CN106399291A - Galactosyl grafted-modified alginate microspheres and applications thereof - Google Patents
Galactosyl grafted-modified alginate microspheres and applications thereof Download PDFInfo
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Abstract
The present invention relates to a galactosyl grafted-modified alginate hydrogel microsphere carrier preparation method. According to the present invention, the occupation on the hydroxyl site of alginic acid does not affect the gelation performance of the alginic acid, such that the embedding type hydrogel microsphere carrier with characteristics of good sphericity and good stability can be prepared from the galactosyl alginic acid being subjected to the grafting reaction, and can further be subjected to the film forming reaction with polycations to form the galactosyl alginate/polycation microcapsule.
Description
Technical field
The present invention relates to a kind of alginate microsphere vector product, specifically a kind of galactose group connects
The modified alginate microsphere carrier of branch.
Background technology
The substance encapsulation with biological activity is passed through in film in selectivity, the glomerate microcapsule of shape, referred to as
For " bio-microcapsule " [Chang TMS.Hemoglobin corpuscles.Research report for
honours physiology.Medical Library,McGill University,1957].With biology
Microcapsule is isolated and vehicle as the immunity of cell, using the metabolism of gene-recombinated cell or primary cell
Product adjusting body physiological function, therapy-related disease be biomedical worker study hotspot [Basic D,
Vacek I,Sun A M.Microencapsulation and transplantation of engineered
cells：a new approach to somatic gene therapy.Art Cells Blood Subs
ImmobBiotechnol,1996,24(3)：219-255].Alginate has excellent physics because of it
Chemical property and biocompatible property, become the main material of microencapsulation.Existing microencapsulation exists
During culture hepatocyte, due to the growth characteristics of hepatocyte anchorage dependence, microcapsule interior three-dimensional steric environment can not
Enough make the rapid Adaptable growth of hepatocyte.Galactose group is the spy of surface of hepatocytes asialoglycoprotein receptor
Specific ligand, can specific recognition surface of hepatocytes asialoglycoprotein receptor, therefore, in microcapsule material
The galactose group of Liver targeting is introduced on material, can induce and improve hepatocyte sticking within microcapsule and increase
Grow behavior [Jun Yang, Galactosylated alginate as a scaffold for hepatocytes
entrapment,Biomaterials,2002,23:471-479].Gala is introduced at present inside microcapsule
Glycosyl group can be by the blending on alginic acid or covalent modification.Macromolecular substances containing galactose group
[Seog-Jin Seo,Yun-Jaie Choi,Alginate microcapsules prepared with
xyloglucan as a synthetic extracellular matrix for hepatocyte attachment,
Biomaterials,2005,26:3607-3615] mainly sent out by preparing microcapsule with alginic acid blending
Wave effect, the material of blending easily leaks the stability reduction that therefore result in this kind of microcapsule；And be directed to
Covalent modification in alginic acid occurs mainly in the carboxyl position of alginic acid, carboxyl site occupy so that Sargassum
Acid degree of gelation when gelation reaction forms microcapsule reduces, for not excessive influence alginic acid
Gel-forming property, galactosyl substitution value is restricted, and document report substitution value 19% no affects on gelation process
[Ivan Donati,AmedeoVetere,Galactose-Substituted Alginate:Preliminary
Characterization and Study of Gelling Properties,Biomacromolecules,
2003,4:624-631].Due to the restriction of microcapsule stability and galactosyl substitution value, either it is blended
Or covalent modification alginic acid, all cannot introduce the half of higher concentration at present in the method for report inside microcapsule
Lactose group.
Content of the invention
For the problems referred to above, the present invention propose a kind of alginate of new galactose covalent modification/poly- sun from
Sub- microcapsule product, can be on the premise of ensureing strength of microcapsules and function of immune isolation, using alginic acid
Oh group covalence graft galactosyl group devicative is preparing galactosyl alginate/micro- glue of polycation
Capsule, realizes containing the stability not affecting microcapsule while galactose group again inside microcapsule, thus luring
Lead and improve hepatocyte sticking within microcapsule and propagation behavior.
