CN109880151A - A kind of preparation method and porous support materials of hydrogel porous microsphere - Google Patents
A kind of preparation method and porous support materials of hydrogel porous microsphere Download PDFInfo
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Abstract
The present invention provides a kind of preparation methods of hydrogel porous microsphere, comprising the following steps: A) hydrogel material, buffer are mixed with photoinitiator, obtain hydrogel solution；Oily material is mixed with surfactant, obtains oily solution；B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than outlet end internal diameter, and the outlet end internal diameter of the first container is less than the internal diameter of the second container；The hydrogel solution is injected in the first container, the oily solution is injected to the intersection of the first container Yu the second container, the oily solution and the hydrogel solution form Water-In-Oil drop under the action of Osima jacoti, Osima excavata；C the Water-In-Oil drop) is subjected to ultraviolet light, obtains hydrogel microsphere after crosslinking；D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.The application has obtained uniform and stable hydrogel porous microsphere using microflow control technique.
Description
Technical field
The present invention relates to the preparation methods and its porous branch of tissue engineering material more particularly to a kind of hydrogel porous microsphere
Frame material.
Background technique
For human body because of disease, wound or the non-renewable downright bad or missing for damaging caused local organization are that current medical treatment is led
One great difficult problem in domain, these tissues generally require to replace or transplant regeneration.However, autotransplantation or heteroplastic transplantation are controlled
There are significant problems for treatment, first is that graft is expensive, medical burden weight, patient suffering, while in migration process often
There are the repulsions of immuning tissue, cause operative failure.
Tissue engineering technique is intended to by developing biologic replacement materials, to maintain or improve the function of patient, is had important
Application prospect.With the development of tissue engineering technique, tissue engineering bracket is led in regeneration, reparation and stem-cell therapy
Domain plays an important role, and goes to repair substitution to carry the biomaterial scaffolds of cell and has damaged the tissue of disappearance increasingly
It is possible that.
Tradition tissue stenter to implant operation wound is big, therefore, in order to reduce operation risk, reduces patient suffering and raising
The mode of operation satisfaction, minimally invasive injection implantation receives more and more attention.Injectable gel and microballoon are that minimally invasive bracket is planted
The ideal material entered.Hydrogel material is because it is in massive aggregates shape, and the continuous tight of gel, nutrient protein molecule and metabolism are useless
Object can only be spread in inside, and mass exchange limited area, this causes obstruction to the growth and migration of cell part.Traditional
Microballoon is mostly smooth surface, and the cell that bracket is loaded into can only be migrated in rack surface.Since cell attachment is in microsphere surface,
Because the destruction of stream shear force amount, the cell survival rate of microsphere surface is not high, while such during injection is implanted into the patient
The space environment for lacking suitable cell growth inside microballoon, causing the microsphere support for carrying cell to be implanted into, the effect is unsatisfactory, in body
Inside disappear soon.
The method of microballoon production at present has emulsification-evaporation method, phase separation method, spray drying process and membrane emulsification.Cream
After change-solvent evaporation method is material dissolution, be placed in another immiscible solvent, then shaken by mechanical stirring or
Material is dispersed into emulsion droplet by the methods of ultrasonic emulsification, later by the modes such as photo-crosslinking, temperature crosslink or volatilization make microballoon at
Shape is precipitated.This method is easy to operate, is not necessarily to special installation, but balling-up state is uncontrollable, cannot form stabilization, uniform size it is micro-
Ball.
Summary of the invention
Present invention solves the technical problem that be to provide it is a kind of can be formed stablize, the hydrogel porous microsphere of uniform size
Preparation method.
In view of this, this application provides a kind of preparation methods of hydrogel porous microsphere, comprising the following steps:
A hydrogel material, buffer are mixed with photoinitiator), obtain hydrogel solution；Oily material and surface is living
Property agent mixing, obtain oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than in outlet end
Diameter, the outlet end internal diameter of the first container are less than the internal diameter of the second container；Hydrogel solution injection first is held
In device, the oily solution injects to the intersection of the first container Yu the second container, the oily solution with it is described
Hydrogel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C the Water-In-Oil drop) is subjected to ultraviolet light, obtains hydrogel microsphere after crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
Present invention also provides a kind of preparation methods of hydrogel porous microsphere, comprising the following steps:
A hydrogel material is mixed with buffer), obtains hydrogel solution；Oily material is mixed with surfactant,
Obtain oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than in outlet end
Diameter, the outlet end internal diameter of the first container are less than the internal diameter of the second container；Hydrogel solution injection first is held
In device, the oily solution injects to the intersection of the first container Yu the second container, the oily solution with it is described
Hydrogel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C) hydrogel microsphere will be obtained after Water-In-Oil drop crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
It preferably, further include pore creating material in the hydrogel solution.
