CN108743545A - A kind of alginate-drug-carrying nanometer particle-polycation microcapsules and its preparation and application - Google Patents
A kind of alginate-drug-carrying nanometer particle-polycation microcapsules and its preparation and application Download PDFInfo
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Abstract
The present invention relates to a kind of alginate-drug-carrying nanometer particle-polycation microcapsule product and its preparations and application.Microcapsule product is the spherical microcapsule of 100-1000 microns of grain size, its structure is divided into microcapsule membrane and kernel two parts, microcapsule membrane forms polyelectrolyte compound water congealing glued membrane by alginate, drug-carrying nanometer particle, polycation by LBL self-assembly, and kernel is the hydrogel containing zooblast or hydrosol environment.This microcapsule product is used for the occlusion vehicle of cell transplantation as immune isolating tool.Under the premise of ensureing strength of microcapsules and function of immune isolation, the problem of reducing microcapsule membrane surface roughness and surface charge, realize that anti-inflammatory drug is discharged in the original position of surface of microcapsule simultaneously, stablize micro-capsule membrane structure from drug-carrying nanometer particle solves fibrotic encapsulation problem with anti-inflammatory two angles of drug in situ release.
Description
Technical field
The present invention relates to a kind of microcapsule product, specifically a kind of alginic acids for bioactive substance embedding
Salt-drug-carrying nanometer particle-polycation bio-microcapsule product and its preparation method and application.
Background technology
In the 1960s, Chang reports semi-permeable membrane microcapsules, it is indicated that embed the bioactivity such as protein, enzyme with it
Substance and cell can keep biological substance active[1].In the early 1980s, Lim and Sun is directed to tissue/cell functional impairment
Property disease (such as diabetes), is successfully prepared sodium alginate/α-polylysine semi-permeable membrane microcapsules（Abbreviation α-APA microcapsules）,
Encapsulating Wistar rat Islet cells are simultaneously implanted into diabetes WistarLewis rat bodies, and secretion release insulin is to regulate and control
Blood glucose amount[2].Thus the fast development for having pushed microencapsulation associated materials and preparation method research, in cell transplantation, drug
It is used widely in the preclinical study of biomedical sectors such as release and gene therapy[3].In numerous bio-microcapsules
In, sodium alginate-polylysine microcapsules are using more, but main asking of being still suffered from process of clinical application of microcapsules
Topic is in microencapsulated cell body after transplanting, and treatment initial performances are apparent, but curative effect declines after a period of time, essential reason
It is the micro-capsule to implant to cause the immune response of host and form fibrotic encapsulation, influences the substance of nutrition and metabolism product
It transmits, prevents micro-capsule inner tissue/cell because nutriment and oxygen supply are insufficient, intracapsular metabolite is from diffusing out outside capsule, lead
Graft function is caused to lose, graft failure[4]。
For this purpose, there is researcher from the angle for resisting host inflammation generation, anti-inflammatory drug will be loaded with（Such as dexamethasone）
Polylactic acid（Poly (lactic-co-glycolic) acid, PLGA）Carrier contains in microcapsules kernel[5], it is intended to pass through
Drug reaches antiphlogistic effects in locally release, but since PLGA carriers are introduced into microcapsules kernel, it is molten to change sodium alginate
The hydrophilic and hydrophobic of liquid influences the sphericity of calcium alginate gel bead, and microcapsules sphericity difference is to cause fibrotic encapsulation
One of key factor；Acid product after PLGA degradations can inherently excite host inflammation to react, and aggravate the fibrosis of graft
Package；Anti-inflammatory drug is firstly gathered at after PLGA releases in microcapsules kernel, needs to diffuse out microcapsules competence exertion anti-inflammatory
Effect.Therefore, this microcapsules by the PLGA nanoparticles for being loaded with anti-inflammatory drug can not effectively solve the fibrosis after transplanting
Package problem.
Invention content
The present invention mainly solves the problems, such as：Fibrotic encapsulation problem after microcapsules transplanting.
Used technical solution is to provide a kind of alginate-medicament-carried nano to the present invention to solve above-mentioned technical problem
Grain-polycation microcapsule product, the microcapsule product structure are divided into microcapsule membrane and kernel two parts, the microcapsule membrane
For the polyelectrolyte compound water congealing glued membrane formed by alginate, drug-carrying nanometer particle, polycation, kernel is to contain zooblast
Alginate liquid or alginate hydrogel environment.
