CN104841021A - Preparation method for chitosan-based biomimetic membrane material with asymmetric structure - Google Patents
Preparation method for chitosan-based biomimetic membrane material with asymmetric structure Download PDFInfo
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Abstract
The invention discloses a preparation method for a chitosan-based biomimetic membrane material with an asymmetric structure, and the preparation method comprises the following steps of dispersing chitosan into acetic acid aqueous solution, adding absolute methanol and acetic anhydride for reacting after the chitosan is completely dissolved, and then, centrifugally removing sediment; dropping in alkaline solidification liquid under stirring condition, and filtering to obtain chitosan microgel; adding the chitosan microgel in glycerin solution, injecting the solution in a mold, and standing and draining water, freeze-drying one part to obtain a chitosan-based porous sponge material, and drying the other part to obtain a chitosan-based dense membrane material; bonding the chitosan-based porous sponge material and the chitosan-based dense membrane material to obtain the chitosan-based biomimetic membrane material with asymmetric structure. The preparation method provided by the invention has the advantages that the preparation technology is simple, the prepared chitosan-based biomimetic membrane material has a porous sponge structure and a dense membrane material structure, the chitosan-based biomimetic membrane material is not curly when encountering water, guiding bone regeneration performance is good, and the chitosan-based biomimetic membrane material is degradable and controllable and can be widely used as a guided bone tissue regeneration membrane material in treatment of the fields such as dental implant, osteoarthritis, bone defect, and the like.
Description
Technical field
The present invention relates to a kind of preparation method with the chitosan-based bionic membrane material of unsymmetric structure.
Background technology
Chitosan is a kind of biodegradable natural polysaccharide, and its catabolite is glucosamine, in alkalescence, to human body and organize nontoxic, harmless, has excellent biocompatibility, natural pharmaceutically active, anti-tumor activity, antiinflammation etc.Chitosan has been widely used in the fields such as organizational project, drug release, postoperative anti-adhesion membrane, orthopaedics repair materials.
Dental Implant has been risen gradually at dental field and has been popularized, Guide Periodontal Tissue Regeneration technology (Periodontal Guided Tissue Regeneration, GTR) be current state-of-the-art periodontal surgery Therapeutic Method, namely be positioned between root of the tooth and gingiva tissue lobe with a kind of barrier film material, physical barriers gingival connective tissue cell first contacts with root of the tooth with epithelial cell, ensures preferentially occupy root face by the periodontal membrane tissue-derived cell of root side remnants and alveolar bone cell and grow thereon and creep to hat side.For obtaining best soft tissue and the regeneration effect of sclerous tissues, except barrier function, good wound healing properties also becomes more and more important.The comparatively ripe periodontal isolating membrane of current Clinical practice is the Bio-Gide film of Geistlich company, and this is by pig
type and
collagen Type VI through processing synthesis, highly purified after the obtained asymmetric biomembrane of absorbability with one side compact texture, another side loose and porous structure.The compacted zone of film can stop that fibrous connective tissue is grown into, and has good cell iris action; The porosity and looseness layer of film plays the effect of stabilised blood grumeleuse, for osteoblastic adhesion and bone apposition provide the carrier of attachment.But Bio-Gide film there will be, and fin comes off, hemorrhage, local inflammation, bone loss, infection or the complication such as pain, anaphylaxis, and degradation speed is too fast.Thus, develop that a kind of osteoinductive is excellent, biocompatibility is good, the regulatable periodontal tissue of degradability guides regeneration isolating membrane to be current clinical Dental Implant field institute urgent problem.
Summary of the invention
The object of this invention is to provide a kind of preparation method with the chitosan-based bionic membrane material of unsymmetric structure, to obtain the chitosan-based bionic membrane material with unsymmetric structure of inducting osseous tissue regeneration excellent performance, good biocompatibility.
The preparation method with the chitosan-based bionic membrane material of unsymmetric structure of the present invention, step is as follows:
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 2 ~ 5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 4:3 ~ 2, the mass ratio of chitosan and acetic anhydride is 3.5 ~ 6.5, and standing and reacting 1.5-3h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 10-20min of the rotating speed of 5000-8000rpm, after removing precipitate, instill alkaline solidification liquid under agitation wherein to solution alkaline, filter and wash much filtrate to washing liquid in neutral with tri-distilled water, acquisition chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 10% ~ 20%, stir 30 ~ 45min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 40-60 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical bio glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
The present invention adopts acid-soluble system to prepare chitosan solution, by adding the deacetylation of absolute methanol and acetic anhydride adjustment chitosan, add alkaline solidification liquid under agitation and prepare chitosan microgel, chitosan microgel assembling is piled up and is formed chitosan-based aquagel membrane, obtain chitosan-based mandruka material and chitosan-based dense film respectively by the mode of vacuum lyophilization and vacuum drying, the chitosan-based bionic membrane material with unsymmetric structure can be obtained with medical bio glue bond.