Preferably, the first container is connect and is sealed with rectangular tube with the intersection of the second container.
Preferably, the hydrogel material be selected from the gelatin of methacryl group base group modification, collagen, fibroin,
One of chitosan, hyaluronic acid, alginate, fibrin, polylactic acid, polyaminoacid and polyethylene glycol are a variety of；Institute
It states oily material and is selected from one of perfluor oil, mineral oil, paraffin oil and isopropyl myristate or a variety of.
Preferably, step B) described in hydrogel solution flow velocity be 10~40 μ l/min, the flow velocity of the oily solution
For 60~100 μ l/min；The temperature of the freezing is -10~-50 DEG C, and the time of the freeze-drying is 36~72h；The ultraviolet light
The time of irradiation is 10~30s.
Preferably, the surface apertures of the hydrogel porous microsphere are 10~70 μm, the grain of the hydrogel porous microsphere
Diameter is 100~500 μm.
Present invention also provides a kind of porous support materials, porous micro- including hydrogel prepared by the preparation method
Ball and active material, cell, metal material or the bioceramic for being carried on hydrogel porous microsphere surface.
Preferably, the porous support materials further include the active material for being carried on hydrogel porous microsphere surface,
Cell, metal material or bioceramic.
Preferably, the active material is selected from drug, active factors or active peptides.
This application provides a kind of preparation methods of hydrogel porous microsphere, utilize two kinds of immiscible liquid phases-
Hydrogel solution and oily solution generate target grain size, uniform Water-In-Oil microlayer model using two kinds of containers, then pass through
Ultraviolet light Quick cross-linking or directly crosslinking generate hydrogel microsphere, and then by freeze-drying, it is more that rapid build has gone out hydrogel
Hole microballoon.
Hydrogel porous microsphere provided by the present application can load various cells, such as stem cell, moreover it is possible to load various active matters
Matter, such as various drugs, active factors, active peptides, moreover it is possible to load the particles such as various metals, bioceramic；The hydrogel is more
The office of load cells/active material microballoon can be achieved from several microns to several hundred microns in the partial size of hole microballoon after load cells
Portion's injection treatment, to realize minimally-invasive treatment；In addition, the cell of more gel porous microsphere surface loads can more preferably maintain cell
Survival rate is treated, it can be achieved that part release drug to reach local organization regeneration etc..
Detailed description of the invention
Fig. 1 is the apparatus structure schematic diagram that the present invention prepares hydrogel porous microsphere；
Fig. 2 is the specific pattern variation of hydrogel porous microsphere prepared by the embodiment of the present invention 1；
Fig. 3 is body sight, mechanical property and the degradation speed figure of hydrogel porous microsphere prepared by the embodiment of the present invention 1；
Fig. 4 is the growth situation that hydrogel porous microsphere prepared by the embodiment of the present invention 1 and cell co-culture；
Fig. 5 is that hydrogel porous microsphere prepared by the embodiment of the present invention 1 and cell co-culture the stem cell microballoon to be formed
Cell morphology and cell survival photo；
Fig. 6 be the embodiment of the present invention 1 prepare hydrogel porous microsphere and cell co-culture the stem cell microballoon to be formed with
Blank group injects the intracorporal effect contrast figure of rabbit；
Fig. 7 is the electron microscope photo scanning of hydrogel porous microsphere prepared by the embodiment of the present invention 1；
Fig. 8 is the electron microscope photo scanning of hydrogel porous microsphere prepared by the embodiment of the present invention 2；
Fig. 9 is the electron microscope photo scanning of hydrogel porous microsphere prepared by the embodiment of the present invention 4.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, still
It should be appreciated that these descriptions are only further explanation the features and advantages of the present invention, rather than to the claims in the present invention
Limitation.
