CN113713182A - Preparation and application of resveratrol liposome-containing guided bone tissue regeneration membrane - Google Patents
Preparation and application of resveratrol liposome-containing guided bone tissue regeneration membrane Download PDFInfo
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- CN113713182A CN113713182A CN202111105310.6A CN202111105310A CN113713182A CN 113713182 A CN113713182 A CN 113713182A CN 202111105310 A CN202111105310 A CN 202111105310A CN 113713182 A CN113713182 A CN 113713182A
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/146—Porous materials, e.g. foams or sponges
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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Abstract
The invention discloses preparation and application of a resveratrol liposome-containing guided bone tissue regeneration membrane. Belongs to the field of medical materials, and comprises the following steps: 1. dissolving medical degradable material in organic solvent to obtain electrospinning matrix solution; 2. preparing resveratrol into resveratrol liposome solution by ethanol injection method, freeze-drying to obtain resveratrol liposome, and dissolving the resveratrol liposome in electrospinning matrix solution to obtain electrostatic spinning solution; 3. carrying out electrostatic spinning on the prepared electrostatic spinning solution by adopting an electrostatic spinning technology to obtain an electrospun membrane; 4. and drying the electrospun membrane to obtain the functional bone tissue regeneration membrane. The guided bone tissue regeneration membrane containing the resveratrol liposome prepared by the invention has good biocompatibility and controllable degradation performance; the excellent three-dimensional scaffold structure can effectively load and locally slowly release the resveratrol serving as an active substance, and achieves the purposes of promoting migration and proliferation of osteoblasts, promoting osteogenic differentiation and improving osteogenic performance.
Description
Technical Field
The invention belongs to the technical field of medical materials, and relates to a degradable resveratrol modified guided bone tissue regeneration membrane containing an active substance, a preparation method and application thereof.
Background
The Guided Bone tissue Regeneration (GBR) technology is one of the most common and effective technologies for clinically solving the Bone tissue defect problem, and is characterized in that a Guided Bone tissue Regeneration barrier membrane is placed between soft tissue and Bone defect to artificially establish a biological barrier and create an isolation space, prevent fibroblasts and epithelial cells in the soft tissue from growing into the Bone defect area, ensure that the osteogenesis process has no interference of the fibroblasts, and finally realize complete Bone repair of the defect area; the technology has important clinical significance in the fields of oral periodontal and implantation; the guided bone tissue regeneration membrane plays an important role in GBR, and the ideal GBR membrane not only plays a good barrier role, but also can promote the adhesion and proliferation of osteoblasts and the regeneration of bone tissues.
In order to better meet the performance requirements of the GBR membrane, the improvement of the preparation technology is particularly important; the electrostatic spinning technology is a method for preparing a nanofiber membrane by using an electric field force generated by a high-voltage electrostatic field, and the electrostatic spinning membrane prepared by the method has certain advantages compared with the traditional GBR membrane, such as: the membrane has a three-dimensional space structure, and can provide enough growth, extension and proliferation space for the growth of osteocytes; such a film may also achieve versatility of the material.
Besides selecting a proper GBR membrane preparation method, the selection aspect of preparation materials is also important; although the GBR membrane prepared from the medical degradable material can provide a foundation for further clinical application of the bone implant material and realization of multi-functionalization of the guided bone tissue regeneration membrane to a certain extent, the GBR membrane still has great defects in the aspect of guided bone tissue regeneration, so that the clinical application of the electrostatic spinning membrane is limited to a certain extent; in addition, in clinical research, large-area bone tissue damage is generally accompanied by the threat of inflammation, and further repair of the bone tissue is prevented; research shows that the resveratrol can be widely applied to the treatment of osteoporosis, the enhancement of bone formation, the repair of fracture or the defect of bone reconstruction, can promote the differentiation of bone marrow stromal cells into osteoblasts, and improves the activities of alkaline phosphatase and prolyl hydroxylase in a dose-dependent mode; however, the chemical property of the resveratrol is unstable, the resveratrol is easily oxidized and has poor water solubility under the action of light, heat and an oxidant, and the defects of poor oral absorption, low bioavailability, non-lasting effect and the like exist, so that the application of the resveratrol in some fields is limited to a great extent; therefore, research is carried out by preparing resveratrol into nano liposome; can effectively protect the wrapped medicine from the oxidation of the functional and some components of the medicine, thereby improving the stability and the bioavailability of the medicine, and can also be administrated by various ways, so that the medicine has the characteristics of targeting property, slow release property, cell affinity and the like.
