CN101444642A - Nano biomimetic biomaterial capable of controlling and releasing Chinese medicine and preparation method thereof - Google Patents
Nano biomimetic biomaterial capable of controlling and releasing Chinese medicine and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a nano biomimetic biomaterial capable of controlling and releasing Chinese medicine and a preparation method thereof. The biomaterial can be used as support material for cartilage tissue engineering restoration and reconstruction. The biomaterial takes biomedical material of poly-L-lactide which is frequently used in tissue engineering as base material and adopts phase separation, solvent displacement and freeze-drying method to prepare nano fiber three-dimensional network support material; from the biomimetic perspective, the support material is modified by glutin and chitosan, thereby forming the support material which is similar to cartilage extracellular matrix; tetrandrine is loaded on the support material to improve cartilage cell reproduction, enhance cartilage cell activity and maintain cartilage cell function expression as a cell growth factor analog. The medicine-releasing characteristic of the nano biomimetic biomaterial capable of controlling and releasing Chinese medicine is as follows: in phosphate buffer solution with the pH value of 7-8 and under the 37 DEG C biomimetic condition, the tetrandrine can be steadily released along with the degradation process of the material; and the accumulative drug release rate in six months is approximately 70-80% of drug loading.
Description
Technical field
The present invention relates to a kind of nano biomimetic biomaterial capable that is used for cartilage tissue engineered sustained release Chinese medicine and preparation method thereof, belong to the tissue engineering bracket material technology of preparing.
Background technology
The damage or the afunction of tissue and organ still now all are a difficult problem that threatens human health and puzzlement medical circle in the past, and the whole world is annual all to have the millions of people to die from the damage or the afunction of tissue and organ.Be used to organize the treatment means with organ injury or afunction to mainly contain at present: transplanting, reconstruction in vitro or use medical apparatus are as supplementary means.But, transplant and to exist the donor source difficulty and be difficult to satisfy that treatment needs, operation are complicated, needs of patients is accepted shortcomings such as immunosuppressant therapy and expense costliness throughout one's life.And auxiliary device can not replace all functions of organ, can not cure diseases, and can only relief of symptoms or short-term continuity patient's life.Organizational project is that the living cells that the function of the high concentration of In vitro culture is relevant is planted on extracellular matrix natural or synthetic (timbering material), through cultivation after a while, then it is transplanted in the body, reaching the purpose that forms the new tissue that function is arranged, is the most rising treatment means of generally acknowledging at present.
Should copy the microenvironment of cell as the timbering material of organizational project, make cell adhesion on support, the synthetic cell epimatrix, transmit chemical messenger (somatomedin and cytokine) and mechanical signal, implement the transmission of nutrient and metabolite, the energy delivery and the information transmission to keep the suitable balance of cell proliferation, differentiation and apoptosis, could form the biological substituent of through engineering approaches.
In cartilage tissue engineered, chondrocyte can dedifferente gradually, forfeiture phenotype of chondrocytes feature.Can utilize this moment derive morphogenetic proteins (CDMPs), transforming growth factor (TGF-β) and insulin like growth factor (IGFs) of some somatomedin such as cartilage to keep cell to break up, regulate cell proliferation again and to change the product of cell synthetic and act on the process of tissue formation.Therefore in biomaterial research,, promote tissue repair to rebuild, keep the effect of cell phenotype and function to reach with these factors of timbering material sustained release.If rebuild the defective tissue of bulk, just wish substrate be the sustained release carrier of somatomedin be again support simultaneously.Somatomedin mostly is and derives from the intravital water soluble protein of human or animal, costs an arm and a leg, and biological property is very unstable, makes it be carried on timbering material and acquires a certain degree of difficulty.Studies show that somatomedin dosage is excessive may lure cell generation tumorization into.Antibiotics also is used to tissue engineering bracket material, is used for infection in in-vitro antibacterial and the body.And antibiotic pharmacological action is single, and is just simply antibacterial, lacks physiologically active.
