CN117838913B

CN117838913B - A kind of zedoary oil/oxidized regenerated cellulose embolic microsphere and its preparation method and application

Info

Publication number: CN117838913B
Application number: CN202410251122.1A
Authority: CN
Inventors: 张维芬; 曹大栋; 张竞竞; 张德蒙; 张希全; 蹇兆成; 刘峰
Original assignee: Shandong Second Medical University
Current assignee: Shandong Second Medical University
Priority date: 2024-03-06
Filing date: 2024-03-06
Publication date: 2024-05-24
Anticipated expiration: 2044-03-06
Also published as: CN117838913A; WO2025184983A1; WO2025184983A8

Abstract

The present invention relates to the technical field of pharmaceutical preparations, and provides a zedoary oil/oxidized regenerated cellulose embolic microsphere, and a preparation method and application thereof. The present invention uses zedoary oil as a core solution, a mixed solution of oxidized regenerated cellulose, sodium alginate and gelatin as a shell solution, and a divalent or trivalent metal salt solution as a receiving solution, and prepares zedoary oil/oxidized regenerated cellulose embolic microspheres by a coaxial electrostatic spray method. The zedoary oil/oxidized regenerated cellulose embolic microspheres prepared by the present invention have an obvious core-shell structure, uniform size, good sphericity, can effectively mask the pungent smell of zedoary oil, improve the stability of zedoary oil, achieve directional embolization in tumor blood vessels, and reduce toxic side effects; and the embolic microspheres prepared by the present invention have good mechanical properties, effectively resist the extrusion deformation of the microspheres, avoid crushing in the body, and enable zedoary oil to firmly embolize in the tumor blood vessel site.

Description

Zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere and preparation method and application thereof

Technical Field

The invention relates to the technical field of pharmaceutical preparations, in particular to zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere, and a preparation method and application thereof.

Background

The zedoary turmeric oil is an oily substance obtained by mixing multiple compounds. The chemical components of the medicine comprise curcumenol, curcuminoid, elemene, curcumenol, isocurcumenol, dehydrocurcuminoid, kaempferide, curcumenone, bisabolol, curcumenol, etc. up to tens. The zedoary turmeric oil has wide anti-tumor effects, mainly comprises inhibiting proliferation of tumor cells, inducing apoptosis of tumor cells, directly destroying tumor cells, inhibiting metastasis and invasion of tumor cells, inhibiting angiogenesis of tumor, enhancing sensitivity of chemotherapeutics, reversing drug resistance, regulating immunity, and the like, and is effective on ovarian cancer, endometrial cancer, cervical cancer, breast cancer, lung cancer, gastric cancer, rectal cancer, liver cancer, and the like.

The hepatic artery chemoembolization (TACE) is the first choice treatment method for middle and late stage liver cancer, and the method is characterized in that femoral artery cannulation is used for selectively inserting blood supply arteries of the liver cancer, embolizing and infusing chemotherapeutic drugs, embolizing main tumor blood vessels to block tumor blood supply, making the tumor ischemic necrosis, infusing the chemotherapeutic drugs to kill tumor cells, and the method is suitable for liver cancer with rich blood supply. The TACE has the advantages of small wound, light side effect, repeated treatment, improvement of life quality and life extension, and is an effective means for treating tumors.

The medicine carrying microsphere is one new kind of embolism agent capable of carrying medicine and releasing chemical medicine to kill tumor cell while embolizing blood vessel to provide tumor with nutrients, so as to play the role of double antitumor. At present, most of researches on zedoary turmeric oil embolic microspheres are zedoary turmeric oil gelatin microspheres, which are generally prepared by an emulsion crosslinking method, the preparation process is complex, and the prepared microspheres are uneven in size and poor in mechanical property.

Disclosure of Invention

In view of the above, the invention provides zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres, and a preparation method and application thereof. The preparation method provided by the invention is simple to operate, and the obtained microsphere is uniform in size, good in mechanical property and high in drug loading rate.