In view of the above, there is a need to provide a guided bone tissue regeneration membrane modified by an osteogenic active substance (resveratrol-containing liposome), a preparation method and applications thereof, so as to solve the above problems.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a method for preparing a guided bone tissue regeneration membrane containing resveratrol liposome, which is used for preparing the guided bone tissue regeneration membrane with medical degradable materials and osteogenic active substances and has the advantages of promoting bone regeneration and good biocompatibility.
The technical scheme is as follows: the invention relates to a preparation method of a resveratrol liposome-containing guided bone tissue regeneration membrane, which comprises the following specific operation steps:
(1) dissolving the medical degradable material in an organic solvent to obtain an electrospinning matrix solution, and standing for later use;
(2) preparing the prepared resveratrol into resveratrol liposome solution by adopting an ethanol injection method in advance; preparing the prepared resveratrol liposome solution into resveratrol liposomes by a freeze drying method, and finally dissolving the resveratrol liposomes in the prepared electrospinning matrix solution to prepare an electrostatic spinning solution;
(3) carrying out electrostatic spinning on the prepared electrostatic spinning solution by adopting an electrostatic spinning technology to obtain an electrospun membrane;
(4) drying the prepared electrospun membrane to finally obtain a functional bone tissue regeneration membrane; namely a guided bone tissue regeneration membrane containing resveratrol liposome.
Further, in the step (1), the medical degradable material is at least one of polylactic acid-glycolic acid copolymer, polycaprolactone, polylactic acid and poly beta-hydroxybutyrate;
the organic solvent is at least one of trifluoroethanol, acetone, hexafluoroisopropanol, dimethylformamide and tetrahydrofuran;
the dosage of the medical degradable material is 0.1-2g, and the dosage of the organic solvent is 1-10 mL.
Further, in the step (2), the specific preparation process of the resveratrol liposome is as follows:
firstly, weighing a certain amount of lecithin, glycerol and glyceryl stearate as mixed oil; weighing a certain amount of Tween80, cetostearyl alcohol and PGPR as a mixed emulsifier; weighing a certain amount of rhamnose and resveratrol, and dissolving the substances in absolute ethyl alcohol to obtain an oil phase;
secondly, sucking 1 ml of oil phase, dripping into 20 ml of water phase heated in a water bath, stirring at constant speed for 1-2h in a water bath kettle, preserving heat for 1h in the water bath, and performing ultrasonic treatment in an ice-water bath for 3-8min to obtain resveratrol liposome liquid;
thirdly, adding a proper amount of resveratrol liposome into the BMP-2 protein aqueous solution, and carrying out constant temperature shaking for 24 hours to obtain the resveratrol liposome with the BMP-2 protein embedded on the surface;
and (IV) finally, preparing the resveratrol liposome by adopting a freeze-drying protective agent through a freeze-drying method.
Further, in the step (one), the mass ratio of the lecithin to the glycerol stearate is as follows: 6:3: 1;
the mass ratio of the Tween80 to the cetostearyl alcohol to the PGPR is as follows: 8:1: 1;
the mass ratio of the mixed oil to the mixed emulsifier to the rhamnose to the resveratrol is as follows: 30:30:6: 1;
in step (ii), the aqueous phase is: PBS aqueous solution with pH value of 6.5 and 30 mmol/L;
in the third step, the concentration of the BMP-2 protein aqueous solution is 5%;
in the step (IV), the freeze-drying protective agent is mannitol, and the concentration of the mannitol dissolved in the resveratrol liposome solution is 3%.