Some effective ingredient of Chinese medicine has and similar function of cytokine and corresponding pharmacological action, and cheap, and physicochemical property is relatively stable, has the good prospect of using as the cytokine substitute.Radix Stephaniae Tetrandrae is a Chinese medicine, and the beginning is stated from Shennong's Herbal, wind-damp dispelling is arranged, pain relieving, the effect of diuretic.Modern study shows, the topmost effective ingredient of Radix Stephaniae Tetrandrae is a tetrandrine, belong to bisbenzylisoquinoline alkaloid, have antiinflammatory and immunosuppressive action, a plurality of links that can the inflammation-inhibiting process comprise inflammation-inhibiting cell function, free radical resisting damage, the inflammation-inhibiting medium discharges and the effect of antagonism inflammatory mediator.Radix stephaniae tetrandrae is usually used in treating rheumatic arthralgia in the Chinese medicine prescription, the existing treatment that is used for rheumatism, rheumatoid arthritis more.The tetrandrine that studies show that low concentration can promote the propagation of chondrocyte, improves the activity of chondrocyte, promotes chondrocyte secretion glycosaminoglycans and II Collagen Type VI, helps external cartilaginous tissue reconstruction.As growth factor analogs, be used for cartilage tissue engineeredly tetrandrine, have broad application prospects.
Summary of the invention
The object of the invention is to overcome existing shortcoming of carrying somatomedin and medicine carrying tissue biological engineering material, and a kind of nano biomimetic biomaterial capable that Chinese medicine is released in cartilage tissue engineered control that is used for is provided.Employing is separated, solvent exchange, freeze-drying have prepared nanofiber three-dimensional network timbering material; From bionical angle, use gelatin and chitosan that timbering material is modified, made up the timbering material that is similar to cartilage cell epimatrix; On timbering material, carried tetrandrine,, promoted the propagation of chondrocyte, improved the activity of chondrocyte, kept the functional expression of chondrocyte as the cell growth factor substitute.Just because of the reasonable combination of above-mentioned means, solved the problem that present cartilage tissue engineered technical field exists, have broad application prospects.The middle pharmaceutically active ingredient that has similar cell growth factor physiologically active with the surface biomimetic decorative layer sustained release of nanofiber biomaterial, be applied to cartilage tissue engineered field, part replaces the effect of cell growth factor, regulating cell proliferation, functional expression and phenotype keeps, the seed cell that solves the puzzlement field of tissue engineering technology dedifferentes problem, has cheap cost, less equipment input and simple preparation process condition.
In order to achieve the above object, the present invention is achieved through the following technical solutions:
1) preparation 1～7% concentration (mass volume ratio) poly--L-lactic acid tetrahydrofuran solution, add 1～50g/L tetrandrine chloroform soln, put under-20～8 ℃ the temperature and left standstill 5～10 hours, make solution gelization;
2) gelatin solution, chitosan solution or both mixed solutions of preparation 0.5～5% concentration (mass volume ratio) concentration are put pre-cooling in 2～8 ℃ the refrigerator; Poly--L-lactic acid solution of the gelation of step 1) is put in gelatin solution, chitosan solution or both mixed solutions after the pre-cooling displacement oxolane wherein; Replace oxolane with the deionized water of pre-cooling then;
3) gel that will replace tetrahydrofuran solvent removes the deionized water of attachment removal, places-40～-20 ℃ of cryogenic refrigerator pre-freezes, and it is dry to put into freezer dryer then, obtains nano biomimetic biomaterial capable.
4) materialogy characterizes: the nano biomimetic biomaterial capable of preparation is carried out photoelectricity energy spectrum analysis, infrared spectrum analysis, X-ray diffraction analysis, differential scanning calorimetric analysis, and adopt electron scanning Electronic Speculum (SEM) to carry out morphology analysis.