The thickness of above-mentioned microcapsule membrane is at 1-50 microns；The alginate of component film, drug-carrying nanometer particle, polycation three
Mass ratio is 10:1:1-1:10:10.
Drug-carrying nanometer particle is the nanometer formulation for carrying anti-inflammatory drug.Anti-inflammatory drug includes dexamethasone, betamethasone, times chlorine rice
Pine, Triamcinolone acetonide, hydrocortisone, prednisolone, prednisone, curcumin, resveratrol.
The sodium alginate derivative and drug molecule that the chitosan derivatives or amphiphilic that nanoparticle is modified by amphiphilic are modified are certainly
Assemble nanoparticle, grain size is in 50-500nm.
Alginate in the film component of microcapsules is one in alginic acid calcium salt or alginic acid barium salt or alginic acid zinc salt
Kind or two kinds, also can include or one or both of not comprising alginic acid sodium salt or sylvite simultaneously.
Polycation in the film component of microcapsules is in α or ε polylysines, poly ornithine, poly arginine, chitosan
It is one or more kinds of.
Alginate hydrogel is one or more kinds of seas in divalent metal calcium, barium or zinc in the kernel of microcapsules
Alginate hydrogel, alginate solution are the sylvite or sodium salt solution of alginic acid, a concentration of 0.1-50g/L of alginate.
The method for preparing a kind of above-mentioned alginate-drug-carrying nanometer particle-polycation microcapsules, this method includes following step
Suddenly：
1）Prepare the alginate microballoon for being embedded with zooblast, referred to as microballoon A；
2）By step 1）In microballoon A immerse in said polycation solution, microballoon A is 1 with said polycation solution volume ratio:1~1:
40, it reacts 1-60 minutes, obtains alginate-polycation microcapsules at this time, referred to as microballoon B, taking-up is washed with physiology salt
It washs；
3）By step 2）In microballoon B be immersed in the nanoparticle solution for carrying medicine, microballoon B and nanoparticle liquor capacity ratio are 1：1~
1：40, it reacts 1-60 minutes, obtains the microcapsules of internal gel core at this time, referred to as microballoon C, taking-up is washed with physiology salt
It washs；
4）By step 3）In microballoon C be immersed in alkali metal alginate solution, microballoon C is with alginate solution volume ratio
1：1~1：40, it reacts 1-60 minutes, the microcapsules for obtaining the inside gel core of surface neutralization are microballoon D；
5）By step 4）In microballoon D immerse in organic metal chelating agent solution, the alginate inside the microcapsules that liquefy,
Microballoon D is 1 with organic metal chelating agent solution volume ratio：1~1：40, it reacts 1-60 minutes, taking-up brine, this
When obtain interior liquid core microcapsules be microballoon E.
A kind of alginate-drug-carrying nanometer particle-polycation microcapsules are applied to the embedding of zooblast.The animal
Cell includes the cell with secretion bioactive substance function, cell line cell, genetically engineered cell, stem cell or stem cell
The cell of differentiation.
The in vitro islet cells of the cell behaviour that there is secretion bioactive substance function or mammal source,
Liver cell, thyroid cell, parathyroid cells, adrenal medullary cell.
The advantageous effects of the present invention：
Due to containing load anti-inflammatory drug on the microcapsule membrane of product of the present invention（Such as curcumin）Nanoparticle, realize drug molecule exist
The local release in situ of transplanting directly acts on inflammatory cell, reduction inflammatory factor generates, the fiber after solution microcapsules implantation
Change package problem, not only enhanced drug in transplanting local anti-inflammatory effect but also avoids the side effect of Formulations for systemic administration.
With traditional sodium alginate-polylysine microcapsules（APA microcapsules）It compares, this novel alga of the invention acid
Salt-drug-carrying nanometer particle-polycation bio-microcapsule product, the surface roughness of microcapsule membrane are substantially less than APA microcapsules
With ACA microcapsules, more preferably biocompatibility is shown.
Introducing of the microcapsule membrane of product of the present invention due to nanoparticle especially amphiphilic chitosan nano, it is possible to reduce tradition
Polycation（Such as polylysine, poly ornithine, poly arginine）Dosage, improve polycation（Such as polylysine, poly- bird ammonia
Acid, poly arginine etc.）Stability on microcapsule membrane, polycation falls off the host immune of initiation after avoiding microcapsules from transplanting
Reaction, to reduce fibrotic encapsulation.