Further, in the step (2), the preparation steps of the electrospinning solution are as follows: dissolving a certain amount of resveratrol liposome in the electrospinning matrix solution, and stirring at constant speed for 2-3 h; finally preparing an electrostatic spinning solution;
wherein the resveratrol liposome is 0.01-0.03 g.
Further, in the step (3), the preparation conditions for electrospinning the electrospinning solution to obtain the electrospun membrane are as follows: negative pressure of-2.5 kV, positive pressure of 10-20kV, receiving distance of 10-20cm, sample introduction speed of 2-3mL/h, and humidity of less than 60%.
Further, in the step (4), the preparation conditions for obtaining the functional bone tissue regeneration membrane by drying the electrospun membrane are as follows: the drying oven temperature was 37 ℃ and the drying time was 24 h.
Further, a guided bone tissue regeneration membrane prepared by the preparation method of any one of claims 1 to 7.
Further, the application of the guided bone tissue regeneration membrane in bone repair.
Has the advantages that: compared with the prior art, the guided bone tissue regeneration membrane prepared by the invention has the advantages that the guided bone tissue regeneration membrane has medical degradable materials, resveratrol and resveratrol liposome; on one hand, the bone tissue regeneration membrane has good degradability and biocompatibility based on medical degradable materials, and is guided to have a three-dimensional porous structure simulating an extracellular matrix, so that migration and proliferation of osteoblasts are facilitated; on the other hand, the addition of resveratrol promotes osteogenic differentiation and improves osteogenic performance; on the other hand, the liposome can protect the biological characteristics of active substances in the preparation process, and based on the addition of the resveratrol liposome, the resveratrol is slowly released, thereby further promoting the cell differentiation.
Drawings
FIG. 1 is a flow chart of the preparation of the present invention.
Detailed Description
The invention is further described below with reference to the following figures and specific examples.
In the invention, the medical degradable material has better biocompatibility and plasticity, the GBR membrane is prepared by simply adopting the medical degradable material, and a foundation can be provided for further clinical application of the bone implant material and realization of the multifunctionalization of the guided bone tissue regeneration membrane to a certain extent, but the medical degradable material still has great defects in the aspect of guided bone tissue regeneration, so that the clinical application of the electrostatic spinning membrane is limited to a certain extent; therefore, through research, a preparation method and application of the resveratrol liposome-containing guided bone tissue regeneration membrane are provided, and the prepared guided bone tissue regeneration membrane is modified and modified by adding an osteogenic active substance on the basis of a medical degradable material electrostatic spinning membrane, so that the osteogenesis effect is realized.
Specifically, Resveratrol (RSV) is a non-flavonoid polyphenolic compound, and is also a polyphenolic phytoestrogen and phytoalexin found in plants; the biological effects of resveratrol include cardiovascular protection and anti-tumor, anti-inflammatory, antioxidant, anti-aging and bone protective activities, and have various health benefits for mammals; several biological effects have been reported for resveratrol, including attenuation of platelet aggregation, protection of cardiovascular, anticancer activity and neuroprotective effects, resveratrol can inhibit oxidation of low density lipoprotein cholesterol and is expected to prevent atherosclerotic changes, reduction of platelet aggregation by inhibiting metabolism of arachidonic acid, and vasodilatory activity in isolated aorta.
The resveratrol liposome mainly uses stearin, lecithin and the like as membrane materials to encapsulate resveratrol so as to form a bilayer vesicle with a structure similar to that of a biological membrane; can effectively protect the wrapped medicine from the oxidation of the functional and some components of the medicine, thereby improving the stability and the bioavailability of the medicine, and can also be administrated by various ways, so that the medicine has the characteristics of targeting property, slow release property, cell affinity and the like.
