Nanofiber yarn 3D printing composite scaffold as well as preparation method and application thereof

Abstract

The invention discloses a preparation method of a nanofiber yarn 3D printing composite scaffold, and the preparation method comprises the following steps: S1, dissolving a polymer and / or an inorganic source required by spinning in a corresponding solvent to obtain a spinning precursor solution; s2, spinning by using the spinning precursor solution in the step S1 through nanofiber spinning equipment to obtain nanofiber yarns; s3, printing by utilizing a 3D printing technology to obtain a polymer framework; and S4, weaving the nanofiber yarns in the step S2 in the polymer framework in the step S3 to obtain the nanofiber yarn 3D printing composite scaffold. The scaffold prepared by the preparation method not only has a 3D printed geometric structure, but also has high specific surface area and biological activity of nanofibers, can better simulate the microenvironment of natural tissues, can overcome the limitation of a traditional scaffold, and provides a more ideal tissue engineering scaffold.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomaterials, and particularly relates to a 3D printing composite scaffold of nanofiber yarns, a preparation method thereof, and an application thereof. Background Art

[0002] Traditional bone repair methods, such as autologous bone transplantation, allogeneic bone transplantation, and implantation of artificial substitute materials, etc., have certain limitations. Bone tissue engineering is expected to overcome these limitations by using the patient's own cells and tissues, and provide a safer and more effective repair method.

[0003] For the traditional technology of preparing nanofiber scaffolds by fiber homogeneous dispersion - freeze drying, the obtained nanofiber scaffolds have poor mechanical properties and poor controllability of the scaffold structure, while the scaffolds printed by 3D printing technology can well solve the above - mentioned disadvantages. The 3D printing technology is simple and efficient, and has unique advantages in terms of precision, mechanical strength, porosity, mechanical properties, and the design and control of complex spatial structures, and can provide a good environment for cell growth; while nanofibers can endow the scaffold with better biocompatibility, multifunctionality, etc. Combining 3D - printed scaffolds with nanofibers can make the scaffold have the advantages of both 3D - printed materials and nanofiber materials, and enable the scaffold to obtain good application performance. Summary of the Invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above - mentioned prior art. For this reason, the present invention provides a preparation method of a 3D printing composite scaffold of nanofiber yarns. The scaffold prepared by this preparation method not only has a complex geometric structure of 3D printing, but also has a high specific surface area and biological activity of nanofibers, can better simulate the microenvironment of natural tissues, can overcome the limitations of traditional scaffolds, and provides a more ideal tissue engineering scaffold.

[0005] The present invention also provides a 3D printing composite scaffold of nanofiber yarns prepared by the above - mentioned preparation method.

[0006] The present invention also provides an application.

[0007] According to the first aspect of the present invention, a preparation method of a 3D printing composite scaffold of nanofiber yarns is provided. The preparation method includes the following steps:

[0008] S1: Dissolve the polymer and / or inorganic source required for spinning in the corresponding solvent to obtain a spinning precursor solution;

[0009] S2: Use the spinning precursor solution described in step S1 to spin with a nanofiber spinning device to obtain nanofiber yarns;

[0010] S3: Use 3D printing technology to print a polymer framework;

[0011] S4: Weave the nanofiber yarn described in S2 in the polymer framework described in S3 to obtain a 3D printed composite scaffold of nanofiber yarn.

[0012] In some embodiments of the present invention, the polymer includes at least one of polylactic acid (PLA), polycaprolactone (PCL), polyvinyl alcohol (PVA), polyacrylonitrile (PAN), polyvinylpyrrolidone (PVP), sodium alginate, polyethylene oxide (PEO), poly (lactic - co - glycolic acid) (PLGA), polyurethane (PU), chitosan, collagen, and gelatin.

[0013] In some embodiments of the present invention, the solvent includes at least one of water, dichloromethane, ethanol, trifluoroethanol, hexafluoroisopropanol (HFIP), acetone, tert - butanol, DMF, DMSO, DMAc, THF, and acetic acid.

