CN103993425A - Preparing method of polycaprolactone-keratin composite nano fiber film - Google Patents

Abstract

The invention relates to a preparation method of a polycaprolactone-keratin composite nanofiber film, comprising: dissolving polycaprolactone in a solvent, stirring and dissolving to obtain a polycaprolactone solution; dissolving keratin in a solvent, Stir and dissolve to obtain a keratin solution; mix the polycaprolactone solution and the keratin solution at a mass ratio of 95:5-5:95, stir and mix evenly to obtain a spinning solution, and then perform electrostatic spinning to obtain the product. In the present invention, the natural biomacromolecular material keratin and polycaprolactone are compounded into a nanofiber membrane through an electrospinning method, so that the nanofiber membrane exhibits good mechanical and biological properties while having a high porosity. performance.

Description

A kind of preparation method of polycaprolactone-keratin composite nanofiber film

technical field

The invention belongs to the field of preparation of nanofiber membranes, in particular to a preparation method of polycaprolactone-keratin composite nanofiber membranes.

Background technique

Keratin is an insoluble fibrous animal protein, a structural protein of ectodermal cells, widely present in animal skin and skin appendages, such as hair, hooves, shells, claws, horns, scales, etc. A large number of studies in recent years have shown that keratin is a high-quality biomedical material with good biocompatibility and not rejected by the body's immune system, and has broad application prospects. At present, a large number of basic research and animal experiment research on keratin biomaterials have been carried out at home and abroad. :141-153] and nerve repair [Biomaterials34 (2013) 5907-5914] and other aspects have achieved good results, and some products have been used clinically.

However, existing studies have shown that due to the low molecular weight of keratin, the membrane material made of a single keratin material is usually brittle and has low mechanical strength, which limits its application. Therefore, at present, most keratin-based biomaterials often use keratin and natural polymers or artificial polymers, which can maintain and improve the poor mechanical properties of single keratin, such as silk fibroin [Biomacromolecules, 2008 ,9,1299–1305], PVA[Advances in Materials Science and Engineering,2014,Article ID163678], PLGA[Journal of Bioactive and Compatible Polymers,2013,28:141-153], PLLA[Biomed.Mater.2013,8:1 -9] etc. Polycaprolactone (PCL) is a commercialized; medical polymer material, has good biocompatibility and degradability, has been approved by the US Food and Drug Administration, and is widely used in drug carriers, soft tissue repair, Tissue engineering materials and other biomedical fields. Therefore, the present invention develops a composite material based on two materials, polycaprolactone and keratin. At the same time, in order to obtain a film-like material with high porosity, the present invention adopts an electrospinning method to prepare a composite film of polycaprolactone and keratin. Electrospinning technology refers to the use of a high-voltage electric field environment to form a charged jet flow from the polymer spinning solution. The jet flow is elongated under the action of the electric field, the solvent is volatilized, and finally nanofibers of a certain shape are formed on the receiving device. In the past ten years, this technology has become one of the effective ways to prepare nanofiber materials. The obtained nanofibers have high porosity and controllable shape, and are widely used in medical dressings, tissue engineering scaffolds and other fields. The polycaprolactone-keratin composite nanofiber membrane prepared by the invention not only maintains good biological activity of keratin, but also has good toughness and mechanical strength of polycaprolactone, and is expected to be applied to fields such as medical dressings.

Contents of the invention

The technical problem to be solved by the present invention is to provide a preparation method of polycaprolactone-keratin composite nanofiber membrane, which is prepared by compounding natural biomacromolecular material keratin and polycaprolactone by electrospinning The nanofibrous membrane is formed into a nanofibrous membrane, so that the nanofibrous membrane exhibits good mechanical and biological properties while having high porosity.

A kind of preparation method of polycaprolactone-keratin composite nanofiber film of the present invention comprises:

(1) dissolving polycaprolactone in a solvent, stirring and dissolving to obtain a polycaprolactone solution;

(2) dissolving keratin in a solvent, stirring and dissolving to obtain a keratin solution;

(3) Mix polycaprolactone solution and keratin solution at a mass ratio of 95:5-5:95, stir and mix to obtain spinning solution, and then perform electrospinning to obtain polycaprolactone-keratin Composite nanofiber membrane.

