CN115478328A - A nanofiber three-dimensional scaffold preparation system and method - Google Patents

Abstract

The invention discloses a system and a method for preparing a nanofiber three-dimensional support, belonging to the technical field of nanofiber supports, and comprising a forming box, an electrostatic spinning device, a low-temperature collecting device and a spraying device, wherein the electrostatic spinning device, the low-temperature collecting device and the spraying device are positioned in the forming box; one side of the forming box is provided with a spray inlet; a spinning solution nozzle of the electrostatic spinning device is fixed at the top of the forming box, the spraying device is arranged outside the forming box, and a mist outlet of the spraying device is communicated with a spraying inlet; the low-temperature collecting device comprises a collecting table and a semiconductor refrigerator, wherein the collecting table is fixed at the upper end of the semiconductor refrigerator and corresponds to the lower part of the spinning solution spray head; an alternating current high voltage generator is loaded between the spinning solution nozzle and the collecting platform to form an alternating electric field; the spinning solution or the melt ejected by the spinning solution nozzle forms the nano-fiber under the action of the alternating electric field. The size and the density of fog drops can be changed by regulating and controlling the spraying device, and the three-dimensional nanofiber support with a fluffy structure and controllable pore size can be prepared by the combined action of the spraying device and the low-temperature device.

Description

A nanofiber three-dimensional scaffold preparation system and method

technical field

The invention relates to the technical field of nanofiber scaffolds, and more specifically relates to a preparation system and method for a nanofiber three-dimensional scaffold.

Background technique

At present, the principle of tissue engineering is to induce and promote a series of physiological activities such as cell growth, migration and proliferation in vitro or in vivo, and finally form organs or tissues with three-dimensional structures. Researchers use engineering methods to create biomimetic structures that mimic the physiological environment of natural tissues, including structural, physical, and morphological characteristics. Among them, the microstructure plays a vital role in the proliferation, adhesion, inward growth of cells in the scaffold to form tissues, the good transportation of nutrients and metabolites, and the final tissue construction. At present, electrospinning is mostly used to prepare nanofibrous tissue engineering scaffolds. Electrospun nanofibers have been widely used in tissue engineering due to their morphological similarity to extracellular matrix (ECM) and chemical and physical properties that can modulate cell behavior and function. Nanofibrous tissue engineering scaffolds prepared by electrospinning have great potential for applications in regenerative medicine and wound healing. The electrospinning method has the advantages of simplicity, quickness, and low cost, and is expected to become an ideal method for preparing biomimetic scaffolds for tissue engineering. However, due to their own inherent defects, conventional electrospun nanofibrous membranes are usually packed very densely resulting in too small pore size, which limits the penetration of cells and the regeneration of three-dimensional tissues.

In traditional electrospinning, the spinning solution or melt forms a Taylor cone under the directional action of a DC electric field force, and finally breaks through the surface tension to form a jet. The jet is stretched into nanofibers under the action of Coulomb force, and the fibers are attracted and piled up by the grounded receiving plate layer by layer, forming a thin film or a nearly three-dimensional scaffold. However, the fibers of the scaffold are piled up by the attraction of electric charge, the arrangement is tight, and the gap between the fibers is too small, which makes it difficult for cells to grow in, and it is impossible to construct a satisfactory three-dimensional tissue.

At present, there is an invention patent with the application number CN201510778850.9, which discloses a nanofiber scaffold preparation device and method with a three-dimensional structure, including an AC high-voltage generator, a liquid supply device, a gas supply device, a coaxial spinning liquid nozzle and a rotating receiver. The coaxial spinning liquid nozzle has a liquid outlet and an air outlet. The liquid supply device can send the spinning solution or melt into the coaxial spinning liquid nozzle and flow out from the liquid outlet. The air supply device can generate The airflow blown from the liquid outlet to the rotating receiver, the AC high voltage generator can form an alternating electric field between the coaxial spinning liquid nozzle and the rotating receiver. The rotating receiver includes a rotating shaft, a transmission device and a supporting arm, and the supporting arm follows the rotating shaft. When rotating, a bowl-shaped rotary surface can be formed, and the spinning solution or melt flowing out of the liquid outlet forms a jet under the action of airflow and alternating electric field. The three-dimensional scaffold formed by this method has a small aperture, and cells cannot grow inside it.