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

a preparation method of zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere comprises the following steps:

Dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain sodium alginate gelatin solution; mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution;

zedoary turmeric oil is used as core solution; spraying the shell solution and the core solution into receiving liquid by adopting a coaxial electrostatic spraying method to obtain zedoary turmeric oil/oxidized regenerated cellulose embolism microspheres; the receiving liquid is a metal salt solution; the metal salt is a divalent metal salt or a trivalent metal salt.

Preferably, in the oxidized regenerated cellulose solution, the mass ratio of oxidized regenerated cellulose, sodium hydroxide, urea and water is (1-5): (1-5): (4-10): (30-60).

Preferably, in the sodium alginate gelatin solution, the concentration of sodium alginate is 0.5-5wt% and the concentration of gelatin is 0.1-3wt%.

Preferably, sodium chloride is also added into the sodium alginate gelatin solution, and the concentration of the sodium chloride in the sodium alginate gelatin solution is 0.1-2wt%.

Preferably, the volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is (1-5).

Preferably, the divalent metal salt is one or more of calcium salt, barium salt, strontium salt, zinc salt and copper salt; the trivalent metal salt is one or more of ferric salt and aluminum salt.

Preferably, the operating conditions of the coaxial electrostatic spray include: the pushing speed of the shell layer solution is 0.1-1 mm/min, the pushing speed of the core layer solution is 0.1-1 mm/min, the voltage is 10-20V, and the receiving distance is 2-8 cm.

The invention also provides zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere prepared by the preparation method; the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere has a core-shell structure, wherein a core is zedoary turmeric oil, and shell components comprise gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid.

The invention also provides application of the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres in preparation of hepatic artery chemoembolic agents.

The invention provides a preparation method of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres, which comprises the following steps: dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain sodium alginate gelatin solution; mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution; zedoary turmeric oil is used as core solution; spraying the shell solution and the core solution into receiving liquid by adopting a coaxial electrostatic spraying method to obtain zedoary turmeric oil/oxidized regenerated cellulose embolism microspheres; the receiving liquid is a metal salt solution; the metal salt is a divalent metal salt or a trivalent metal salt. The invention adopts the coaxial electrostatic spraying method to prepare the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere, which can completely wrap the zedoary turmeric oil in a shell material, and has simple operation and uniform particle size of the obtained microsphere.

In addition, the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere prepared by the invention has obvious core-shell structure, uniform size and good sphericity, can effectively mask the pungent smell of the zedoary turmeric oil, improve the stability of the zedoary turmeric oil, can realize directional embolism in tumor blood vessels, improve the concentration of the zedoary turmeric oil at tumor sites, reduce the concentration of the zedoary turmeric oil in normal tissues, reduce toxic and side effects, play a targeted treatment effect, and the microsphere with the core-shell structure has a slow release effect and high bioavailability; the curcuma zedoary turmeric oil contains antitumor active ingredients such as curcumenol, beta-elemene, zedoary turmeric ketone and the like, and has an effective treatment effect on tumors.

The invention also adds oxidized regenerated cellulose into the shell layer, and the oxidized regenerated cellulose can form a cross-linked network in the microsphere shell layer, thereby improving the mechanical property of the microsphere, effectively resisting the extrusion deformation of the microsphere, avoiding the occurrence of crushing in vivo and ensuring that the zedoary turmeric oil is firmly embolized at the tumor vascular position.

In conclusion, the preparation method provided by the invention has the advantages of simple steps, low cost, good mechanical property and uniform particle size of the obtained microsphere, can protect zedoary turmeric oil, mask the unpleasant smell of zedoary turmeric oil, has a slow release effect, and has a wide application prospect in the field of hepatic artery chemoembolization.

Drawings

FIG. 1 is a physical diagram of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using a coaxial needle of 28G#20G;

FIG. 2 is an optical micrograph of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial spray heads in example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, and D is 32G#22G;

FIG. 3 is a graph showing the particle size distribution of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial spray heads in example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, and D is 32G#22G;

FIG. 4 is a scanning electron microscope image of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using a coaxial needle of 28G#20G in example 1;

FIG. 5 is an infrared spectrum of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using a coaxial needle of 28G#20G in example 1;

FIG. 6 shows the test results of drug loading and encapsulation efficiency of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using different types of coaxial spray heads;

FIG. 7 is an in vitro release profile of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial sprinklers in example 1;

FIG. 8 is a force-compression curve of zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere and zedoary turmeric oil/sodium alginate gelatin embolic microsphere prepared in example 1.

