at a constant rate for 3min, and finally raised from 25 ℃ to 37 ℃ at a constant rate for 3 min.
7. The method for manufacturing the artificial bone scaffold based on the 3D printing process as claimed in claim 1, wherein the preliminarily formed artificial bone scaffold is sterilized by gamma ray after being refrigerated and freeze-dried to obtain the artificial bone scaffold.
8. The method for manufacturing an artificial bone scaffold based on 3D printing process as claimed in claim 1, wherein the 3D printing process uses an extrusion biological 3D printer.
9. An artificial bone scaffold obtained by the method for manufacturing an artificial bone scaffold based on the 3D printing process according to any one of claims 1 to 8.
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