[0014] In some embodiments of the present invention, the inorganic source in step S1 includes at least one of tetraethyl orthosilicate, polysilicate tetraethyl ester, tetrabutyl titanate, tin tetrachloride, calcium nitrate, sodium chloride (NaCl), sodium carbonate (Na2CO3), and potassium chloride (KCl).

[0015] In some embodiments of the present invention, the spinning precursor solution further includes an additive.

[0016] In some embodiments of the present invention, the additive includes at least one of acetic acid, oxalic acid, phosphoric acid, hydrochloric acid, nitric acid, and sulfuric acid.

[0017] In some embodiments of the present invention, the ambient temperature of the electrospinning is 20°C to 30°C.

[0018] In some embodiments of the present invention, the ambient relative humidity of the electrospinning is 30% to 60%.

[0019] In some embodiments of the present invention, the perfusion rate of the electrospinning is 0.5 to 5 mL / h.

[0020] In some embodiments of the present invention, the spinning voltage of the electrospinning is 10 to 30 kV.

[0021] In some embodiments of the present invention, in step S2, the spinning uses a wire drum collecting device to collect the yarn, and a vertical rotating twisting and winding machine is used for twisting and winding.

[0022] In some embodiments of the present invention, the distance between the spinning needle and the wire drum during spinning is 10 to 25 cm.

[0023] In some embodiments of the present invention, the rotational speed of the twisting device in the twisting and winding machine is 100 - 500 r / min.

[0024] In some embodiments of the present invention, the rotational speed of the winding device in the twisting and winding machine is 50 - 300 r / min.

[0025] In some embodiments of the present invention, the ambient temperature for 3D printing in step S3 is 20°C - 30°C.

[0026] In some embodiments of the present invention, the ambient relative humidity for 3D printing in step S3 is 30% - 60%.

[0027] In some embodiments of the present invention, the moving speed of the printer needle for 3D printing in step S3 is 0.5 - 2 mm / s.

[0028] In some embodiments of the present invention, the material spraying speed for 3D printing in step S3 is 0.01 - 0.03 mm / s.

[0029] In some embodiments of the present invention, the temperature of the printer lumen for 3D printing in step S3 is 50°C - 200°C.

[0030] In some embodiments of the present invention, the temperature of the printer needle for 3D printing in step S3 is 60°C - 200°C.

[0031] In some embodiments of the present invention, the diameter of the printer needle for 3D printing in step S3 is 0.2 - 0.6 mm.

[0032] In some embodiments of the present invention, the planar line distance for 3D printing in step S3 is 2 - 5 mm.

[0033] In some embodiments of the present invention, step S4 further includes the calcination or carbonization treatment of the nanofiber yarn or precursor nanofiber yarn.

[0034] In some embodiments of the present invention, the temperature of the calcination treatment is 60°C - 800°C.

[0035] In some embodiments of the present invention, the temperature of the carbonization is 600°C - 1000°C.

[0036] In some embodiments of the present invention, the time of the carbonization is 1 - 5 h.

[0037] According to the second aspect of the present invention, there is provided a 3D printed composite scaffold of nanofiber yarn prepared by the preparation method described in the first aspect of the present invention.

[0038] According to the third aspect of the present invention, there is provided an application of the nanofiber yarn 3D printing composite scaffold prepared by the preparation method described in the first aspect of the present invention in the preparation of bone repair materials.