Preparation technology of the present invention is simple, by regulating the amount adding acetic anhydride to regulate and control the deacetylation of chitosan, thus realizes chitosan-based bionic membrane material degradation property, the controllable adjustment guiding regenerability and mechanical property.Obtained chitosan-based bionic membrane material has mandruka structure and dense membrane structure concurrently, meet water not curling, have that self-bone grafting regenerability is good, the advantage such as controlled of degrading, can be used as the treatment that inducting osseous tissue regeneration membrane material is widely used in the fields such as Dental Implant, osteoarthritis, Cranial defect.
Accompanying drawing explanation
Fig. 1 is the stereoscan photograph of chitosan-based mandruka material;
Fig. 2 is the stereoscan photograph of chitosan-based dense film.
Detailed description of the invention
The present invention is further illustrated below in conjunction with accompanying drawing and instantiation.
embodiment 1
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 2 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 4:3, the mass ratio of chitosan and acetic anhydride is 3.5, and standing and reacting 1.5h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 10min of the rotating speed of 5000rpm, after removing precipitate, instilling mass fraction wherein is under agitation that the NaOH aqueous solution of 5% is to solution alkaline, filter and wash much filtrate to washing liquid in neutral with tri-distilled water, acquisition chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 10%, stir 30min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material, and its scanning electron microscope (SEM) photograph as shown in Figure 1, can find out that its internal structure is the through loose shape of porous; Another group puts into vacuum drying oven, 40 DEG C of oven dry, obtains chitosan-based dense film material, and its scanning electron microscope (SEM) photograph as shown in Figure 2, can find out that membrane superficial tissue is comparatively fine and close;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material Algin bond, and obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 2
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 2.5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 5:3, the mass ratio of chitosan and acetic anhydride is 3.8, and standing and reacting 2h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 12min of the rotating speed of 6000rpm, after removing precipitate, instill wherein under agitation mass fraction be the KOH aqueous solution of 5% to solution alkaline, filter also by tri-distilled water washing much filtrate to washing liquid in neutral, obtain chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 12%, stir 30min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 45 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material Algin bond, and obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 3
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 3 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 4:3, the mass ratio of chitosan and acetic anhydride is 4.0, and standing and reacting 2h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 15min of the rotating speed of 6500rpm, after removing precipitate, instill wherein under agitation mass fraction be the KOH aqueous solution of 5% to solution alkaline, filter also by tri-distilled water washing much filtrate to washing liquid in neutral, obtain chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 15%, stir 35min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 50 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material Algin bond, and obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 4
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 3.5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 5:3, the mass ratio of chitosan and acetic anhydride is 4.5, and standing and reacting 2.5h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 18min of the rotating speed of 7000rpm, after removing precipitate, instill the NaOH aqueous solution of 5% under agitation wherein to solution alkaline, it is neutral for filtering and washing much filtrate to washing liquid with tri-distilled water, obtains chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 16%, stir 35min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 60 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical e alpha-cyanoacrylate 6B system glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 5
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 4 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 2, the mass ratio of chitosan and acetic anhydride is 5.0, and standing and reacting 2.5h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 20min of the rotating speed of 8000rpm, after removing precipitate, instill saturated ammonia aqueous solution under agitation wherein to solution alkaline, it is neutral for filtering and washing much filtrate to washing liquid with tri-distilled water, obtains chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 18%, stir 40min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 60 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical e alpha-cyanoacrylate 6B system glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 6
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 4.5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 3:2, the mass ratio of chitosan and acetic anhydride is 5.5, and standing and reacting 2.5h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 20min of the rotating speed of 8000rpm, after removing precipitate, instill saturated ammonia aqueous solution under agitation wherein to solution alkaline, it is neutral for filtering and washing much filtrate to washing liquid with tri-distilled water, obtains chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 20%, stir 45min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 55 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical e alpha-cyanoacrylate 6B system glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 7
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 3, the mass ratio of chitosan and acetic anhydride is 6.5, and standing and reacting 3h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 20min of the rotating speed of 8000rpm, after removing precipitate, instill the NaOH aqueous solution of 5% under agitation wherein to solution alkaline, it is neutral for filtering and washing much filtrate to washing liquid with tri-distilled water, obtains chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 20%, stir 45min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 60 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical e alpha-cyanoacrylate 6B system glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
embodiment 8
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 4.8wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 2, the mass ratio of chitosan and acetic anhydride is 6.5, and standing and reacting 3h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 15min of the rotating speed of 7500rpm, after removing precipitate, instill saturated ammonia aqueous solution under agitation wherein to solution alkaline, it is neutral for filtering and washing much filtrate to washing liquid with tri-distilled water, obtains chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 19%, stir 42min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 55 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material Algin bond, and obtain the chitosan-based bionic membrane material with unsymmetric structure.