Specifically, the preparation method of the resveratrol liposome-containing guided bone tissue regeneration membrane comprises the following specific operation steps:
(1) dissolving the medical degradable material in an organic solvent to obtain an electrospinning matrix solution, and standing for later use;
(2) preparing the prepared resveratrol into resveratrol liposome solution by adopting an ethanol injection method in advance; preparing the prepared resveratrol liposome solution into resveratrol liposomes by a freeze drying method, and finally dissolving the resveratrol liposomes in the prepared electrospinning matrix solution to prepare an electrostatic spinning solution;
(3) carrying out electrostatic spinning on the prepared electrostatic spinning solution by adopting an electrostatic spinning technology to obtain an electrospun membrane;
(4) drying the prepared electrospun membrane to finally obtain a functional bone tissue regeneration membrane; namely a guided bone tissue regeneration membrane containing resveratrol liposome.
Further, in the step (1), the medical degradable material is at least one of polylactic acid-glycolic acid copolymer, polycaprolactone, polylactic acid, poly beta-hydroxybutyrate and the like;
the organic solvent is at least one of trifluoroethanol, acetone, hexafluoroisopropanol, dimethylformamide, tetrahydrofuran and the like;
the dosage of the medical degradable material is 0.1-2g, and the dosage of the organic solvent is 1-10 mL.
Further, in the step (2), the specific preparation process of the resveratrol liposome is as follows:
firstly, weighing a certain amount of lecithin, glycerol and glyceryl stearate as mixed oil; weighing a certain amount of Tween80, cetostearyl alcohol and PGPR as a mixed emulsifier; weighing a certain amount of rhamnose and resveratrol, and dissolving the substances in absolute ethyl alcohol to obtain an oil phase;
secondly, sucking 1 ml of oil phase, dripping into 20 ml of water phase heated in a water bath, stirring at constant speed for 1-2h in a water bath kettle, preserving heat for 1h in the water bath, and performing ultrasonic treatment in an ice-water bath for 3-8min to obtain resveratrol liposome liquid;
thirdly, adding a proper amount of resveratrol liposome into the BMP-2 protein aqueous solution, and carrying out constant temperature shaking for 24 hours to obtain the resveratrol liposome with the BMP-2 protein embedded on the surface;
and (IV) finally, preparing the resveratrol liposome by adopting a freeze-drying protective agent through a freeze-drying method.
Further, in the step (one), the mass ratio of the lecithin to the glycerol stearate is as follows: 6:3: 1;
the mass ratio of the Tween80 to the cetostearyl alcohol to the PGPR is as follows: 8:1: 1;
the mass ratio of the mixed oil to the mixed emulsifier to the rhamnose to the resveratrol is as follows: 30:30:6: 1;
in step (ii), the aqueous phase is: PBS aqueous solution with pH value of 6.5 and 30 mmol/L;
in the third step, the concentration of the BMP-2 protein aqueous solution is 5%;
in the step (IV), the freeze-drying protective agent is mannitol, and the concentration of the mannitol dissolved in the resveratrol liposome solution is 3%.
Further, in the step (2), the preparation steps of the electrospinning solution are as follows: dissolving a certain amount of resveratrol liposome in the electrospinning matrix solution, and stirring at constant speed for 2-3 h; finally preparing an electrostatic spinning solution;
wherein the resveratrol liposome is 0.01-0.03 g.
Further, in the step (3), the preparation conditions for electrospinning the electrospinning solution to obtain the electrospun membrane are as follows: negative pressure of-2.5 kV, positive pressure of 10-20kV, receiving distance of 10-20cm, sample introduction speed of 2-3mL/h, and humidity of less than 60%.
Further, in the step (4), the preparation conditions for obtaining the functional bone tissue regeneration membrane by drying the electrospun membrane are as follows: the drying oven temperature was 37 ℃ and the drying time was 24 h.