[0039] The present invention has at least the following beneficial effects:

[0040] The preparation method of the nanofiber yarn 3D printing composite scaffold provided by the present invention combines nanofiber yarns with 3D printing technology to manufacture a multi-structured scaffold with a nano-morphology structure. This scaffold not only has a complex geometric structure formed by 3D printing but also has a nano-fiber morphology structure, which can better simulate the microenvironment of natural tissues, thereby overcoming the limitations of traditional scaffolds and providing a more ideal tissue engineering scaffold. The nanofiber yarn 3D printing composite scaffold provided by the present invention has a wide range of applications in the field of tissue engineering, has excellent biocompatibility, high porosity, and biodegradability, is beneficial to cell adhesion, migration, and proliferation, and is suitable for fields such as bone defect repair and tissue engineering. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] The following further describes the present invention in conjunction with the drawings and embodiments, where:

[0042] Figure 1 is a morphological structure diagram of nanofibers under a super-depth-of-field microscope for the nanofibers prepared in Example 1 of the present invention, where the scale bar is 1:50;

[0043] Figure 2 is a morphological structure diagram of the nanofiber yarn prepared in Example 1 of the present invention under an electron scanning microscope;

[0044] Figure 3 is a physical diagram of the nanofiber yarn scaffold prepared in Example 1 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0045] The following will clearly and completely describe the concept and technical effects generated by the present invention in conjunction with the embodiments to fully understand the purpose, features, and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of protection of the present invention.

[0046] Example 1

[0047] In this example, a nanofiber yarn 3D printing composite scaffold was prepared, and its preparation method specifically included the following steps:

[0048] 1) Weigh 10 g of polyvinyl butyral (PVB) powder and 90 mL of absolute ethanol. Add the absolute ethanol to a container with a magnetic stirring rotor and place it on a magnetic stirrer for continuous stirring. At this time, slowly add the pre-weighed 10 g of PVB powder and mix evenly to prepare a 10% PVB solution.

[0049] 2) Weigh 8 g of absolute ethanol, 2.4 g of ultrapure water, 0.8 g of glacial acetic acid, and 7.48 g of tetraethyl orthosilicate, mix them evenly and stir for more than 0.5 h to obtain a mixed solution containing Si element.

[0050] 3) Mix the 10% PVB solution prepared in step 1) and the mixed solution containing Si element prepared in step 2) according to a volume ratio of 1:1, and place it on a magnetic stirrer for continuous stirring to obtain a spinning precursor solution.

[0051] 4) Perform electrospinning with the spinning precursor solution prepared in step 3), adjust the temperature of the spinning environment to 27 ± 2 °C, the relative humidity of the environment to 55 ± 5%, the perfusion speed to 2 mL / h; the spinning voltage to 20 kV to obtain precursor nanofibers, and their morphological structure under the electron microscope is as Figure 1 shown.

[0052] 5) Place the precursor nanofibers obtained in step 4) in a drawing and twisting device, adjust the distance between the spinning needle and the wire roller to 16.5 cm, the injection speed of the micro-injection pump to 2 mL / h, the rotation speed of the wire roller to 0.4 r / min, and the voltage of the high-voltage electrostatic generator to 20 kV to spin nanofiber yarns, and their morphological structure under the scanning electron microscope is as Figure 2 shown, Figure 2 as shown below, the fibers in the above nanofiber yarns are mainly vertically arranged along the radial direction of the yarn, and have a high degree of orientation compared with the fibers prepared by traditional electrospinning devices ( Figure 2 the fiber structure shown above).

[0053] 6) Set up the required three-dimensional model support, print it with PLA as the material to obtain a 3D printed support; adjust the parameters: the temperature of the printing environment should be 27 ± 2 °C, the relative humidity of the environment should be 55 ± 5%, the moving speed of the printer needle should be 1.0 mm / s, the spraying speed should be 0.02 mm / s, the temperature of the printer lumen should be 80 °C, the temperature of the printer needle should be 77 °C, the needle diameter should be 0.4 mm, and the plane line distance should be 3.5 mm.

[0054] 7) Weave the nanofiber yarns prepared in step 5) in the three-dimensional model support obtained in step 6) to construct a nanofiber yarn 3D printed composite support, so that the surface and inside of the support have a nanofiber structure.