In summary, there is still no simple and effective solution to prepare nanofibrous 3D scaffolds with a wide range of materials, extremely small fiber diameters, uniform filament diameter distribution, fluffy structure, large pore size, and superior mechanical properties.

Contents of the invention

In view of this, the present invention aims to provide a nanofiber three-dimensional scaffold preparation system and method to at least solve one of the above-mentioned technical problems in the prior art to a certain extent.

In order to achieve the above object, the present invention adopts the following technical solutions:

A nanofiber three-dimensional scaffold preparation system, comprising: a forming box, an electrospinning device located in the forming box, a low-temperature collecting device and a spraying device;

One side of the forming box has a spray inlet; the electrospinning device includes a spinning liquid nozzle, a liquid feeder and an AC high-voltage generator, and the liquid feeder pipeline of the electrostatic spinning device is connected to the spinning A liquid spray head, the spinning liquid spray head is fixed on the inner top of the forming box, and the outlet of the liquid supply device is connected to the spinning liquid spray head through a pipeline;

The spray device is placed outside the molding box, and the mist outlet of the spray device communicates with the spray inlet; the low-temperature collection device includes a collection table and a semiconductor refrigerator, and the collection table is fixed on the semiconductor refrigerator. The upper end of the refrigerator corresponds to the lower part of the spinning solution nozzle; the AC high-voltage generator is loaded between the spinning solution nozzle and the collecting table to form an alternating electric field; the spinning solution ejected from the spinning solution nozzle Silk solution or melt forms nanofibers under the action of alternating electric field.

Turn on the AC high-voltage generator, an alternating electric field is formed between the spinning solution nozzle and the low-temperature collecting device, and the spinning solution or melt provided by the liquid supply device flows out from the outlet of the spinning solution nozzle to form a jet, and the jet splits under the action of the alternating electric field The nanofibers are split and stretched to form nanofibers and gather on the collection platform. The spray device can generate micron-sized mist droplets. The mist droplets and nanofibers can form a frozen three-dimensional nanofiber scaffold on the collection platform under the action of a semiconductor refrigerator.

Preferably, the spray device includes a liquid loader and an atomizer, the liquid loader is located above the atomizer and is detachably connected to its interface, and the mist outlet is set on the atomizer housing In addition, the upper housing of the atomizer is also provided with a switch and an adjustment button, and the switch and the adjustment button are in contact with the control module inside the atomizer. The spray device can be adjusted according to the needs, spraying droplets of different densities and sizes.

Preferably, the low temperature collection device also includes a lifting support platform and a fixed plate, and the lifting support platform adopts a lifting type lifting support platform, and the low temperature collection device also includes a lifting support platform and a fixed plate, and the fixed plate is placed on the On the lifting support platform, the lower end of the semiconductor refrigerator is fixedly connected to the fixing plate and electrically connected to the power supply outside the forming box. The height of the lifting support table can be adjusted, and the distance between the collection table and the spinning liquid nozzle can be changed according to the experimental needs. The semiconductor refrigerator provides a low-temperature environment.

Preferably, an air outlet tube is formed inside the box on the side of the spray inlet, the outlet tube corresponds to the top of the collection platform, and the spray inlet is horizontally aligned with the mist outlet of the spray device. The air outlet tube guides the mist to the low-temperature collection platform, so that the fog droplets fall on the collection platform more accurately.

Preferably, an air outlet is provided at the bottom of the box, and a porous plate is fixedly connected to the inner wall of the air outlet. The mist that does not fall on the collection table is discharged from the air outlet at the bottom of the box to the outside of the box. The air outlet is designed with a perforated plate to balance the pressure difference between the spinning space inside the box and the outside.

Preferably, the low-temperature collection device is made of conductive metal, and the lifting support platform is connected with a grounding wire. It is convenient for conducting electricity and collecting station electricity.

Preferably, the forming box body is made of insulating material. A closed spinning space can be formed inside, which is not affected by other factors.

The vertical distance from the spinning dope nozzle to the collection platform is 10cm-20cm; the horizontal distance from the air outlet tube to the collection platform is 5cm-15cm.