Technical scheme
A kind of novel alga hydrochlorate/polycation microcapsule of the present invention, alginate hydroxyl is through containing amino base
The galactosyl Derivatives Modified of group, is the Sargassum containing living cells through galactose graft modification in microcapsule
The liquid of hydrochlorate or hydrogel environment.Surface of microcapsule can form poly- electricity by polycation and alginate
Solution matter compound water congealing glued membrane, such polycationic material includes：Shitosan, its deacetylation is 80-98%,
Molecular weight is 1kDa-800kDa；α-polylysine, molecular weight is 2kDa-500kDa；Epsilon-polylysine,
Molecular weight is 2kDa-500kDa；Poly arginine, molecular weight is 1kDa-500kDa；Poly ornithine, point
Son is measured as 1kDa-500kDa；Polyhistidyl, molecular weight is 1kDa-500kDa.Microcapsule can immerse organic
In metal-chelating agent solution, the internal alginate of liquefaction microcapsule, participate in the organic metal of liquefaction reaction
Chelating agent solution is the sodium citrate of 40-70mmol/L or the EDTA solution of 50-200mmol/L, microcapsule
It is 1 with organic metal chelating agent solution volume range:1～1:40, react 1-60 minute, taking-up physiology salt
Water washing, now obtains the microcapsule of interior liquid core.
In the present invention, galactosyl alginate/polycation Microcapsules Size is in 100-1000 micron；Institute
It is 10kDa-10000kDa with alginate molecular weight, the galactosyl that alginate is grafted amido-containing group spreads out
Biological percent grafting is 1%-199%.In the galactosyl derivant of amino group, with Lactose amine (monoamine
Terminated lactobionic lactone) as a example, its course of reaction is as follows：
Wherein, alginate represents alginate；M representation metal ion is (for example：Sodium, potassium)；CN-alginate
Represent the alginate molecule after CNBr activated hydroxyl groups；L-NH2Represent Lactose amine (monoamine
Terminated lactobionic lactone) molecule；L-NH2- alginate represents galactose grafting and changes
Alginate after property；Group representated by X is：
L-NH containing galactose group2By lactobionic acid (LA) in dimethyl sulfoxide solvent (DMSO) with second two
Amine (NH2CH2CH2NH2) it is total to hot preparation, reaction equation is as follows：
In microcapsule, galactosyl alginate is divalent metal calcium, the galactosyl alginic acid of barium or zinc
Hydrogel, galactosyl alginate solution is sodium salt or the potassium salt soln of galactosyl alginic acid.
It is micro- that above-mentioned microcapsule directly can prepare embedding type hydrogel by galactosyl alginate through chelating
Balloon borne body, then react formation polycation thin film formation microcapsule, the concrete preparation step of product with polycation
Suddenly it is：
1st, the synthesis of galactosyl alginic acid
1) lactobionic acid and ethylenediamine common hot preparation L-NH in dimethyl sulphoxide solution2.
2) prepare Sargassum acid solution, by the Sargassum acid solution being prepared into 1-10g/L soluble in water for alginic acid.Its
In, alginic acid molecular weight is 10KDa-10000kDa.
3) filter Sargassum acid solution and remove impurity, NaOH solution adjusts alginic acid pH value of solution to alkalescence.
4) weigh the CNBr solid of certain mass, with the dissolving of a small amount of organic solvent, Deca is in Sargassum acid solution
In and control ph constant.
5) solution after cleaning activation, until remove unreacted CNBr small molecule.
6) add the L-NH of certain mass in solution after cleaning2, reaction 2 day is stirred at room temperature.
7) clean reacted solution, until removing unreacted L-NH2.
8) solution after lyophilization cleaning, obtains galactosyl alginic acid.
2nd, galactosyl alginate/polycation microcapsule preparation
1) weigh galactosyl alginic acid solid, be dissolved in and in normal saline, form the solution that concentration is 5-50g/L.
2) prepare alginate embedding type hydrogel microsphere carrier, by orifice extrusion molding, electrostatic drop generation,
Emulsion process, rotating pan etc. are prepared into galactosyl alginate hydrogel microsphere.
3) by step 2) in embedding type gel micro-ball immersion said polycation solution in, embedding type gel micro-ball with
Said polycation solution volume ratio is 1:1-1:40, the response time be 1-60 minute, reaction temperature at 0-37 DEG C,
Now obtain galactosyl alginate/polycation microcapsule.
