CN100441755C - Method for preparing gelatin/chitosan blend for use in bionic extracellular matrix fiber stent - Google Patents
Method for preparing gelatin/chitosan blend for use in bionic extracellular matrix fiber stent Download PDFInfo
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- CN100441755C CN100441755C CNB2006101169900A CN200610116990A CN100441755C CN 100441755 C CN100441755 C CN 100441755C CN B2006101169900 A CNB2006101169900 A CN B2006101169900A CN 200610116990 A CN200610116990 A CN 200610116990A CN 100441755 C CN100441755 C CN 100441755C
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Abstract
The invention relates to a gelatin-chitosan complex manufacture method for fiber scaffold of bionic extra-cellular matrix, comprising (1) chitosan solution with 4-10% (g/ml) concentration is obtained after it is dissolved into the mixed solvent of HFIP and TFA; (2) gelatin solution with 4-10% concentration is obtained after it is dissolved into HFIP; (3) gelatin-chitosan complex solution with 4-10% concentration is obtained by mixing the chitosan and gelatin solution in the same cubage and gelatin-chitosan nano composite fiber coat is manufactured by electrostatic spinning.
Description
Technical field
The invention belongs to the electrostatic spinning field, be specifically related to a kind of preparation method who is used for the gelatin/chitosan blend of bionic extracellular matrix fiber stent.
Background technology
The depletion of histoorgan, damaged or dysfunction are one of main harm of facing of human health, there are defectives such as donor source deficiency in traditional treatment pattern " with the wound repair wound ", the proposition of organizational engineering provides a kind of technological means of regeneration, promotes the development of regenerative medicine.The compound existing many reports of extracellular timbering material that are used for of chitosan-gelatin, [1] such as Mao J S is by freeze drying pore method, preparation chitosan-gelatin porous compound support frame, the introducing of gelatin can enter the normal cell cycle by inducing mouse fibroblast L929 cell, promote cell proliferation, reduce the apoptosis of cell.Xia Wanyao etc. [2] plant the credulous osteocyte of pig with the chitosan-gelatin compound support of freeze-drying preparation, behind amplification in vitro, be transplanted to the subcutaneous abdomen of pig, 16 weeks back cartilage formation support is degraded and the NIP reaction fully, illustrate that porous chitosan-gelatin-compounded thing support is the suitable support of organization engineered cartilage, the chitosan-gelatin composite of constructing with the gelatin of positively charged shitosan and both sexes becomes the material that the application future is arranged very much.The collagen fabric size range of n cell epimatrix is 50-500nm[3], it is one of technology of nano material processing that static spins, fibre diameter from several nanometers to several microns [4,5], collagen structure that can farthest bionical n cell epimatrix, shitosan [6,7,8] and gelatin [9,10,11] successfully static be spun into nanofiber.But up to now, the preparation that static spins the chitosan-gelatin intermingling material yet there are no report, and the present invention prepares bionic extracellular matrix fiber stent to gelatine-chitosan co-mixing system electrostatic spinning exactly.List of references is as follows:
[11]Mao?J?S，Cui?Y?L，Wang?X?H，et?al.A?preliminary?study?on?chitosanand?gelation?polyelectrolyte?complex?cytocompatibility?by?cell?cycle?andapoptosis?analysis.Biomaterials.2004；25：3973-3981
[2] Xia Wanyao, Liu Wei, Cui Lei etc. the chitosan-gelatin porous compound support frame makes up the experimental study of autologous tissue's through engineering approaches cartilaginous tissue. Chinese Medical Journal .2003; 83 (7): 577-579
[3]Min?B?M，You?Y，Kim?J?M，et?al.Formation?of?nanostructuredpoly(lactic-co-glycolic?acid)/chitin?matrix?and?its?cellular?response?tonormal?human?keratinocytes?and?fibroblasts.Carbohydrate?Polymers2004；57：285-292
