阅读说明：本技术 一种丝素蛋白纤维中空神经导管及其制备方法和应用 (hollow silk fibroin fiber nerve conduit and preparation method and application thereof ) 是由 魏亮 孙润军 刘呈坤 张昭环 毛雪 董洁 于 2019-11-04 设计创作，主要内容包括：本发明公开的一种丝素蛋白纤维中空神经导管及其制备方法和应用,属于生物医用材料技术领域。采用天然的丝素蛋白丝作为原料,具有良好的生物相容性和生物可降解性。利用聚二氧环己酮/甲酸溶液对丝素蛋白纤维进行溶胀制备出的中空神经导管具有良好的压缩回弹性。原料价格低廉,来源广泛,操作方法简便,工艺流程短,能够快速方便的制备不同尺寸要求的中空神经导管,能够批量生产。制得的丝素蛋白纤维中空神经导管具有非常好的力学性能和压缩回弹性能,不易造成支架材料的坍塌,可以保证神经细胞的正常生长。支架材料孔隙率高且有较多的连通孔道,神经细胞能够在神经导管材料上粘附、增殖、迁移和生长,能够促进损伤神经的再生修复。(The invention discloses silk fibroin fiber hollow nerve conduits, a preparation method and application thereof, belonging to the technical field of biomedical materials.)

1, kinds of silk fibroin fiber hollow nerve conduit preparation method, characterized by, divide into the following steps:

step 1: drying the degummed silk to obtain degummed and dried silk fibroin fiber for later use;

step 2: weighing 0.1-1 part by mass of polydioxanone, adding into 1-20 parts by mass of 98% formic acid solution, and stirring until polydioxanone is completely dissolved to obtain polydioxanone/formic acid solution;

and step 3: adding 0.5-5 parts by mass of silk fibroin fibers into 1.1-21 parts by mass of polydioxanone/formic acid solution, and stirring to obtain a gelatinous silk fibroin fiber swelling substance;

and 4, step 4: and (3) putting the silk fibroin fiber swelling matter into a mold for freezing and freeze drying, removing the mold and then washing to obtain the silk fibroin fiber hollow nerve conduit.

2. The method for preparing the silk fibroin fiber hollow nerve conduit of claim 1, wherein the specific steps of step 1 are as follows:

1.1 shearing the silkworm cocoons with the silkworm chrysalis removed for standby;

1.2 heating 0.5-2 parts of deionized water by mass until boiling, putting 1-10 parts of cut silkworm cocoons into the boiled deionized water, and stirring; adding 0.1-10 parts of sodium carbonate in the stirring process, and boiling for 10-60 min to obtain degummed silk fibroin fibers;

1.3 taking out the silk fibroin fibers and extruding water, and washing 1-3 parts of deionized water for 1-5 times in parts by mass;

and 1.4, extruding water in the silk fibroin fibers, dispersing, flatly spreading and naturally airing to obtain the degummed and dried silk fibroin fibers.

3. The preparation method of the silk fibroin fiber hollow nerve conduit according to claim 1, wherein in the step 2, the stirring speed is 100-1000 r/min, and the stirring time is 1-5 h.

4. The preparation method of the silk fibroin fiber hollow nerve conduit as claimed in claim 1, wherein in step 3, the stirring speed is 600-800 r/min, and the stirring time is 1-5 h.

5. The preparation method of the silk fibroin fiber hollow nerve conduit according to claim 1, wherein in the step 4, the freezing temperature is-80 ℃ to-60 ℃, and the freezing time is 24-72 hours.

6. The preparation method of the silk fibroin fiber hollow nerve conduit according to claim 1, wherein in the step 4, the temperature of freeze drying is-50 ℃ to-70 ℃, and the time of freeze drying is 24-72 hours.

7. The method for preparing the silk fibroin fiber hollow nerve conduit according to claim 1, wherein in the step 4, washing is performed for 10-60 min each time for 1-5 times in a phosphate buffer solution.

8. The silk fibroin fiber hollow nerve conduit prepared by the method for preparing the silk fibroin fiber hollow nerve conduit of any of claims 1-7.

