Preparation method of layered regulation fiber membrane
Through the preparation method of layered control of fiber membranes, the pores are adjusted by spinning liquid traction and fiber accumulation methods, which solves the problem of single pores of electrospinning membranes, and realizes the application of multifunctional tissue engineering scaffolds.
Patent Information
- Application Number
- CN202510770674.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-10
- Publication Date
- 2025-09-05
AI Technical Summary
The pore distribution of existing fiber membranes prepared by electrospinning is relatively single, making it difficult to realize the composite function of tissue engineering scaffolds.
The preparation method of layered control fiber membranes is adopted, and the pore changes are adjusted through spinning liquid traction and fiber accumulation to form different pore areas. A double- or triple-layer fiber membrane is prepared using polymers and hydrophilic materials. The inner layer is a high-pore layer and the outer layer is a poreless layer or a low-pore layer to achieve layered control of porosity.
The porosity layered regulation of fiber membranes is achieved, cell adhesion and tissue repair effects are improved, and mechanical enhancement and anti-adhesion functions are provided to meet the multifunctional needs of tissue engineering scaffolds.
Smart Images
Figure CN120591965A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomaterials, and in particular relates to a method for preparing a layered regulated fiber membrane. Background Art
[0002] Tissue engineering scaffolds are widely used in modern medicine, promoting tissue repair, providing adequate mechanical support, and preventing adhesion. Their porous structure allows for cell adhesion, binding, and proliferation, while also facilitating the diffusion of nutrients, oxygen, and waste products. Studies have shown that large-pore tissue engineering scaffolds facilitate nutrient supply and waste removal, while small-pore tissue engineering scaffolds provide more surface area for cell adhesion.
[0003] Fiber membranes produced by electrospinning have a high specific surface area and suitable porosity, which facilitates the provision of cell binding sites and promotes tissue repair. Therefore, electrospinning technology is considered an ideal process for mimicking the extracellular matrix and is widely used in the preparation of tissue engineering scaffolds. However, current electrospinning processes use a single polymer as a raw material, resulting in a relatively monotonous pore distribution in the resulting electrospun fiber membranes, making it difficult to achieve the composite functions of tissue engineering scaffolds. Summary of the Invention
[0004] In response to the problems in the prior art, the present invention provides a method for preparing layered controlled fiber membranes, which solves the problem of single pore size in the existing electrospinning method for preparing fiber membranes. The spinning solution traction method and the fiber stacking method are used to adjust the pore changes, thereby forming different pore areas in electrospinning, thereby achieving the effect of layered control of porosity.
[0005] In order to achieve the above technical objectives, the technical solution of the present invention is: A method for preparing a layered regulated fiber membrane, using one or more polymers as raw materials and an electrospinning process to prepare the layered fiber membrane, wherein the layered fiber membrane is a double-layer fiber membrane or a triple-layer fiber membrane, and the pores between the layers of the layered fiber membrane are regulated to achieve the functions of cell growth, mechanical enhancement, anti-adhesion and step-by-step degradation, wherein a hydrophilic material is added to the inner fiber membrane.
[0006] The raw materials are one or more of polyglycolic acid, polylactide-caprolactone copolymer, polyglycolide-trimethylene carbonate block copolymer, polylactic acid-polyethylene glycol block copolymer, polylactic acid, polycaprolactone, and polyglycolide-lactide copolymer.
[0007] The solvent is one or more of N,N-dimethylformamide, acetone, hexafluoroisopropanol, dichloromethane or chloroform.
[0008] The hydrophilic material is polyethylene glycol or glycerol, and the addition amount is 2-8% w / v. Adding the hydrophilic material to the spinning solution of the inner fiber membrane can increase the hydrophilicity of the inner fiber membrane and achieve a better adhesion effect.
[0009] The inner fiber membrane of the double-layer fiber membrane is a highly porous layer with a porosity of 60-90%, preferably 65-85%; the thickness is 50-100 μm, preferably 50-80 μm; this highly porous layer can stably adhere to tissue. The outer fiber membrane of the double-layer fiber membrane is a non-porous layer with a thickness of 20-50 μm, preferably 20-30 μm. This non-porous layer provides mechanical reinforcement and anti-adhesion properties.