Further, a guided bone tissue regeneration membrane prepared by the preparation method of any one of the above.
Further, the application of the guided bone tissue regeneration membrane in bone repair is described.
The preparation process of the present invention will be further described with reference to specific examples.
The first embodiment is as follows:
weighing 2g of polycaprolactone, adding the polycaprolactone into 10mL of hexafluoroisopropanol solvent, and uniformly stirring the polycaprolactone on a magnetic stirrer to obtain a polymer solution;
precisely weighing 180mg (270/360/450) of lecithin, 90mg (135/180/225) of glycerol and 30mg (45/60/75) of glyceryl stearate, weighing 240mg (360/480/600) of Tween80, 30mg (45/60/75) of hexadecanol and 30mg (45/60/75) of PGPR, weighing 60mg (90/120/150) of rhamnose and 10mg (15/20/25) of resveratrol, dissolving in 10mL of absolute ethyl alcohol, slowly dropwise adding into 20 mL of PBS solution heated in a water bath, uniformly stirring in a water bath kettle for 1-2h, preserving heat in the water bath for 1h, carrying out ultrasonic treatment in an ice water bath for 3-8min to obtain a resveratrol liposome solution, naturally cooling to room temperature, adding 3% of mannitol, and carrying out dark freeze drying to obtain a solid liposome; adding 0.03g of resveratrol liposome into the polymer solution, and uniformly mixing to obtain an electrostatic spinning solution;
adding the prepared electrostatic spinning solution into an electrostatic spinning device for electrostatic spinning to obtain an electrostatic spinning membrane with a three-dimensional structure, and obtaining the electrostatic spinning membrane under the conditions of negative pressure of-2.5 kV, positive pressure of +13.5kV, receiving distance of 18cm, sample injection speed of 2.2mL/h and humidity of less than 60%; at the moment, the electrostatic spinning contains resveratrol liposome, and has good effect of promoting osteoblast proliferation and differentiation.
Example two:
weighing 1g of polycaprolactone and 1g of poly beta-hydroxybutyrate copolymer, adding the polycaprolactone and the poly beta-hydroxybutyrate copolymer into 10mL of trifluoroethanol solvent, and uniformly stirring the mixture on a magnetic stirrer to prepare polymer solution;
precisely weighing 180mg (270/360/450) of lecithin, 90mg (135/180/225) of glycerol and 30mg (45/60/75) of glyceryl stearate, weighing 240mg (360/480/600) of Tween80, 30mg (45/60/75) of hexadecanol and 30mg (45/60/75) of PGPR, weighing 60mg (90/120/150) of rhamnose and 10mg (15/20/25) of resveratrol, dissolving in 10mL of absolute ethyl alcohol, slowly dropwise adding into 20 mL of PBS solution heated in a water bath, uniformly stirring in a water bath kettle for 1-2h, preserving heat in the water bath for 1h, carrying out ultrasonic treatment in an ice water bath for 3-8min to obtain a resveratrol liposome solution, naturally cooling to room temperature, adding 3% of mannitol, and carrying out dark freeze drying to obtain a solid liposome; adding 0.02g of resveratrol liposome into the polymer solution, and uniformly mixing to obtain an electrostatic spinning solution;
and adding the prepared electrostatic spinning solution into an electrostatic spinning device for electrostatic spinning to obtain an electrostatic spinning membrane with a three-dimensional structure, wherein the electrostatic spinning membrane is obtained under the conditions of negative pressure of-2.5 kV, positive pressure of +15.5kV, receiving distance of 15cm, sample injection speed of 2.5mL/h and humidity of less than 60%, and the electrostatic spinning membrane has medical degradable materials and resveratrol liposome.