A method for preparing a nanofiber three-dimensional scaffold, comprising the following steps:

(1) The liquid supply device can be filled with spinning solution or melt, open the liquid supply device, the spinning solution or melt enters the spinning solution nozzle and flows out from the outlet of the spinning solution nozzle;

(2) Adjust the lifting support platform to a suitable height, ground the ground wire, fix the semiconductor refrigerator on the fixed plate, turn on the power, adjust the semiconductor refrigerator to a suitable cooling temperature, and finally the surface temperature of the collecting table reaches the cooling temperature;

(3) Adjust the position of the mist outlet to be flush with the collection platform, turn on the switch of the spray device, adjust the button to the appropriate parameter, the atomizer will generate mist, spray from the mist outlet, and the mist can pass through the outlet of the forming box The air cavity tube is evenly deposited on the collection table; the uncured mist is discharged from the air outlet;

(4) Turn on the AC high-voltage generator, output the voltage with appropriate waveform, frequency, amplitude, and bias value, and form an alternating electric field between the spinning liquid nozzle and the low-temperature collection device, and under the action of the alternating electric field, the spinning The solution or melt droplets are stretched to form a jet, and under the action of the alternating electric field force, the jet is split and stretched to obtain nanofibers;

(5) Under the joint action of the mist formed by the spray device and the low-temperature collection device, the nanofibers and mist are deposited on the collection platform for instantaneous freezing and solidification, and after continuous deposition, they become a porous and fluffy frozen three-dimensional scaffold structure;

(6) The porous and fluffy frozen three-dimensional scaffold structure is freeze-dried in a freeze dryer to finally obtain a three-dimensional nanofiber tissue engineering scaffold.

It can be seen from the above technical solutions that, compared with the prior art, the present invention discloses a system and method for preparing a nanofiber three-dimensional scaffold, which has the following beneficial effects:

1. The present invention can change the droplet size and droplet density by adjusting the spray device, thereby controlling the aperture ratio and aperture size; different types of nanofibers can be prepared by using different electrospinning raw materials, and the nanofibers can be changed by setting the parameters of the AC high voltage generator diameter of.

2. The present invention uses a low-temperature device to deposit the nanofibers formed by the mist and the spinning solution or melt on the collection platform together to form a frozen three-dimensional nanofiber scaffold, which is then dried by a freeze-drying agent to form a porous and fluffy three-dimensional nanofiber scaffold. Nanofibrous Scaffolds for Tissue Engineering. The nanofibers obtained through this process have the advantages of controllable fiber diameter, uniform pore size distribution, fluffy structure, controllable pore size, large thickness and superior mechanical properties, and the nanofiber scaffold has a stable process and good versatility.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only It is an embodiment of the present invention, and those skilled in the art can also obtain other drawings according to the provided drawings without creative work.

Fig. 1 is a schematic diagram of a cross-sectional structure provided by the present invention.

1-electrospinning device, 1a-spinning solution nozzle, 1b-liquid supply device, 1c-AC high voltage generator, 1d-nanofiber;

2-spray device, 2a-interface, 2b-liquid filler, 2c-atomizer, 2d-switch, 2e-adjust button, 2f-mist outlet, 2g-droplet;

3-low temperature collection device, 3a-power supply, 3b-semiconductor refrigerator, 3c-collection platform, 3d-fixed plate, 3e-grounding wire, 3f-lifting support platform;

4-forming box, 4a-outlet cavity pipe, 4b-air outlet, 4c-casing.

detailed description

The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

In describing the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", The orientation or positional relationship indicated by "bottom", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying the referred device or positional relationship. Elements must have certain orientations, be constructed and operate in certain orientations, and therefore should not be construed as limitations on the invention.

In the present invention, unless otherwise clearly specified and limited, terms such as "installation", "connection", "connection" and "fixation" should be understood in a broad sense, for example, it can be a fixed connection or a detachable connection , or integrated; it can be mechanically connected or electrically connected; it can be directly connected or indirectly connected through an intermediary, and it can be the internal communication of two components or the interaction relationship between two components. Those of ordinary skill in the art can understand the specific meanings of the above terms in the present invention according to specific situations.