In view of in the prior art, the balling-up state of hydrogel microsphere is uncontrollable, cannot form stable, uniform big minimicrosphere
Status, the application provide a kind of preparation method of hydrogel porous microsphere using Microfluidic droplet technology, the application preparation
Hydrogel porous microsphere surface has micropore, and microballoon is stable, uniform in size.Specifically, herein described hydrogel is porous micro-
The preparation method of ball specifically:
A hydrogel material, buffer are mixed with photoinitiator), obtain hydrogel solution；Oily material and surface is living
Property agent mixing, obtain oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than in outlet end
Diameter, the outlet end internal diameter of the first container are less than the internal diameter of the second container；Hydrogel solution injection first is held
In device, the oily solution injects to the intersection of the first container Yu the second container, the oily solution with it is described
Hydrogel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C the Water-In-Oil drop) is subjected to ultraviolet light, obtains hydrogel microsphere after crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
According to the difference of hydrogel material, present invention also provides a kind of preparation methods of hydrogel porous microsphere, including
Following steps:
A hydrogel material is mixed with buffer), obtains hydrogel solution；Oily material is mixed with surfactant,
Obtain oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than in outlet end
Diameter, the outlet end internal diameter of the first container are less than the internal diameter of the second container；Hydrogel solution injection first is held
In device, the oily solution injects to the intersection of the first container Yu the second container, the oily solution with it is described
Hydrogel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C) hydrogel microsphere will be obtained after Water-In-Oil drop crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
According to the present invention, first preparation raw material, i.e., hydrogel material, buffer is mixed with photoinitiator, obtain water-setting
Oily material is mixed with surfactant, obtains oily solution by sol solution；In the process, the hydrogel material is this
Hydrogel material known to the technical staff of field, specific gelatin, collagen, the silk for being selected from methacryl group base group modification
One of albumen, chitosan, hyaluronic acid, alginate, fibrin, polylactic acid, polyaminoacid and polyethylene glycol are more
Kind；The buffer is buffer well known to those skilled in the art, is chosen in particular from phosphate buffer；The photoinitiator choosing
From photoinitiator well known to those skilled in the art, in this application, optionally it is added according to the classification of hydrogel material
Photoinitiator, i.e., according to the difference of hydrogel material, different photoinitiators is may be selected in the photoinitiator, for certain special
Hydrogel material, such as: collagen, fibrin, the photoinitiator can also be added without, and not have the case where photoinitiator
Under, the cross-linking reaction of subsequent progress does not need ultraviolet light irradiation then；The oily material is specific to select as a kind of oil-based solvent
From one of perfluor oil, mineral oil, paraffin oil and isopropyl myristate or a variety of；The surfactant is selected from this field skill
Surfactant known to art personnel.In above-mentioned raw materials, the quality percentage of hydrogel material described in the hydrogel solution
Than for 5wt%, the mass percent of surfactant described in the oily solution is 5wt%, the hydrogel material and institute
The volume ratio for stating oily solution is 10:(1~2).
After raw material preparation is complete, the application will be then respectively placed in different containers in above two raw material, utilize this
The type of flow of the set-up mode of container and two kinds of raw materials prepares uniform and stable Water-In-Oil drop；Specifically: first is held
In the nesting to second container of device part, the arrival end internal diameter of the first container is greater than outlet end internal diameter, the first container
Outlet end internal diameter is less than the internal diameter of the second container；The hydrogel solution is injected in the first container, the oiliness is molten
Liquid injects the intersection of the first container Yu the second container, and the oily solution is cut with the hydrogel solution in flowing
Water-In-Oil drop is formed under the action of shear force.As shown in FIG. 1, FIG. 1 is the flowings of the set-up mode and two kinds of liquid of two kinds of containers
Schematic diagram；Pipe C is the first container in figure, and pipe D is second container, and solution A is hydrogel solution, and solution B is oily solution；Such as
Shown in figure, solution A enters from pipe C, and pipe C is inserted in pipe D, is entangled in the intersection of pipe C and pipe D with rectangular tube, and with sealing
Glue sealing, solution B enter from seal pipe, are entered in pipe D by seal, and the swiftly flowing solution B of pipe D is cut by flowing
Shear force shears the solution A slowly flowed out, to form Water-In-Oil drop.During above-mentioned formation Water-In-Oil drop, described
The internal diameter of one container and the second container is adjustable, and the diameter of hydrogel porous microsphere that can be as needed adjusts
The internal diameter of one container and second container.In this application, the first container and second container are selected from glass capillary, and
The arrival end of one glass capillary and the internal diameter of outlet end successively decrease, and the internal diameter of the second glass capillary is consistent, the second sweater glass
The internal diameter of pipe can be identical as the internal diameter of the first glass capillary arrival end.In the application, the flow velocity of the hydrogel solution is 10
~40 μ l/min, the flow velocity of the oily solution are 60~100 μ l/min；Above-mentioned flow velocity can guarantee that oily solution forms high speed
Osima jacoti, Osima excavata shears the hydrogel solution slowly flowed out, to form oily solution covering on hydrogel solution surface, finally
Form Water-In-Oil drop.
