CN104672484A - Cross-linked polysaccharide tissue engineering porous scaffold preparation method - Google Patents
Cross-linked polysaccharide tissue engineering porous scaffold preparation method Download PDFInfo
- Publication number
- CN104672484A CN104672484A CN201310609764.6A CN201310609764A CN104672484A CN 104672484 A CN104672484 A CN 104672484A CN 201310609764 A CN201310609764 A CN 201310609764A CN 104672484 A CN104672484 A CN 104672484A
- Authority
- CN
- China
- Prior art keywords
- chitosan
- sodium alginate
- cyclooctyne
- room temperature
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The present invention discloses a cross-linked polysaccharide tissue engineering porous scaffold preparation method, wherein chitosan, sodium alginate and other natural polysaccharides are adopted as matrix materials, and the chitosan and the sodium alginate are respectively treated by chemical modification, such that the chitosan and the sodium alginate can have active groups capable of being subjected to conjugated cross-linking so as to carry out automatic cross-linking forming under mild conditions. According to the present invention, the copper-free catalysis cross-linking is adopted, the use of the toxic metal catalyst is avoided, the cell compatibility of the porous scaffold can be significantly improved, and the use safety of the porous scaffold is increased; the freeze drying way is combined to obtain the continuous porous tissue engineering cell scaffold materials so as to easily achieve cell adhesion, migration and proliferation; and the preparation method of the invention has characteristics of simple process, low cost, and low cross-linking temperature, and is suitable for bone regeneration, cartilage regeneration, nerve regeneration, skin regeneration and other tissue engineering.
Description
Technical field
The invention belongs to the technology of preparing of biomaterial, particularly a kind of preparation method of crosslinked poly sugar tissue engineered porous scaffold.
Background technology
Tissue engineering bracket provides adapt circumstance for biological cells and tissues growth, degrades gradually and disappears, thus new space is supplied to tissue and cell, and make newly-generated tissue and organ possess the geometric shape identical with cytoskeleton along with the division of cell.The Main Function of support in the engineered tissue of structure or organ comprises: the adhesion that (1) is cell provides physical support, and cell is delivered to damaged part exactly; (2) for the propagation of cell, metabolism provide space; (3) provide specific Macrocosm and microcosm structure, guide tissue or the organ of cell construction specific function; (4) chemistry or mechanical signal is transmitted, the phenotype of regulating cell.The design of tissue engineering bracket and structure relate to the problem of three yardsticks, be respectively macrostructure (more than centimetre yardstick), i.e. profile, microscopic aperture, porosity and rack surface topological framework (micron order yardstick), the impact (nano-scale dimension) of rack surface attachment proteins and gene pairs cell.
Desirable scaffold for tissue engineering has following basic demand: the cell compatibility that (1) is good.Except the general requirement meeting biomaterial is as nontoxic, not teratogenesis, degraded product, to the effect of cytotoxic evil, does not cause inflammation outside reaction, also will be conducive to the adhesion of seed cell, propagation, the more important thing is and the genetic expression that energy activating cells is special maintain the phenotypic expression of cell.(2) good biological degradability.Timbering material should be able to be degraded after repair of damaged tissues, and degradation speed should match with the speed of tissue regeneration.(3) there is 3 D stereo vesicular structure.There is suitable aperture, high porosity and large specific surface area.(4) suitable physical strength.Support should possess the physical strength matched with institute repair tissue, for cambium provides support, can maintain original form of histoorgan.(5) in order to protect from infection, support also must be easy to sterilization and preserve.
The body material being generally used for tissue engineering bracket has two classes: natural macromolecular material and synthesized polymer material.Natural macromolecular material mainly comprises protein and saccharan, as collagen, gelatin, chitosan, chondroitin sulfate, Lalgine, Mierocrystalline cellulose etc.Conventional artificial macromolecular material comprises poly(lactic acid) (PLA), polycaprolactone (PCL), polyoxyethylene glycol (PEG), urethane (PU) etc.In general, natural polymer has excellent biocompatibility and degradation property, and synthesizes polymer and have good physical and mechanical properties, processing characteristics and chemical stability.These material controllabilitys are good, and practicality is high, have respective advantage and disadvantage, therefore to obtain in organizational project and regenerative medicine field as biomaterial and apply widely.
Natural polysaccharides molecule (as chitosan and Lalgine etc.) is the ideal material preparing tissue engineering bracket, they have good biocompatibility on the one hand, on the other hand its molecular chain has the multiple active function groups reacted, the modified methods such as grafting can be easily passed through and carry out chemically modified, effectively can improve the Performance and quality of material.At present, the method that crosslinked poly sugar porous support is conventional is chemical reagent crosslinking and uv cross-linking method.Chemical reagent crosslinking is by adding chemical cross-linking agent in the base if paraformaldehyde, glutaraldehyde and water-soluble carbodiimide etc. are as condensing agent, and its effect is as amino or carboxyl etc. reach crosslinked object through condensation reaction by the active group in substrate molecule.Uv cross-linking method be first by matrix through chemical modification, make it with unsaturated double-bond, then add initiator, caused the adduction of unsaturated double-bond by ultraviolet light irradiation, thus it is crosslinked that matrix is occurred.The material adopting these methods crosslinked is more stable in chemical structure, the processing requirement that material makes can be met, but owing to producing the residual of poisonous chemical reagent or initiator unavoidably, therefore material has stimulation in various degree to human body, and security can not be protected.Therefore, avoiding using poisonous small-molecule chemical linking agent, is the effective way improving support cell compatibility.But, in prior art, there is not the method adopting nontoxic chemical reagent or initiator crosslinked poly sugar porous support.
Summary of the invention
For current saccharan porous support many employings chemical cross-linking agent or metal catalyst, reagent is easily caused to remain and reduce cell compatibility and the safety in utilization of material.The object of the present invention is to provide the preparation method of a kind of easy and simple to handle and nontoxic crosslinked poly sugar tissue engineered porous scaffold.
Realizing technical solution of the present invention is: the preparation method of a kind of crosslinked poly sugar tissue engineered porous scaffold, and with chitosan and sodium alginate for base material, by forming without copper catalytic crosslinking, employing lyophilize means, specifically comprise the following steps:
Under step 1, room temperature by chitosan and I-hydroxybenzotriazole soluble in water;
Under step 2, room temperature, cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of tetrahydrofuran (THF)/water;
Add diisopropylethylamine, stirring reaction by after above two kinds of solution mixing under step 3, room temperature, dialysis freeze-drying obtains cyclooctyne chitosan;
Under step 4, room temperature by sodium alginate and water-soluble carbodiimide soluble in water, add 11-nitrine-3,6,9-tri-ether-1-amine, stirring reaction, dialysis freeze-drying obtain azide sodium alginate;
Above-mentioned steps is processed respectively chitosan and Lalgine by chemical modification, makes them respectively carry and can carry out the crosslinked active group of conjugation, make it automatically cross-linked shaping in a mild condition.
Step 5, prepare the phosphate buffer soln of certain density cyclooctyne chitosan and azide sodium alginate respectively, be mixed in proportion reaction under room temperature, last freeze-drying obtains porous support.
Wherein, in step 1, prepare cyclooctyne chitosan, described chitosan and I-hydroxybenzotriazole for etc. mass ratio, their mass concentration is 0.05 ~ 0.15%.
Prepare cyclooctyne chitosan in step 2, the tetrahydrofuran (THF) in described mixed solvent and the volume ratio of water are 1:3, and the mass concentration of cyclooctyne-3-oxyacetic acid is 0.5 ~ 1%.
Prepare cyclooctyne chitosan in step 3, described in add diisopropylethylamine mass concentration be 0.05 ~ 0.2%.
Prepare azide sodium alginate in step 4, the mass ratio of described sodium alginate and water-soluble carbodiimide is 1:1 ~ 1:3, and the mass ratio of sodium alginate and 11-nitrine-3,6,9-tri-ether-1-amine is 1:1 ~ 1:3.
Cross-linked porous support in step 5, the aqueous solution of described cyclooctyne chitosan and the reactant aqueous solution volume ratio of azide sodium alginate are 1:1, and wherein, the aqueous solution of cyclooctyne chitosan and the aqueous solution mass concentration of azide sodium alginate are 0.8% ~ 2.0%.
Amino and carboxyl is contained respectively in the chitosan adopted and the molecular structure of Lalgine, preparation method of the present invention is that amino by containing in the molecular structure of chitosan and Lalgine and carboxyl carry out modified-reaction, in like manner also can react containing natural macromolecular material that is amino and carboxyl.
Compared with prior art, its remarkable advantage is in the present invention: (1) this porous support adopts and is cross-linked without copper catalysis conjugation reaction.Respectively modification is carried out to chitosan and sodium alginate, makes them respectively carry can to carry out the active group reacted, can be automatically cross-linked shaping in a mild condition after making it mixing, without the need to adding poisonous metal catalyst, ensure that the security of material; (2) the present invention avoids employing toxicity copper catalyst, therefore there is not the residual of cupric ion in support, ensure that cell compatibility and the safety in utilization of support, be applicable to the organizational projects such as bone, cartilage, nerve, skin; (2) the technology of the present invention has the advantages such as crosslinking temperature is low, curing speed fast, operational safety, material cost are cheap, is applicable to commercially producing.
Raw material and reagent: chitosan, sodium alginate, I-hydroxybenzotriazole, diisopropylethylamine, water-soluble carbodiimide, 11-nitrine-3,6,9-tri-ether-1-amine, 3-(4,5-dimethylthiazole)-2,5-diphenyltetrazolium bromide bromine salt (MTT), is purchased from Sigma company; Sodium-chlor, Repone K, analytical pure, Shanghai reagent three factory; Sodium phosphate dibasic, potassium primary phosphate, analytical pure, Hangzhou chemical reagent company limited; Dimethyl sulfoxide (DMSO), sodium laurylsulfonate, analytical pure, Solution on Chemical Reagents in Shanghai factory.DMEM substratum, is purchased from Giboco company.
The preparation of phosphate buffer soln (PBS): take 8 grams, analytical pure sodium-chlor, 0.2 gram, Repone K, Sodium phosphate dibasic 2.9 grams, potassium primary phosphate 0.2 gram, be dissolved in 1000 ml distilled waters, regulates pH to be 7.4.
The structure observation of support: after 24 hours, by support metal spraying (Cressington 108 Auto), then observe internal microstructure in scanning electronic microscope (JSM-6330F, JEOL) through-50 DEG C of lyophilizes (FD-1A-50, Beijing rich doctor health).
The external weightlessness of support: accurately take weight support frame (W
0), then sample is immersed in 37
oin the PBS of C, weigh every for some time sample thief.By formula (W
0– W
t)/W
0the percentage loss of weight of porous support under × 100% calculating certain hour, W in formula
0the initial weight of porous support, W
tit is the dry weight after porous support hatches process in PBS.
The external water-absorbent of support: first weigh support (W
d), then sample is immersed in 37
oin the PBS of C, weigh every for some time sample thief.During test, from solution, take out support, suck the water of rack surface with filter paper, weigh (W at once
w), each sample parallel tests 5 times.The formula calculating support water absorbing properties is (W
w– W
d)/W
d.
The cell compatibility of support: first by support 75% alcohol immersion sterilization in 2 hours, then with phosphate buffer soln rinsing removing ethanol repeatedly; Inoculating quantity is in the bracket 5 × 10
6calf knee cartilage cell, be put in 37
ohatch under C, add the phosphate buffer soln of 0.5wt%MTT after 7 days, be placed in 37
oadd dimethyl sulfoxide (DMSO) after hatching 4 hours under C, after shaken well, adopt microplate reader (Biorad, Model 550) to measure the absorbancy of purple material in 570 nanometers.
The present invention adopts ANOVA analysis of variance, significant difference value
pbe set to≤0.05.
Below in conjunction with accompanying drawing, further detailed description is done to the present invention.
Accompanying drawing explanation
Fig. 1 is the schematic diagram that the present invention adopts without copper catalytic crosslinking saccharan tissue engineered porous scaffold.
Fig. 2 is the scanning electron microscope (SEM) photograph of crosslinked poly of the present invention sugar tissue engineered porous scaffold internal structure.
Fig. 3 is the time dependent water-intake rate of the embodiment of the present invention 1 porous support.
Fig. 4 is the time dependent rate of weight loss of the embodiment of the present invention 1 porous support.
Fig. 5 is the embodiment of the present invention 1 porous support and the traditional copper catalytic support absorbancy that the 7th day records after repopulating cell.
Embodiment
Cross-linking method of the present invention be with chitosan and sodium alginate for base material, by forming without copper catalytic crosslinking, specifically comprise the following steps:
Under step 1, room temperature by chitosan and I-hydroxybenzotriazole soluble in water;
Under step 2, room temperature, cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of tetrahydrofuran (THF)/water;
Add diisopropylethylamine, stirring reaction by after above two kinds of solution mixing under step 3, room temperature, dialysis freeze-drying obtains cyclooctyne chitosan;
Under step 4, room temperature by sodium alginate and water-soluble carbodiimide soluble in water, add 11-nitrine-3,6,9-tri-ether-1-amine, stirring reaction, dialysis freeze-drying obtain azide sodium alginate;
Step 5, prepare the phosphate buffer soln of certain density cyclooctyne chitosan and azide sodium alginate respectively, be mixed in proportion reaction under room temperature, last freeze-drying obtains porous support.
embodiment 1:
Concrete operation step is:
(1) under room temperature, 0.05 gram of chitosan and 0.05 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 0.5 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.1 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtain cyclooctyne chitosan, molecular structure is shown in Fig. 1;
(4) under room temperature, 0.2 gram of sodium alginate and 0.2 gram of water-soluble carbodiimide are dissolved in 100 ml waters, then add 0.2 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, freeze-drying after 3 days of dialysing, obtain azide sodium alginate, molecular structure is shown in Fig. 1;
(5) preparing mass concentration is respectively the cyclooctyne chitosan of 2% (w/v) and the phosphate buffer soln of azide sodium alginate, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.Its microscopic appearance is shown in Fig. 2, and figure is the stereoscan photograph of chitin-sodium alginate internal stent structure.This support is vesicular structure, and mean pore size is 150 microns and mutually runs through, and is conducive to adhesion and the migration of cell.
Measure the water-intake rate of gained porous support, time dependent water-intake rate is shown in Fig. 3.As can be seen from the figure the water-retaining capacity of this support is strong, and Absorbable rod is equivalent to the liquid of own wt 24 ~ 32 times, illustrates that support can be cells with nutrient preferably.
Measure the external weightlessness of gained porous support, time dependent rate of weight loss is shown in Fig. 4.As can be seen from the figure cultivate after 14 days, the rate of weight loss of this support is about 13%, and this wt-lossing rates can meet the requirement of the tissue growth such as bone, cartilage preferably.
Measure the cell compatibility of gained porous support, the absorbancy of the 7th day is shown in Fig. 5, as can be seen from the figure the absorbancy without copper catalytic crosslinking support is significantly higher than employing copper catalytic crosslinking support, absorbancy directly characterizes consistency, it is good without the cell compatibility of copper catalytic crosslinking support that absorbancy height illustrates, thus be beneficial to the growth of cell and promote the regeneration of tissue.
embodiment 2:
Concrete operation step is:
(1) under room temperature, 0.08 gram of chitosan and 0.08 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 0.6 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.15 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtains cyclooctyne chitosan;
(4) 0.2 gram of sodium alginate and 0.5 gram of water-soluble carbodiimide are dissolved in 100 ml waters under room temperature, then add 0.35 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, and freeze-drying after 3 days of dialysing, obtains azide sodium alginate;
(5) preparing mass concentration is respectively the cyclooctyne chitosan of 1.8% (w/v) and the phosphate buffer soln of azide sodium alginate, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.
embodiment 3:
Concrete operation step is:
(1) under room temperature, 0.1 gram of chitosan and 0.1 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 0.7 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.2 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtains cyclooctyne chitosan;
(4) 0.2 gram of sodium alginate and 0.6 gram of water-soluble carbodiimide are dissolved in 100 ml waters under room temperature, then add 0.6 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, and freeze-drying after 3 days of dialysing, obtains azide sodium alginate;
(5) preparing mass concentration is respectively the cyclooctyne chitosan of 1.5% (w/v) and the phosphate buffer soln of azide sodium alginate, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.
embodiment 4:
Concrete operation step is:
(1) under room temperature, 0.12 gram of chitosan and 0.12 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 0.8 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.25 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtains cyclooctyne chitosan;
(4) 0.2 gram of sodium alginate and 0.4 gram of water-soluble carbodiimide are dissolved in 100 ml waters under room temperature, then add 0.5 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, and freeze-drying after 3 days of dialysing, obtains azide sodium alginate;
(5) preparing mass concentration is respectively the cyclooctyne chitosan of 1.2% (w/v) and the phosphate buffer soln of azide sodium alginate, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.
embodiment 5:
Concrete operation step is:
(1) under room temperature, 0.12 gram of chitosan and 0.12 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 0.9 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.35 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtains cyclooctyne chitosan;
(4) 0.2 gram of sodium alginate and 0.3 gram of water-soluble carbodiimide are dissolved in 100 ml waters under room temperature, then add 0.3 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, and freeze-drying after 3 days of dialysing, obtains azide sodium alginate;
(5) aglycon amount concentration is the cyclooctyne chitosan of 1% (w/v) and the phosphate buffer soln of azide sodium alginate respectively, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.
embodiment 6:
Concrete operation step is:
(1) under room temperature, 0.15 gram of chitosan and 0.15 gram of I-hydroxybenzotriazole are dissolved in 100 ml waters;
(2) under room temperature, 1 gram of cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of 100 milliliters of tetrahydrofuran (THF)/water (volume ratio 1:3);
(3) under room temperature, above two kinds of solution are fully mixed, add 0.4 gram of diisopropylethylamine, 0
ostirring reaction 12 hours under C condition, freeze-drying after 3 days of dialysing, obtains cyclooctyne chitosan;
(4) 0.2 gram of sodium alginate and 0.35 gram of water-soluble carbodiimide are dissolved in 100 ml waters under room temperature, then add 0.4 milliliter of 11-nitrine-3,6,9-tri-ether-1-amine, stirring at room temperature reacts 24 hours, and freeze-drying after 3 days of dialysing, obtains azide sodium alginate;
(5) preparing mass concentration is respectively the cyclooctyne chitosan of 0.8% (w/v) and the phosphate buffer soln of azide sodium alginate, fully mixes, 37 under room temperature by equal-volume
oplace 6 hours under C, finally-50
ounder C, lyophilize obtains porous support in 24 hours.
Above embodiment covers the representational experimental data of most.
Claims (7)
1. a preparation method for crosslinked poly sugar tissue engineered porous scaffold, is characterized in that, with chitosan and sodium alginate for body material, by shaping without copper catalytic crosslinking, employing Freeze Drying Technique obtains vesicular structure, specifically comprises the following steps:
Under step 1, room temperature by chitosan and I-hydroxybenzotriazole soluble in water;
Under step 2, room temperature, cyclooctyne-3-oxyacetic acid is dissolved in the mixed solvent of tetrahydrofuran (THF)/water;
Add diisopropylethylamine, stirring reaction by after above two kinds of solution mixing under step 3, room temperature, dialysis freeze-drying obtains cyclooctyne chitosan;
Under step 4, room temperature by sodium alginate and water-soluble carbodiimide soluble in water, add 11-nitrine-3,6,9-tri-ether-1-amine, stirring reaction, dialysis freeze-drying obtain azide sodium alginate;
Step 5, prepare the phosphate buffer soln of certain density cyclooctyne chitosan and azide sodium alginate respectively, be mixed in proportion reaction under room temperature, last freeze-drying obtains porous support.
2. the preparation method of porous support according to claim 1, is characterized in that, contains amino and carboxyl respectively in the chitosan of employing and the molecular structure of Lalgine.
3. the preparation method of porous support according to claim 1, is characterized in that, prepares cyclooctyne chitosan in step 1, described chitosan and I-hydroxybenzotriazole for etc. mass ratio, their mass concentration is 0.05 ~ 0.15%.
4. the preparation method of porous support according to claim 1, is characterized in that, prepares cyclooctyne chitosan in step 2, and the tetrahydrofuran (THF) in described mixed solvent and the volume ratio of water are 1:3, and the mass concentration of cyclooctyne-3-oxyacetic acid is 0.5 ~ 1%.
5. the preparation method of porous support according to claim 1, is characterized in that, prepares cyclooctyne chitosan in step 3, described in add diisopropylethylamine mass concentration be 0.05 ~ 0.2%.
6. the preparation method of porous support according to claim 1, it is characterized in that, azide sodium alginate is prepared in step 4, the mass ratio of described sodium alginate and water-soluble carbodiimide is 1:1 ~ 1:3, sodium alginate and 11-nitrine-3, the mass ratio of 6,9-tri-ether-1-amine is 1:1 ~ 1:3.
7. the preparation method of porous support according to claim 1, it is characterized in that, cross-linked porous support in step 5, the aqueous solution of described cyclooctyne chitosan and the reactant aqueous solution volume ratio of azide sodium alginate are 1:1, wherein, the aqueous solution of cyclooctyne chitosan and the aqueous solution mass concentration of azide sodium alginate are 0.8% ~ 2.0%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310609764.6A CN104672484A (en) | 2013-11-27 | 2013-11-27 | Cross-linked polysaccharide tissue engineering porous scaffold preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310609764.6A CN104672484A (en) | 2013-11-27 | 2013-11-27 | Cross-linked polysaccharide tissue engineering porous scaffold preparation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104672484A true CN104672484A (en) | 2015-06-03 |
Family
ID=53308103
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310609764.6A Pending CN104672484A (en) | 2013-11-27 | 2013-11-27 | Cross-linked polysaccharide tissue engineering porous scaffold preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104672484A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106693067A (en) * | 2016-07-15 | 2017-05-24 | 温州生物材料与工程研究所 | Preparation of self-healing and template-free porous scaffold |
| CN107137752A (en) * | 2016-03-01 | 2017-09-08 | 南京理工大学 | A kind of preparation method of luffa resist blocking and that Wound dressing |
| CN114929754A (en) * | 2019-12-18 | 2022-08-19 | 持田制药株式会社 | Novel cross-linked alginic acid |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080242850A1 (en) * | 2006-03-28 | 2008-10-02 | Korea Atomic Energy Research Institute | Method of producing chitosan scaffold having high tensile strength and chitosan scaffold produced using the method |
| CN102940903A (en) * | 2012-11-20 | 2013-02-27 | 南京理工大学 | Method for preparing medical dressing of polysaccharide cavernous body |
-
2013
- 2013-11-27 CN CN201310609764.6A patent/CN104672484A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080242850A1 (en) * | 2006-03-28 | 2008-10-02 | Korea Atomic Energy Research Institute | Method of producing chitosan scaffold having high tensile strength and chitosan scaffold produced using the method |
| CN102940903A (en) * | 2012-11-20 | 2013-02-27 | 南京理工大学 | Method for preparing medical dressing of polysaccharide cavernous body |
Non-Patent Citations (2)
| Title |
|---|
| AKIRA TAKAHASHI等: ""In Situ Cross-Linkable Hydrogel of Hyaluronan Produced via Copper-Free Click Chemistry"", 《BIOMACROMOLECULES》 * |
| XIAOHONG HU等: ""Biological hydrogel synthesized from hyaluronic acid, gelatin and chondroitin sulfate by click chemistry"", 《ACTA BIOMATERIALIA》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107137752A (en) * | 2016-03-01 | 2017-09-08 | 南京理工大学 | A kind of preparation method of luffa resist blocking and that Wound dressing |
| CN106693067A (en) * | 2016-07-15 | 2017-05-24 | 温州生物材料与工程研究所 | Preparation of self-healing and template-free porous scaffold |
| CN114929754A (en) * | 2019-12-18 | 2022-08-19 | 持田制药株式会社 | Novel cross-linked alginic acid |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11511016B2 (en) | Method for preparing porous scaffold for tissue engineering, cell culture and cell delivery | |
| Pourjavadi et al. | Injectable chitosan/κ-carrageenan hydrogel designed with au nanoparticles: A conductive scaffold for tissue engineering demands | |
| US9555164B2 (en) | Method for preparing porous scaffold for tissue engineering | |
| CN104892962A (en) | Preparation method and application of sulfhydryl/disulfide bond controllable self-crosslinked hyaluronic acid hydrogel | |
| CN112587726B (en) | Composite hydrogel stent and preparation method and application thereof | |
| Weng et al. | Self‐crosslinkable hydrogels composed of partially oxidized hyaluronan and gelatin: In vitro and in vivo responses | |
| Lv et al. | Progress in preparation and properties of chitosan-based hydrogels | |
| CN102604149B (en) | Three-dimensional chitosan hydrogel and preparation method thereof | |
| Kil’deeva et al. | Biodegradablescaffolds based on chitosan: Preparation, properties, and use for the cultivation of animal cells | |
| CN104672484A (en) | Cross-linked polysaccharide tissue engineering porous scaffold preparation method | |
| CN105797211A (en) | Preparation method of hydrogel, osteoblast containing hydrogel and preparation method of osteoblast containing hydrogel | |
| CN108478874B (en) | Preparation method of hydroxyethyl chitosan nano composite bone scaffold material | |
| CN109852574B (en) | Cell scaffold and preparation method thereof | |
| CA2442868A1 (en) | Chitosan and hydroxy carboxylic acid based porous and non-porous matrices | |
| Jose et al. | Recent advances in the design and development of bioink formulations for various biomedical applications | |
| TW201016722A (en) | Porous composite biomaterials and production method of the same | |
| Kesharwani et al. | Tissue regeneration properties of hydrogels derived from biological macromolecules: A review | |
| WO2023012333A2 (en) | Preparation of composite gels, polymer scaffolds, aggregates and films comprising soluble cross-linked chitosan & uses thereof | |
| US20050191356A1 (en) | Porous and non-porous matrices based on chitosan and hydroxy carboxylic acids | |
| JP2006070055A (en) | Manufacturing method of polymer composite and polymer composite obtained by manufacturing method, and culturing method of animal cell using polymer composite |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150603 |