The microcapsule membrane of product of the present invention has played the effect of similar " concrete " due to the introducing of nanoparticle, preparation
Higher mechanical strength and mechanical stability is presented in microcapsule membrane, and it is sudden and violent to reduce after microcapsules are implanted into the cell that stress is broken, transplants
Reveal situation, to reduce fibrotic encapsulation.
The microcapsule membrane of product of the present invention has superior function of immune isolation, when being used for heteroplasm's cell transplantation, energy
Keep function of immune isolation, i.e., the cell embedded in micro-capsule that cannot go out microcapsules, antibody molecule, complement molecule outside micro-capsule are exempted from
Epidemic disease cell, which cannot enter, kills cell in microcapsules, while the active constituent of cell metabolism secretion can free in and out microcapsules.
The preparation process mild condition of product of the present invention, the activity for being conducive to cell are kept.
Description of the drawings
Fig. 1 is to illustrate a kind of alginic acid of the present invention by taking the amphiphilic chitosan nano of polylysine and load curcumin as an example
Salt-drug-carrying nanometer particle-polycation bio-microcapsule product structure schematic diagram.
Specific implementation mode
The chitosan nano for being loaded with anti-inflammatory drug or alginate nano grain are used for bio-microcapsule film by the present invention
It prepares, has invented a kind of novel alginate-drug-carrying nanometer particle for zooblast embedding-polycation microcapsules production
Product reduce microcapsule membrane surface roughness and surface charge ensureing strength of microcapsules and under the premise of function of immune isolation
Problem, while realizing that anti-inflammatory drug is discharged in the original position of surface of microcapsule, stablize micro-capsule membrane structure and drug from drug-carrying nanometer particle
It is in situ to discharge anti-inflammatory two angles solution fibrotic encapsulation problem.
A kind of alginate-drug-carrying nanometer particle-polycation microcapsules, structure are divided into microcapsule membrane and kernel two parts,
The microcapsule membrane is the polyelectrolyte compound water congealing glued membrane formed by alginate, drug-carrying nanometer particle, polycation, kernel
For alginate liquid or alginate hydrogel environment containing zooblast.The thickness of microcapsule membrane is at 1-50 microns；Group
Alginate, drug-carrying nanometer particle, the polycation three's mass ratio of film forming are 10:1:1-1:10:10.
Drug in drug-carrying nanometer particle refers to anti-inflammatory drug, including dexamethasone, betamethasone, beclomethasone, Triamcinolone acetonide,
Hydrocortisone, prednisolone, prednisone, curcumin, resveratrol.The chitosan derivatives or two that nanoparticle is modified by amphiphilic
The modified sodium alginate derivative of parent forms nanoparticle with drug molecule self assembly, and grain size is in 50-500nm.
Alginate in the film component of microcapsules is one in alginic acid calcium salt or alginic acid barium salt or alginic acid zinc salt
Kind or two kinds, also can include or one or both of not comprising alginic acid sodium salt or sylvite simultaneously.
Polycation in the film component of microcapsules is in α or ε polylysines, poly ornithine, poly arginine, chitosan
It is one or more kinds of.
Alginate hydrogel is one or more kinds of seas in divalent metal calcium, barium or zinc in the kernel of microcapsules
Alginate hydrogel, alginate solution are the sylvite or sodium salt solution of alginic acid, a concentration of 0.1-50g/L of alginate.
The method for preparing a kind of above-mentioned alginate-drug-carrying nanometer particle-polycation microcapsules, this method includes following step
Suddenly：
1）Prepare the alginate microballoon for being embedded with zooblast, referred to as microballoon A；
2）By step 1）In microballoon A immerse in said polycation solution, microballoon A is 1 with said polycation solution volume ratio:1~1:
40, it reacts 1-60 minutes, obtains alginate-polycation microcapsules at this time, referred to as microballoon B, taking-up is washed with physiology salt
It washs；
3）By step 2）In microballoon B be immersed in the nanoparticle solution for carrying medicine, microballoon B and nanoparticle liquor capacity ratio are 1：1~
1：40, it reacts 1-60 minutes, obtains the microcapsules of internal gel core at this time, referred to as microballoon C, taking-up is washed with physiology salt
It washs；
4）By step 3）In microballoon C be immersed in alkali metal alginate solution, microballoon C is with alginate solution volume ratio
1：1~1：40, it reacts 1-60 minutes, the microcapsules for obtaining the inside gel core of surface neutralization are microballoon D；
Wherein, alkali metal alginate solution is sylvite or sodium salt, and molecular weight distribution is 10KDa~2000KDa, and alginate is dense
Degree is 0.1-5g/L；
5）By step 4）In microballoon D immerse in organic metal chelating agent solution, the alginate inside the microcapsules that liquefy,
Microballoon D is 1 with organic metal chelating agent solution volume ratio：1~1：40, it reacts 1-60 minutes, taking-up brine, this
When obtain interior liquid core microcapsules be microballoon E.
