CN111973795A - Dressing for stopping bleeding and preventing cancer recurrence after liver cancer resection - Google Patents

Abstract

The present application relates to a hemostasis and anti-cancer recurrence dressing after liver cancer resection. Glycolide and/or lactide are used as monomers to carry out a polymerization reaction to prepare a polymer, the above polymer is used as a shell material, and verteporfin is used as a material. Raw materials, using polyethylene glycol derivatized phosphatidylethanolamine (PEG-PE) as a carrier, the film dispersion-hydration method is used to prepare verteporfin sustained-release nanomicelles; The core fluid and the shell fluid are coaxially electrospun to obtain hemostatic and anti-cancer recurrence dressings after liver cancer resection. The hemostatic and anti-cancer recurrence dressing of the present application provides good conditions for hemostasis and inhibition of tumor microenvironment after liver resection, and realizes that the release of verteporfin can have a sufficient concentration in the initial stage, and can maintain a long-term sustained release Effect.

Description

A kind of dressing for hemostasis and cancer recurrence prevention after liver cancer resection

technical field

The invention belongs to the technical field of medical materials, and mainly relates to a dressing for hemostasis and cancer recurrence prevention after liver cancer resection applied in the clinical field.

Background technique

Bleeding in liver cancer surgery is a major problem in hepatobiliary surgery. On the one hand, liver blood flow is abundant, and it is relatively difficult to stop bleeding after bleeding. On the other hand, bleeding may contribute to tumor recurrence. At present, only relying on surgical methods cannot effectively control bleeding during surgery, and the use of high-efficiency hemostatic materials can significantly reduce the amount of bleeding. At present, the most studied hemostatic materials at home and abroad are mainly divided into the following categories: fibrin, gelatin, oxidized cellulose, chitosan, etc. In recent years, electrospinning technology has attracted extensive attention as a simple and effective method to prepare functionalized nanofibers. Compared with traditional hemostatic materials, nanofiber wound dressings prepared by electrospinning have the advantages of larger specific surface area, tunable porosity and better ductility, and can also be loaded with drugs or other biomolecules. The functional hemostatic material of science and technology has good development prospects in the medical field, and is expected to achieve effective hemostasis after liver cancer resection and inhibit the tumor microenvironment at the same time.

SUMMARY OF THE INVENTION

In the present application, the coaxial electrospinning method is used to arrange the sustained-release nano-micelles containing the activating factor verteporfin on the core layer of the fiber, and a hemostatic dressing with a composite pore size of 10-500 microns is obtained, thereby realizing the verteporfin. The release can not only have a sufficient concentration at the initial stage, but also maintain a sustained release effect for a long time. The specific technical solutions of this application are as follows:

A dressing for hemostasis and cancer recurrence prevention after liver cancer resection is prepared by the following steps:

1) Preparation of verteporfin sustained-release nanomicelles

Using glycolide and/or lactide as a monomer to carry out a polymerization reaction to prepare a polymer, using the above polymer as a shell material, using verteporfin as a raw material, using polyethylene glycol derivatized phosphatidylethanolamine (PEG-PE ) as the carrier, and the verteporfin sustained-release nanomicelles were prepared by thin film dispersion-hydration method;

2) Preparation of Verteporfin Sustained Release Nanomicelle Modified Electrospinning Dressing

Coaxial electrospinning was carried out by using the core liquid and shell liquid containing verteporfin sustained-release nanomicelles to obtain verteporfin sustained-release nanomicelle modified electrospinning dressings;

Preferably, in step 1), polyethylene glycol is added in the monomer polymerization reaction for block modification to obtain a polymer, wherein the molecular weight of polyethylene glycol is 400-4000, and the mass of polyethylene glycol and monomer is The ratio is 0.5-2:10.

Preferably, in step 1), verteporfin and polyethylene glycol derivatized phosphatidylethanolamine (PEG-PE) are added to the methanol solution, mixed and stirred until completely dissolved; a low-pressure rotary evaporator is used to obtain a thin film, and then vacuum Dry, remove the remaining organic solvent, and generate a dry polymer drug film; add pH 7.4 phosphate buffer to hydrate the film, then hydrate in a water bath at 40 °C, and after ultrasonication, filter through a 0.22 μm microporous membrane to obtain a clear The clear micellar solution was vacuum frozen with the addition of 2% mannitol as a lyoprotectant.