Example three:
weighing 0.5g of poly beta-hydroxybutyrate copolymer and 0.5g of polylactic acid-glycolic acid copolymer, adding into 8mL of trifluoroethanol solvent, and uniformly stirring on a magnetic stirrer to prepare a polymer solution;
precisely weighing 180mg (270/360/450) of lecithin, 90mg (135/180/225) of glycerol and 30mg (45/60/75) of glyceryl stearate, weighing 240mg (360/480/600) of Tween80, 30mg (45/60/75) of hexadecanol and 30mg (45/60/75) of PGPR, weighing 60mg (90/120/150) of rhamnose and 10mg (15/20/25) of resveratrol, dissolving in 10mL of absolute ethyl alcohol, slowly dropwise adding into 20 mL of PBS solution heated in a water bath, uniformly stirring in a water bath kettle for 1-2h, preserving heat in the water bath for 1h, carrying out ultrasonic treatment in an ice water bath for 3-8min to obtain a resveratrol liposome solution, naturally cooling to room temperature, adding 3% of mannitol, and carrying out dark freeze drying to obtain a solid liposome; adding 0.02g of resveratrol liposome into the polymer solution, and uniformly mixing to obtain an electrostatic spinning solution;
and adding the prepared electrostatic spinning solution into an electrostatic spinning device for electrostatic spinning to obtain an electrostatic spinning membrane with a three-dimensional structure, wherein the electrostatic spinning membrane is obtained under the conditions of negative pressure of-2.5 kV, positive pressure of +16.5kV, receiving distance of 16cm, sample injection speed of 2.7mL/h and humidity of less than 60%, and the electrostatic spinning membrane has medical degradable materials and resveratrol liposome.
Example four:
weighing 0.5g of polycaprolactone and 0.5g of polylactic acid-glycolic acid copolymer, adding into 5mL of trifluoroethanol solvent, and uniformly stirring on a magnetic stirrer to obtain a polymer solution;
precisely weighing 180mg (270/360/450) of lecithin, 90mg (135/180/225) of glycerol and 30mg (45/60/75) of glyceryl stearate, weighing 240mg (360/480/600) of Tween80, 30mg (45/60/75) of hexadecanol and 30mg (45/60/75) of PGPR, weighing 60mg (90/120/150) of rhamnose and 10mg (15/20/25) of resveratrol, dissolving in 10mL of absolute ethyl alcohol, slowly dropwise adding into 20 mL of PBS solution heated in a water bath, uniformly stirring in a water bath kettle for 1-2h, preserving heat in the water bath for 1h, carrying out ultrasonic treatment in an ice water bath for 3-8min to obtain a resveratrol liposome solution, naturally cooling to room temperature, adding 3% of mannitol, and carrying out dark freeze drying to obtain a solid liposome; adding 0.01g of resveratrol liposome into the polymer solution, and uniformly mixing to obtain an electrostatic spinning solution;
and adding the prepared electrostatic spinning solution into an electrostatic spinning device for electrostatic spinning to obtain an electrostatic spinning membrane with a three-dimensional structure, wherein the electrostatic spinning membrane is obtained under the conditions of negative pressure of-2.5 kV, positive pressure of +18kV, receiving distance of 14cm, sample injection speed of 2.6mL/h and humidity of less than 60%, and the electrostatic spinning membrane has medical degradable materials and resveratrol liposome.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may be made by those skilled in the art without departing from the principle of the invention.
Claims (9)
1. A preparation method of a resveratrol liposome-containing guided bone tissue regeneration membrane is characterized by comprising the following specific operation steps:
(1) dissolving the medical degradable material in an organic solvent to obtain an electrospinning matrix solution, and standing for later use;
(2) preparing the prepared resveratrol into resveratrol liposome solution by adopting an ethanol injection method in advance; preparing the prepared resveratrol liposome solution into resveratrol liposomes by a freeze drying method, and finally dissolving the resveratrol liposomes in the prepared electrospinning matrix solution to prepare an electrostatic spinning solution;
(3) carrying out electrostatic spinning on the prepared electrostatic spinning solution by adopting an electrostatic spinning technology to obtain an electrospun membrane;
(4) drying the prepared electrospun membrane to finally obtain a functional bone tissue regeneration membrane; namely a guided bone tissue regeneration membrane containing resveratrol liposome.