[4]Huang?Z?M，Zhang?Y?Z，kotakic?M，et?al.A?review?on?polymer?nanofibersby?electrospinning?and?their?applications?in?nanocomposites.CompositesScience?and?Technology?2003；63：2223-2253
[5]Subbiah?T，Bhat?G?S，Tock?R?W，et?al.Electrospinning?of?Nanofibers.Journal?of?Applied?Polymer?Science.2005；96：557-569
[6]Mina?B?M，Leeb?S?W，Limb?J?N，et?al.Chitin?and?chitosan?nanofibers：electrospinning?of?chitin?and?deacetylation?of?chitin?nanofibers.Polymer2004；45：7137-7142
[7]K?Ohkawa，D?Cha，H?Kim，et?al.Electrospinning?of?Chitosan.Macromol.Rapid?Commun.2004；25：1600-1605
[8]Xinying?Geng，Oh-Hyeong?Kwon，Jinho?Jang.Electrospinning?of?chitosandissolved?in?concentrated?acetic?acid?solution.Biomaterials?2005；26：5427-5432
[9]Z?M.Huang，Y?Z.Zhang，S?Ramakrishna，et?al.Electrospinning?andmechanical?characterization?of?gelatin?nanofibers.Polymer.2004；45：5361-5368
[10]C?S.Ki，D?H.Baek，K?D.Gang，et?al.Characterization?of?gelatinnanofiber?prepared?from?gelatin-formic?acid?solution.Polymer.2005；46：5094-5102
[11]M?Y.Li，J?Mondrinos，M?R，Gandhi，et?al.Electrospun?protein?fibersas?matrices?for?tissue?engineering.Biomaterials.2005；26：5999-6008
Summary of the invention
Preparation method of the present invention comprises the steps:
(1) preparation chitosan solution: shitosan is dissolved in the mixed solvent of hexafluoroisopropanol (HFIP) and trifluoroacetic acid (TFA), and the HFIP/TFA volume ratio is 8/2, and low-grade fever is stirred to transparent, obtains the chitosan solution that concentration is 4-10% (grams per milliliter);
(2) preparation gelatin solution: gelatin is dissolved in hexafluoroisopropanol, and low-grade fever is stirred to dissolving fully, obtains the gelatin solution that concentration is 4-10% (grams per milliliter);
(3) preparation gelatin and shitosan mixed solution: chitosan solution and gelatin solution are mixed with equal-volume, and obtaining total concentration is gelatin and the chitosan blend solution of 4%-10% (grams per milliliter);
(4) method of spinning by static can prepare the gelatin/chitosan composite nano-fiber membrane.
Above method adopts and disposes shitosan and gelatin solution respectively, after treating to dissolve fully, respectively get equal-volume solution and mix, left standstill after stirring ten minutes, carry out spinning then, the electrostatic spinning process parameter of chitosan solution and gelatin mixed solution: voltage: 12-40 kilovolt; Electric field distance: 60-200 millimeter; Spinning speed, the 0.1-1 milliliter/hour.
The present invention is that electro-spinning is equipped with the gelatin/chitosan blend micro/nano-fibre non-woven material, and the chitosan-gelatin composite of constructing with the gelatin of positively charged shitosan and both sexes becomes the material that the application future is arranged very much, can be used as the bionic extracellular matrix material.
Description of drawings
Fig. 1 is the stereoscan photograph of gelatin/chitosan blend electrostatic spinning
Fig. 2 is the film forming photo of glue/chitosan blend electrostatic spinning
The specific embodiment
Example one
Take by weighing 0.12 gram shitosan (taking off acetyl 85%) with electronic analytical balance and be dissolved in 2 milliliters of hexafluoroisopropanols and trifluoroacetic acid (volume ratio the is 8/2) mixed solvent, obtain the chitosan solution that concentration is 6% (grams per milliliter); Taking by weighing 0.12 gram gelatin is dissolved in and obtains the gelatin solution that concentration is 6% (grams per milliliter) in 2 milliliters of hexafluoroisopropanols; After the normal temperature lower magnetic force stirs and treats to dissolve fully, get that equal-volume mixes and stir that to obtain total concentration be shitosan/gelatin cross-blend solution of 6%, leave standstill that static spins blend solution after ten minutes, voltage is 24kv, and the syringe pump fltting speed is 0.6ml/h, receiving range is 13cm, select syringe needle for use No. 7, aluminium foil receives or thick cotton base cloth receives, and obtains static and spins the nonwoven fabric matrix that gelatin/chitosan blend nanofiber average diameter is 101nm.