Preferably, in step 1), the prepared polymer is tested for hydrophilicity with a contact angle <45°, a viscosity <1.5, and an elongation at break <5%.

Preferably, in step 1), the particle size of verteporfin sustained-release nanomicelles is 0.5-8 microns, the particle size distribution DPI<0.1, the drug loading rate>50%, and the drug loading amount is 3-50%.

Preferably, in step 2), the core solution of electrospinning is composed of verteporfin sustained-release nanomicelles, chitosan/gelatin/collagen mixture, ultrapure water, and verteporfin sustained-release nanomicelles The mass content is 1-10%, and the mass content of the chitosan/gelatin/collagen mixture is 5-15%.

Preferably, in step 2), the electrospinning shell liquid is composed of a chitosan/gelatin/collagen mixture and ultrapure water, and the mass content of the chitosan/gelatin/collagen mixture is 15-25%.

Preferably, in step 2), the electrostatic high voltage is 1-50 kV, the receiving distance is 50-300 mm, and the electrospinning rate is 0.05-0.2 ml/min.

Beneficial effects of this application:

(1) The present application organically combines the two technologies of verteporfin sustained-release nanomicelles and electrospinning, and the electrospinning dressing achieves the hemostatic effect and also the release characteristics of tumor suppressors. Most of the activating factors in the prior art are released explosively in the early stage, and cannot achieve long-term effects. It is not easy to achieve a good sustained-release effect in one technology, although the combination of multiple activator release technologies (such as the use of embedding technology and surface immobilization technology) can achieve both early and late activators. However, the use of multiple technologies has many and complicated preparation steps, and there is no mutual relationship between multiple technologies, which is only a simple superposition of technologies. In the present application, a good sustained-release effect is achieved by arranging the verteporfin sustained-release nanomicelle on the core layer of the electrospinning. The main function of arranging verteporfin sustained-release nanomicelles in the core layer of the electrospinning fibers is to make the verteporfin activator multiple encapsulated by the nanomicelles and the fiber shell to have an excellent sustained-release effect. The technology has a simple preparation process, and can simply regulate the release process of the activator by adjusting parameters such as the content of the sustained-release verteporfin nanomicelles. The application of sustained-release nanomicelles arranged in the electrospinning core layer is the key to solving the problem of sustained release of verteporfin activators. Many other sustained-release technologies are not easy to achieve the effect of simultaneous slow release of tumor suppressors in this application.

(2) This application uses polyethylene glycol to modify the polymer of glycolide and/or lactide, mainly to improve the hydrophilicity of the polymer, so that the shell layer of the sustained-release nanomicelle has a good Hydrophilic, which not only increases the good biocompatibility, but also further improves the release effect of the activator. Extended release effect.

Detailed ways

The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

Example 1

1) Preparation of verteporfin sustained-release nanomicelles

Add 8g lactide, 2g glycolide, 1g polyethylene glycol 2000 to the polymerization tube, then add 0.5g of 0.3% stannous octoate in dichloromethane solution as a catalyst, and heat the polymerization tube to 150°C under vacuum to mix The material is melted and mixed uniformly, and kept for 8h. After natural cooling, it was dissolved in acetone, precipitated and washed with water to obtain a polymer as a shell material. The hydrophilicity test contact angle of the polymer prepared above was 30°, the viscosity was 1.2, and the elongation at break was 4.5%.

50 mg of verteporfin and 200 mg of PEG-PE were added to a flask containing 50 ml of methanol, and the mixture was stirred until completely homogeneously dissolved. Use a low-pressure rotary evaporator at 60 °C and 50 r/min to form a thin film, then place the rotary bottle in ice water, vacuum dry to remove excess organic solvent, and form a dry polymer drug film. Add 5 ml of phosphate buffered saline (PBS), stir for 60 min under hydration conditions at 40°C, and finally filter through a 0.22 μm microporous membrane for 4 to 5 times to remove the precipitate to obtain a clear and transparent micelle solution. Mannitol was added as a freeze-drying protective agent, and vacuum-frozen for 24 hours to obtain verteporfin nanomicelle freeze-dried powder.

