CN101288778A - Preparation method of porous bacteria cellulose sponges - Google Patents
Preparation method of porous bacteria cellulose sponges Download PDFInfo
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- CN101288778A CN101288778A CNA2008100535523A CN200810053552A CN101288778A CN 101288778 A CN101288778 A CN 101288778A CN A2008100535523 A CNA2008100535523 A CN A2008100535523A CN 200810053552 A CN200810053552 A CN 200810053552A CN 101288778 A CN101288778 A CN 101288778A
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Abstract
The invention relates to a preparation method of porous bacteria cellulose sponge. A bacterial cellulose hydrogel membrane and an ice crystal are completely mixed, pre-frozen is carried out at subzero 20 to subzero 50 DEG C and are prepared into a homogeneous dispersed system. At last, a freeze-drying method is used for preparing the porous bacteria cellulose sponge. Through the invention, bacteria cellulose sponge material containing large holes is simply, rapidly and efficiently manufactured by the invention, and the application of nanometer bacteria cellulose in the field of the tissue engineering stent can be greatly developed.
Description
Technical field
The present invention relates to a kind of preparation method of porous bacteria cellulose sponges.
Background technology
Along with the development of biomedical engineering research, organizational project has become a big focus of current scientific research and clinical practice.The research of tissue engineering bracket material also enjoys the concern of research worker.At present, people have utilized electrostatic spinning, method such as be separated prepares numerous macromolecule tissue engineering brackets such as collagen, chitosan, shell element, PLGA.
Bacterial cellulose is compared with above-mentioned biopolymer, has special structure and physical and chemical performance.Character such as Bacterial cellulose good mechanical performance, permeability, biocompatibility make it have very wide range of commercial application potential in the bio-medical field.The modern wound dressing that Bacterial cellulose is prepared has just had in 1987 and successfully has been used to cure burn, scald and dermatoplastic example.Up to now, the medical commodity of Bacterial cellulose have occurred a lot, as Biofil
, Gengiflex
Deng.
Bacterial cellulose has also had progress as the research of soft tissue engineering scaffold recently.Though chondrocyte the reproduction speed on the Bacterial cellulose matrix only be on collagen I I 50%, under identical external immunoreation degree, chondrocyte will be higher than the speed of growth on the alginate matrix in the speed of growth on the Bacterial cellulose matrix; Chondrocyte does not significantly improve in the speed of growth on the bacteria cellulose film of bonderizing and vulcanizing treatment.On this support that is made of Bacterial cellulose, chondrocyte has kept differentiated.
(though 3～10nm) have natural meticulous network structure, excellent mechanical property and biocompatibility, the cellulosic aperture of natural bacteria mostly is about 1 μ m Bacterial cellulose.Therefore cell can only can not well be grown into to material internal in its surface adhesion and propagation.Therefore, become suitable more tissue engineering bracket material for making Bacterial cellulose, the Bacterial cellulose porous support that preparation contains macropore (50～400 μ m) seems particularly crucial.
Summary of the invention
The preparation method that the purpose of this invention is to provide a kind of porous bacteria cellulose sponges.Adopt phase disengagement method to prepare the bacteria cellulose sponges material that contains macropore, can be fit to sticking and, using of cell thereby make Bacterial cellulose be suitable as tissue engineering bracket more to the growing into of material internal.Efficient, simple, fast characteristics that the present invention has.
The preparation method of a kind of porous bacteria cellulose sponges provided by the invention comprises the steps:
1) utilizes microbial fermentation processes to prepare the nanometer bacteria cellulose hydrogel,, obtain purified bacteria cellulose aquagel through the alkali liquor purification process and after fully cleaning.
2) with above-mentioned bacteria cellulose aquagel and porogen mix homogeneously; Porogen is ice crystal or sodium bicarbonate.
3) under-20～-50 ℃, carry out pre-freeze and to make concentration be 0.1～1% homodisperse system.