2. The preparation method of the membrane for guiding bone tissue regeneration containing resveratrol liposome according to claim 1,
in the step (1), the medical degradable material is at least one of polylactic acid-glycolic acid copolymer, polycaprolactone, polylactic acid and poly beta-hydroxybutyrate;
the organic solvent is at least one of trifluoroethanol, acetone, hexafluoroisopropanol, dimethylformamide and tetrahydrofuran;
the dosage of the medical degradable material is 0.1-2g, and the dosage of the organic solvent is 1-10 mL.
3. The preparation method of the membrane for guiding bone tissue regeneration containing resveratrol liposome according to claim 1,
in the step (2), the specific preparation process of the resveratrol liposome is as follows:
firstly, weighing a certain amount of lecithin, glycerol and glyceryl stearate as mixed oil; weighing a certain amount of Tween80, cetostearyl alcohol and PGPR as a mixed emulsifier; weighing a certain amount of rhamnose and resveratrol, and dissolving the substances in absolute ethyl alcohol to obtain an oil phase;
secondly, sucking 1 ml of oil phase, dripping into 20 ml of water phase heated in a water bath, stirring at constant speed for 1-2h in a water bath kettle, preserving heat for 1h in the water bath, and performing ultrasonic treatment in an ice-water bath for 3-8min to obtain resveratrol liposome liquid;
thirdly, adding a proper amount of resveratrol liposome into the BMP-2 protein aqueous solution, and carrying out constant temperature shaking for 24 hours to obtain the resveratrol liposome with the BMP-2 protein embedded on the surface;
and (IV) finally, preparing the resveratrol liposome by adopting a freeze-drying protective agent through a freeze-drying method.
4. The preparation method of the membrane containing resveratrol liposome for guiding bone tissue regeneration according to claim 3,
in the step (one), the mass ratio of the lecithin to the glycerol and the glycerol stearate is as follows: 6:3: 1;
the mass ratio of the Tween80 to the cetostearyl alcohol to the PGPR is as follows: 8:1: 1;
the mass ratio of the mixed oil to the mixed emulsifier to the rhamnose to the resveratrol is as follows: 30:30:6: 1;
in step (ii), the aqueous phase is: PBS aqueous solution with pH value of 6.5 and 30 mmol/L;
in the third step, the concentration of the BMP-2 protein aqueous solution is 5%;
in the step (IV), the freeze-drying protective agent is mannitol, and the concentration of the mannitol dissolved in the resveratrol liposome solution is 3%.
5. The preparation method of the membrane for guiding bone tissue regeneration containing resveratrol liposome according to claim 1,
in the step (2), the preparation steps of the electrospinning solution are as follows: dissolving a certain amount of resveratrol liposome in the electrospinning matrix solution, and stirring at constant speed for 2-3 h; finally preparing an electrostatic spinning solution;
wherein the resveratrol liposome is 0.01-0.03 g.
6. The preparation method of the membrane for guiding bone tissue regeneration containing resveratrol liposome according to claim 1,
in the step (3), the preparation conditions for electrospinning the electrospinning solution to obtain the electrospun membrane are as follows: negative pressure of-2.5 kV, positive pressure of 10-20kV, receiving distance of 10-20cm, sample introduction speed of 2-3mL/h, and humidity of less than 60%.
7. The preparation method of the membrane for guiding bone tissue regeneration containing resveratrol liposome according to claim 1,
in the step (4), the preparation conditions of the functional bone tissue regeneration membrane obtained by drying the electrospun membrane are as follows: the drying oven temperature was 37 ℃ and the drying time was 24 h.
8. A guided bone tissue regeneration membrane produced by the production method according to any one of claims 1 to 7.
9. Use of the guided bone tissue regeneration membrane of claim 8 in bone repair.
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