Wherein, the organic metal chelating agent solution for participating in liquefaction reaction is the sodium citrate or/and 50- of 40-70mmol/L
The EDTA of 200mmol/L.
The microcapsules are used for the embedding of zooblast, and the zooblast is behaved or the in vitro pancreas islet of mammal source
Cell, liver cell, thyroid cell, parathyroid cells, adrenal medullary cell etc. have the function of to secrete bioactive substance
Cell, cell line cell, genetically engineered cell, stem cell or stem cell differentiation various cells.
The mode for forming alginate microballoon is electrostatic drop generation[6], orifice extrusion molding[7], emulsification-exterior gel
Method[8], emulsion-internal gelatification method[9]Or membrane emulsification[10]。
It is prepared by drug-carrying nanometer particle：Amphiphilic Chitosan Nanoparticles for Drug Delivery is self-assembly method[11]；Amphiphilic sodium alginate medicament-carried nano
Grain is self-assembly method[12]。
By taking the amphiphilic chitosan nano of polylysine and load curcumin as an example, the present invention is further illustrated.
Embodiment 1
Prepare hydrophobic substitution 4.5, the amphiphilic chitosan material that cationic hydrophilic groups degree of substitution is 60.
The cationic amphiphic chitosan nano for carrying curcumin is prepared under aseptic condition.
Calcium alginate gel bead is prepared by high-pressure electrostatic method under aseptic condition.
By (polylysine molecule amount 20kDa, solution is by normal saline, concentration in microballoon immersion polylysin solution
0.5g/L), microballoon and polylysin solution volume ratio are 1:10, it reacts 5 minutes, brine three times.
Step 3）The microballoon of preparation is immersed in step 1 again）The cationic amphiphic chitosan nano of the load curcumin of preparation
In grain solution, microballoon is 1 with nanoparticle liquor capacity ratio:10, it reacts 10 minutes, brine three times.
Step 4）The microballoon of preparation reacts 10 minutes again with 0.2% sodium alginate soln, and brine is prepared into sea
Mosanom-polylysine-load curcumin chitosan nano（APCcur）Microcapsules.
The APCcur microcapsule membrane surface profilers being prepared into measure the surface roughness of film, as a result show that film surface is thick
Rugosity is 25 ± 12nm.
The APCcur microcapsules being prepared into are in 55mmol/L sodium citrate solutions, microballoon and sodium citrate solution volume ratio
It is 1:10,50rpm concussions 60 minutes, count the grain size of 50 microballoons under microscope, it is 60% to calculate microsphere volume expansion rate.
APCcur microcapsules are implanted into C57BL/6 mouse peritoneals with syringe for 250 microlitres, mouse is put to death after 4 weeks, abdominal cavity fills
Wash recycling microcapsules, the rate of recovery of free microcapsules is up to 95%, and the microcapsules good sphericity recycled, and surface is smooth, surface without
Fibrotic encapsulation phenomenon, the cell encapsulation rate 1.25 ± 0.22% for the micro-capsule that dissociates.
Comparative example 1
By the calcium alginate gel bead prepared in embodiment 1 immerse polylysin solution in (polylysine molecule amount 20kDa,
Solution is by normal saline, concentration 0.5g/L), microballoon is 1 with polylysin solution volume ratio:10, it reacts 5 minutes, physiology
It is reacted 10 minutes with 0.2% sodium alginate soln again after salt water washing, brine is prepared into AP1 microcapsules.
The AP1 microcapsule membrane surface profilers being prepared into measure the surface roughness of film, as a result show that film surface is coarse
Degree is 100 ± 26nm, hence it is evident that is higher than 1 result APCcur microcapsule membranes of embodiment.