2) Preparation of Verteporfin Sustained Release Nanomicelle Modified Electrospinning Dressing

The core solution of electrospinning is composed of verteporfin sustained-release nanomicelles, chitosan/gelatin/collagen mixture, ultrapure water, the mass content of verteporfin sustained-release nanomicelles is 5%, chitosan The mass content of gelatin/collagen mixture is 10%; the shell liquid of electrospinning is composed of chitosan/gelatin/collagen mixture and ultrapure water, and the mass content of chitosan/gelatin/collagen mixture is 15%; using Prepared by coaxial electrospinning, wherein the electrostatic high voltage is 15 kV, the receiving distance is 100 mm, and the electrospinning rate is 0.1 ml/min, and the verteporfin slow-release nanomicelle modified electrospinning dressing is prepared.

Example 2

1) Preparation of verteporfin sustained-release nanomicelles: This procedure is the same as that in Example 1.

2) Preparation of Verteporfin Sustained Release Nanomicelle Modified Electrospinning Dressing

The core solution of electrospinning is composed of verteporfin sustained-release nanomicelles, chitosan/gelatin/collagen mixture, ultrapure water, the mass content of verteporfin sustained-release nanomicelles is 10%, chitosan The mass content of gelatin/collagen mixture is 5%; the electrospinning shell liquid is composed of chitosan/gelatin/collagen mixture and ultrapure water, and the mass content of chitosan/gelatin/collagen mixture is 20%; using Prepared by coaxial electrospinning method, wherein the electrostatic high voltage is 50 kV, the receiving distance is 200 mm, and the electrospinning rate is 0.05 ml/min, and the verteporfin sustained-release nanomicelle modified electrospinning dressing is prepared.

Example 3

1) Preparation of verteporfin sustained-release nanomicelles: This procedure is the same as that in Example 1.

2) Preparation of Verteporfin Sustained Release Nanomicelle Modified Electrospinning Dressing

The core solution of electrospinning is composed of verteporfin sustained-release nanomicelles, chitosan/gelatin/collagen mixture, ultrapure water, the mass content of verteporfin sustained-release nanomicelles is 1%, chitosan The mass content of gelatin/collagen mixture is 15%; the shell liquid of electrospinning is composed of chitosan/gelatin/collagen mixture and ultrapure water, and the mass content of chitosan/gelatin/collagen mixture is 25%; using Prepared by coaxial electrospinning method, wherein the electrostatic high voltage is 1 kV, the receiving distance is 50 mm, and the electrospinning rate is 0.2 ml/min, and the verteporfin slow-release nanomicelle modified electrospinning dressing is prepared.

Example 4

Same as Example 1, but no polyethylene glycol was added in step 1).

Comparative Example 1

It is the same as Example 1, but the sustained-release nanomicelles are not added to the core layer, and the sustained-release nanomicelles are added to the shell layer.

Comparative Example 2

Same as Example 1, but the core liquid of electrospinning is composed of verteporfin sustained-release nanomicelles, chitosan/gelatin/collagen mixture, ultrapure water, and the mass of verteporfin sustained-release nanomicelles The content is 0.5%, and the mass content of the chitosan/gelatin/collagen mixture is 10%.

The cumulative release of verteporfin detected on the fifth day was used to represent the initial concentration of the activator, and the days required for the cumulative release of verteporfin to be 80% were used to represent the sustained release ability of the activator. The test results of each sample are shown in Table 1.