[0055] Example 2

[0056] In this example, a 3D printed composite scaffold of nanofiber yarn was prepared, and its preparation method specifically includes the following steps:

[0057] 1) Weigh 1 g of polycaprolactone (PCL) powder and 9 g of hexafluoroisopropanol (HFIP). Add the hexafluoroisopropanol into a container with a magnetic stirring rotor and place it on a magnetic stirrer for continuous stirring. At this time, slowly add the pre-weighed 1 g of PCL powder and mix evenly until completely dissolved to prepare a PCL solution;

[0058] 2) Weigh 0.5 g of gelatin powder and 9.5 g of acetic acid, mix evenly and stir for more than 2 h until a transparent solution is obtained to get a gelatin solution;

[0059] 3) Mix the PCL solution prepared in step 1) and the gelatin solution prepared in step 2) according to a volume ratio of 7:3, and place it on a magnetic stirrer for continuous stirring to obtain a spinning precursor solution;

[0060] 4) Perform electrospinning with the spinning precursor solution prepared in step 3), adjust the temperature of the spinning environment to 25 ± 2 °C, the relative humidity of the environment to 40 ± 5%, the perfusion speed to 1.5 mL / h; the spinning voltage to 20 kV to obtain precursor nanofibers;

[0061] 5) Place the precursor nanofibers obtained in step 4) in a drawing and twisting device, adjust the distance between the spinning needle and the wire roller to 16.5 cm, the injection speed of the micro-injection pump to 1.5 mL / h, the rotation speed of the wire roller to 0.5 r / min, and the voltage of the high-voltage electrostatic generator to 20 kV to spin and obtain nanofiber yarn;

[0062] 6) Prepare a three-dimensional model scaffold:

[0063] Set the required three-dimensional model scaffold, use PVA as the material for printing to obtain a 3D printed scaffold; adjust the parameters: the temperature of the printing environment should be 25 ± 2 °C, the relative humidity of the environment should be 50 ± 5%, the moving speed of the printer needle should be 1.0 mm / s, the material spraying speed should be 0.02 mm / s, the temperature of the printer lumen should be 180 °C, the temperature of the printer needle should be 170 °C, the diameter of the selected needle should be 0.4 mm, and the plane line distance should be 3.5 mm.

[0064] 7) Prepare a nanofiber scaffold:

[0065] Weave the nanofiber yarn prepared in step 5) in the three-dimensional model scaffold obtained in step 6) to construct a 3D printed composite scaffold of nanofiber yarn, so that the surface and interior of the scaffold have a nanofiber structure.

[0066] Example 3

[0067] In this example, a nanofiber yarn 3D printing composite scaffold was prepared. The difference in its preparation method from that of Example 1 is only that the formulation of the mixed solution in step 2) was adjusted to 4 g of absolute ethanol, 0.6 g of ultrapure water, 0.2 g of acetic acid, and 5.6 g of tetrabutyl titanate to obtain a mixed solution containing Si element, and the remaining steps are the same as those in Example 1.

[0068] Example 4

[0069] In this example, a nanofiber yarn 3D printing composite scaffold was prepared. The specific preparation method includes the following steps:

[0070] 1) Weigh 10 g of polyvinyl alcohol (PVA) powder and 90 mL of deionized water. Add the deionized water to a container with a magnetic stirring rotor and place it on a magnetic stirrer for continuous stirring. At this time, slowly add the pre-weighed 10 g of PVA powder and mix evenly to prepare a 10% PVA solution.

[0071] 2) Weigh 2 g of collagen solution and 10 mL of acetic acid, mix evenly and stir continuously for 2 h until the solution dissolves to form a clear solution to prepare a collagen solution.

[0072] 3) Mix the 10% PVA solution prepared in step 1) and the collagen solution prepared in step 2) according to a volume ratio of 1:1, and place it on a magnetic stirrer for continuous stirring to obtain a spinning precursor solution.

[0073] 4) Use the spinning suspension prepared in step 3) for electrospinning. Adjust the temperature of the spinning environment to 25 ± 2 °C, the environmental relative humidity to 50 ± 5%, and the perfusion speed to 1 mL / h; the spinning voltage is 20 kV to obtain precursor nanofibers.