In 55mmol/L sodium citrate solutions, microballoon is the AP1 microcapsules being prepared into sodium citrate solution volume ratio
1:10,50rpm concussions 60 minutes, count the grain size of 50 microballoons under microscope, it is 300% to calculate microsphere volume expansion rate.
AP1 microcapsules are implanted into C57BL/6 mouse peritoneals with syringe for 250 microlitres, mouse, peritoneal lavage are put to death after 4 weeks
Microcapsules are recycled, the rate of recovery for the microcapsules that dissociate is 38%, and the microcapsules of recycling, which have, to be obviously crushed, and there is fibrotic encapsulation on surface
Phenomenon, the cell encapsulation rate 9.67 ± 0.38% for the micro-capsule that dissociates.
Comparative example 2
By the calcium alginate gel bead prepared in embodiment 1 immerse polylysin solution in (polylysine molecule amount 20kDa,
Solution is by normal saline, concentration 0.5g/L), microballoon is 1 with polylysin solution volume ratio:10, it reacts 15 minutes, physiology
It is reacted 10 minutes with 0.2% sodium alginate soln again after salt water washing, brine is prepared into AP2 microcapsules.
The AP2 microcapsule membrane surface profilers being prepared into measure the surface roughness of film, as a result show that film surface is coarse
Degree is 165 ± 41nm, hence it is evident that is higher than 1 result APCcur microcapsule membranes of embodiment.
In 55mmol/L sodium citrate solutions, microballoon is the AP2 microcapsules being prepared into sodium citrate solution volume ratio
1:10,50rpm concussions 60 minutes, count the grain size of 50 microballoons under microscope, it is 140% to calculate microsphere volume expansion rate.
AP2 microcapsules are implanted into C57BL/6 mouse peritoneals with syringe for 250 microlitres, mouse, peritoneal lavage are put to death after 4 weeks
Microcapsules are recycled, the rate of recovery for the microcapsules that dissociate is 31%, and the microcapsules sphericity of recycling is preferable, and surface has fibrotic encapsulation existing
As the cell encapsulation rate 8.92 ± 0.26% for the micro-capsule that dissociates.
Embodiment 2
Bibliography method prepares the amphiphilic sodium alginate material of hydrophobically modified.
The amphiphilic sodium alginate nanoparticles solution for carrying curcumin is prepared under aseptic condition.
Prepared by orifice extrusion molding is embedded with the calcium alginate gel bead of porcine hepatocyte, cell content 5 × 107/ in microballoon
Ml microballoons.
Microballoon is successively immersed in polylysin solution (polylysine molecule amount 20kDa, solution by normal saline,
Concentration 0.5g/L), microballoon is 1 with polylysin solution volume ratio:10, it reacts 10 minutes, brine three times.
Step 4）The microballoon of preparation is immersed in step 2 again）The amphiphilic sodium alginate nanoparticles solution of the load curcumin of preparation
In, microballoon is 1 with nanoparticle liquor capacity ratio:10, it reacts 15 minutes, brine three times.
After the liquefaction of 55mM sodium citrates, brine, then 10 minutes are reacted with 0.15% sodium alginate soln, physiology
Salt water washing is prepared into the APAcur microcapsules for being embedded with liver cell.
The APAcur microcapsules for being embedded with porcine hepatocyte are prepared into external bioartificial liver system, the animal mould for the monkey that declines for liver
Type, the results show that after extracorporal circulatory system 4 days, the decline glutamic-pyruvic transaminase of monkey, glutamic-oxalacetic transaminease level of liver is restored to normal level,
Blood ammonia index restores normal, and the liver of the monkey symptom that declines is corrected, and APAcur microcapsules keep form complete in bioartificial liver system,
Protein adsorption phenomenon is not found after blood perfusion.
Embodiment 3
Prepare hydrophobic substitution 3.5, the amphiphilic chitosan material that cationic hydrophilic groups degree of substitution is 62.
The cationic amphiphic chitosan nano solution for carrying curcumin is prepared under aseptic condition.
High-pressure electrostatic method prepares the calcium alginate gel bead for being embedded with porcine islet, contains 1-2 in each microballoon
Pancreas islet.