Detailed Description

The invention provides a preparation method of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres, which comprises the following steps:

Zedoary turmeric oil is used as core solution;

spraying the shell solution and the core solution into receiving liquid by adopting a coaxial electrostatic spraying method to obtain zedoary turmeric oil/oxidized regenerated cellulose embolism microspheres; the receiving liquid is a metal salt solution; the metal salt is a divalent metal salt or a trivalent metal salt.

The invention dissolves oxidized regenerated cellulose in sodium hydroxide urea solution to obtain oxidized regenerated cellulose solution. In the oxidized regenerated cellulose solution, the mass ratio of the oxidized regenerated cellulose to the sodium hydroxide to the urea to the water is preferably (1-5): (4-10): (30-60), more preferably (2-3): (5-8): (40-50).

In the specific embodiment of the invention, sodium hydroxide and urea are preferably dissolved in water, the obtained sodium hydroxide urea solution is put into a refrigerator at the temperature of minus 20 ℃ to be frozen into an ice-water mixed state, oxidized regenerated cellulose is taken and added into the sodium hydroxide urea solution, and the solution is magnetically stirred for 10 min and stored at the temperature of minus 20 ℃ after being dissolved. When in use, the oxidized regenerated cellulose solution is preferably filtered three times by a filter membrane with the diameter of 0.22 mu m to obtain a clear and transparent oxidized regenerated cellulose solution; in a specific embodiment of the present invention, preferably, oxidized cellulose membranes are used as raw materials, and the oxidized cellulose membranes are sheared and then added into the sodium hydroxide urea solution for dissolution.

The sodium alginate and the gelatin are dissolved in water to obtain sodium alginate gelatin solution. In the sodium alginate gelatin solution, the concentration of sodium alginate is preferably 0.5-5wt%, more preferably 1-3wt%, and the concentration of gelatin is preferably 0.1-3wt%, more preferably 1-2wt%; the water used to dissolve the sodium alginate and gelatin is preferably double distilled water. In the specific embodiment of the invention, preferably, sodium alginate and gelatin are added into double distilled water, and magnetic stirring is carried out for more than 12 h to obtain transparent solution, namely the sodium alginate gelatin solution.

In the invention, sodium chloride is also added into the sodium alginate gelatin solution, and the concentration of the sodium chloride in the sodium alginate gelatin solution is 0.1-2wt%; when the sodium alginate gelatin solution is prepared, the sodium chloride, sodium alginate and gelatin are added into double distilled water together to be stirred and dissolved.

After the oxidized regenerated cellulose solution and the sodium alginate gelatin solution are obtained, the oxidized regenerated cellulose solution and the sodium alginate gelatin solution are mixed to obtain a shell layer solution. In the invention, the volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is preferably (1-5): (1-5), more preferably 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5:1.

After obtaining a shell solution, the invention takes zedoary turmeric oil as a core solution, and sprays the shell solution and the core solution into a receiving solution by adopting a coaxial electrostatic spraying method to obtain zedoary turmeric oil/oxidized regenerated cellulose embolism microspheres; the receiving liquid is a metal salt solution. In the present invention, the metal salt is a divalent metal salt or a trivalent metal salt; the divalent metal salt is preferably one or more of calcium salt, barium salt, strontium salt, zinc salt and copper salt; the trivalent metal salt is ferric salt or aluminum salt; the divalent metal salt is preferably a hydrochloride salt of a divalent metal, and the trivalent metal salt is preferably a hydrochloride salt of a trivalent metal; in a specific embodiment of the present invention, the metal salt is preferably calcium chloride, barium sulfate, zinc chloride or copper sulfate; the concentration of the metal salt solution is preferably 0.2-0.5 mol/L.