The compound method of said polycation solution is：
Shitosan is dissolved in the Acetic acid-sodium acetate buffer that pH is 5.0-7.0, and chitosan concentration is 0.1-15g/L；
α-polylysine is dissolved in 3-9g/L NaCl solution, and α-polylysine concentration is 0.01-10g/L；
Epsilon-polylysine is dissolved in 3-9g/L NaCl solution, and epsilon-polylysine concentration is 0.01-10g/L；
Poly arginine is dissolved in 3-9g/L NaCl solution, and poly arginine concentration is 0.01-10g/L；
Poly ornithine is dissolved in 3-9g/L NaCl solution, and poly ornithine concentration is 0.01-10g/L；
Polyhistidyl is dissolved in 3-9g/L NaCl solution, and polyhistidyl concentration is 0.01-10g/L.
Above-mentioned embedding type hydrogel microsphere carrier can be used for the embedding of living cells, and cell content is 105-2*107Individual
/ mL, activity keeps more than 80%.Described living cells behaviour or the in vitro hepatocyte of animal origin, stem cell,
Stem cell differentiation the cell with hepatocyte function, hepatic cell line cell, transdifferentiation there is hepatocyte work(
The cell of energy, endotheliocyte, Kupffer cell, stellate cells, fibroblast, medulla mesenchyma
In stem cell one or two or more kinds.
Beneficial effects of the present invention
1st, compared with traditional alginic acid graft modification, the reactive group of the inventive method is limited at alginic acid
The gel-forming property of alginic acid on oh group, will not be affected because carboxyl reaction site is occupied.
2nd, compared with traditional galactosyl alginate/polycation microcapsule preparation method, the method energy
Enough guarantees form microcapsule mechanical strength not because alginic acid graft modification is impacted.
3rd, the galactosyl alginate/polycation microcapsule of the method preparation, can be thrown by reactant
The change of material ratio adjusts the content of the internal galactose group of microcapsule, discloseder in alginate carboxyl
The method of upper grafting galactose is compared, and the grafting substitution value on hydroxyl reaches as high as 199%, and does not affect sea
Alginate jelly microsphere performance.
4th, the graft modification course of reaction mild condition of the inventive method, be conducive to hepatocellular activity keep and
Function improves.
5th, the microcapsule of the inventive method preparation keeps excellent biocompatibility, can guarantee that and moves as histiocyte
In plant, cell culture application process, microcapsule keeps integrity.
6th, the microcapsule function of immune isolation of product of the present invention is not affected by graft modification, for heteroplasm
During cell transplantation, function of immune isolation can be kept, that is, in microcapsule, the cell of embedding can not go out microcapsule, micro-
Antibody molecule outside capsule, complement molecule, immunocyte can not enter in microcapsule and kill cell, again simultaneously
Maintain the due permeability of microcapsule itself, i.e. the active component of cellular metabolism secretion, the battalion outside microcapsule
Foster small molecule can free in and out microcapsule.
Specific embodiment
The method preparing galactosyl alginate/polycation microcapsule is electrostatic drop generation (list of references：
In Vivo Culture of Encapsulated Endostatin-Secreting Chinese Hamster
Ovary Cells for Systemic Tumor Inhibition,Human Gene Therapy.2007,18:
474-481).
Embodiment 1
1) prepare Sargassum acid solution：The alginic acid Sargassum acid solution being prepared into 1g/L soluble in water, volume is
500mL.Wherein, alginic acid molecular weight is 350kDa.
2) filter 1) the solution removal of impurity, adjust alginic acid pH value of solution with mass concentration 240g/L NaOH solution
To 10.
3) by 0.06gCNBr solid with 0.5mL acetonitrile dissolving after be added dropwise to step 2) Sargassum acid solution in,
And control ph is constant.
4) cleaning step 3) solution after the activation prepared, until removing unreacted CNBr small molecule.
5) add 0.4g Lactose amine solid in solution after cleaning, reaction 2 day is stirred at room temperature.
6) cleaning step 5) reacted solution, until removing unreacted Lactose amine molecule.
7) by step 6) cleaning after solution lyophilization, obtain galactosyl alginic acid, grafting degree 30%.
8) by 7) be obtained galactosyl alginic acid be dissolved in the galactosyl being prepared into 15g/L in normal saline
Sodium alginate soln.
9) prepare α-polylysin solution：α-polylysine is dissolved in normal saline and is prepared into 5g/L's
α-polylysin solution.Wherein, α-polylysine molecule amount 30kDa.
10) prepare coagulation bath solution：Anhydrous calcium chloride is dissolved in the calcium chloride being prepared into 11g/L in deionized water
Solution.