The nano biomimetic biomaterial capable of sustained release Chinese medicine of the present invention, it is the nanofiber three-dimensional network timbering material that has the Chinese medicine ingredients tetrandrine, the porosity of material is 92.4～98.2%, and distribution of fiber diameters is in 157～433nm scope, and pore size distribution is at 3.6～25.4 mu m ranges.
In preparation process, with the oxolane in gelatin and the chitosan solution replacement deionized water displacement gel, when the molecule segment of poly--L-lactic acid condenses gathering, in the gap of poly--L-lactic acid molecules that gelatin and chitosan molecule are trapped within simultaneously, in fibroplastic process, it is carried out biomimetic modification.Photoelectron spectroscopy analysis and infrared spectrum analysis proof gelatin and chitosan molecule are fixed among the poly--L-lactic acid timbering material of modified; The body of X-ray diffraction analysis and differential scanning calorimetric analysis proof timbering material remains poly--L-lactic acid, and gelatin and chitosan molecule mainly are distributed in the surface of timbering material.
Unique distinction of the present invention is, carries tetrandrine on the nano biomimetic biomaterial capable, and it can promote the propagation of chondrocyte as somatomedin, improves the activity of chondrocyte, keeps the functional expression of chondrocyte; Tetrandrine is along with the degradation process of timbering material discharges for a long time, and long duration of action is kept phenotype of chondrocytes and cell function in the cartilaginous tissue of reconstruction.The alkalescence of tetrandrine and antiinflammatory pharmacological action can be alleviated poly--L-lactic acid degraded and generate the local organization pH variation stimulation that lactic acid causes, and prevent the generation of aseptic inflammation.The biomimetic modification layer can improve the biomaterial biocompatibility, not only promotes cell adhesion and propagation, improves the biomaterial cell compatibility, and can play the sustained release effect to the tetrandrine that the timbering material body contains.
Description of drawings
Fig. 1: the nano biomimetic biomaterial capable SEM photo of the sustained release Chinese medicine of 1% polylactic acid formulations prepared from solutions;
Fig. 2: the nano biomimetic biomaterial capable SEM photo of the sustained release Chinese medicine of 3% polylactic acid formulations prepared from solutions;
Fig. 3: the nano biomimetic biomaterial capable SEM photo of the sustained release Chinese medicine of 5% polylactic acid formulations prepared from solutions;
Fig. 4: the nano biomimetic biomaterial capable of chondrocyte implantation sustained release Chinese medicine is cultivated the SEM photo behind the 7d;
Fig. 5: the nano biomimetic biomaterial capable of chondrocyte implantation sustained release Chinese medicine is cultivated the SEM photo behind the 14d;
Fig. 6: the nano biomimetic biomaterial capable of chondrocyte implantation sustained release Chinese medicine is cultivated the SEM photo behind the 21d;
Fig. 7: the through engineering approaches cartilaginous tissue of external structure is learned photo in the nano biomimetic biomaterial capable of sustained release Chinese medicine.
The specific embodiment
The preparation method of the sustained release Chinese medicine nano biomimetic biomaterial capable that the present invention relates to mainly comprises the gelation of the poly--L-lactic acid solution that contains tetrandrine; Solvent exchange and biomimetic modification; Vacuum lyophilization.The material microstructure scanning electron microscope analysis, medicine sustained release high-efficient liquid phase chromatogram technique analysis.
Embodiment 1:
1) get the 1mg tetrandrine, be dissolved in (being 1g/L tetrandrine chloroform soln) in the 1ml chloroform, standby.Precision takes by weighing 0.5g and gathers-L-lactic acid, put into 200ml ground triangular flask, add 50ml oxolane (promptly 1% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 15 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.The polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 8h, makes solution gelization.
2) take by weighing the 5.0g gelatin, put into the 1000ml beaker, add the 1000ml deionized water, be heated to 40 ℃ gelatin is dissolved fully, be made into 0.5% aqueous gelatin solution.Put into 4 ℃ of refrigerator pre-coolings.