According to the present invention, above-mentioned Water-In-Oil drop is then subjected to ultraviolet light, hydrogel microsphere is obtained after crosslinking；
Under ultraviolet source irradiation, the Water-In-Oil drop Quick cross-linking quickly generates hydrogel microsphere.In this application, described ultraviolet
The time of light irradiation is 10~30s, and the Water-In-Oil drop can crosslink during this period.
Above-mentioned hydrogel microsphere is finally freezed and is lyophilized to get porous hydrogel microsphere is arrived by the application.Herein
In the process, the freezing and the freeze-drying are technical approach well known to those skilled in the art, herein without particularly superfluous
It states；The temperature of the freezing is -10~-50 DEG C, and the time of the freeze-drying is 36~72h；In a particular embodiment, the freezing
Temperature be -20 DEG C, time of the freeze-drying is 72h.
The application has obtained the porous microsphere of stabilization, uniform size, during the preparation process, hydrogel by microflow control technique
Concentration, the concentration of oily solution, the flow velocity of liquid and the time of freeze-drying of solution all can guarantee microballoon state is controllable, stablize and
It is uniform.The pore size of hydrogel porous microsphere and the diameter of microballoon of the application preparation can observe directly, in scanning electron microscope
Under, it can disposably observe hundreds of microballoons, and size uniformity.
Present invention also provides a kind of porous support materials comprising hydrogel porous microsphere described in above scheme.It is right
In this kind of porous support materials, active material, cell, metal, bioceramic etc. can be carried on institute according to actual needs
State hydrogel porous microsphere surface.Specifically, active material can be selected from drug, active factors or active peptides etc., the cell
It can be living cells, be also possible to stem cell, the metal is some standard biologic medical metal materials in the prior art.?
It is specifically to co-culture cell and above-mentioned hydrogel porous microsphere to form water-setting when hydrogel porous microsphere area load cell
Cellula adhesiae microballoon is applied to position to be repaired, to realize the quick reparation of local organization.
Preparation for a further understanding of the present invention, below with reference to embodiment to hydrogel porous microsphere provided by the invention
Method is described in detail, and protection scope of the present invention is not limited by the following examples.
Embodiment 1
1) solution A: 2ml phosphate buffer solution will be added in the gelatin of 100mg methacryl group modification, will add simultaneously
Enter 20mg photoinitiator, be placed in 60 DEG C of water-bath 45 minutes, interruption concussion shakes up, and sufficiently dissolves, becomes colorless to solution
It is stand-by after clear；
2) solution B: take perfluor oil 50ml that the surfactant span80 of mass ratio 1%, concussion dissolution, ultrasound removal is added
Bubble；
3) glass capillary C: taking internal diameter is 100 μm of glass capillary, and distal end wire drawing is cut in outlet under the microscope
The capillary that diameter is 50 μm；
4) glass capillary D: the glass capillary that internal diameter is 100 μm；
5) pipe D is inserted in the outlet end pipe C, is placed on glass slide, it is fixed with natural gum；It is closed with 21G syringe needle nesting
The interface port of pipe C and pipe D, while in the arrival end nesting 21G size syringe needle (as shown in Figure 1) of pipe C；
6) pipe C is interior phase, by injecting solution A inside syringe needle；
7) pipe D is foreign minister, is solution B by injection needle outflow；
8) flow velocity of the flow velocity of regulation pipe C internal solution A and pipe D internal solution B, solution A is 20 μ l/min, and solution B is
80 μ l/min, it is final to obtain by coated 300 μm uniform solution A drops of solution B；
9) the ultraviolet light of 365nm is used, the drop 20s of generation is irradiated, becomes solid-state-microspherical to drop, collect the micro- of acquisition
Ball, with 75% alcohol repeated flushing, finally wash with distilled water 3 times；
10) microballoon that will acquire is placed in distilled water, is placed in -20 DEG C of freezings；
11) freezing is placed on freeze-drying 72 hours in freeze dryer, rear to collect the microballoon obtained.