Table 1

It can be seen from Examples 1-2 that the verteporfin sustained-release nanomicelle-modified electrospinning dressing of the present application has a sufficient concentration in the initial verteporfin release, which can basically reach 10%-20% , and the sustained sustained release ability is also very significant, basically to about 30 days. It shows that the dressing of the present application not only achieves hemostasis, but also improves the release characteristics of the activating factor, so that the release of verteporfin can not only have a sufficient concentration at the initial stage, but also maintain a sustained release effect for a long time. By comparing Example 1 and Comparative Example 1, it can be found that the release of verteporfin sustained-release nanomicelles in the fiber shell layer can obtain a higher release amount of activating factors than that in the fiber core layer, which is consistent with more slow-release nanomicelles. The release capsule is related to the dressing surface, but this setting has a significant impact on the effect of sustained release, and too many activating factors released in the early stage may also cause the activation efficiency to decrease. It shows that setting the sustained-release capsule in the fiber core layer can have a better effect. By comparing Example 1 and Comparative Examples 1-2, it can be seen that the organic combination of the two technologies of verteporfin sustained-release nanomicelle and electrospinning in the present application is the key to obtaining excellent results. By comparing Example 1 and Example 4, it can be found that if the shell material of the sustained-release capsule is hydrophilically modified with polyethylene glycol, it can reduce the release amount of the initial activator, but at the same time increase the sustained-release capacity, and its It shows that the hydrophilic modification of polyethylene glycol hinders the release of verteporfin or the introduction of polyethylene glycol slows down the degradation of the polymer and thus slows down the release of verteporfin.

The present application combines two technologies of sustained-release nanomicelle and electrospinning. By placing the sustained-release capsule on the core layer of the electrospinning fiber, it has good hemostatic properties and also achieves a good release effect of tumor suppressors. The preparation method of the present application is simple, all FDA-approved and degradable materials are used, and it has good biocompatibility, and provides an alternative solution for hemostasis and reduction of tumor recurrence after liver cancer resection.

The above-mentioned embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or transformations made by those skilled in the art on the basis of the present invention are all within the protection scope of the present invention. The protection scope of the present invention is subject to the claims.

Claims (8)

1. a hemostasis and anti-cancer recurrence dressing after liver resection, is characterized in that, is prepared by the following steps: 1) Preparation of verteporfin sustained-release nanomicelles Using glycolide and/or lactide as a monomer to carry out a polymerization reaction to prepare a polymer, using the above polymer as a shell material, using verteporfin as a raw material, using polyethylene glycol derivatized phosphatidylethanolamine (PEG-PE ) as the carrier, and the verteporfin sustained-release nanomicelles were prepared by thin film dispersion-hydration method; 2) Preparation of Verteporfin Sustained Release Nanomicelle Modified Electrospinning Dressing The core liquid and shell liquid containing verteporfin nanomicelles are used for coaxial electrospinning to obtain verteporfin sustained-release nanomicelle modified electrospinning dressings. 2. hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 1), in monomer polymerization reaction, add polyethylene glycol to carry out block modification to obtain polymer, The molecular weight of polyethylene glycol is 400-4000, and the mass ratio of polyethylene glycol to monomer is 0.5-2:10. 3. hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 1), by verteporfin and polyethylene glycol derivatized phosphatidylethanolamine (PEG-PE) Add to methanol solution, mix and stir until completely dissolved; use a low-pressure rotary evaporator to obtain a film, and then vacuum dry to remove the remaining organic solvent to generate a dry polymer drug film; add pH 7.4 phosphate buffer to promote film hydration, and then in Hydrate in a 40°C water bath, and after ultrasonication, filter through a 0.22 μm microporous membrane to obtain a clear and transparent micellar solution, which was added with 2% mannitol as a freeze-drying protective agent and then vacuum-frozen. 4. The hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, characterized in that, in step 1), the prepared polymer hydrophilicity test contact angle<45o, viscosity<1.5, elongation at break rate < 5%. 5. hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 1) in, verteporfin sustained-release nano-micelle particle size is 20-55 nanometers, and particle size distribution DPI<0.1, drug loading rate>50%, and drug loading capacity of 3-50%. 6. The hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 2), the core liquid of electrospinning is composed of verteporfin sustained-release nanomicelle, chitosan /gelatin/collagen mixture, ultrapure water, the mass content of the verteporfin sustained-release nanomicelle is 1-10%, and the mass content of the chitosan/gelatin/collagen mixture is 5-15%. 7. The hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 2), the shell liquid of electrospinning is composed of chitosan/gelatin/collagen mixture, ultrapure water composition, the mass content of chitosan/gelatin/collagen mixture is 15-25%. 8. hemostasis and anti-cancer recurrence dressing after liver cancer resection according to claim 1, is characterized in that, in step 2) in, electrostatic high voltage is 1-50 kV, receiving distance is 50-300mm, electrospinning speed 0.05-0.2ml/min.