3) take out the ground weighing botle, uncap is put into and is pre-chilled to 4 ℃ aqueous gelatin solution, room temperature dissolving displacement oxolane 16h.Outwell aqueous gelatin solution then, change to deionized water, replace 48h continuously.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 24h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.Through measuring, the porosity of material is 98.2%.Scanning electron microscope (SEM) is analyzed, and measures the fibre diameter and the pore-size of material and also does statistical analysis, and distribution of fiber diameters is in 194.6～350.2nm scope, pore size distribution 10.5～25.4 mu m ranges (Fig. 1).
Embodiment 2:
1) precision takes by weighing the 50mg tetrandrine, is dissolved in (being 50g/L tetrandrine chloroform soln) in the 1ml chloroform, and is standby.Precision takes by weighing 3.5g and gathers-L-lactic acid, put into 200ml ground triangular flask, add 50ml oxolane (promptly 7% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 15 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.The polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 8h, makes solution gelization.
2) take by weighing the 50g gelatin, put into the 1000ml beaker, add the 1000ml deionized water, be heated to 40 ℃ gelatin is dissolved fully, be made into 5% aqueous gelatin solution.Put into 4 ℃ of refrigerator pre-coolings.
3) take out the ground weighing botle, uncap is put into and is pre-chilled to 4 ℃ aqueous gelatin solution, room temperature dissolving displacement oxolane 16h.Outwell aqueous gelatin solution then, change to deionized water, replace 48h continuously.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 24h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.Through measuring, the porosity of material is 92.4%.Scanning electron microscope (SEM) is analyzed, and measures the fibre diameter and the pore-size of material and also does statistical analysis, and distribution of fiber diameters is in 221～431.8nm scope, pore size distribution 3.6～4.5 mu m ranges.
Embodiment 3:
1) precision takes by weighing the 10mg tetrandrine, is dissolved in (being 20g/L tetrandrine chloroform soln) in the 0.5ml chloroform, and is standby.Precision takes by weighing 3g and gathers-L-lactic acid, put into 250ml ground triangular flask, add 100ml oxolane (promptly 3% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 30 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.
2) the polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 10h, makes solution gelization.
3) take by weighing the 15.0g gelatin, put into the 1000ml beaker, add the 1000ml deionized water, be heated to 40 ℃ gelatin is dissolved fully, be made into 1.5% aqueous gelatin solution.Put into 4 ℃ of refrigerator pre-coolings.
4) take out the ground weighing botle, uncap is put into and is pre-chilled to 4 ℃ aqueous gelatin solution, room temperature dissolving displacement oxolane 12h.Outwell aqueous gelatin solution then, change to deionized water, replace 60h continuously.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 32h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.Through measuring, the porosity of material is 97%.Scanning electron microscope (SEM) is analyzed, and measures the fibre diameter and the pore-size of material and also does statistical analysis, and distribution of fiber diameters is in 212.4～406.6nm scope, pore size distribution 8.7～15.2 mu m ranges (Fig. 2).
Embodiment 4:
1) precision takes by weighing the 1mg tetrandrine, is dissolved in (being 1g/L tetrandrine chloroform soln) in the 1ml chloroform, and is standby.Precision takes by weighing 5g and gathers-L-lactic acid, put into 200ml ground triangular flask, add 100ml oxolane (promptly 5% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 15 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.
2) the polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 8h, makes solution gelization.
3) take by weighing the 5.0g gelatin, put into the 1000ml beaker, add the 1000ml deionized water, be heated to 40 ℃ gelatin is dissolved fully, be made into 0.5% aqueous gelatin solution.Put into 4 ℃ of refrigerator pre-coolings.