12) a small amount of sample, quality inspection under scanning electron microscope are taken, confirmation obtains product quality.
Fig. 2 is the specific pattern variation diagram of hydrogel porous microsphere manufactured in the present embodiment；Figure A is microfluidic device material object
Figure, figure B is the picture for being locally generated microballoon, and figure C is the character under the microscope of the hydrogel porous microsphere prepared, and figure D is system
Standby Water-In-Oil droplet size distribution figure, figure E are the state diagrams after Water-In-Oil drop is crosslinked in water, and figure F is Water-In-Oil drop
Size distribution figure after crosslinking；As can be seen from FIG. 2, it is porous micro- can to prepare uniform controllable hydrogel for method provided by the present application
Ball.
Fig. 3 is body sight, mechanical property and the degradation speed figure of hydrogel porous microsphere prepared by the embodiment of the present invention 1；Scheme A
Piece is taken into consideration for the body of hydrogel porous microsphere, and figure B is the mechanical curves figure of hydrogel porous microsphere, and figure C is that hydrogel is more
The degradation speed curve graph of hole microballoon.
The hydrogel porous microsphere of above-mentioned preparation and cell are co-cultured, Fig. 4 is the growing state figure of the cell after culture,
Figure A, B, C, D are represented as the extension cell number of time gradually increases；Figure E shows with the extension of time, cell survival rate
It is identical；Figure F shows with the extension of time, DNA content is more and more inside porous microsphere, the side light proliferation of cell.
Fig. 5 is the cell morphology and cell survival of the stem cell microballoon formed after hydrogel porous microsphere is co-cultured with cell
Photo, figure A is picture under the microscope containing stem cell microballoon, and figure B is the cell inside microballoon under laser confocal microscope
Three-dimensional distribution situation；Figure C, D are the cell survival pictures of local microballoon.
Different groups of cells is injected into the position of rabbit part bone defect by way of injection, to observe rabbit bone
Growing state, as shown in fig. 6, figure A is respectively blank group, blank microballoon group, carries stem cell microballoon group and load containing BMP is dry
Cell microsphere group, the effect for injecting the promotion bone uptake of the load stem cell microballoon group containing BMP as shown in Figure 6 contain better than injection
Stem cell microballoon group is carried, injection is better than blank microballoon group, direct injection blank containing stem cell microballoon group Bone formation effect is carried
Microballoon group Bone formation effect is better than blank group.
Embodiment 2
1) solution A: 2ml deionized water will be added in the gelatin of 80mg methacryl group modification, adds the sea 20mg
Mosanom, while 20mg photoinitiator is added, it being placed in 60 DEG C of water-bath 45 minutes, interruption concussion shakes up, sufficiently dissolves, to
Solution becomes colorless stand-by after clear；
2) solution B: take perfluor oil 50ml that the surfactant span80 of mass ratio 1%, concussion dissolution, ultrasound removal is added
Bubble；
3) glass capillary C: taking internal diameter is 100 μm of glass capillary, and distal end wire drawing is cut in outlet under the microscope
The capillary that diameter is 50 μm；
4) glass capillary D: the glass capillary that internal diameter is 100 μm；
5) pipe D is inserted in the outlet end pipe C, is placed on glass slide, it is fixed with natural gum, it is closed with 21G syringe needle nesting
The interface port of pipe C and pipe D, while in the arrival end nesting 21G size syringe needle of pipe C；
6) pipe C is interior phase, by injecting solution A inside syringe needle；
7) pipe D is foreign minister, is solution B by injection needle outflow；
8) flow velocity of the flow velocity of regulation pipe C internal solution A and pipe D internal solution B, solution A is 20 μ l/min, and solution B is
80 μ l/min, it is final to obtain by coated 300 μm uniform solution A drops of solution B；
9) the ultraviolet light of 365nm is used, the drop 20s of generation is irradiated, becomes solid-state-microspherical to drop, collect the micro- of acquisition
Ball, calcium chloride solution and 75% alcohol repeated flushing with 1%, finally wash with distilled water 3 times；
10) microballoon that will acquire is placed in distilled water, is placed in -20 DEG C of freezings；
11) after freezing, freeze-drying 72 hours in freeze dryer are placed in, it is rear to collect the microballoon obtained；
12) a small amount of sample, quality inspection under scanning electron microscope are taken, confirmation obtains product quality.
Embodiment 3
1) solution A: 2ml phosphate buffer solution and 20mg will be added in the gelatin of 100mg methacryl group modification
Nanometer hydroxyapatite, while 20mg photoinitiator is added, it is placed in 60 DEG C of water-bath 45 minutes, interruption concussion shakes up, and fills
Divide dissolution, it is stand-by after solution becomes colorless clear；
2) solution B: taking perfluor oil 50ml, and the surfactant span80 of mass ratio 1%, concussion dissolution is added, and ultrasound is gone
Bubble removing；
3) glass capillary C: taking internal diameter is 100 μm of glass capillary, and distal end wire drawing is cut in outlet under the microscope
The capillary that diameter is 50 μm；
4) glass capillary D: the glass capillary that internal diameter is 100 μm；
5) pipe D is inserted in the outlet end pipe C, is placed on glass slide, it is fixed with natural gum, it is closed with 21G syringe needle nesting
The interface port of pipe C and pipe D, while in the arrival end nesting 21G size syringe needle of pipe C；
6) pipe C is interior phase, by injecting solution A inside syringe needle；
7) pipe D is foreign minister, is solution B by injection needle outflow；
8) flow velocity of the flow velocity of regulation pipe C internal solution A and pipe D internal solution B, solution A is 20 μ l/min, and solution B is
80 μ l/min, it is final to obtain by coated 300 μm uniform solution A drops of solution B；
9) the ultraviolet light of 365nm is used, the drop 20s of generation is irradiated, becomes solid-state-microspherical to drop, collect the micro- of acquisition
Ball, with 75% alcohol repeated flushing, finally wash with distilled water 3 times；
10) microballoon that will acquire is placed in distilled water, is placed in -20 DEG C of freezings；
11) after freezing, freeze-drying 72 hours in freeze dryer are placed in, it is rear to collect the microballoon obtained；
12) a small amount of sample, quality inspection under scanning electron microscope are taken, confirmation obtains product quality.
Fig. 7 is the electron microscope photo scanning of porous microsphere prepared by embodiment 1,
Fig. 8 is the electron-microscope scanning figure and light microscopic figure of porous microsphere prepared by embodiment 2, and as seen from the figure, microsphere surface is thin
Pine is porous, which is 300 μm, and for surface apertures at 50 μm or so, right figure is the microballoon lyophilized preceding pattern under light microscopic.
Embodiment 4
Preparation method is same as Example 1, and difference is: photoinitiator is not added, does not carry out illumination, finally obtains
Porous microsphere as shown in Figure 9；The size of shown porous microsphere is 400 μm, and aperture is 30 μm.
The above description of the embodiment is only used to help understand the method for the present invention and its core ideas.It should be pointed out that pair
For those skilled in the art, without departing from the principle of the present invention, the present invention can also be carried out
Some improvements and modifications, these improvements and modifications also fall within the scope of protection of the claims of the present invention.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (10)
1. a kind of preparation method of hydrogel porous microsphere, comprising the following steps:
A hydrogel material, buffer are mixed with photoinitiator), obtain hydrogel solution；By oily material and surfactant
Mixing, obtains oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than outlet end internal diameter,
The outlet end internal diameter of the first container is less than the internal diameter of the second container；The hydrogel solution is injected into the first container
It is interior, the oily solution is injected to the intersection of the first container Yu the second container, the oily solution and the water
Gel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C the Water-In-Oil drop) is subjected to ultraviolet light, obtains hydrogel microsphere after crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
2. a kind of preparation method of hydrogel porous microsphere, comprising the following steps:
A hydrogel material is mixed with buffer), obtains hydrogel solution；Oily material is mixed with surfactant, is obtained
Oily solution；
B) by the nesting to second container of the first container part, the arrival end internal diameter of the first container is greater than outlet end internal diameter,
The outlet end internal diameter of the first container is less than the internal diameter of the second container；The hydrogel solution is injected into the first container
It is interior, the oily solution is injected to the intersection of the first container Yu the second container, the oily solution and the water
Gel solution forms Water-In-Oil drop under the action of Osima jacoti, Osima excavata；
C) hydrogel microsphere will be obtained after Water-In-Oil drop crosslinking；
D) hydrogel microsphere is freezed, is lyophilized, hydrogel porous microsphere is obtained.
3. preparation method according to claim 1 or 2, which is characterized in that further include pore creating material in the hydrogel solution.
4. preparation method according to claim 1 or 2, which is characterized in that the first container and the second container
Intersection is with rectangular tube connection and seals.
5. preparation method according to claim 1 or 2, which is characterized in that the hydrogel material is selected from methacryl
The gelatin of group base group modification, fibroin, chitosan, hyaluronic acid, alginate, fibrin, polylactic acid, gathers collagen
One of amino acid and polyethylene glycol are a variety of；It is different that the oily material is selected from perfluor oil, mineral oil, paraffin oil and myristic acid
One of propyl ester is a variety of.
6. preparation method according to claim 1 or 2, which is characterized in that step B) described in hydrogel solution flow velocity
For 10~40 μ l/min, the flow velocity of the oily solution is 60~100 μ l/min；The temperature of the freezing is -10~-50 DEG C,
The time of the freeze-drying is 36~72h；The time of the ultraviolet light is 10~30s.
7. preparation method according to claim 1 or 2, which is characterized in that the surface apertures of the hydrogel porous microsphere
It is 10~70 μm, the partial size of the hydrogel porous microsphere is 100~500 μm.
8. a kind of porous support materials, porous including hydrogel prepared by the described in any item preparation methods of claim 1~7
Microballoon and active material, cell, metal material or the bioceramic for being carried on hydrogel porous microsphere surface.
9. porous support materials according to claim 8, which is characterized in that the porous support materials further include being carried on
Active material, cell, metal material or the bioceramic on hydrogel porous microsphere surface.
10. porous support materials according to claim 8, which is characterized in that the active material be selected from drug, activity because
Son or active peptides.
Priority Applications (1)
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| CN110251725A (en) * | 2019-08-02 | 2019-09-20 | 科先医疗科技(苏州)有限公司 | A kind of sodium alginate micro ball packing material and preparation method thereof |
| CN111298196A (en) * | 2020-03-27 | 2020-06-19 | 常州药物研究所有限公司 | Polylactic acid porous microsphere, preparation method and application thereof |
| CN111389318A (en) * | 2020-04-17 | 2020-07-10 | 南京鼓楼医院 | Preparation method of interpenetrating network microcapsule with molecular sieve effect |
| CN111978588A (en) * | 2020-08-05 | 2020-11-24 | 广东省医疗器械研究所 | Macroporous hydrogel and preparation method and application thereof |
| CN112245658A (en) * | 2020-10-09 | 2021-01-22 | 北京大学 | Injectable crystal gel microsphere cell amplification carrier and preparation method thereof |
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| CN112618571A (en) * | 2020-09-30 | 2021-04-09 | 上海市伤骨科研究所 | Injectable hydrogel microspheres for treating orthopedic diseases and preparation method and application thereof |
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| CN111978588B (en) * | 2020-08-05 | 2022-05-03 | 广东省医疗器械研究所 | Macroporous hydrogel and preparation method and application thereof |
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| WO2023284107A1 (en) * | 2021-07-16 | 2023-01-19 | 江南大学 | Hydrogel microsphere for adsorbing growth factor in stem cell supernatant, and preparation thereof |
| CN113546157A (en) * | 2021-07-16 | 2021-10-26 | 江南大学 | Hydrogel microsphere for adsorbing growth factors in supernatant of stem cells and preparation method thereof |
| CN113546157B (en) * | 2021-07-16 | 2023-11-10 | 江南大学 | Hydrogel microsphere for adsorbing growth factors in stem cell supernatant and preparation method thereof |
| CN115926198A (en) * | 2021-09-23 | 2023-04-07 | 四川大学 | Injectable tissue regeneration type filler and preparation method thereof |
| CN113995891A (en) * | 2021-10-12 | 2022-02-01 | 重庆医科大学附属第一医院 | Self-renewing hydrated lubrication drug-loaded hydrogel microsphere and preparation method and application thereof |
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| CN114085394A (en) * | 2021-12-16 | 2022-02-25 | 西安德诺海思医疗科技有限公司 | Recombinant collagen two-phase gel and preparation method and application thereof |
| CN114085394B (en) * | 2021-12-16 | 2023-04-18 | 西安德诺海思医疗科技有限公司 | Recombinant collagen two-phase gel and preparation method and application thereof |
| CN114377212A (en) * | 2022-01-20 | 2022-04-22 | 上海交通大学医学院附属第九人民医院 | BMP-2 synergistic induction system for bone regeneration and construction method thereof |
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