11) prepare galactosyl calcium alginate hydrogel microsphere：By step 8) the galactosyl alginic acid prepared
Sodium solution forms jet off field in high-pressure electrostatic, and drop falls into step 10) the calcium chloride coagulation bath solution prepared
In, solidify 30 minutes, that is, obtain galactosyl calcium alginate hydrogel microsphere.
12) prepare galactosyl calcium alginate/α-polylysine microcapsule：By this galactosyl calcium alginate
Hydrogel microsphere immerses step 9) in α-polylysin solution of preparing, hydrogel microsphere is molten with polylysine
Liquid volume ratio is 1:10, react 10 minutes, brine, be prepared into galactosyl calcium alginate/α-
Polylysine microcapsule, basis of microscopic observation, 450 μm of microcapsule diameter, good sphericity, 10 microns of film thickness.
13) by step 12) be obtained galactosyl calcium alginate/α-polylysine microcapsule, by microcapsule：
IgG=1：20 volume ratios are put in IgG (γ-immunoglobulin) solution of FITC labelling, oscillation incubation 24
Hour, observe under laser confocal microscope, in microcapsule, there is no fluorescence signal, show that microcapsule keeps good
Function of immune isolation.
14) by step 12) the galactosyl calcium alginate/α-polylysine microcapsule and the agate ball that are obtained,
Normal saline is positioned in triangular flask jointly, at 37 degree, concussion ball milling 24 hours under the conditions of 170r/min,
The percentage of head rice of microcapsule is maintained at more than 95%, shows that microcapsule has good mechanical strength.
Comparative example 1
1) prepare Sargassum acid solution：Alginic acid is dissolved in the sodium alginate soln being prepared into 15g/L in normal saline,
Wherein, alginic acid molecular weight is 350kDa.
2) prepare α-polylysin solution：α-polylysine is dissolved in normal saline and is prepared into 5g/L's
α-polylysin solution.Wherein, α-polylysine molecule amount 30kDa.
3) prepare coagulation bath solution：Anhydrous calcium chloride be dissolved in be prepared in deionized water 11g/L calcium chloride molten
Liquid.
4) prepare calcium alginate hydrogel microsphere：By step 1) sodium alginate soln prepared is in high-pressure electrostatic
Form jet off field, drop falls into step 3) in the calcium chloride coagulation bath solution prepared, solidify 30 minutes,
Obtain calcium alginate hydrogel microsphere.
5) prepare calcium alginate/α-polylysine microcapsule：This calcium alginate hydrogel microsphere is immersed step
2), in the α-polylysin solution prepared, hydrogel microsphere and polylysin solution volume ratio are 1:10, instead
Answer 10 minutes, brine, be prepared into calcium alginate/α-polylysine microcapsule, see under microscope
Examine, 450 μm of microcapsule diameter, good sphericity, 10 microns of film thickness.
6) by step 5) be obtained calcium alginate/α-polylysine microcapsule, by microcapsule：IgG=1：20
Volume ratio is put in the IgG solution of FITC labelling, and oscillation incubation 24 hours, under laser confocal microscope
Observe, there is no fluorescence signal in microcapsule, show that microcapsule has good function of immune isolation.
7) by step 5) be obtained calcium alginate/α-polylysine microcapsule common with agate ball, normal saline
With being positioned in triangular flask, at 37 degree, under the conditions of 170r/min, shake ball milling 24 hours, microcapsule complete
Whole rate is maintained at more than 95%, shows that microcapsule has good mechanical strength.
Comparative example 2
1) prepare Sargassum acid solution：The alginic acid Sargassum acid solution being prepared into 1g/L soluble in water, volume is
500mL.Wherein, alginic acid molecular weight is 350kDa.
2) filter 1) the solution removal of impurity, add TEMED (N ＇, N ＇, N ＇ in the solution
- tetramethylethylenediamine), sodium chloride, EDC (1-ethyl- (dimethylaminopropyl)
Carbodiimide), sulfo-NHS (N-hydroxy-sulfosuccinimide), room temperature reaction 24h.
3) cleaning step 2) reacted solution.
4) by step 3) cleaning after solution lyophilization, obtain carboxyl grafting galactosyl alginic acid, connect
Branch rate 30%.
5) by 4) the carboxyl grafting galactosyl alginic acid that is obtained is dissolved in normal saline and is prepared into 15g/L's
Carboxyl is grafted galactosyl sodium alginate soln.
6) prepare α-polylysin solution：α-polylysine is dissolved in normal saline and is prepared into 5g/L's
α-polylysin solution.Wherein, α-polylysine molecule amount 30kDa.