4) take out the ground weighing botle, uncap is put into and is pre-chilled to 4 ℃ aqueous gelatin solution, room temperature dissolving displacement oxolane 16h.Outwell aqueous gelatin solution then, change to deionized water, replace 36h continuously.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 24h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.Through measuring, the porosity of material is 95.9%.Scanning electron microscope (SEM) is analyzed, and measures the fibre diameter and the pore-size of material and also does statistical analysis, and distribution of fiber diameters is in 157.8～433.8nm scope, pore size distribution 3.6～7.5 mu m ranges (Fig. 3).
Embodiment 5:
1) precision takes by weighing the 10mg tetrandrine, is dissolved in (being 20g/L tetrandrine chloroform soln) in the 0.5ml chloroform, and is standby.Precision takes by weighing 7g and gathers-L-lactic acid, put into 250ml ground triangular flask, add 100ml oxolane (promptly 7% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 30 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.The polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 10h, makes solution gelization.
2) take by weighing the 50.0g chitosan, put into the 1000ml beaker, add 1% acetum of 100ml with deionized water and glacial acetic acid preparation, magnetic agitation makes dissolving fully, adds the 900ml deionized water, is made into 5% chitosan aqueous solution.Put into 5 ℃ of refrigerator pre-coolings.
3) take out the ground weighing botle, uncap is put into and is pre-chilled to 5 ℃ chitosan aqueous solution, room temperature dissolving displacement oxolane 10h.Outwell chitosan aqueous solution then, change to deionized water, change water once, replace 48h continuously every 8h.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 20h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.
Embodiment 6:
1) precision takes by weighing the 25mg tetrandrine, is dissolved in (being 50g/L tetrandrine chloroform soln) in the 0.5ml chloroform, and is standby.Precision takes by weighing 1g and gathers-L-lactic acid, put into 250ml ground triangular flask, add 100ml oxolane (promptly 1% concentration is gathered-L-lactic acid tetrahydrofuran solution), measure 30 μ l tetrandrine chloroform solns with the pipettor precision then, add in the triangular flask jam-pack bottle stopper, constantly jolting in 60 ℃ of water-baths, it is dissolved fully, be mixed with the polylactic acid solution that contains tetrandrine.The polylactic acid solution that 4ml is contained tetrandrine joins in the grournd glass weighing botle, covers lid completely, puts into 5 ℃ refrigerator 10h, makes solution gelization.
2) take by weighing 5g chitosan and 5g gelatin, together put into the 1000ml beaker, add 1% acetum of 100ml with deionized water and glacial acetic acid preparation, magnetic agitation makes dissolving fully, add the 900ml deionized water, be made into each aqueous solution of 0.5% of chitosan-containing/gelatin.Put into 5 ℃ of refrigerator pre-coolings.
3) take out the ground weighing botle, uncap is put into and is pre-chilled to chitosan/aqueous gelatin solution of 5 ℃, room temperature dissolving displacement oxolane 18h.Outwell chitosan/aqueous gelatin solution then, change to deionized water, change water once, replace 50h continuously every 5h.Gel is taken out together with the glass weighing botle, remove the deionized water of attachment removal as far as possible, place-40 ℃ of cryogenic refrigerators freezing, it is freezed.Take out the glass weighing botle, seal bottleneck with cotton paper, put into freezer dryer, the 24h that starts shooting continuously makes its bone dry.Take out the timbering material in the weighing botle, cut into suitable shape.The gamma-radiation sterilization, exposure dose 25kGy obtains aseptic sustained release Chinese medicine nano biomimetic biomaterial capable.
4) drug loading is measured and the analysis of medicine sustained release: the drug loading that utilizes high-pressure liquid phase chromatogram therapy determining carried stent material.Chromatographic column: Hypersil ODS-2Columns 250 x 4.6mm5 μ m; Mobile phase: methanol-0.3% dihexylamine (80:20, v/v); Flow velocity: 0.8ml/min; Detect wavelength: 282nm; Column temperature: 25 ℃.Tetrandrine is in 0～2.30 μ g scope internal linear relation good (r=0.9999), and average recovery is 98.05%, and RSD is 3.46%.The content of tetrandrine is between 0.04-2mg/g (tetrandrine/polylactic acid) in the nano biomimetic biomaterial capable of mensuration sustained release Chinese medicine.