9. The silk fibroin fiber hollow nerve conduit of claim 8, wherein the silk fibroin fiber hollow nerve conduit is composed of fibers with a diameter of 1-2 μm, the porosity is 70%, the pore diameter is 20 μm, and the compression rebound rate is greater than 95%.

10. Use of the silk fibroin fiber hollow nerve conduit of claim 8 or 9 as a nerve conduit.

Technical Field

The invention belongs to the technical field of biomedical materials, and particularly relates to silk fibroin fiber hollow nerve conduits, and a preparation method and application thereof.

Background

The peripheral nerve system has the functions of regeneration and self-repairing, so that the small segment nerve damage can be automatically repaired without manual operation, but for the long segment nerve damage, the peripheral nerve system can not be automatically repaired, and meanwhile, the growth of the nerve can be blocked by the glial tissue and the connective tissue around the damaged nerve section, and finally the loss of the motion and perception functions of the patient can be caused.

At present, three main methods for regenerating and repairing long-section damaged nerves include autografting, allotransplantation and artificial nerve scaffold materials. Autografting is considered as the gold standard for nerve injury treatment, but has problems of donor nerve injury, donor site function loss, secondary operation, limited source of donor nerves and the like; the problem of immunological rejection reaction of an organism exists after the damaged nerve is repaired by adopting a xenotransplantation method, and a large amount of medicines are required to be taken to reduce the immunological reaction of the regenerated and repaired nerve, so that great economic burden is caused to a patient; compared with the former two methods, the artificial nerve scaffold material is an ideal method for regenerating and repairing long-section nerve injury. The artificial nerve scaffold material mainly bridges a designed nerve conduit at the near end and the far end of a long-section damaged nerve, nerve cells can be adhered, proliferated and migrated on the scaffold material, the regenerated nerve can lap the near end and the far end of the long-section damaged nerve to realize the regeneration and repair of the damaged nerve, meanwhile, the scaffold material of the nerve conduit is gradually degraded, the nerve cells gradually migrate and grow to complete the whole repair process, and finally the recovery of the motion and perception functions of a patient is realized. However, the existing nerve conduit stent material has the problems of poor mechanical property and compression resilience, low porosity, difficult cell growth, complex preparation process and the like.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention aims to provide silk fibroin fiber hollow nerve conduits, a preparation method and application thereof, wherein the raw materials are easy to obtain, the preparation method is simple, and very ideal biomedical scaffold materials can be obtained.

The invention is realized by the following technical scheme:

the invention discloses a preparation method of a silk fibroin fiber hollow nerve conduit, which comprises the following steps:

step 1: drying the degummed silk to obtain degummed and dried silk fibroin fiber for later use;

step 2: weighing 0.1-1 part by mass of polydioxanone, adding into 1-20 parts by mass of 98% formic acid solution, and stirring until polydioxanone is completely dissolved to obtain polydioxanone/formic acid solution;

and step 3: adding 0.5-5 parts by mass of silk fibroin fibers into 1.1-21 parts by mass of polydioxanone/formic acid solution, and stirring to obtain a gelatinous silk fibroin fiber swelling substance;

and 4, step 4: and (3) putting the silk fibroin fiber swelling matter into a mold for freezing and freeze drying, removing the mold and then washing to obtain the silk fibroin fiber hollow nerve conduit.

Preferably, the specific steps of step 1 are:

1.1 shearing the silkworm cocoons with the silkworm chrysalis removed for standby;

1.2 heating 0.5-2 parts of deionized water by mass until boiling, putting 1-10 parts of cut silkworm cocoons into the boiled deionized water, and stirring; adding 0.1-10 parts of sodium carbonate in the stirring process, and boiling for 10-60 min to obtain degummed silk fibroin fibers;

1.3 taking out the silk fibroin fibers and extruding water, and washing 1-3 parts of deionized water for 1-5 times in parts by mass;

and 1.4, extruding water in the silk fibroin fibers, dispersing, flatly spreading and naturally airing to obtain the degummed and dried silk fibroin fibers.

Preferably, in the step 2, the stirring speed is 100-1000 r/min, and the stirring time is 1-5 h.