With the method for embodiment 1, microballoon is successively immersed into (polylysine molecule amount 20kDa, solution in polylysin solution
By normal saline, concentration 0.5g/L), and carry curcumin cationic amphiphic chitosan nano solution in, it is secondary at
It is reacted 10 minutes with 0.2% sodium alginate soln again after film reaction, brine, it is raw after the liquefaction of 55mM sodium citrates
Salt water washing is managed, the APCcur microcapsules for being embedded with islet cells are prepared into.
The APCcur microcapsules being prepared into are used for the cell therapy of diabetes rat model, pass through intraperitoneal transplantation, rat serum
Sugar level restores normal level in 1 day after the transfer, and diabetic symptom is significantly improved, and transplanting in vivo is recycled after 6 months, is sent out
Existing microcapsules are complete, and surface of microcapsule is smooth, and surface is without fibrotic encapsulation phenomenon, and islet cells keeps insulin double in micro-capsule
Sulphur hydrazone stained positive.
Embodiment 4
Prepare hydrophobic substitution 4, the amphiphilic chitosan material that cationic hydrophilic groups degree of substitution is 60.
The cationic amphiphic chitosan nano solution for carrying curcumin is prepared under aseptic condition.
Prepared by high-pressure electrostatic method is embedded with the calcium alginate gel bead of rat thyroid cell, cell content 3 in microballoon
× 107/ml microballoons.
With the method for embodiment 1, microballoon is successively immersed into (polylysine molecule amount 20kDa, solution in polylysin solution
By normal saline, concentration 0.5g/L), and carry curcumin cationic amphiphic chitosan nano solution in, it is secondary at
It is reacted 10 minutes with 0.2% sodium alginate soln again after film reaction, brine, it is raw after the liquefaction of 55mM sodium citrates
Salt water washing is managed, the APCcur microcapsules for being embedded with thyroid cell are prepared into.
The APCcur microcapsules for being embedded with rat thyroid cell being prepared into are used for the low model deltoid muscle of xenogenesis rat first
Transplanting, the low symptom of first of disease rat are corrected, T3, and T4 levels restore normal, and APCcur microcapsules return after transplanting three months
It receives, is recovered to form and keeps complete APCcur microcapsules, surface does not have fibrosis phenomenon.
Embodiment 5
With the cationic amphiphic chitosan nano solution of load curcumin prepared by the method for embodiment 4.
High-pressure electrostatic method prepares the calcium alginate gel bead for being embedded with bovine adrenal medullary epithelium, and cell contains in microballoon
Measure 2 × 107/ml microballoons.
With the method for embodiment 4, microballoon is successively immersed into (polylysine molecule amount 20kDa, solution in polylysin solution
By normal saline, concentration 0.5g/L), and carry curcumin cationic amphiphic chitosan nano solution in, it is secondary at
It is reacted 10 minutes with 0.2% sodium alginate soln again after film reaction, brine, it is raw after the liquefaction of 55mM sodium citrates
Salt water washing is managed, the APCcur microcapsules for being embedded with bovine adrenal medullary epithelium are prepared into.
The APCcur microcapsules for being embedded with bovine adrenal medullary epithelium being prepared into are used for the cranium of Parkinson disease model monkey
Interior fixed point transplanting, the parkinson symptoms such as hemiplegia of disease monkey are corrected, and APCcur microcapsules recycle after transplanting six months, recycle
Complete APCcur microcapsules are kept to form, surface does not have fibrosis phenomenon.
Embodiment 6
The APCcur microcapsules for being embedded with bovine adrenal medullary epithelium that embodiment 5 is prepared into are big for intractable pain model
The cavitas subarachnoidealis spinalis of mouse is transplanted, and the pain symptom of disease rat is corrected, and limbs are twitched number and significantly reduced, APCcur
Microcapsules recycle after transplanting six months, are recovered to form and keep complete APCcur microcapsules, surface does not have fibrosis phenomenon.
Embodiment 7
With the cationic amphiphic chitosan nano solution of load curcumin prepared by the method for embodiment 4.
High-pressure electrostatic method prepares the Chinese hamster ovary celI for being embedded with recombinant vascular endothelial cell growth inhibitory factor (endostatin)
Calcium alginate gel bead, cell content 5 × 107/ml microballoons in microballoon.
With the method for embodiment 4, microballoon is successively immersed into (polylysine molecule amount 20kDa, solution in polylysin solution
By normal saline, concentration 0.5g/L), and carry curcumin cationic amphiphic chitosan nano solution in, it is secondary at
It is reacted 10 minutes with 0.2% sodium alginate soln again after film reaction, brine, it is raw after the liquefaction of 55mM sodium citrates
Salt water washing is managed, the APCcur microcapsules for being embedded with endostatin-CHO cells are prepared into.