7) prepare coagulation bath solution：Anhydrous calcium chloride be dissolved in be prepared in deionized water 11g/L calcium chloride molten
Liquid.
8) prepare carboxyl grafting galactosyl calcium alginate hydrogel microsphere：By step 5) carboxyl prepared connects
Branch galactosyl sodium alginate soln forms jet off field in high-pressure electrostatic, and drop falls into step 7) chlorine prepared
Change in calcium coagulation bath solution, solidify 30 minutes, that is, obtain carboxyl grafting galactosyl calcium alginate hydrogel micro-
Ball.
9) prepare carboxyl grafting galactosyl calcium alginate/α-polylysine microcapsule：By this carboxyl grafting half
In Lactose base calcium alginate hydrogel microsphere immersion α-polylysin solution, hydrogel microsphere is molten with polylysine
Liquid volume ratio is 1:10, react 10 minutes, brine, be prepared into carboxyl grafting galactosyl Sargassum
Sour calcium/α-polylysine microcapsule, basis of microscopic observation, 450 μm of microcapsule diameter, good sphericity, thickness
10 microns of degree.
10) by step 9) be obtained carboxyl grafting galactosyl calcium alginate/α-polylysine microcapsule, press
1：20 volume ratios are put in the IgG solution of FITC labelling, oscillation incubation 24 hours, and laser co-focusing is micro-
Microscopic observation, in microcapsule, fluorescence signal reaches the 10% of the outer fluorescence signal intensity of microcapsule, shows the immunity of microcapsule
Isolation performance is destroyed.
11) by step 9) carboxyl grafting galactosyl calcium alginate/α-polylysine microcapsule and agate of being obtained
Nao ball, normal saline are positioned in triangular flask jointly, at 37 degree, shake ball milling 24 under the conditions of 170r/min
Hour, the percentage of head rice of microcapsule is less than 80%, shows carboxyl grafting galactosyl alginic acid microcapsule mechanical strength
Weaker.
Embodiment 2
1) prepare galactosyl sodium alginate soln：Galactosyl alginic acid is dissolved in normal saline and being prepared into
The galactosyl sodium alginate soln of 15g/L, wherein, galactosyl alginic acid molecular weight is 350kDa.
2) prepare chitosan solution：Shitosan is dissolved in the Acetic acid-sodium acetate buffer that pH is 6.0, shitosan
Concentration is 5g/L.Wherein, chitosan molecule amount 30kDa, deacetylation 90%.
3) prepare coagulation bath solution：Anhydrous calcium chloride be dissolved in be prepared in deionized water 11g/L calcium chloride molten
Liquid.
4) galactosyl calcium alginate/chitosan microcapsules are prepared：High-pressure electrostatic method prepares galactosyl Sargassum
Sour calcium embedding type hydrogel microsphere, this embedding type hydrogel microsphere is immersed in chitosan solution, embedding type water
Gel micro-ball and chitosan solution volume ratio are 1:10, react 10 minutes, brine, be prepared into half
Lactose base calcium alginate/chitosan microcapsules.
5) basis of microscopic observation embedding type hydrogel microsphere and galactosyl calcium alginate/chitosan microcapsules,
The sphericity of the two is all good, film formation reaction 10 minutes, and film thickness is more than 10 microns.
Embodiment 3
1) prepare galactosyl sodium alginate soln：Galactosyl alginic acid is dissolved in normal saline and being prepared into
The galactosyl sodium alginate soln of 15g/L, aseptic filtration.Wherein, galactosyl alginic acid molecular weight is
350kDa.
2) prepare α-polylysin solution：α-polylysine is dissolved in normal saline and is prepared into 5g/L's
α-polylysin solution, aseptic filtration.Wherein, α-polylysine molecule amount 30kDa.
3) prepare coagulation bath solution：Anhydrous calcium chloride be dissolved in be prepared in deionized water 11g/L calcium chloride molten
Liquid, filtration sterilization.
4) preparation is wrapped up hepatocellular galactosyl calcium alginate/α-polylysine embedding type hydrogel microsphere and is carried
Body：Take 3mL galactosyl sodium alginate soln, add 6*10^6Individual Rat Primary Hepatocytes, after mix homogeneously,
It is embedded with the galactosyl calcium alginate embedded type hydrogel of primary rat hepatocyte using electrostatic drop generation preparation
Microsphere.