The solvent in the step (1) is acetic acid or acetic acid/formic acid mixed solution; wherein the mass volume percent concentration of acetic acid is greater than or equal to 80%, and the volume ratio of acetic acid and formic acid in the acetic acid/formic acid mixed solution is 1:9-9: 1.

The average molecular weight of the polycaprolactone in the step (1) is 70,000-90,000.

The concentration of the polycaprolactone solution in the step (1) is 80-100mg/ml.

The solvent in the step (2) is formic acid or acetic acid/formic acid mixed solution; wherein the mass volume percent concentration of formic acid is greater than or equal to 80%, and the volume ratio of acetic acid and formic acid in the acetic acid/formic acid mixed solution is 1:9-9: 1.

The concentration of the keratin solution in the step (2) is 10-20 mg/ml.

The keratin in the step (2) is the keratin obtained by reduction method, oxidation method and hydrolysis method.

The keratin in the step (2) is keratin extracted from animals or obtained by gene recombination technology.

The keratin can be extracted from human or animal hair such as human hair, wool, poultry feathers, cow hair, etc., or can be derived from other reported animals such as hagfish, or can be keratin obtained by genetic recombination technology. product.

The molecular weight of keratin in the step (2) is 3-300kDa.

Electrospinning process parameters in the step (3) are: voltage 13-35kV, receiving distance 8-22cm, spinning rate 0.5-1.5ml/h, spinneret inner diameter 0.7-0.9mm, spinning temperature 20 -30°C, spinning humidity 45-65%; receive nanofilaments with aluminum foil or cloth.

The polycaprolactone-keratin composite nanofiber membrane obtained in the step (3) is used for medical wound dressing.

The nanofibrous membrane obtained in the step (3) has a three-dimensional network structure, and the fiber diameter is 50-800nm.

Beneficial effect

In the present invention, the natural biomacromolecular material keratin and polycaprolactone are compounded into a nanofiber membrane through an electrospinning method, so that the nanofiber membrane exhibits good mechanical and biological properties while having a high porosity. performance.

Detailed ways

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Example 1

Dissolve 0.8 g of polycaprolactone with a molecular weight range of 70,000-90,000 in 10 ml of acetic acid/formic acid mixture with a volume ratio of 30/70, and magnetically stir at room temperature until completely dissolved to obtain a concentration of 80 mg/ ml of polycaprolactone solution. 0.2 g of solid keratin was dissolved in 10 ml of formic acid, and magnetically stirred at room temperature until completely dissolved to obtain a keratin solution with a concentration of 20 mg/ml. The above polycaprolactone and keratin solution are mixed according to the mass ratio of 60:40, and magnetically stirred until the mixture is uniform. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 15kV, receiving distance 12cm, spinning rate 0.5ml/h, spinneret inner diameter 0.7mm, spinning temperature 25°C , Spinning humidity 50%. The aluminum foil is used as a receiver to receive the nano-filaments produced by the jet, and the polycaprolactone-keratin composite nano-fiber membrane is obtained.

Example 2

Dissolve 1.0 g of polycaprolactone with a molecular weight of 70,000-90,000 in 10 ml of acetic acid, and magnetically stir at room temperature until completely dissolved to obtain a polycaprolactone solution with a concentration of 100 mg/ml. 0.15 g of solid keratin was dissolved in pure formic acid, and magnetically stirred at room temperature until completely dissolved to obtain a keratin solution with a concentration of 15 mg/ml. The above polycaprolactone and keratin solution were mixed according to the mass ratio of 50:50, and magnetically stirred until uniformly mixed. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 13kV, receiving distance 10cm, spinning rate 0.8ml/h, spinneret inner diameter 0.7mm, spinning temperature 30°C , Spinning humidity 65%. The aluminum foil is used as a receiver to receive the nano-filaments produced by the jet, and the polycaprolactone-keratin composite nano-fiber membrane is obtained.