Example two
Take by weighing 0.16 gram shitosan (taking off acetyl 85%) with electronic analytical balance and be dissolved in 2 milliliters of hexafluoroisopropanols and trifluoroacetic acid (volume ratio the is 8/2) mixed solvent, obtain the chitosan solution that concentration is 8% (grams per milliliter); Taking by weighing 0.16 gram gelatin is dissolved in and obtains the gelatin solution that concentration is 8% (grams per milliliter) in 2 milliliters of hexafluoroisopropanols; After the normal temperature lower magnetic force stirs and treats to dissolve fully, get that equal-volume mixes and stir that to obtain total concentration be shitosan/gelatin cross-blend solution of 8%, leave standstill that static spins blend solution after ten minutes, voltage is 24kv, and the syringe pump fltting speed is 0.6ml/h, receiving range is 13cm, select syringe needle for use No. 7, aluminium foil receives or thick cotton base cloth receives, and obtains static and spins the nonwoven fabric matrix that gelatin/chitosan blend nanofiber average diameter is 245nm.
Example three
Take by weighing 0.20 gram shitosan (taking off acetyl 85%) with electronic analytical balance and be dissolved in 2 milliliters of hexafluoroisopropanols and trifluoroacetic acid (volume ratio the is 8/2) mixed solvent, obtain the chitosan solution that concentration is 10% (grams per milliliter); Taking by weighing 0.20 gram gelatin is dissolved in and obtains the gelatin solution that concentration is 10% (grams per milliliter) in 2 milliliters of hexafluoroisopropanols; After the normal temperature lower magnetic force stirs and treats to dissolve fully, get that equal-volume mixes and stir that to obtain total concentration be shitosan/gelatin cross-blend solution of 10%, leave standstill that static spins blend solution after ten minutes, voltage is 24kv, and the syringe pump fltting speed is 0.6ml/h, receiving range is 13cm, select syringe needle for use No. 7, aluminium foil receives or thick cotton base cloth receives, and obtains static and spins the nonwoven fabric matrix that gelatin/chitosan blend nanofiber average diameter is 335nm.
Example four
Take by weighing 0.08 gram shitosan (taking off acetyl 85%) with electronic analytical balance and be dissolved in 2 milliliters of hexafluoroisopropanols and trifluoroacetic acid (volume ratio the is 8/2) mixed solvent, obtain the chitosan solution that concentration is 4% (grams per milliliter); Taking by weighing 0.24 gram gelatin is dissolved in and obtains the gelatin solution that concentration is 12% (grams per milliliter) in 2 milliliters of hexafluoroisopropanols; After the normal temperature lower magnetic force stirs and treats to dissolve fully, get that equal-volume mixes and stir that to obtain total concentration be shitosan/collagen blending solution of 8%, leave standstill that static spins blend solution after ten minutes, voltage is 24kv, and the syringe pump fltting speed is 0.6ml/h, receiving range is 13cm, select syringe needle for use No. 7, aluminium foil receives or thick cotton base cloth receives, and obtains static and spins the nonwoven fabric matrix that gelatin/chitosan blend nanofiber average diameter is 112nm.