In the present invention, the operating conditions of the coaxial electrostatic spray preferably include: the pushing speed of the shell layer solution is 0.1-1 mm/min, preferably 0.3-0.5 mm/min; the pushing speed of the core solution is 0.1-1 mm/min, preferably 0.1-0.5 mm/min; the voltage is 10-25V, preferably 15-20V, and the receiving distance is 2-10 cm, preferably 5-9 cm; the type of the coaxial needle adopted by the coaxial electrostatic spraying is preferably 32G#22G, 30G#21G, 28G#20G or 25G#18G, wherein 32G represents the type of the needle sleeved on the inner side of the coaxial needle, and 22G represents the type of the needle on the outer side; g represents the size of the needle tube, specifically, the 18G inner diameter is 0.9mm, the outer diameter is 1.3mm, the 20G inner diameter is 0.6mm, the outer diameter is 0.9mm, the 21G inner diameter is 0.5mm, the outer diameter is 0.8mm, the 22G inner diameter is 0.4mm, the outer diameter is 0.7mm, the 25G inner diameter is 0.2mm, the outer diameter is 0.5mm, the 28G inner diameter is 0.15mm, the outer diameter is 0.35mm, the 30G inner diameter is 0.12mm, the outer diameter is 0.32mm, the 32G inner diameter is 0.06mm and the outer diameter is 0.24mm. In a specific embodiment of the present invention, the shell solution is specifically injected into the shell flow chamber of the coaxial nozzle, and the core solution is injected into the core flow chamber of the coaxial nozzle, followed by coaxial electrostatic spraying under the above conditions. The invention has no special requirement on the device adopted by the coaxial electrostatic spraying, and the coaxial electrostatic spraying is carried out by adopting an electrostatic spinning instrument which is well known to the person skilled in the art.

In the invention, after a shell layer solution and a core solution are sprayed into receiving liquid through a coaxial electrostatic spraying method, sodium alginate and metal ions react to form gel, gelatin and alginate gel form a shell layer, and oxidized regenerated cellulose is doped in the shell layer to form a skeleton structure, and the shell layer coats zedoary turmeric oil to form core-shell microspheres; after the core-shell microspheres are obtained, the preferred microspheres are collected and washed by double distilled water, the receiving liquid on the surface is removed, and then the microspheres are stored in the double distilled water.

The invention also provides zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres (also called zedoary turmeric oil core-shell microspheres) prepared by the preparation method; the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere has a core-shell structure, wherein a core is zedoary turmeric oil, and shell components comprise gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid, preferably calcium alginate; the drug loading rate of the zedoary turmeric oil in the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres is preferably 28.62-43.20%.

The invention also provides application of the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres in preparation of hepatic artery chemoembolic agents. The zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere provided by the invention has the advantages of slow release performance, high bioavailability and good mechanical property, can effectively resist extrusion deformation of the microsphere, and avoid crushing in vivo, so that the zedoary turmeric oil can be firmly embolized at tumor vascular positions, and has wide application prospects.

The following description of the embodiments of the present invention will clearly and fully describe the technical solutions of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Example 1

1. Preparation of reagents

1) Preparing a sodium alginate gelatin solution: sodium alginate 0.6 g, gelatin 0.2 g, sodium chloride 0.18 g were weighed and double distilled water was added to 20 mL. Stir on a magnetic stirrer overnight until a clear solution forms.

Preparing oxidized regenerated cellulose solution: naOH 2.8 g and urea 4.8 g were dissolved in 40 mL water and the resulting solution was frozen in a-20℃refrigerator to ice-water mix. Cutting oxidized regenerated cellulose fiber membrane of 2g into pieces, dissolving in frozen alkaline urine system, magnetically stirring for 10min to obtain clear solution, and storing in a refrigerator at-20deg.C. When in use, the filter membrane with the diameter of 0.22 mu m is needed to be used twice, so that the needle head is prevented from being blocked.

Mixing 6 mL of sodium alginate gelatin solution and 2 mL of oxidized regenerated cellulose solution, and magnetically stirring to obtain a shell solution.

2) 5 ML zedoary turmeric oil is taken as core solution.