Example 3

Dissolve 1.0 g of polycaprolactone with a molecular weight range of 70,000-90,000 in 10 ml of acetic acid/formic acid mixture with a volume ratio of 10/90, and stir magnetically at room temperature until it is completely dissolved to obtain a concentration of 100 mg/ ml of polycaprolactone solution. Dissolve 0.15 g of solid keratin in 10 ml of acetic acid/formic acid mixture with a volume ratio of 10/90, and magnetically stir at room temperature until completely dissolved to obtain a keratin solution with a concentration of 15 mg/ml. The above polycaprolactone and keratin solution were mixed according to the mass ratio of 95:5, and magnetically stirred until the mixture was uniform. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 13kV, receiving distance 8cm, spinning rate 1.5ml/h, spinneret inner diameter 0.9mm, spinning temperature 20°C , Spinning humidity 45%. The aluminum foil is used as a receiver to receive the nano-filaments produced by the jet, and the polycaprolactone-keratin composite nano-fiber membrane is obtained.

Example 4

Dissolve 0.8 g of polycaprolactone with a molecular weight range of 70,000-90,000 in 10 ml of acetic acid/formic acid mixture with a volume ratio of 90/10, and magnetically stir at room temperature until completely dissolved to obtain a concentration of 80 mg/ ml of polycaprolactone solution. Dissolve 0.2 g of solid keratin in 10 ml of acetic acid/formic acid mixture with a volume ratio of 90/10, and magnetically stir at room temperature until completely dissolved to obtain a keratin solution with a concentration of 20 mg/ml. The above polycaprolactone and keratin solution were mixed according to the mass ratio of 5:95, and magnetically stirred until the mixture was uniform. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 35kV, receiving distance 22cm, spinning rate 1.5ml/h, spinneret inner diameter 0.9mm, spinning temperature 20°C , Spinning humidity 55%. The cloth is used as a receiver to receive the jet-generated nanofilaments, and a polycaprolactone-keratin composite nanofiber membrane is obtained.

Example 5

Dissolve 0.8 g of polycaprolactone with a molecular weight of 70,000-90,000 in 10 ml of acetic acid, and magnetically stir at room temperature until completely dissolved to obtain a polycaprolactone solution with a concentration of 80 mg/ml. 0.15 g of solid keratin was dissolved in 10 ml of acetic acid, and magnetically stirred at room temperature until completely dissolved to obtain a keratin solution with a concentration of 15 mg/ml. The above polycaprolactone and keratin solution were mixed according to the mass ratio of 95:5, and magnetically stirred until the mixture was uniform. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 15kV, receiving distance 12cm, spinning rate 1.0ml/h, spinneret inner diameter 0.7mm, spinning temperature 15°C , Spinning humidity 55%. The aluminum foil is used as a receiver to receive the nano-filaments produced by the jet, and the polycaprolactone-keratin composite nano-fiber membrane is obtained.

Example 6

Dissolve polycaprolactone with a mass of 1.0g and a molecular weight range of 70,000-90,000 in 10ml at a volume ratio of 30/70

The acetic acid/formic acid mixture was magnetically stirred at room temperature until it was completely dissolved to obtain a polycaprolactone solution with a concentration of 100 mg/ml. 0.2 g of solid keratin was dissolved in 10 ml of acetic acid, and magnetically stirred at room temperature until completely dissolved to obtain a keratin solution with a concentration of 20 mg/ml. The above polycaprolactone and keratin solution are mixed according to the mass ratio of 90:10, and magnetically stirred until the mixture is uniform. The polycaprolactone/keratin solution obtained above was subjected to electrospinning, the electrospinning parameters were voltage 15kV, receiving distance 12cm, spinning rate 1.2ml/h, spinneret inner diameter 0.7mm, spinning temperature 15°C , Spinning humidity 55%. The aluminum foil is used as a receiver to receive the nano-filaments produced by the jet, and the polycaprolactone-keratin composite nano-fiber membrane is obtained.