[0074] 5) Place the precursor nanofibers obtained in step 4) in a drawing and twisting device. Adjust the distance between the spinning needle and the wire drum to 16.5 cm, the pushing speed of the micro-injection pump to 2 mL / h, the rotation speed of the wire drum to 0.4 r / min, and the voltage of the high-voltage electrostatic generator to 20 kV to spin nanofiber yarns.

[0075] 6) Prepare a 3D printed scaffold:

[0076] Set the required 3D model scaffold, use PLA as the material for printing to obtain a 3D printed scaffold; adjust the parameters: the temperature of the printing environment should be 27 ± 2 °C, the environmental relative humidity should be 55 ± 5%, the moving speed of the printer needle is 1.0 mm / s, the material spraying speed is 0.02 mm / s, the temperature of the printer lumen is 80 °C, the temperature of the printer needle is 77 °C, the diameter of the selected needle is 0.4 mm, and the plane line distance is 3.5 mm.

[0077] 7) Preparation of nanofiber scaffold:

[0078] Weave the nanofiber yarn obtained in step 5) in the 3D model scaffold obtained in step 6) to construct a 3D printed composite scaffold of nanofiber yarn, so that the surface and interior of the scaffold have a nanofiber structure.

[0079] Example 5

[0080] In this example, a 3D printed composite scaffold of nanofiber yarn was prepared, and its preparation method specifically includes the following steps:

[0081] 1) Weigh 2.5 g of polylactic acid (PLA) powder and 25 mL of dichloromethane (DCM). Add dichloromethane to a container with a magnetic stirring rotor and place it on a magnetic stirrer for continuous stirring. At this time, slowly add the pre-weighed 2.5 g of polylactic acid powder and mix evenly to obtain a PLA solution;

[0082] 2) Put the solution obtained in step 1) into a constant temperature water bath, set the water bath temperature to 25 °C - 30 °C, and continuously stir for 4 - 6 hours. As time goes by, the solution gradually becomes transparent to obtain a spinning precursor solution;

[0083] 3) Perform electrospinning with the spinning suspension prepared in step 2), adjust the temperature of the spinning environment to 22 ± 2 °C, the relative humidity of the environment to 45 ± 5%, the perfusion speed to 1.5 mL / h; the spinning voltage to 18 kV to obtain precursor nanofibers;

[0084] 4) Place the precursor nanofibers obtained in step 4) in a drawing and twisting device, adjust the distance between the spinning needle and the wire roller to 16.5 cm, the injection speed of the micro-injection pump to 2 mL / h, the rotation speed of the wire roller to 0.4 r / min, and the voltage of the high-voltage electrostatic generator to 20 kV to spin nanofiber yarn;

[0085] 5) Preparation of 3D printed scaffold:

[0086] Set the required 3D model scaffold, print with PLA as the material to obtain a 3D printed scaffold; adjust the parameters: the temperature of the printing environment should be 27 ± 2 °C, the relative humidity of the environment should be 55 ± 5%, the moving speed of the printer needle should be 1.0 mm / s, the material spraying speed should be 0.02 mm / s, the temperature of the printer lumen should be 80 °C, the temperature of the printer needle should be 77 °C, the diameter of the selected needle should be 0.4 mm, and the plane line distance should be 3.5 mm;

[0087] 6) Preparation of nanofiber scaffold:

[0088] The nanofiber yarn prepared in step 5) is woven in the 3D model scaffold obtained in step 6) to construct a nanofiber yarn 3D printing composite scaffold, so that the surface and interior of the scaffold have a nanofiber structure.