2. Synthesis of zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere

The zedoary turmeric oil is used as an oil phase, and oxidized regenerated cellulose sodium alginate gelatin solution is used as a water phase to prepare the microsphere with an oil/water type (O/W) core-shell structure. The coaxial electric spraying method is adopted, and the preparation is carried out by relying on an electrostatic spinning instrument, and the specific steps are as follows: respectively sucking the shell solution and the core solution by using a syringe at each 5mL, placing the shell solution and the core solution in a pushing part of an electrostatic spinning instrument, setting the pushing speed of the shell solution to be 0.3 mm/min, the pushing speed of the core solution to be 0.1 mm/min, and the voltage to be 20V, wherein the environment parameters are required to be stable, taking 0.2 mol/L of calcium chloride solution 100mL as receiving solution, and the receiving distance to be 8 cm. The types of coaxial needles used were 32G#22G, 30G#21G, 28G#20G, 25G#18G, respectively. Obtaining the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere with a core-shell structure. Washing the microspheres with double distilled water for multiple times, removing redundant receiving liquid on the surfaces, and then placing the microspheres into a refrigerator at 4 ℃ to store the microspheres in the double distilled water.

Example 2

1. Preparation of reagents

1) Preparing a sodium alginate gelatin solution: sodium alginate 0.4 g, gelatin 0.2 g, sodium chloride 0.18 g were weighed out. Double distilled water was added to 20 mL and stirred on a magnetic stirrer overnight until a clear solution formed.

2) 5 ML zedoary turmeric oil is taken as core solution.

The zedoary turmeric oil is used as an oil phase, and oxidized regenerated cellulose sodium alginate gelatin solution is used as a water phase to prepare the microsphere with an oil/water type (O/W) core-shell structure. The coaxial electric spraying method is adopted, and the preparation is carried out by relying on an electrostatic spinning instrument, and the specific steps are as follows: respectively sucking the shell solution and the core solution by using a syringe to form 5mL parts, placing the shell solution and the core solution in a pushing part of an electrostatic spinning instrument, setting the pushing speed of the shell solution to be 0.3 mm/min, the pushing speed of the core solution to be 0.1 mm/min, the voltage to be 20V, requiring stable environmental parameters, taking 0.4 mol/L barium sulfate solution 100 mL as receiving solution, and receiving the receiving distance to be 8 cm, wherein the types of the coaxial needles are respectively 32G#22G, 30G#21G, 28G#20G and 25G#18G. Obtaining the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere with a core-shell structure. Washing the microspheres with double distilled water for multiple times, removing redundant receiving liquid on the surfaces, and then placing the microspheres into a refrigerator at 4 ℃ to store the microspheres in the double distilled water.

Example 3

1. Preparation of reagents

1) Preparing a sodium alginate gelatin solution: sodium alginate 0.5 g, gelatin 0.2 g, sodium chloride 0.18 g were weighed out. Double distilled water was added to 20 mL and stirred on a magnetic stirrer overnight until a clear solution formed.

Mixing sodium alginate gelatin solution 2 mL and oxidized regenerated cellulose solution 2 mL, and magnetically stirring to obtain shell solution.

2) 5 ML zedoary turmeric oil is taken as core solution.

The zedoary turmeric oil is used as an oil phase, and oxidized regenerated cellulose sodium alginate gelatin solution is used as a water phase to prepare the microsphere with an oil/water type (O/W) core-shell structure. The coaxial electric spraying method is adopted, and the preparation is carried out by relying on an electrostatic spinning instrument, and the specific steps are as follows: respectively sucking the shell solution and the core solution by using a syringe to form 5mL parts, placing the shell solution and the core solution in a pushing part of an electrostatic spinning instrument, setting the pushing speed of the shell solution to be 0.5 mm/min, setting the pushing speed of the core solution to be 0.1 mm/min, setting the voltage to be 12V, requiring stable environmental parameters, taking 0.2 mol/L zinc chloride solution 100mL as receiving solution, and taking the receiving distance to be 8 cm, wherein the types of the coaxial needles are respectively 32G#22G, 30G#21G, 28G#20G and 25G#18G. Obtaining the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere with a core-shell structure. Washing the microspheres with double distilled water for multiple times, removing redundant receiving liquid on the surfaces, and then placing the microspheres into a refrigerator at 4 ℃ to store the microspheres in the double distilled water.