The APCcur microcapsules for being embedded with recombination endostatin-CHO cells being prepared into are used for melanoma tumor model mouse
Intraperitoneal transplantation, the tumour of disease rat is obviously reduced, and the transplanting of APCcur microcapsules is recycled after two months, is recovered to form holding
Complete APCcur microcapsules, surface does not have fibrosis phenomenon.
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Claims (12)
1. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules, which is characterized in that microcapsule structure is divided into microcapsules
Film and kernel two parts, the microcapsule membrane is that the polyelectrolyte formed by alginate, drug-carrying nanometer particle, polycation is answered
Heshui gel mould, kernel are alginate liquid or alginate hydrogel environment containing zooblast.
2. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1, which is characterized in that described
Microcapsule membrane thickness at 1-50 microns；Alginate, drug-carrying nanometer particle, the polycation three's mass ratio of component film be
10:1:1-1:10:10。
3. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1, which is characterized in that described
Drug-carrying nanometer particle be carry anti-inflammatory drug nanometer formulation.
4. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as claimed in claim 3, which is characterized in that described
Anti-inflammatory drug includes dexamethasone, betamethasone, beclomethasone, Triamcinolone acetonide, hydrocortisone, prednisolone, prednisone, ginger
Flavine, resveratrol.
5. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as claimed in claim 3, which is characterized in that described
The sodium alginate derivative that the chitosan derivatives or amphiphilic that nanoparticle is modified by amphiphilic are modified is formed with drug molecule self assembly
Nanoparticle, grain size is in 50-500nm.
6. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1, which is characterized in that described
Microcapsules film component in alginate be alginic acid calcium salt or alginic acid barium salt or one kind in alginic acid zinc salt or two
One or both of kind, also can include or not include alginic acid sodium salt or sylvite simultaneously.
7. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1, which is characterized in that described
Microcapsules film component in polycation be α or ε polylysines, poly ornithine, poly arginine, one kind in chitosan or
More than one.
8. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1, which is characterized in that described
Microcapsules kernel in alginate hydrogel be one or more kinds of alginates in divalent metal calcium, barium or zinc
Hydrogel, alginate solution are the sylvite or sodium salt solution of alginic acid, a concentration of 0.1-50g/L of alginate.
9. a kind of method of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1 is prepared, it is special
Sign is that this approach includes the following steps：
1）Prepare the alginate microballoon for being embedded with zooblast, referred to as microballoon A；
2）By step 1）In microballoon A immerse in said polycation solution, microballoon A is 1 with said polycation solution volume ratio:1~1:
40, it reacts 1-60 minutes, obtains alginate-polycation microcapsules at this time, referred to as microballoon B, taking-up is washed with physiology salt
It washs；
3）By step 2）In microballoon B be immersed in the nanoparticle solution for carrying medicine, microballoon B and nanoparticle liquor capacity ratio are 1：1~
1：40, it reacts 1-60 minutes, obtains the microcapsules of internal gel core at this time, referred to as microballoon C, taking-up is washed with physiology salt
It washs；
4）By step 3）In microballoon C be immersed in alkali metal alginate solution, microballoon C is with alginate solution volume ratio
1：1~1：40, it reacts 1-60 minutes, the microcapsules for obtaining the inside gel core of surface neutralization are microballoon D；
5）By step 4）In microballoon D immerse in organic metal chelating agent solution, the alginate inside the microcapsules that liquefy,
Microballoon D is 1 with organic metal chelating agent solution volume ratio：1~1：40, it reacts 1-60 minutes, taking-up brine, this
When obtain interior liquid core microcapsules be microballoon E.
10. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as described in claim 1 are applied to zooblast
Embedding.
11. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as claimed in claim 10, which is characterized in that institute
The zooblast stated includes the cell with secretion bioactive substance function, cell line cell, genetically engineered cell, stem cell
Or the cell of stem cell differentiation.
12. a kind of alginate-drug-carrying nanometer particle-polycation microcapsules as claimed in claim 11, which is characterized in that institute
State the cell behaviour with secretion bioactive substance function or the in vitro islet cells of mammal source, liver cell, first
Shape gland cell, parathyroid cells, adrenal medullary cell.
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