[0089] The embodiments of the present invention have been described in detail above in conjunction with the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made without departing from the gist of the present invention within the scope of knowledge possessed by those of ordinary skill in the art. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A preparation method of a 3D printing composite scaffold made of nanofiber yarns, characterized in that, The preparation method includes the following steps: S1: Dissolve the polymer and / or inorganic source required for spinning in the corresponding solvent to obtain a spinning precursor solution; S2: Use the spinning precursor solution described in step S1 to spin yarn using a nanofiber spinning device to obtain nanofiber yarn; S3: Use 3D printing technology to print a polymer framework; S4: Weave the nanofiber yarn described in S2 in the polymer framework described in S3 to obtain a 3D printed composite scaffold of nanofiber yarn.

2. The preparation method according to claim 1, characterized in that, The polymer described in step S1 includes at least one of polylactic acid, polycaprolactone, polyvinyl alcohol, polyacrylonitrile, polyvinyl pyrrolidone, sodium alginate, polyethylene oxide, poly(lactic-co-glycolic acid), polyurethane, chitosan, collagen, and gelatin; Preferably, the inorganic source includes at least one of tetraethyl orthosilicate, polysilicate ethyl, tetrabutyl titanate, stannic chloride, calcium nitrate, sodium chloride, sodium carbonate, and potassium chloride; Preferably, the solvent includes at least one of water, dichloromethane, hexafluoroisopropanol, ethanol, trifluoroethanol, acetone, tert-butanol, DMF, DMSO, DMAc, THF, and acetic acid; Preferably, the spinning precursor solution further includes an additive; More preferably, the additive is at least one of acetic acid, oxalic acid, phosphoric acid, hydrochloric acid, nitric acid, and sulfuric acid.

3. The preparation method according to claim 1, characterized in that, The ambient temperature of the electrospinning in step S2 is 20°C to 30°C; Preferably, the relative humidity of the environment for the electrospinning is 30% to 60%; Preferably, the perfusion rate of the electrospinning is 0.5 to 5 mL / h; Preferably, the spinning voltage of the electrospinning is 10 to 30 kV.

4. The preparation method according to claim 1, wherein In step S2, the spinning uses a wire drum collecting device to collect the yarn, and a vertically rotating twisting and winding machine is used for twisting and winding.

5. The preparation method according to claim 4, characterized in that, The distance between the spinning needle and the wire drum during spinning is 10 to 25 cm; Preferably, the rotation speed of the wire drum during spinning is 0.1 to 0.7 r / min; Preferably, the rotation speed of the twisting device in the nanofiber spinning device is 100 to 500 r / min; Preferably, the rotation speed of the winding device in the nanofiber spinning device is 50 to 300 r / min.

6. The preparation method according to claim 1, wherein The ambient temperature of the 3D printing in step S3 is 20°C to 35°C; Preferably, the relative humidity of the environment for the 3D printing in step S3 is 30% to 60%; Preferably, the moving speed of the printer needle for the 3D printing in step S3 is 0.5 to 2 mm / s; Preferably, the spraying speed of the 3D printing in step S3 is 0.01 to 0.03 mm / s.

7. The preparation method according to claim 1, wherein The temperature of the printer lumen for the 3D printing in step S3 is 50°C to 200°C; Preferably, the temperature of the printer needle for the 3D printing in step S3 is 50°C to 200°C; Preferably, the diameter of the printer needle for the 3D printing in step S3 is 0.2 to 0.6 mm; Preferably, the plane line distance for the 3D printing in step S3 is 2 to 5 mm.

8. The preparation method according to claim 1, wherein, The raw materials of the polymer framework described in step S3 include at least one of polylactic acid, polycaprolactone, poly(lactic-co-glycolic acid), polymethyl methacrylate, gelatin, collagen, chitosan, pectin, carrageenan, sodium alginate, hydroxyapatite, tricalcium phosphate, and calcium sulfate.

9. A 3D printed composite scaffold of nanofiber yarns prepared by the preparation method according to any one of claims 1 to 8.

10. Use of a 3D printed composite scaffold of nanofiber yarns prepared by the preparation method according to any one of claims 1 to 8 in the preparation of bone repair materials.

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