Claims (2)
1. an electro-spinning Preparation Method that is used for the gelatin/chitosan blend nano fibrous membrane of bionic extracellular matrix fiber stent comprises the steps:
(1) preparation chitosan solution: shitosan is dissolved in the mixed solvent of hexafluoroisopropanol HFIP and trifluoroacetic acid TFA, and its HFIP/TFA volume ratio is 8/2; Low-grade fever is stirred to transparent, obtains the chitosan solution that concentration is the 4-10% grams per milliliter;
(2) preparation gelatin solution: gelatin is dissolved in hexafluoroisopropanol, and low-grade fever is stirred to dissolving fully, obtains the gelatin solution that concentration is the 4-10% grams per milliliter;
(3) preparation gelatin and shitosan mixed solution: chitosan solution and gelatin solution are mixed with equal-volume, obtain gelatin and chitosan blend solution that total concentration is the 4%-10% grams per milliliter;
(4) method of spinning by static prepares the gelatin/chitosan composite nano-fiber membrane.
2. the electro-spinning Preparation Method that is used for the gelatin/chitosan blend nano fibrous membrane of bionic extracellular matrix fiber stent as claimed in claim 1 is characterized in that: the total concentration of gelatin and chitosan blend solution is 6% grams per milliliter, 8% grams per milliliter or 10% grams per milliliter.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101078134B (en) * | 2007-06-27 | 2011-11-09 | 东华大学 | Preparation of natural material/polymer material coaxial electrostatic spinning nano fibre |
| CN101187111B (en) * | 2007-11-29 | 2011-04-27 | 东华大学 | Composite nanometer fiber felt containing nano silver gelatin/chitosan for medical dressing and its preparation |
| CN101695585B (en) * | 2009-10-27 | 2013-02-20 | 吉林大学 | Degradation controlled tissue engineering cornea fibrous scaffold and preparation method thereof |
| US8389498B2 (en) * | 2010-03-26 | 2013-03-05 | Taiwan Textile Research Institute | Spinning solution and method for manufacturing biomaterial fibers |
| CN102691114B (en) * | 2011-03-25 | 2015-08-26 | 财团法人纺织产业综合研究所 | Spinning solution and manufacture raw doctor fiber process |
| CN105254917B (en) * | 2015-11-03 | 2018-10-02 | 中山大学 | A method of preparing cell patch using Sodium Alginate Hydrogel Films |
| CN106757496B (en) * | 2016-12-05 | 2020-09-22 | 华东理工大学 | Double-component polymer superfine fiber containing synthetic polypeptide and chitosan and preparation thereof |
Citations (5)
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|---|---|---|---|---|
| JPH04285565A (en) * | 1991-03-13 | 1992-10-09 | Unitika Ltd | Regenerative and inductive material for periodontium |
| US20020090725A1 (en) * | 2000-11-17 | 2002-07-11 | Simpson David G. | Electroprocessed collagen |
| WO2003099230A2 (en) * | 2002-05-28 | 2003-12-04 | Virginia Commonwealth University Intellectual Property Foundation | Electroprocessed collagen and tissue engineering |
| CN1670272A (en) * | 2005-02-06 | 2005-09-21 | 苏州大学 | Nanometer level regenerated spider silk fiber and its preparation method |
| WO2006036130A1 (en) * | 2004-09-29 | 2006-04-06 | National University Of Singapore | A composite, method of producing the composite and uses of the same |
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- 2006-10-11 CN CNB2006101169900A patent/CN100441755C/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04285565A (en) * | 1991-03-13 | 1992-10-09 | Unitika Ltd | Regenerative and inductive material for periodontium |
| US20020090725A1 (en) * | 2000-11-17 | 2002-07-11 | Simpson David G. | Electroprocessed collagen |
| WO2003099230A2 (en) * | 2002-05-28 | 2003-12-04 | Virginia Commonwealth University Intellectual Property Foundation | Electroprocessed collagen and tissue engineering |
| WO2006036130A1 (en) * | 2004-09-29 | 2006-04-06 | National University Of Singapore | A composite, method of producing the composite and uses of the same |
| CN1670272A (en) * | 2005-02-06 | 2005-09-21 | 苏州大学 | Nanometer level regenerated spider silk fiber and its preparation method |
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