Example 4

1. Preparation of reagents

1) Preparing a sodium alginate gelatin solution: sodium alginate 0.5 g, gelatin 0.2 g, sodium chloride 0.18 g were weighed and double distilled water was added to 20 mL. Stir on a magnetic stirrer overnight until a clear solution forms.

Mixing 4 mL of sodium alginate gelatin solution and 1 mL of oxidized regenerated cellulose solution, and magnetically stirring to obtain a shell solution.

2) 5 ML zedoary turmeric oil is taken as core solution.

The zedoary turmeric oil is used as an oil phase, and oxidized regenerated cellulose sodium alginate gelatin solution is used as a water phase to prepare the microsphere with an oil/water type (O/W) core-shell structure. The coaxial electric spraying method is adopted, and the preparation is carried out by relying on an electrostatic spinning instrument, and the specific steps are as follows: respectively sucking the shell solution and the core solution by using a syringe to form 5mL parts, placing the shell solution and the core solution in a pushing part of an electrostatic spinning instrument, setting the pushing speed of the shell solution to be 0.5 mm/min, setting the pushing speed of the core solution to be 0.1 mm/min, setting the voltage to be 15V, requiring stable environmental parameters, taking 0.2 mol/L copper sulfate solution 100mL as receiving liquid, and taking the receiving distance to be 8 cm, wherein the types of the coaxial needles are respectively 32G#22G, 30G#21G, 28G#20G and 25G#18G. Obtaining the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere with a core-shell structure. Washing the microspheres with double distilled water for multiple times, removing redundant receiving liquid on the surfaces, and then placing the microspheres into a refrigerator at 4 ℃ to store the microspheres in the double distilled water.

Example 5

Other conditions were the same as in example 1, except that the preparation of the sodium alginate gelatin solution was changed to: sodium alginate 0.1 g, gelatin 0.02 g, sodium chloride 0.02 g were weighed and double distilled water was added to 20 mL. Stir on a magnetic stirrer overnight until a clear solution forms.

The preparation was carried out by using a 28G#20G coaxial needle under the conditions of example 1 to obtain zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres of core-shell structure.

Example 6

Other conditions were the same as in example 1, except that the preparation of the sodium alginate gelatin solution was changed to: sodium alginate 1 g, gelatin 0.6 g, sodium chloride 0.4 g were weighed and double distilled water was added to 20 mL. Stir on a magnetic stirrer overnight until a clear solution forms.

Example 7

Other conditions were the same as in example 1, and sodium chloride was omitted only when preparing the sodium alginate gelatin solution. And, when preparing the shell layer solution, 1 mL sodium alginate gelatin solution and 3 mL oxidized regenerated cellulose solution are taken and mixed.

Example 8

Other conditions were the same as in example 1, except that the amount of NaOH was changed to 1g, the amount of urea was changed to 4g, the amount of water was changed to 30mL, and the amount of oxidized regenerated cellulose was changed to 1g when preparing the oxidized regenerated cellulose solution.

In addition, 5mL of sodium alginate gelatin solution and 1mL of oxidized regenerated cellulose solution are taken and mixed when preparing the shell solution.

Example 9

Other conditions were the same as in example 1, except that the amount of NaOH was changed to 5g, the amount of urea was changed to 10g, the amount of water was changed to 60mL, and the amount of oxidized regenerated cellulose was changed to 5g when preparing the oxidized regenerated cellulose solution.

In addition, 1mL of sodium alginate gelatin solution and 5mL of oxidized regenerated cellulose solution are taken and mixed when preparing the shell solution.

Performance test:

1. topography test and criteria

FIG. 1 is a physical diagram of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using a 28G#20G coaxial needle. As can be seen from figure 1, the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere prepared by the invention has uniform particle size and good dispersibility. In addition, the observation shows that the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres with uniform particle size and good dispersibility can be prepared in each of the examples 2-9.