Please refer to Fig. 1, the embodiment of the present invention discloses a nanofiber 1d three-dimensional scaffold preparation system based on a low-temperature collection device 3, including: a forming box 4, an electrospinning device 1 located in the forming box, a low-temperature collection device 3 and a spray device 2;

One side of the molding box 4 has a spray inlet; the electrospinning device 1 includes a spinning solution nozzle 1a, a liquid supply device 1b and an AC high-voltage generator 1c, and the liquid supply device 1b of the electrospinning device 1 is connected to the spinning solution nozzle 1a , the spinning solution nozzle 1a is fixed on the inner top of the forming box 4, and the outlet of the liquid supply device 1b is connected to the spinning solution nozzle 1a through a pipeline;

The spray device 2 is placed outside the molding box 4, and the mist outlet 2f of the spray device 2 communicates with the spray inlet; the low-temperature collection device 3 includes a collection platform 3c and a semiconductor refrigerator 3b, and the collection platform 3c is fixed on the upper end of the semiconductor refrigerator 3b and corresponds to Below the spinning solution nozzle 1a; AC high-voltage generator 1c is loaded between the spinning solution nozzle 1a and the collecting table 3c to form an alternating electric field; the spinning solution or melt ejected from the spinning solution nozzle 1a acts on the alternating electric field The nanofibers 1d are formed below.

The electrospinning device 1 is an existing technology, so the spinning liquid spray head 1a and the liquid supply device 1b can be selected according to requirements. Turn on the AC high-voltage generator 1c, an alternating electric field is formed between the spinning solution nozzle 1a and the low-temperature collecting device 3 of the metal material, and the spinning solution or melt provided by the liquid supply device 1b flows out from the outlet of the spinning solution nozzle 1a to form a jet, Under the action of an alternating electric field, the jet splits and stretches to form nanofibers 1d, which are gathered on the collection platform 3c. The spray device 2 can produce micron-sized droplets 2g, and the mist droplets 2g and nanofibers 1d can act on the semiconductor refrigerator 3b. Next, a frozen three-dimensional nanofiber scaffold is formed on the collection table 3c.

Wherein, the spray device 2 can produce micron-sized mist droplets 2g, the spray device 2 includes a liquid loader 2b and an atomizer 2c, the liquid loader 2b is located above the atomizer 2c and the interface 2a is threaded, and the mist outlet is set On the atomizer 2c, a switch 2d and an adjustment button 2e are also provided on the upper shell of the atomizer 2c, and both the switch 2d and the adjustment button 2e are in contact with the control module inside the atomizer 2c. The spraying device 2 can be adjusted according to needs, spraying out 2g of mist droplets with different densities and sizes.

The cryogenic collection device 3 also includes a lifting support platform 3f and a fixed plate 3d, the fixed plate 3d is placed on the lifting support platform 3f, the semiconductor refrigerator 3b is fixed on the fixed plate 3f, and is electrically connected to the power supply 3a outside the molding box 4. In a specific embodiment, the semiconductor refrigerator 3b is fixed on the fixed plate 3d by an adhesive, and the lifting support platform 3f adopts the L2241 model laboratory lifting platform of Core Silicon Valley.

Further, an air outlet tube 4a is formed inside the box on the spray inlet side, and the outlet tube 4a corresponds to the top of the collecting platform 3c, and the spray inlet is horizontally aligned with the mist outlet 2f of the spray device 2 .

Further, the bottom of the box body 4c is provided with an air outlet 4b, and the inner wall of the air outlet 4b is fixedly connected with a perforated plate. The air outlet tube 4a accurately guides the mist 2g to the collection platform 3c, and the mist 2g that does not fall on the collection platform 3c is discharged from the air outlet 4b at the bottom of the box 4c to the outside of the box 4c. There are a plurality of micropores, which can balance the pressure difference between the spinning space inside the casing 4c and the outside world.

Further, the cryogenic collection device 3 is made of conductive metal, and the lifting support platform 3f is connected with a ground wire 3e.

Further, the AC high voltage generator 1c may be a function signal generator or a high voltage amplifier. The AC high-voltage generator 1c can provide voltages of different frequencies and amplitudes to form an alternating electric field between the spinning solution nozzle 1a and the low-temperature collecting device 3 .

Further, the box body 4c of the molding box 4 is made of insulating material. A closed spinning space can be formed inside, which is not affected by other factors.

The vertical distance between the spinning dope nozzle 1a and the collection platform 3c is 10cm-20cm; the horizontal distance between the spray inlet and outlet and the collection platform 3c is 5cm-15cm.