[0010] The inner fiber membrane of the three-layer fiber membrane is a high-porosity layer with a porosity of 60-90%, preferably 65-85%; the thickness is 50-100 μm, preferably 50-80 μm, and the high-porosity layer can stably adhere to tissue. The middle fiber membrane is a non-porous layer, and the outer fiber membrane is a low-porosity layer with a low porosity of 20-60%, preferably 20-40%, and a thickness of 20-50 μm, preferably 30-50 μm.
[0011] The layered fiber membrane achieves pore change by suppressing the spinning solution drawing mode and increasing the unit density fiber stacking mode.
[0012] The method of suppressing the stretching of the spinning solution is achieved by adjusting the spinning height, the spinning solution concentration, the spinning voltage and the spinning solution advancing speed.
[0013] Furthermore, when the low-porosity layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 1-3 cm, preferably 1.5-3 cm. Furthermore, when the non-porous layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-2 cm, preferably 0.5-1.5 cm. The method of increasing the unit density of fiber stacking is achieved by adjusting the moving speed of the receiving device.
[0014] Furthermore, when the low-porosity layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-3 cm, preferably 1-2 cm; the moving speed of the receiving device is set to L1 / (1-3), where L1 is the width of the spinning area.
[0015] Furthermore, when the non-porous layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-3 cm, preferably 1-2 cm; the moving speed of the receiving device is set to L2 / (2-5), where L2 is the width of the spinning area.
[0016] It can be seen from the above description that the present invention has the following advantages: 1. The present invention solves the problem of single pore size in existing electrospinning fiber membrane preparation. It uses the spinning solution traction method and fiber stacking method to adjust the pore changes, thereby forming different pore areas in electrospinning and achieving the effect of layered porosity regulation.
[0017] 2. The present invention utilizes the continuous electrospinning method of the same material to effectively reduce the interface difference of fiber membranes with different porosities, ensuring the integrated characteristics of the layered regulation fiber membrane. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1-3 is a scanning electron microscope image of the high-porosity layer, the low-porosity layer and the non-porous layer in Example 1 of the present invention; Figure 4-6 is a scanning electron microscope image of the high-porosity layer, the low-porosity layer and the non-porous layer in Example 2 of the present invention; Figure 7 This is the cell ingrowth into the low-porosity layer in Example 5 of the present invention.
[0019] Figure 8 This is the cell ingrowth into the high-porosity layer in Example 5 of the present invention.
[0020] Figure 9 This is the attachment condition of the fiber membrane prepared in Example 1 among Example 6 of the present invention.
[0021] Figure 10 This is the attachment condition of the fiber membrane prepared in Example 6 of Example 6 of the present invention. DETAILED DESCRIPTION
[0022] Combine Figure 1-10 , which describes the present invention in detail but does not limit the claims of the present invention.
[0023] Example 1 A method for preparing a three-layer fiber membrane comprises preparing a 40 w / v% spinning solution of polyglycolide-lactide copolymer and polylactic acid-polyethylene glycol block copolymer, wherein the solvent is a mixture of N,N-dimethylformamide and acetone in a volume ratio of 7:3, and the mass ratio of polyglycolide-lactide copolymer to polylactic acid-polyethylene glycol block copolymer is 70:30.
[0024] The spinning parameters of the high-porosity layer in the three-layer fiber membrane are as follows: spinning height 25 cm, spinning voltage 30 kV, and spinning solution advancing speed 20 μL / min; The spinning parameters of the low-porosity layer were as follows: spinning height 18 cm, spinning voltage 20 kV, and spinning liquid propulsion speed 30 μL / min; The spinning parameters of the non-porous layer are as follows: spinning height of 12 cm, spinning voltage of 20 kV, propulsion speed of the killing liquid of 30 μL / min; the receiving device is a roller, and the moving speed is 0.5 cm / s.
[0025] The micromorphology of each layer of the three-layer fiber membrane was observed using a scanning electron microscope. Figure 1-3 shown.