5) hepatocellular galactosyl calcium alginate/α-polylysine microcapsule is wrapped up in preparation：To be embedded with former
Galactosyl calcium alginate embedded type hydrogel microsphere for rat hepatocytes immerses step 2) α for preparing-poly-
In lysine solution, embedding type hydrogel microsphere and α-polylysin solution volume ratio are 1:10, react 10
Minute, brine, it is prepared into galactosyl calcium alginate/α-polylysine microcapsule.
6) primary rat hepatocyte in this microcapsule is cultivated and hepatocyte function is characterized, after one week,
Microcapsule keeps intact form, and sphericity is good；Microcapsule inner cell activity keeps the 75% of initial activity, liver
Cell albumin secretion amount is g/10^6 cell of 2.24 ± 0.68 μ, urea synthesiss amount for 388 ±
11 μ g/10^6 cells.
Comparative example 3
1) prepare sodium alginate soln：Alginic acid be dissolved in be prepared in normal saline 15g/L sodium alginate molten
Liquid, aseptic filtration.Wherein, alginic acid molecular weight is 350kDa.
2) prepare α-polylysin solution：α-polylysine is dissolved in normal saline and is prepared into 5g/L's
α-polylysin solution, aseptic filtration.Wherein, α-polylysine molecule amount 30kDa.
3) prepare coagulation bath solution, anhydrous calcium chloride be dissolved in be prepared in deionized water 11g/L calcium chloride molten
Liquid, filtration sterilization.
4) preparation parcel hepatocellular calcium alginate/α-polylysine embedding type hydrogel microsphere carrier：Take 3mL
Sodium alginate soln, adds 6*10^6 Rat Primary Hepatocytes, after mix homogeneously, using electrostatic drop generation
Preparation is embedded with the calcium alginate embedded type hydrogel microsphere of primary rat hepatocyte.
5) hepatocellular calcium alginate/α-polylysine microcapsule is wrapped up in preparation：Primary rat hepatic will be embedded with
The calcium alginate embedded type hydrogel microsphere of cell immerses step 2) in α-polylysin solution of preparing, bag
Burying type hydrogel microsphere with α-polylysin solution volume ratio is 1:10, react 10 minutes, physiology salt is washed
Wash, be prepared into calcium alginate/α-polylysine microcapsule.
6) primary rat hepatocyte in this microcapsule is cultivated and hepatocyte function is characterized, after one week,
Microcapsule keeps intact form, and sphericity is good；Microcapsule inner cell activity keeps the 44% of initial activity, liver
Cell albumin secretion amount is g/10^6 cell of 0.46 ± 0.05 μ, urea synthesiss amount for 231 ±
29 μ g/10^6 cells.
Claims (9)
1. a kind of galactosyl alginate hydrogel microsphere supported it is characterised in that：Galactose group is covalent
Modify on alginate molecules hydroxyl groups, galactose group is present in microsphere supported interior of alginate hydrogel
Portion.
2. according to the galactosyl alginate hydrogel described in right 1 microsphere supported it is characterised in that：Institute
State microsphere supported preparation process as follows：
1) under alkalescence condition, alginate oh group after CNBr activation under room temperature with contain amino group
Galactosyl derivant occur coupling reaction generate galactosyl alginate, wherein, alginate be grafted
(every 100 alginate monomers galactosyl containing amino group for the grafting derives the percent grafting of galactose group
The percent of thing molecule) it is 1%-199%；
2) by step 1) material prepared is configured to 5-50g/L galactosyl alginate solution, by sharp
It is micro- that hole extrusion molding, electrostatic drop generation, emulsion process or rotating pan are prepared into galactosyl alginate hydrogel
Ball；Wherein galactosyl alginate is the galactosyl alginic acid saline of divalent metal calcium, barium or zinc
Gel.
3. microsphere supported according to the galactosyl alginate hydrogel described in claim 1 or 2, its feature
It is：Described microsphere supported particle diameter is 100-1000 micron.
4. microsphere supported according to the galactosyl alginate hydrogel described in claim 1 or 2, its feature
It is：Described alginate hydrogel is microsphere supported to be continued to react with polycation, in microsphere surface complexation
Form polyelectrolyte structure of composite membrane, i.e. microcapsule, microcapsule membrane thickness is in 1-100 micron；Wherein, gather
Cation include following any one or two or more：Polyamino acid class (as polylysine, poly ornithine,
Poly arginine, polyhistidyl etc.), polyamine class (as polyethyleneimine, polymethylene guanidine, poly- N vinyl oneself
Lactams, Carboxy-propy-acrylamide copolymer, DEAE-dextran, amino-polyethyleneglycols etc.), shell gather
Sugar.