5) the nano biomimetic biomaterial capable precision of sustained release Chinese medicine is weighed, be placed in the phosphate buffered solution of pH7～8, place under 37 ℃ of constant temperatures, the phosphate buffered solution of timing sampling and additional respective amount continues 6 months.Release rate with the high effective liquid chromatography for measuring medicine.The drug release characteristics of the nano biomimetic biomaterial capable of sustained release Chinese medicine is that tetrandrine can steadily discharge with the material degradation process among the present invention, and the accumulative total drug release rate is about 70～80% of drug loading in 6 months.
6) cartilage tissue engineered reconstruction: chondrocyte is planted in the nano biomimetic biomaterial capable of sustained release Chinese medicine, fiber in the timbering material is similar to the collagen fiber diameter in the natural cartilage extracellular matrix, collagen fiber in the excretory extracellular matrix of chondrocyte make above can stick to well, and be orientated along scaffold fibers, connect mutually, ordered distribution is built into the extracellular matrix (Fig. 4 that is similar to natural cartilage, 5,6).The histological observation result shows (Fig. 7), and the through engineering approaches cartilage of external structure has to natural cartilage and organizes similar characteristics of organizational structure.By the specific stain and the immunohistochemical staining of cartilaginous tissue, confirmed that the extracellular matrix components of through engineering approaches cartilaginous tissue is also organized identical with natural cartilage.The polylactic acid bracket material that contains tetrandrine, the fibre gap structure that not only has fibre diameter and 5～17 μ m of 100～500nm, the tetrandrine that wherein contains has facilitation to chondrocyte proliferation and functional expression, more help attaching, the growth of chondrocyte, make up through engineering approaches cartilage with natural three dimensional structure and biomechanical property.
Nano biomimetic biomaterial capable of the sustained release Chinese medicine that the present invention proposes and preparation method thereof, be described by preferred embodiment, person skilled obviously can be in not breaking away from content of the present invention, spirit and scope to material as herein described with manufacture method is changed or suitably change and combination, realize the technology of the present invention.Special needs to be pointed out is, the replacement that all are similar and change apparent to those skilled in the artly, they are regarded as being included in spirit of the present invention, scope and the content.
Claims (2)
1. the nano biomimetic biomaterial capable of a sustained release Chinese medicine, it is characterized in that having the nanofiber three-dimensional network timbering material of tetrandrine Chinese medicine ingredients, the porosity of material is 92.4～98.2%, distribution of fiber diameters is in 157～433nm scope, and pore size distribution is at 3.6～25.4 mu m ranges.
2. the preparation method of the sustained release Chinese medicine nano biomimetic biomaterial capable of claim 1 is characterized in that step is as follows:
1). poly--L-lactic acid tetrahydrofuran solution of preparation mass volume ratio 1～7% concentration, add 1～50g/L tetrandrine chloroform soln, put under-20～8 ℃ the temperature and left standstill 5～10 hours, make solution gelization;
2). gelatin solution, chitosan solution or both mixed solutions of preparation mass volume ratio 0.5～5% concentration, put pre-cooling in 2～8 ℃ the refrigerator; Poly--L-lactic acid solution of the gelation of step 1) is put in gelatin solution, chitosan solution or both mixed solutions after the pre-cooling displacement oxolane wherein; Replace oxolane with the deionized water of pre-cooling then;
3). the gel that will replace tetrahydrofuran solvent removes the deionized water of attachment removal, places-40～-20 ℃ of cryogenic refrigerator pre-freezes, and it is dry to put into freezer dryer then, obtains nano biomimetic biomaterial capable.
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| CN103933909B (en) * | 2014-03-06 | 2015-08-12 | 天津大学 | Rutin-trivalent chromium sodium alginate chitosan microcapsules system and preparation method and application |
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