FIG. 2 is an optical micrograph of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using coaxial spray heads of different types in example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, D is 32G#22G, all on a scale of 200 μm. According to fig. 2, it can be seen that zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres with different sizes can be prepared by adopting coaxial spray heads with different types, and the obtained microspheres have the characteristic of uniform particle size.

FIG. 3 is a graph showing the particle size distribution of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial spray heads in example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, and D is 32G#22G. FIG. 3 is a histogram of the particle size distribution obtained by calculating the particle sizes of 100 microspheres using ImageJ software. As can be seen from FIG. 3, the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere prepared by the invention has narrower particle size distribution.

Zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using 28G#20G coaxial needles were freeze-dried and subjected to scanning electron microscopy, the results of which are shown in FIG. 4. According to fig. 4, a remarkable core-shell structure can be observed, and the zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere has a shrinkage phenomenon on the surface after freeze-drying.

2. Infrared spectroscopy testing

FIG. 5 is an infrared spectrum of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using a 28G#20G coaxial needle. As can be seen from FIG. 5, the peak near 3436 cm ^-1 in the sodium alginate spectrum corresponds to the stretching vibration of-OH. 1606 Peaks at cm ^-1 and 1415 cm ^-1 belong to the c=o and C-O stretching vibrations, respectively, belonging to the-COO-group of sodium alginate. For the gelatin spectrum, a peak at about 3436 cm ^-1 is assigned to an-OH or-NH group. Peaks at 1630 cm ^-1 and 1518 cm ^-1 correspond to c=o and C-N groups, respectively. The significant 1722 cm ^-1 peak of oxidized regenerated cellulose is the-COOH group. Zedoary turmeric oil exhibits a characteristic peak at 1697 cm ^-1, which is caused by stretching vibration of carbonyl group (c=o) in zedoary turmeric oil, and a peak near 2926 cm ^-1, which is caused by stretching vibration of hydroxyl group, and a peak near 1444 cm ^-1 and 1365 cm ^-1, which is caused by bending vibration in saturated C-H bond plane. The zedoary turmeric oil core-shell microsphere contains obvious 2926 cm ^-1 characteristic peak, which shows that the zedoary turmeric oil is stably coated in the microsphere and has stable property. The characteristic peaks of the microspheres at 1701 cm ^-1 and 1452 cm ^-1 are significantly enhanced compared to sodium alginate gelatin, possibly explained by the formation of a cross-linked network between the carboxyl groups of oxidized regenerated cellulose and the adjacent alginate gelatin molecules. These results indicate that there is ionic interaction and molecular compatibility between sodium alginate gelatin and oxidized regenerated cellulose.

3. Drug loading and encapsulation efficiency test

Fig. 6 shows the test results of drug loading and encapsulation efficiency of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 using different types of coaxial spray heads. Specific test data are shown in table 1.

Table 1 drug loading and encapsulation efficiency of zedoary turmeric oil core-shell microspheres (mean ± SD, n=3,%)

According to fig. 6 and table 1, the drug loading rate of the zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere prepared by the invention is 28.62-43.20%, and the encapsulation rate is 92.84-99.72%.

4. In vitro Release test

The in vitro drug release characteristics of the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 were determined by a dialysis bag method, and the specific steps are as follows: precisely transferring 10 mL of wet zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared by 4 coaxial spray heads of different types, dividing the microspheres into three parts, and placing the three parts in a pretreated dialysis bag (the molecular weight cut-off is 3500). The dialysis bags were placed in conical flasks containing 40 mL% release medium (PBS, pH7.4, 0.5% Tween-80) and placed in a constant temperature water bath shaker at 37.+ -. 0.5 ℃ C., 100 r/min, respectively. Dialysate 2 mL in the conical flask was taken at 0, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 12, 24, 48, 72, 96 h, respectively, while an equal amount of fresh release medium was replenished. Repeating for 3 times, collecting the above solution 0.2 mL in 2 mL EP tubes, and shaking with vanillin sulfuric acid solution to constant volume of 2 mL. The absorbance was measured at 508: 508 nm by light-shielding color development 1h, and the cumulative release rate was calculated to examine the release behavior. Meanwhile, free zedoary turmeric oil was used for the same experiment as a comparison.