5. according to the embedding type hydrogel microcapsule described in claim 4 it is characterised in that：Described micro- glue
Capsule can immerse in organic metal chelating agent solution, the internal alginate of liquefaction microcapsule；Participate in liquefaction anti-
The organic metal chelating agent solution answered is the sodium citrate of 40-70mmol/L or the EDTA of 50-200mmol/L
Solution, microcapsule and organic metal chelating agent solution volume range are 1:1～1:40, react 1-60 minute,
Taking-up brine, now obtains the microcapsule of interior liquid core.
6. microsphere supported according to the galactosyl alginate hydrogel described in claim 1 or 2, its feature
It is：Described alginate includes one of calcium alginate, barium alginate or the alginic acid zinc that must contain
Or more than two kinds, do not contain or nonessential containing sodium alginate, one of potassium alginate or more than two kinds
Alginate, alginate mean molecule quantity guluronic acid in 10kDa-10000kDa, alginate
Monomer molar content is in 20-98%.
7. microsphere supported according to the galactosyl alginate hydrogel described in claim 1 or 2, its feature
It is：The galactose group used in described galactosyl alginate is：Any one contains amino base
The galactosyl derivant of group, including Lactose amine (monoamine terminated lactobionic
Lactone), or 1- amino -1- β-D- deoxy-galactose (1-amino-1-deoxy- β-D-galactose),
Obtain galactosyl alginic acid structural formula as follows：
Wherein, M representation metal ion sodium or potassium, X represents the galactosyl that any one contains amino group and spreads out
Biology, wherein, 50<n<50000.
Taking Lactose amine as a example, the structure such as following formula of X：
Taking 1- amino -1- β-D- deoxy-galactose as a example, the structure such as following formula of X：
8. the embedding type hydrogel microsphere carrier described in a kind of claim 1 or 2 application it is characterised in that：
The described microsphere supported embedding being mainly used in living cells.
9. according to the embedding type hydrogel microsphere carrier described in claim 8 application it is characterised in that：Institute
State the in vitro hepatocyte of living cells behaviour or animal origin, stem cell, what stem cell broke up has hepatocyte
The cell of function, hepatic cell line cell, the cell with hepatocyte function of transdifferentiation, endotheliocyte, liver
Kupffer's cells, stellate cells, fibroblast, in mesenchymal stem cells MSCs one or two or more kinds.
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| CN109480274A (en) * | 2018-11-19 | 2019-03-19 | 浙江工商大学 | Boc- histidine-sodium alginate with pH responsiveness independently fills the preparation method and applications of capsule of nano |
| CN111388445A (en) * | 2020-03-30 | 2020-07-10 | 杭州鹿扬科技有限公司 | Microcapsule material with prolonged release of active pharmaceutical ingredient and preparation method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109480274A (en) * | 2018-11-19 | 2019-03-19 | 浙江工商大学 | Boc- histidine-sodium alginate with pH responsiveness independently fills the preparation method and applications of capsule of nano |
| CN109480274B (en) * | 2018-11-19 | 2022-01-21 | 浙江工商大学 | Preparation method and application of Boc-histidine-sodium alginate self-contained nano microcapsule with pH responsiveness |
| CN111388445A (en) * | 2020-03-30 | 2020-07-10 | 杭州鹿扬科技有限公司 | Microcapsule material with prolonged release of active pharmaceutical ingredient and preparation method thereof |
| CN111533825A (en) * | 2020-06-17 | 2020-08-14 | 昆山京昆油田化学科技有限公司 | Glucosamine grafted sodium alginate derivative and preparation method and application thereof |
| CN111533825B (en) * | 2020-06-17 | 2022-03-01 | 昆山京昆油田化学科技有限公司 | Glucosamine grafted sodium alginate derivative and preparation method and application thereof |
| CN112956473A (en) * | 2021-03-10 | 2021-06-15 | 吉林省强参生物技术有限公司 | Artificial transparent belt and preparation method thereof |
| CN112956473B (en) * | 2021-03-10 | 2023-05-26 | 中科强参(吉林)科技有限公司 | Artificial transparent belt and preparation method thereof |
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