Claims (4)
1. there is a preparation method for the chitosan-based bionic membrane material of unsymmetric structure, it is characterized in that, comprise the following steps:
1) Chitosan powder being scattered in volume fraction is in the aqueous acetic acid of 2%, be stirred to and dissolve completely, be mixed with the chitosan solution of 2 ~ 5 wt %, add absolute methanol and acetic anhydride wherein again, the volume ratio of absolute methanol and water is made to be 4:3 ~ 2, the mass ratio of chitosan and acetic anhydride is 3.5 ~ 6.5, and standing and reacting 1.5-3h after stirring obtains acid-soluble chitosan solution;
2) by the acid-soluble chitosan solution of step 1) with the centrifugal 10-20min of the rotating speed of 5000-8000rpm, after removing precipitate, instill alkaline solidification liquid under agitation wherein to solution alkaline, filter and wash much filtrate to washing liquid in neutral with tri-distilled water, acquisition chitosan microgel;
3) by step 2) chitosan microgel to join volume fraction be in the glycerin solution of 10% ~ 20%, stir 30 ~ 45min, obtain mixed liquor, mixed liquor is injected mould, leave standstill under room temperature and drain away the water, obtain chitosan-based aquagel membrane;
4) chitosan-based aquagel membrane step 3) obtained is divided into two groups, puts into vacuum freeze drier lyophilizing for one group, obtains chitosan-based mandruka material; Another group puts into vacuum drying oven, 40-60 DEG C of oven dry, obtains chitosan-based dense film material;
5) chitosan-based mandruka material step 4) obtained and chitosan-based dense film material medical bio glue bond, obtain the chitosan-based bionic membrane material with unsymmetric structure.
2. the preparation method with the chitosan-based bionic membrane material of unsymmetric structure according to claim 1, it is characterized in that the deacetylation of described Chitosan powder is 80% ~ 100%, molecular weight is 300000 ~ 800000.
3. the preparation method with the chitosan-based bionic membrane material of unsymmetric structure according to claim 1, is characterized in that described step 2) in alkaline solidification liquid to be mass fraction be 5% NaOH aqueous solution, mass fraction be 5% KOH aqueous solution or saturated ammonia aqueous solution.
4. the preparation method with the chitosan-based bionic membrane material of unsymmetric structure according to claim 1, is characterized in that the Medical Living Creature Gum in described step 5) is Algin or medical e alpha-cyanoacrylate 6B system glue.
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| CN109745577A (en) * | 2017-11-08 | 2019-05-14 | 南京理工大学 | The preparation method of chitosan base load medicine flexibility sponge |
| CN110180028A (en) * | 2019-06-05 | 2019-08-30 | 湖南师范大学 | A kind of preparation method of the bamboo fiber-based composite membrane for inducting osseous tissue regeneration |
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| CN113351175A (en) * | 2021-07-05 | 2021-09-07 | 江南大学 | Preparation method and application of zinc-aluminum hydrotalcite/chitosan-based sponge |
| CN114652895A (en) * | 2020-12-23 | 2022-06-24 | 上海其胜生物制剂有限公司 | Preparation method of injectable tissue regeneration type chitosan composite gel scaffold |
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| CN106110407A (en) * | 2016-08-12 | 2016-11-16 | 上海交通大学医学院附属第九人民医院 | A kind of inductive bone regeneration composite film material and preparation method thereof |
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| CN110180028B (en) * | 2019-06-05 | 2021-11-26 | 湖南师范大学 | Preparation method of bamboo fiber-based composite membrane for guiding bone tissue regeneration |
| CN110180028A (en) * | 2019-06-05 | 2019-08-30 | 湖南师范大学 | A kind of preparation method of the bamboo fiber-based composite membrane for inducting osseous tissue regeneration |
| CN111420123A (en) * | 2020-03-16 | 2020-07-17 | 江西光至金辉医疗制品有限公司 | Degradable anti-adhesion double-layer dura mater patch and preparation method thereof |
| CN114652895A (en) * | 2020-12-23 | 2022-06-24 | 上海其胜生物制剂有限公司 | Preparation method of injectable tissue regeneration type chitosan composite gel scaffold |
| CN112716643A (en) * | 2020-12-28 | 2021-04-30 | 浙江大学 | Regional function specificity clinical periodontal defect repair module |
| CN113351175A (en) * | 2021-07-05 | 2021-09-07 | 江南大学 | Preparation method and application of zinc-aluminum hydrotalcite/chitosan-based sponge |
| CN113351175B (en) * | 2021-07-05 | 2022-09-09 | 江南大学 | Preparation method and application of zinc-aluminum hydrotalcite-like compound/chitosan-based sponge |
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