Fig. 7 is an in vitro release profile of zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial spray heads in example 1, wherein ZTO represents free zedoary turmeric oil. As is apparent from FIG. 7, the zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere prepared by the invention has better sustained release capability.

5. Mechanical property test

The mechanical properties of the zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres prepared in example 1 were tested; meanwhile, zedoary turmeric oil/sodium alginate gelatin embolism microsphere is prepared and mechanical property test is carried out, so that the mechanical property of the zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere is compared. The preparation method of the zedoary turmeric oil/sodium alginate gelatin embolism microsphere comprises the following steps: other conditions are the same as those in example 1, only oxidized regenerated cellulose in a shell layer is omitted, namely, the sodium alginate gelatin solution is directly taken as a shell layer solution, zedoary turmeric oil is taken as a core solution, and coaxial electrospraying is carried out according to the conditions in example 1, so as to obtain the zedoary turmeric oil/sodium alginate gelatin embolic microsphere.

FIG. 8 is a force-compression curve of zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere and zedoary turmeric oil/sodium alginate gelatin embolic microsphere prepared in example 1. As can be seen from fig. 8, as the elastic deformation gradually increases from 10% to 50%, the pressure required by the zedoary turmeric oil/oxidized regenerated cellulose embolic microsphere is gradually increased under the same deformation condition than that required by the zedoary turmeric oil/sodium alginate gelatin embolic microsphere, the anti-extrusion performance is stronger, and the better elastic performance is exhibited. The zedoary turmeric oil/oxidized regenerated cellulose embolism microsphere prepared by the invention has good mechanical property, effectively resists extrusion deformation of the microsphere, avoids crushing in vivo, and ensures that the zedoary turmeric oil is firmly embolized at tumor blood vessel positions.

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. A method for preparing zedoary turmeric oil/oxidized regenerated cellulose embolic microspheres, comprising the following steps:

dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain a sodium alginate gelatin solution; mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution;

Use zedoary oil as the core solution;

The shell solution and the core solution are sprayed into a receiving solution by a coaxial electrostatic spray method to obtain zedoary oil/oxidized regenerated cellulose embolic microspheres; the receiving solution is a metal salt solution; the metal salt is a divalent metal salt or a trivalent metal salt;

In the oxidized regenerated cellulose solution, the mass ratio of oxidized regenerated cellulose, sodium hydroxide, urea and water is (1-5): (1-5): (4-10): (30-60);

The operating conditions of the coaxial electrostatic spray include: the shell solution injection speed is 0.1-1 mm/min, the core solution injection speed is 0.1-1 mm/min, the voltage is 10-20 V, and the receiving distance is 2-8 cm.

2. The preparation method according to claim 1 is characterized in that, in the sodium alginate gelatin solution, the concentration of sodium alginate is 0.5~5wt%, and the concentration of gelatin is 0.1~3wt%.

3. The preparation method according to claim 1 is characterized in that sodium chloride is also added to the sodium alginate gelatin solution, and the concentration of sodium chloride in the sodium alginate gelatin solution is 0.1~2wt%.

4. The preparation method according to claim 1, characterized in that the volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is (1~5): (1~5).

5. The preparation method according to claim 1, characterized in that the divalent metal salt is one or more of calcium salt, barium salt, strontium salt, zinc salt and copper salt; the trivalent metal salt is one or more of trivalent iron salt and aluminum salt.

6. The zedoary oil/oxidized regenerated cellulose embolic microspheres prepared by the preparation method according to any one of claims 1 to 5, characterized in that the zedoary oil/oxidized regenerated cellulose embolic microspheres have a core-shell structure, the core is zedoary oil, and the shell components include gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid.

7. Use of the zedoary oil/oxidized regenerated cellulose embolic microspheres according to claim 6 in the preparation of hepatic artery chemoembolization agents.