A method for preparing a nanofiber 1d three-dimensional scaffold based on a low-temperature collection device, comprising the following steps:

(1) The liquid supply device 1b can be filled with spinning solution or melt, and the liquid supply device 1b is opened, and the spinning solution or melt enters the spinning solution nozzle 1a and flows out from the outlet of the spinning solution nozzle 1a;

(2) Adjust the lifting support platform 3f to a suitable height, the grounding wire 3e is grounded, the fixed plate 3d fixes the semiconductor refrigerator 3b, the power supply 3a is turned on, the semiconductor refrigerator 3b is adjusted to a suitable cooling temperature, and finally the surface temperature of the collecting platform 3c reaches refrigeration. temperature;

(3) Adjust the position of the mist outlet 2f to be flush with the collection table 3c, turn on the switch 2d of the spray device 2, and adjust the adjustment button 2e to a suitable parameter, the atomizer 2c will generate mist droplets, which will be sprayed from the mist outlet 2f, and the mist The droplets 2g can pass through the air outlet tube 4a of the molding box 4, and are evenly deposited on the collecting table 3c; the uncured mist droplets 2g are discharged from the air outlet 4b.

(4) Open the AC high-voltage generator 1c, output the voltage of appropriate waveform, frequency, amplitude, and bias value, and form an alternating electric field between the spinning solution nozzle 1a and the low-temperature collection device 3, under the action of the alternating electric field , the spinning solution or melt droplets are stretched to form jets, and under the action of alternating electric field force, the jets are split and stretched to obtain nanofibers 1d;

(5) Under the joint action of the mist 2g formed by the spray device 2 and the low-temperature collection device 3, the nanofiber 1d and the mist 2g are deposited on the collection platform 3c to freeze and solidify instantaneously, and after continuous deposition, they become a porous and fluffy frozen three-dimensional scaffold structure.

(6) The porous and fluffy frozen three-dimensional scaffold structure is freeze-dried in a freeze dryer to finally obtain a three-dimensional nanofiber tissue engineering scaffold.

The method of the invention is used to obtain the three-dimensional nanofiber tissue engineering scaffold, the pore diameter is 100 μm to 300 μm, the fiber diameter is 250 nm to 500 nm, and meets the fluffy requirement.

A porous and fluffy three-dimensional nanofibrous tissue engineering scaffold with adjustable pore size can be prepared through the method of this embodiment, which can be applied to the preparation of three-dimensional structures of organs or tissues.

For different spinning solutions and different spray materials, the following examples are provided:

Example 1:

1. Preparation of spinning solution, weigh 1 gram of polycaprolactone (molecular weight = 100,000 Daltons), dissolve it in 10 ml (9:1, CH ₂ Cl ₂ /DMF, v/v) solvent, and prepare 10% polycaprolactone solution, sealed with a parafilm, magnetically stirred for 4 hours, and set aside.

2. Adjust the parameters of the liquid supply device 1b, the capacity is 10 ml, the propulsion speed is 2 ml/hour, and the polycaprolactone solution is delivered to the spinning solution nozzle 1a, and the outlet diameter of the spinning solution nozzle 1a is 0.3 mm.

3. The lifting support platform 3f is adjusted to a suitable height, so that the distance between the spinning liquid nozzle 1a and the collecting platform 3c is 20cm, the grounding wire 3e is grounded, the fixed plate 3d fixes the semiconductor refrigerator 3b, and the power supply 3a is turned on, and the semiconductor refrigerator 3b Adjust to a suitable refrigeration temperature, and finally the surface temperature of the collection platform 3c reaches -20°C;

4. Put the aqueous solution in the spray device 2, adjust the position of the mist outlet 2f to be flush with the collection table 3c, turn on the switch 2d of the spray device 2, adjust the adjustment button 2e to 0.5 ml of liquid per minute, and the atomizer 2c will generate mist droplets 2g, sprayed from the mist outlet 2f.

5. The mist droplets 2g can pass through the air outlet tube 4a of the forming box 4 and be evenly deposited on the collection table 3c; the uncured mist droplets 2g are discharged from the air outlet 4b.

6. Adjust the AC high-voltage generator 1c, the output frequency is 100Hz, and the peak-to-peak value is 7-12KV sinusoidal high-voltage AC; an alternating electric field is formed between the spinning solution nozzle 1a and the low-temperature collection device 3, and under the action of the alternating electric field, Spinning solution droplets are stretched to form a jet, and under the action of an alternating electric field force, the jet is split and stretched to obtain nanofibers 1d.

7. Under the joint action of the mist droplets 2g formed by the spray device 2 and the low-temperature collection device 3, the nanofibers 1d and the mist droplets 2g are deposited on the collection platform 3c for instant freezing and solidification, and after continuous deposition, they become porous and fluffy frozen polycaprolactone nano Fibrous three-dimensional scaffold structure.

8. Place the frozen three-dimensional scaffold structure in a low-temperature vacuum dryer for 24 hours to freeze-dry. The obtained polycaprolactone three-dimensional nanofiber scaffold has a fluffy structure, a pore diameter of 100 μm to 300 μm, and a fiber diameter of 250 nm to 500 nm.

Example 2:

1. Spinning solution preparation, take by weighing 1.6 grams of polyvinyl alcohol (molecular weight=100,000 Daltons), be dissolved in 20 milliliters of ethanol solvents, make 8% polyvinyl alcohol solution, seal with parafilm, magnetically stir 6 hours ,stand-by.

2. Adjust the parameters of the liquid supply device 1b, with a capacity of 8 milliliters and a propulsion speed of 1.5 milliliters per hour to transport the polycaprolactone solution to the spinning solution nozzle 1a, and the outlet diameter of the spinning solution nozzle 1a is 0.3 mm.

3. The lifting support platform 3f is adjusted to a suitable height, so that the distance between the spinning liquid nozzle 1a and the collecting platform 3c is 20cm, the grounding wire 3e is grounded, the fixed plate 3d fixes the semiconductor refrigerator 3b, and the power supply 3a is turned on, and the semiconductor refrigerator 3b Adjust to a suitable refrigeration temperature, and finally the surface temperature of the collection platform 3c reaches -20°C;

4. Put 1% collagen solution in the spray device 2, adjust the position of the mist outlet 2f to be flush with the collection table 3c, turn on the switch 2d of the spray device 2, adjust the adjustment button 2e to 1 ml of liquid per minute, and the atomizer 2c will Generate mist droplets 2g, which are sprayed out from the mist outlet 2f.

5. The mist droplets 2g can pass through the air outlet tube 4a of the forming box 4 and be evenly deposited on the collection table 3c; the uncured mist droplets 2g are discharged from the air outlet 4b.

6. Adjust the AC high-voltage generator 1c, the output frequency is 100Hz, and the peak-to-peak value is 8-15KV sinusoidal high-voltage AC; an alternating electric field is formed between the spinning solution nozzle 1a and the low-temperature collection device 3, and under the action of the alternating electric field, Spinning solution droplets are stretched to form a jet, and under the action of an alternating electric field force, the jet is split and stretched to obtain nanofibers 1d.

7. Under the joint action of the mist droplets 2g formed by the spray device 2 and the low-temperature collection device 3, the nanofibers 1d and the mist droplets 2g are deposited on the collection platform 3c for instant freezing and solidification, and after continuous deposition, they become porous and fluffy frozen polyvinyl alcohol nanofibers Three-dimensional scaffold structure.

8. Place the frozen three-dimensional scaffold structure in a low-temperature vacuum dryer to freeze-dry for 24 hours. The obtained polyvinyl alcohol three-dimensional nanofiber scaffold has a fluffy structure, a pore diameter of 100 μm to 200 μm, and a fiber diameter of 350 nm to 500 nm.

Each embodiment in this specification is described in a progressive manner, each embodiment focuses on the difference from other embodiments, and the same and similar parts of each embodiment can be referred to each other. As for the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and for the related information, please refer to the description of the method part.