Example 7

Prepare polycaprolactone-keratin composite nanofiber membrane according to the preparation method described in embodiment 1, measure through analysis, the fracture strength of composite nanofiber material in dry state is 1.4MPa, and elongation at break is 60%, The porosity is 93%. Taking vascular endothelial cells as the cell model, the adhesion morphology of cells on the material was observed by scanning electron microscope, and the proliferation behavior of cells was evaluated by MTT method. The results showed that endothelial cells showed good adhesion morphology on the composite nanofiber material. And proliferation behavior, compared with single polycaprolactone nanofibrous membrane, there are significant differences. According to the preparation method described in Example 1, the nanofibrous membrane prepared from a single keratin is highly brittle and easily broken in a dry state.

Claims (11)

1. A preparation method of polycaprolactone-keratin composite nanofibrous membrane, comprising: (1) dissolving polycaprolactone in a solvent, stirring and dissolving to obtain a polycaprolactone solution; (2) dissolving keratin in a solvent, stirring and dissolving to obtain a keratin solution; (3) Mix polycaprolactone solution and keratin solution at a mass ratio of 95:5-5:95, stir and mix to obtain spinning solution, and then perform electrospinning to obtain polycaprolactone-keratin Composite nanofiber membrane. 2. the preparation method of a kind of polycaprolactone-keratin composite nanofiber film according to claim 1, is characterized in that: in described step (1), solvent is acetic acid or acetic acid/formic acid mixed solution; Wherein acetic acid The mass volume percent concentration is greater than or equal to 80%, and the volume ratio of acetic acid to formic acid in the acetic acid/formic acid mixture is 1:9-9:1. 3. The preparation method of a polycaprolactone-keratin composite nanofiber membrane according to claim 1, characterized in that: the average molecular weight of polycaprolactone in the step (1) is 70,000-90,000. 4. the preparation method of a kind of polycaprolactone-keratin composite nanofiber membrane according to claim 1 is characterized in that: the concentration of polycaprolactone solution in the described step (1) is 80-100mg/ml . 5. the preparation method of a kind of polycaprolactone-keratin composite nanofiber film according to claim 1, is characterized in that: in described step (2), solvent is formic acid or acetic acid/formic acid mixed solution; Wherein formic acid The mass volume percent concentration is greater than or equal to 80%, and the volume ratio of acetic acid to formic acid in the acetic acid/formic acid mixture is 1:9-9:1. 6. The preparation method of a polycaprolactone-keratin composite nanofiber membrane according to claim 1, characterized in that: the concentration of the keratin solution in the step (2) is 10-20 mg/ml. 7. the preparation method of a kind of polycaprolactone-keratin composite nanofiber film according to claim 1 is characterized in that: in described step (2), keratin is to adopt reduction method, oxidation method, hydrolysis method to obtain of keratin. 8. the preparation method of a kind of polycaprolactone-keratin composite nanofiber film according to claim 1, is characterized in that: in described step (2), keratin is that animal extracts keratin or gene recombination technology obtains Keratin. 9. The preparation method of a polycaprolactone-keratin composite nanofiber membrane according to claim 1, characterized in that: the molecular weight of keratin in the step (2) is 3-300kDa. 10. The preparation method of a polycaprolactone-keratin composite nanofiber membrane according to claim 1, characterized in that: the electrospinning process parameters in the step (3) are: voltage 13-35kV, receiving The distance is 8-22cm, the spinning rate is 0.5-1.5ml/h, the inner diameter of the spinneret hole is 0.7-0.9mm, the spinning temperature is 20-30°C, and the spinning humidity is 45-65%. The nanofilaments are received by aluminum foil or cloth. 11. the preparation method of a kind of polycaprolactone-keratin composite nanofiber membrane according to claim 1, is characterized in that: the polycaprolactone-keratin composite nanofiber membrane gained in described step (3) For medical wound dressings.