[0026] Example 2 A method for preparing a three-layer fiber membrane comprises preparing a 15% w / v spinning solution with a polyglycolide-lactide copolymer, wherein the solvent is a mixture of hexafluoroisopropanol and dichloromethane in a volume ratio of 7:3, and adding 5% w / v glycerol to the spinning solution prepared for the inner fiber membrane.
[0027] The spinning parameters of the high-porosity layer in the three-layer fiber membrane are as follows: spinning height 28 cm, spinning voltage 30 kV, and spinning solution advancing speed 30 μL / min; The spinning parameters of the low-porosity layer were as follows: spinning height of 20 cm, spinning voltage of 25 kV, and spinning solution advancing speed of 30 μL / min; The spinning parameters of the non-porous layer are as follows: spinning height of 15 cm, spinning voltage of 25 kV, spinning solution advancing speed of 30 μL / min; the receiving device is a roller, and the moving speed is 0.4 cm / s.
[0028] The micromorphology of each layer of the three-layer fiber membrane was observed using a scanning electron microscope. Figure 4-6 shown.
[0029] The scanning electron microscope images of the fiber membranes of Example 1 and Example 2 are shown below:
[0030] Example 3 The porosity test data of each layer of fiber membrane prepared in Example 1 is as follows: (1) Determination of dry sample mass: Prepare a 50*60 mm fiber membrane and dry it in a vacuum drying oven at room temperature to constant weight. Measure the dry weight (m1) of each sample to an accuracy of 0.001 g.
[0031] (2) Sample impregnation: Place the sample in a container filled with purified water and place it in a vacuum drying oven, adjusting the pressure to a negative state until the sample is completely soaked in purified water.
[0032] (3) Determination of the suspended mass of saturated samples Use 4-0 absorbable surgical suture (with needle) to tie to the electronic balance, remove the sample from the immersion liquid, place the saturated sample after immersion on the suture needle, and hang it in a container containing the immersion liquid. Weigh the suspended mass (m2) of the saturated sample in the immersion liquid to an accuracy of 0.001g.
[0033] (4) Determination of saturated sample mass (m3) Take out the sample from the immersion liquid, gently wipe off the liquid attached to the sample surface with filter paper, and quickly weigh the saturated sample mass (m3) to an accuracy of 0.001g.
[0034] Porosity calculation: P=(m3-m1) / (m3-m2)*100%.
[0035] The results of the above porosity test are as follows:
[0036] The above data show that the method of suppressing the stretching of the spinning solution and increasing the unit density fiber stacking adopted in this technical solution can effectively improve the porosity of the fiber membrane and realize the pore regulation in the electrospinning process.
[0037] Example 4 The fiber membrane prepared in Example 2 was used as a test sample to test the mechanical degradation data of the high-porosity layer, the low-porosity layer and the non-porous layer.
[0038] The testing method involves preparing a PBS buffer solution (pH 7.4), cutting the fiber membrane into 3 x 10 cm pieces, and immersing each piece in the buffer solution. The experiment is then conducted in a 37°C constant-temperature drying oven. The tensile strength is then tested using a universal testing machine at 5, 7, 10, 14, and 20 days, observing the point at which mechanical properties are lost.
[0039]
[0040] The above data show that the degradation properties of each layer on the fiber membrane are significantly different based on the porosity, which meets the requirements of tissue engineering scaffolds.
[0041] Example 5 The fiber membrane prepared in Example 2 was used as a test sample to test the cell ingrowth experimental data of the high-porosity layer, the low-porosity layer, and the non-porous layer. The results are as follows:
[0042] The above-mentioned cell ingrowth conditions indicate that the different porosities of the various layers on the fiber membrane have a significant impact on cell incorporation. Among them, the inner layer (high porosity) fiber membrane has good cell incorporation and can promote tissue repair. The outer layer (low porosity) fiber membrane has only a very small amount of cell incorporation and can act as an anti-adhesion layer.
[0043] Example 6 A method for preparing a three-layer fiber membrane comprises preparing a 40 w / v % spinning solution with polyglycolide-lactide copolymer, polylactic acid-polyethylene glycol block copolymer, and glycerol, wherein the solvent is a mixture of N,N-dimethylformamide and acetone in a volume ratio of 7:3, and the mass ratio of polyglycolide-lactide copolymer, polylactic acid-polyethylene glycol block copolymer, and glycerol is 70:25:5.