Preferably, in the step 3, the stirring speed is 600-800 r/min, and the stirring time is 1-5 h.

Preferably, in the step 4, the freezing temperature is-80 ℃ to-60 ℃, and the freezing time is 24-72 hours.

Preferably, in the step 4, the temperature of freeze drying is-50 ℃ to-70 ℃, and the time of freeze drying is 24-72 hours.

Preferably, in the step 4, the washing is performed 1 to 5 times in a phosphate buffer solution, each time lasting for 10 to 60 min.

The invention discloses a silk fibroin fiber hollow nerve conduit prepared by the preparation method of the silk fibroin fiber hollow nerve conduit.

Preferably, the silk fibroin fiber hollow nerve conduit is composed of fibers with the diameter of 1-2 μm, the porosity is 70%, the pore diameter is 20 μm, and the compression rebound rate is more than 95%.

The invention discloses application of the silk fibroin fiber hollow nerve conduit as a nerve conduit.

The invention also discloses the silk fibroin fiber hollow nerve conduit prepared by the preparation method of the silk fibroin fiber hollow nerve conduit.

The invention also discloses application of the silk fibroin fiber hollow nerve conduit as a nerve conduit.

Compared with the prior art, the invention has the following beneficial technical effects:

the invention discloses a preparation method of silk fibroin fiber hollow nerve conduits, which adopts natural silk fibroin fibers as raw materials, has good biocompatibility and biodegradability, and the hollow nerve conduits prepared by swelling silk fibroin fibers by polydioxanone/formic acid solution have good compression resilience, low price of the raw materials, wide source , simple and convenient operation method, short process flow, and capability of quickly and conveniently preparing the hollow nerve conduits with different size requirements, and can be produced in batches.

The silk fibroin fiber hollow nerve conduit prepared by the preparation method disclosed by the invention has a three-dimensional porous three-dimensional structure formed by fibers with the diameter of 1-2 mu m, the porosity is 70%, the pore diameter is 20 mu m, a space can be provided for the adhesion growth of cells and the regeneration of tissues, and the transportation of nutrient substances is facilitated; the compression rebound rate is more than 95 percent, the mechanical property is good, the deformation is not easy, and the reliable support can be provided.

The silk fibroin fiber hollow nerve conduit prepared by the preparation method disclosed by the invention is applied as a nerve conduit, natural silk is used as a raw material, the biocompatibility is good, the biological degradation is realized, the silk fibroin fiber hollow nerve conduit is very suitable for the adhesion growth of cells, in addition, the nerve conduit support material has very good mechanical property and compression resilience, the collapse of the support material is not easy to cause in-vivo implantation, the normal growth of nerve cells on the support can be ensured, the porosity of the support material is high, and the support material has more communicating pore channels, the nerve cells can be adhered, proliferated, migrated and grown on the nerve conduit material, the regeneration and repair of injured nerves can be promoted, and the silk fibroin fiber hollow nerve conduit is very ideal biomedical support materials.

Drawings

FIG. 1 is a diagram of a silk fibroin fiber hollow nerve conduit prepared by the present invention;

FIG. 2 is a scanning electron microscope photograph of a cross section of a hollow nerve conduit made of silk fibroin fibers according to the present invention;

FIG. 3 is a scanning electron microscope photograph of a cross section of a hollow nerve conduit made of silk fibroin fibers according to the present invention;

FIG. 4 is a schematic diagram showing the growth and proliferation of Schwann cells on the silk fibroin fiber hollow nerve conduit prepared by the present invention;

FIG. 5 is a schematic view of a mold for preparing a silk fibroin fiber hollow nerve conduit according to the present invention;

fig. 6 is a schematic view of a silk fibroin fiber hollow nerve conduit prepared by using a mold.

Wherein, 1 is a mould shell, 2 is a core column, 3 is a fixed hole, 4 is an air hole, 5 is a blocking plate, and 6 is a silk fibroin fiber swelling substance.

Detailed Description

The invention is described in further detail with reference to the following figures and specific examples, wherein the parts are by weight and are intended to be illustrative and not limiting: