CN106693067A - Preparation of self-healing and template-free porous scaffold - Google Patents
Preparation of self-healing and template-free porous scaffold Download PDFInfo
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- CN106693067A CN106693067A CN201610559445.2A CN201610559445A CN106693067A CN 106693067 A CN106693067 A CN 106693067A CN 201610559445 A CN201610559445 A CN 201610559445A CN 106693067 A CN106693067 A CN 106693067A
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- Prior art keywords
- porous support
- self
- healing
- preparation
- sodium alginate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/222—Gelatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
- C08J3/246—Intercrosslinking of at least two polymers
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
- C08L89/04—Products derived from waste materials, e.g. horn, hoof or hair
- C08L89/06—Products derived from waste materials, e.g. horn, hoof or hair derived from leather or skin, e.g. gelatin
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/04—Alginic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
- C08J2389/04—Products derived from waste materials, e.g. horn, hoof or hair
- C08J2389/06—Products derived from waste materials, e.g. horn, hoof or hair derived from leather or skin
Abstract
The invention provides a self-healing porous scaffold material and a preparing method thereof. The method specifically comprises the following steps: performing aldehyde modification on sodium alginate to obtain oxidized sodium alginate (OSA), dissolving OSA and gelatin respectively in a PBS (poly butylenes succinate) solution, taking a certain amount of the two sets of solutions for mixture, adding a certain amount of PBS solution containing adopyl diacidhydrazine, performing high-speed shearing and mixing for 10 s to 3 min, and crosslinking at 37 to 60 DEG C for 30 s to 3 min, to obtain the sodium alginate-gelatin porous scaffold material. The method has simple preparing processes, controllable conditions without a porous template; the porous scaffold biological material obtained through preparation has a self-healing performance and characteristics such as good biocompatibility, mechanical property, and macropore performances at the same time. the scaffold material can be widely applied to biomedical fields such as biological scaffolds, cell culturing, drug control, and tissue engineering.
Description
Technical field
The present invention relates to a kind of self-healing multi-porous tissue engineering supporting material, specifically provide a kind of mixed by high speed shear
It is legal to prepare the porous support materials based on aldehyde radical-amino crosslinking.
Background technology
American scientist Joseph P. Vacanti and Robert professors Langer propose tissue work first within 1993
Journey concept, i.e., " grow substitute living, be used to repair, maintain and improve people using organizational engineering and life science principle
The function of body tissue and organ ".One of three elements that timbering material is constituted as organizational project, its research is to field of tissue engineering technology
Research have huge meaning.
Biologic bracket material provides the three dimensions survived as cell, it is necessary to meet following condition：1. have
Beneficial to cell adherence, propagation, differentiation and growth, and binding site can be provided for cell, induce biological respinse, inducing cell is just
It is frequently grown；2. as the carrier of nutriment, oxygen and bioactivator, can store, transport these materials, excretion metabolism gives up
Thing, and constantly degraded in tissue growth forming process, be absorbed by organisms and excreted with or through the circulatory system；3. have
Certain mechanical property, certain pattern, structure snd size, guiding tissue is grown by predetermined form.Conditions above is just to support
Material has porous pattern to define, and the method for preparing porous support is varied.So far, preparation method mainly has fiber
Mull technique, particle pore method, phase separation/freeze-drying, foaming（Physical blowing/chemical blowing）, sintering microballoon method, melting
The method of forming, speed forming method etc..But above method preparation technology is relatively complicated, and add pore-foaming agent or using having
The toxic material such as machine solvent, exerts a certain influence to biocompatibility.
Sodium alginate is by α-L- mannuronic acid (M units）With β-D- guluronic acid (G units）By β -1,4- sugar
The Natural linear polysaccharide that glycosidic bond is formed by connecting, contains a carboxyl in each of which uronic acid unit, therefore in neutral or alkaline bar
The property of polyanion electrolyte is presented under part, and sodium alginate is foodstuffs without toxicity, and U.S.'s medicine was just incorporated into early in 1938
Allusion quotation.Sodium alginate can form gel micro-ball, realize medicine sustained and controlled release by medicine or active material parcel wherein；With blood
Compatibility and the characteristics of the removing that can degrade in vivo, are the good carriers of target administration；With certain mechanical property, can keep
Certain pattern, size and structure, the research as tissue engineering bracket material is also rather popular；On medical imaging and detection
Also the application of sodium alginate is engendered.Gelatin is derivative, its chemical composition and collagen phase obtained from collagenous portion hydrolysis
Seemingly, there is homology with collagen, maintains the triple helix structure of collagen, contain similar arginine-glycine-aspartic acid
（RGD）Sequence, with excellent hydrophily and biocompatibility.Gelatin based composites are used as tissue engineering bracket material and letter
Number molecular vehicle is one of study hotspot of current biomaterial.Porous network supporting structure is formed after gelatin is crosslinked, but
One-component gelatin fragile structure, mechanical property are poor, it is impossible to meet the application of bioengineered tissue, need to be with other biological macromolecule
It is used in combination.
The content of the invention
It is an object of the invention to provide a kind of preparation method of self-healing multi-porous tissue engineering supporting material, the method without
Template is needed, by high speed shear hybrid technology, porous support is obtained, for organizational project Regeneration and Repair.Prepared by the present invention many
Hole timbering material has good biocompatibility and an excellent mechanical properties, preparation process is simple, preparation process green non-pollution, into
This is low.Porous support materials prepared by the present invention have self-healing performance, are capable of achieving the repairing and treating to organizing.
To achieve the above object, the present invention can take following technical proposals：
A kind of preparation technology of self-healing multi-porous tissue engineering supporting material, the tissue engineering bracket material with choose gelatin and
Sodium alginate is raw material, but is not limited only to this, and other are with amino and carboxyl or can be by amination and carboxylated and can aldehyde radical
Bioabsorbable polymer material, such as protein, polypeptide, shitosan, hyaluronic acid.The preparation technology is comprised the following steps：
1）Aldehyde grouping modified treatment is carried out to sodium alginate, the aldehyde grouping modified reagent is sodium metaperiodate；2）Will be certain density bright
Sol solution is mixed with aldehyde radical sodium alginate soln with certain proportion, adds a certain amount of adipic dihydrazide, cut at a high speed
Cut mixing；3）After certain hour, it is placed in and solidifies crosslinking in uniform temperature atmosphere, obtains self-healing tissue engineered porous scaffold material
Material.
Porous support materials biological property to preparing is characterized, and shows good biocompatibility, with cell adhesion,
Cell can adhere to propagation on timbering material.
The present invention obtain beneficial effect be：The porous support materials that the present invention is prepared have self-healing performance, raw
Thing compatibility is good, macroporosity, with certain mechanical strength, is capable of achieving tissue replacement therapy.The invention provides a kind of simple
Convenient porous support technology of preparing, preparation process is nontoxic, pollution-free, environmental protection.
Brief description of the drawings
Below in conjunction with the accompanying drawings and implementation method the present invention is further detailed explanation：
Fig. 1 is the aldehyde grouping modified reaction schematic diagram of sodium alginate
Fig. 2 is gelatin and aldehyde radical sodium alginate heat cross-linking reaction schematic diagram
Fig. 3 is tissue engineering bracket material surface and section surface sweeping electron microscope in patent of the present invention
Fig. 4 is the schematic diagram of tissue engineering bracket material self-healing function in patent of the present invention
Fig. 5 is the schematic diagram of tissue engineering bracket material injectable in patent of the present invention
Fig. 6 is growth fluorescence microscope picture of the cell in the porous support materials
Specific embodiment
The invention is further illustrated by the following examples, but this big bright is not limited only to this.
Embodiment 1：The preparation of aldehyde radical sodium alginate
First, take sodium alginate powder 5g to be dissolved in 25mL ethanol solutions, take sodium metaperiodate 4.32g and be dissolved in 25mL deionized waters
In；Secondly, complete sodium periodate solution will be dissolved to be added in sodium alginate soln, 6h is reacted in lucifuge stirring at room temperature；The
Three, the ethylene glycol 1.137mL terminating reactions with sodium metaperiodate equimolar amounts are added, stir 30min；4th, what is be stirred vigorously
Under the conditions of, the reaction solution is added in 200mL absolute ethyl alcohols, reactant Precipitation, suction filtration；Use a certain amount of deionization
Water dissolves, then precipitated with absolute ethyl alcohol；Such cyclic washing three times, obtains white aldehyde radical sodium metaperiodate product；5th, will produce
Thing is poured into culture dish, and 24h is dried in 50 DEG C of vacuum drying chambers, obtains dry state aldehyde radical sodium alginate, is kept in dark place.
Embodiment 2：The preparation of aldehyde radical sodium alginate
First, take sodium alginate powder 5g to be dissolved in 25mL ethanol solutions, take sodium metaperiodate 4.32g and be dissolved in 25mL deionized waters
In；Secondly, complete sodium periodate solution will be dissolved to be added in sodium alginate soln, 6h is reacted in lucifuge stirring at room temperature；The
Three, the ethylene glycol 1.137mL terminating reactions with sodium metaperiodate equimolar amounts are added, stir 30min；4th, product is entered
Row dialysis, the molecular cut off of bag filter used is 3.5KDa, is dialysed 5-7 days, and dialysis solvent is water, to remove unnecessary unreacted
The small molecule such as sodium metaperiodate, freeze, keep in dark place.
Embodiment 3：The preparation of porous support
Take the aldehyde radical sodium alginate for preparing to be dissolved in PBS, be made into the solution of 10w/v%（Ⅰ）, take gelatin and be dissolved in PBS,
It is made into the solution of 20w/v%（Ⅱ）, take adipic dihydrazide and be dissolved in PBS, it is made into the solution of 50 w/v %（Ⅲ）；Take 100uL I
Solution, the solution of 200uL II, the solution of 26uL III are placed in EP pipes, are placed under high speed shear mixing probe, with the speed of 30000rpm
High speed shear mixes 30s, then is placed in solidification crosslinking 3min in 37 DEG C of water-baths, takes out, and obtains porous support materials.
Embodiment 4：The preparation of porous support
I solution 50uL, the II solution 200uL configured in Example 3, III solution 21.74uL is placed in EP pipes, in high speed shear
Under mixing probe, 30s is mixed with the speed high speed shear of 30000rpm, then be placed in solidification crosslinking 3min in 37 DEG C of water-baths, taken
Go out, obtain porous support materials.
Embodiment 5：The preparation of porous support
I solution 100uL, the II solution 200uL configured in Example 3, III solution 26uL is placed in EP pipes, mixed in high speed shear
Close under probe, 3min mixed with the speed high speed shear of 8000rpm, then be placed in solidification crosslinking 3min in 37 DEG C of water-baths, take out,
Obtain porous support materials.
Embodiment 6
By the preparation method of the porous support materials in embodiment 3,4,5, the porous support materials of different ratio are prepared, by it
It is laid in 96 orifice plates, sterilization treatment is soaked with containing dual anti-PBS solution, uses complete medium（90%F12:DMED=1:1,10%
FBS,1%anti-anti）Preculture 24h is carried out, to remove impurity, by mouse fibroblast（l929）Plant in the timbering material
On, external static culture electron microscopic observation and CCK-8 test analysis cell adherence proliferative conditions after 7 days are shown in that cell can be on support
Good growth, and cell survival rate is up to more than 70%.
Embodiment 7
Take first step gained aldehyde radical sodium alginate to be dissolved in PBS, be made into 10w/v% solution, take gelatin and be dissolved in PBS, be made into
20w/v% solution, takes ADH and is dissolved in PBS, is made into 50w/v% solution, three solution is respectively placed in 100 DEG C of boiling water and boils 5min.Nothing
Under the conditions of bacterium, three is mixed with necessarily matching high speed shear.100uL mixtures are taken, by itself and l929 mixing with cells, cell is dense
It is 10000/hole to spend, and be vortexed concussion 2min；It is added in 96 orifice plates.Culture 24h, detects that cell exists by life or death detection means
Growing state on material, as illustrated, cell well-grown on the multi-porous tissue engineering supporting material.
Specific embodiment is served only for being further illustrated to of the invention, it is impossible to used as the restriction of the scope of the present invention,
Person skilled in art makes some nonessential modifications and adaptations to the present invention according to foregoing invention content simultaneously, is all located at
In protection scope of the present invention, protection scope of the present invention is defined by claims.
Claims (8)
1. preparation of a kind of self-healing without templated porous support, it is characterised in that：The porous support is mixed by high speed shear
The legal cross linked porous structure for preparing.
2. porous support materials as claimed in claim 1, it is characterised in that the timbering material raw material is with amino and work
The bioabsorbable polymer material of property carbonyl, such as gelatin, aldehyde radical sodium alginate, aldehyde radical hyaluronic acid etc..
3. porous support materials as claimed in claim 1, it is characterised in that the porous support includes bifunctional group crosslinking
Small molecule, the small molecule is the small molecules containing bifunctional group such as adipic dihydrazide, and its addition is 4%---8 wt.%.
4. porous support raw material as claimed in claim 2, it is characterised in that described bioabsorbable polymer material containing amino with contain
Active carbonyl group bioabsorbable polymer material, its mass ratio is 20%--500%.
5. preparation of the self-healing as claimed in claim 1 without templated porous support, it is characterised in that carry out cutting at a high speed to raw material
After cutting mixed processing, further to its thermal crosslinking treatment.
6. self-healing porous support is prepared by high speed shear mixing method as claimed in claim 1, it is characterised in that cut at a high speed
Incorporation time is cut for 10-60s, shear rate is 8000-30000 rpm.
7. the preparation of porous support materials as claimed in claim 5, its heat cross-linking temperature is 37-50 DEG C, and the time is 30s-
3min。
8. preparation of a kind of self-healing without templated porous support as claimed in claim 1, it is characterised in that：The porous support
Preparation there is self-healing performance as template, and after external force is cut without pore-foaming agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610559445.2A CN106693067A (en) | 2016-07-15 | 2016-07-15 | Preparation of self-healing and template-free porous scaffold |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610559445.2A CN106693067A (en) | 2016-07-15 | 2016-07-15 | Preparation of self-healing and template-free porous scaffold |
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| CN106693067A true CN106693067A (en) | 2017-05-24 |
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| CN201610559445.2A Pending CN106693067A (en) | 2016-07-15 | 2016-07-15 | Preparation of self-healing and template-free porous scaffold |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109453420A (en) * | 2018-11-29 | 2019-03-12 | 成都美益达医疗科技有限公司 | Hemostatic composition and its preparation method and application |
| CN110498936A (en) * | 2019-07-15 | 2019-11-26 | 北京化工大学 | A kind of preparation method of Sodium Hyaluronate/sodium alginate injection-type composite hydrogel |
| WO2022052150A1 (en) * | 2020-09-10 | 2022-03-17 | 温州医科大学 | Method for preparing composite scaffold for directional guided optic nerve axon regeneration, and application thereof |
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| CN101773683A (en) * | 2010-03-03 | 2010-07-14 | 天津大学 | Chitosan modified alginate hydrogel three-dimensional porous bracket and preparation method thereof |
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| CN1468262A (en) * | 2000-10-10 | 2004-01-14 | LG��ѧ��ʽ���� | Crosslinked amide derivatives of hyaluronic acid and manufacturing method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109453420A (en) * | 2018-11-29 | 2019-03-12 | 成都美益达医疗科技有限公司 | Hemostatic composition and its preparation method and application |
| CN109453420B (en) * | 2018-11-29 | 2021-01-08 | 成都美益达医疗科技有限公司 | Hemostatic composition, preparation method and application thereof |
| CN110498936A (en) * | 2019-07-15 | 2019-11-26 | 北京化工大学 | A kind of preparation method of Sodium Hyaluronate/sodium alginate injection-type composite hydrogel |
| WO2022052150A1 (en) * | 2020-09-10 | 2022-03-17 | 温州医科大学 | Method for preparing composite scaffold for directional guided optic nerve axon regeneration, and application thereof |
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Application publication date: 20170524 |