The above description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention will not be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (9)

1. A nanofiber three-dimensional scaffold preparation system, comprising: the electrostatic spinning device is positioned in the forming box;

one side of the forming box is provided with a spray inlet; the electrostatic spinning device comprises a spinning solution nozzle, a solution supply device and an alternating-current high-voltage generator, wherein the solution supply device of the electrostatic spinning device is connected to the spinning solution nozzle through a pipeline, the spinning solution nozzle is fixed at the top end inside the forming box, and an outlet of the solution supply device is communicated with the spinning solution nozzle through a pipeline;

the spraying device is arranged outside the forming box, and a mist outlet of the spraying device is communicated with the spraying inlet; the low-temperature collecting device comprises a collecting table and a semiconductor refrigerator, and the collecting table is fixed at the upper end of the semiconductor refrigerator and corresponds to the lower part of the spinning solution spray head; the alternating current high voltage generator is loaded between the spinning solution nozzle and the collecting platform to form an alternating electric field; and the spinning solution or the melt jetted by the spinning solution nozzle forms the nano-fiber under the action of the alternating electric field.

2. The nanofiber three-dimensional support preparation system according to claim 1, wherein the spraying device comprises a liquid loading device and an atomizer, the liquid loading device is located above the atomizer and detachably connected with an interface of the atomizer, the mist outlet is formed in the atomizer shell, a switch and an adjusting button are further arranged on the atomizer shell, and the switch and the adjusting button are both abutted to a control module inside the atomizer.

3. The nanofiber three-dimensional scaffold preparing system as claimed in claim 1, wherein the low temperature collecting device further comprises a lifting support table and a fixing plate, the fixing plate is placed on the lifting support table, and the lower end of the semiconductor refrigerator is fixedly connected to the fixing plate and electrically connected to a power supply outside the forming box.

4. The system for preparing three-dimensional nanofiber scaffolds as claimed in claim 1, wherein the spraying inlet side is formed with an air outlet pipe inside the box body, the air outlet pipe corresponds to the upper part of the collection platform, and the spraying inlet is horizontally flush with the mist outlet of the spraying device.

5. The nanofiber three-dimensional support preparation system according to claim 1, wherein a gas outlet is formed in the bottom of the box body, and a porous plate is fixedly connected to the inner wall of the gas outlet.

6. The system for preparing the nanofiber three-dimensional scaffold as claimed in any one of claims 1 to 5, wherein the low-temperature collecting device is made of conductive metal, and the lifting support table is connected with a grounding wire.

7. The nanofiber three-dimensional scaffold preparing system as claimed in any one of claims 1 to 5, wherein the forming box is made of an insulating material.

8. The nanofiber three-dimensional scaffold preparing system as claimed in any one of claims 1 to 5, wherein the vertical distance of the spinning solution nozzle from the collecting stage is in the range of 10cm to 20cm; the horizontal distance range of the air outlet cavity pipe from the collecting platform is 5cm-15cm.

9. A preparation method of a nanofiber three-dimensional scaffold is characterized by comprising the following steps:

(1) The liquid supply device can be added with spinning solution or melt, the liquid supply device is opened, and the spinning solution or melt enters the spinning solution nozzle and flows out from the outlet of the spinning solution nozzle;

(2) Adjusting the lifting support platform to a proper height, grounding a grounding wire, fixing the semiconductor refrigerator by the fixing plate, turning on a power supply, adjusting the semiconductor refrigerator to a proper refrigeration temperature, and finally enabling the surface temperature of the collection platform to reach the refrigeration temperature;

(3) Adjusting the position of the fog outlet to be flush with the collecting platform, turning on a switch of the spraying device, adjusting a button to a proper parameter, enabling the atomizer to generate fog drops, spraying out the fog drops from the fog outlet, and enabling the fog drops to be uniformly deposited on the collecting platform through an air outlet cavity tube of the forming box; discharging the uncured fog drops from the air outlet;

(4) Opening an alternating current high voltage generator, outputting voltage with proper waveform, frequency, amplitude and offset value, forming an alternating electric field between a spinning solution nozzle and a low-temperature collecting device, stretching spinning solution or melt droplets to form jet flow under the action of the alternating electric field, and splitting and stretching the jet flow under the action of the alternating electric field to obtain nano fibers;

(5) Under the combined action of fog drops formed by a spraying device and a low-temperature collecting device, the nano fibers and the fog drops are deposited on a collecting platform for instantaneous freezing and solidification, and after continuous deposition, the nano fibers and the fog drops become a porous fluffy freezing three-dimensional support structure;

(6) And (3) placing the porous fluffy frozen three-dimensional scaffold structure in a freeze dryer for freeze drying to finally obtain the three-dimensional nanofiber tissue engineering scaffold.

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