[0044] The spinning parameters of the high-porosity layer in the three-layer fiber membrane are as follows: spinning height 25 cm, spinning voltage 30 kV, and spinning solution advancing speed 20 μL / min; The spinning parameters of the low-porosity layer were as follows: spinning height 18 cm, spinning voltage 20 kV, and spinning solution advancing speed 30 μL / min; The spinning parameters of the non-porous layer are as follows: spinning height of 12 cm, spinning voltage of 20 kV, spinning solution advancing speed of 30 μL / min; the receiving device is a roller, and the moving speed is 0.5 cm / s.
[0045] The fiber membranes of Example 1 and Example 6 were used in rat cecum experiments to observe the adhesion status of the fiber membranes 5 seconds after implantation. The results were as follows:
[0046] The above comparison experiments show that the addition of hydrophilic materials significantly improves the adhesion effect of the materials.
[0047] It is understood that the above specific description of the present invention is only used to illustrate the present invention and is not limited to the technical solutions described in the embodiments of the present invention. Those skilled in the art should understand that the present invention can still be modified or replaced with equivalents to achieve the same technical effects; as long as the use requirements are met, they are all within the scope of protection of the present invention.
Claims
1. A method for preparing a layered controlled fiber membrane, characterized in that: Using one or more polymers as raw materials, an electrospinning process is adopted to prepare a layered fiber membrane. The layered fiber membrane is a double-layer fiber membrane or a triple-layer fiber membrane, and the pores between each layer of the fiber membrane are regulated to achieve the functions of cell growth, mechanical enhancement, anti-adhesion and step-by-step degradation, wherein a hydrophilic material is added to the inner layer of the fiber membrane.
2. The method for preparing a layered control fiber membrane according to claim 1, wherein: The raw materials are one or more of polyglycolic acid, polylactide-caprolactone copolymer, polyglycolide-trimethylene carbonate block copolymer, polylactic acid-polyethylene glycol block copolymer, polylactic acid, polycaprolactone, and polyglycolide-lactide copolymer.
3. The method for preparing a layered control fiber membrane according to claim 1, wherein: The inner fiber membrane of the double-layer fiber membrane is a high-porosity layer with a porosity of 60-90%; the thickness is 50-100 μm.
4. The method for preparing a layered control fiber membrane according to claim 3, characterized in that: The outer fiber membrane of the double-layer fiber membrane is a non-porous layer with a thickness of 20-50 μm.
5. The method for preparing a layered control fiber membrane according to claim 1, characterized in that: The inner fiber membrane of the three-layer fiber membrane is a high-porosity layer with a porosity of 60-90%; the thickness is 50-100 μm; The middle fiber membrane adopts a non-porous layer, and the outer fiber membrane adopts a low-porosity layer with a low porosity of 20-60% and a thickness of 20-50μm.
6. The method for preparing a layered control fiber membrane according to claim 1, characterized in that: The layered fiber membrane achieves pore change by suppressing the spinning solution drawing mode and increasing the unit density fiber stacking mode.
7. The method for preparing a layered control fiber membrane according to claim 6, characterized in that: The method of suppressing the stretching of the spinning solution is achieved by adjusting the spinning height, the spinning solution concentration, the spinning voltage and the spinning solution advancing speed.
8. The method for preparing a layered control fiber membrane according to claim 7, characterized in that: When the low-porosity layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 1-3 cm, preferably 1.5-3 cm; When the non-porous layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-2 cm, preferably 0.5-1.5 cm.
9. The method for preparing a layered control fiber membrane according to claim 6, characterized in that: The method of increasing the unit density of fiber stacking is achieved by adjusting the moving speed of the receiving device.
10. The method for preparing a layered control fiber membrane according to claim 9, characterized in that: When the low-porosity layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-3 cm, preferably 1-2 cm; the moving speed of the receiving device is set to L1 / (1-3), where L1 is the width of the spinning area; When the non-porous layer is spun using a single nozzle, the width of the spinning area on the flat receiving device is 0.5-3 cm, preferably 1-2 cm; the moving speed of the receiving device is set to L2 / (2-5), where L2 is the width of the spinning area.