4) utilize phase separation method to prepare porous bacteria cellulose sponges.
Described microorganism is one or more of rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter, azotobacter, acetobacter xylinum.
Described alkali liquor purification process is to handle 2h with the sodium hydroxide solution of 0.5M under the boiling condition.
Described bacteria cellulose aquagel and ice crystal mass ratio are 1: 3-5: the concentration of described Bacterial cellulose and ice crystal homodisperse system is 0.1～1%; The temperature of pre-freeze is-20～-50 ℃; Described phase disengagement method is lyophilization, and freeze-drying time is 12～36h.
The preparation method of a kind of porous bacteria cellulose sponges provided by the invention comprises the steps:
1) utilizes acetobacter xylinum as bioreactor, prepare the bacteria cellulose aquagel film by fermentation;
2) above-mentioned bacteria cellulose aquagel film is fully mixed with ice crystal;
3) under-20～-50 ℃, carry out pre-freeze and make the homodisperse system, utilize freeze-drying to prepare porous bacteria cellulose sponges at last.The cryodesiccated time is 12～48h.
The porous bacteria cellulose sponges product that method of the present invention obtains has 50～500 microns macropore.
The porous bacteria cellulose sponges that the present invention prepares has macropore that suitable cell grows into, very high specific surface area and suitable porosity.Efficient, simple, fast characteristics that the present invention has, thus optimized the performance of Bacterial cellulose greatly as tissue engineering bracket material.
Description of drawings
The macro morphology figure of Fig. 1 porous bacteria cellulose sponges.
The environmental scanning electronic microscope of Fig. 2 porous bacteria cellulose sponges (ESEM) figure.
The specific embodiment
Example
Acetobacter xylinum (Acetobacter xylinum) is a starting strain, utilize acetobacter xylinum as bioreactor, prepare milky bacteria cellulose aquagel film [L.Hong by fermentation, Y.L.Wang, S.R.Jia, Y.Huang, C.Gao, Y.Z.Wan Hydroxyapatite/bacterial cellulose composites synthesized via a biomimeticroute, Materials Letters 60 (2006) 1710-1713.].
The bacteria cellulose aquagel film is the translucent lamellar colloid of milky.Bacteria cellulose aquagel film and ice crystal (mass ratio is 1: 3) uniform mixing with the acetobacter xylinum generation, and to be prepared into concentration be 0.125% homodisperse system, this system being placed diameter then is 1.5cm, height is at-40 ℃ of following pre-freezes, lyophilization 36h then in the cylindrical politef mould of 2cm.At last, the sample taking-up is placed 4 ℃ of following stored refrigerated.
The porous bacteria cellulose sponges that obtains can be estimated by following mode:
Microstructure
Reacting bacteria cellulose utilization environmental scanning electronic microscope (ESEM) is observed its microstructure as shown in Figure 1.Porous bacteria cellulose sponges has 50～500 microns macropore, so it can be fit to sticking of cell and inwardly grow into.
Claims (9)
1, a kind of preparation method of porous bacteria cellulose sponges is characterized in that comprising the steps:
1) utilizes microbial fermentation processes to prepare the nanometer bacteria cellulose hydrogel,, obtain purified bacteria cellulose aquagel through the alkali liquor purification process and after fully cleaning;
2) with above-mentioned bacteria cellulose aquagel and ice crystal or sodium bicarbonate porogen mix homogeneously;
3) under-20～-50 ℃, carry out pre-freeze and to make concentration be 0.1～1% homodisperse system;
4) utilize phase separation method to prepare porous bacteria cellulose sponges.
2, method according to claim 1 is characterized in that: described microorganism is one or more of rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter, azotobacter, acetobacter xylinum.
3, method according to claim 1 is characterized in that: described alkali liquor purification process is to handle 2h with the sodium hydroxide solution of 0.5M under the boiling condition.
4, method according to claim 1 is characterized in that: described bacteria cellulose aquagel and ice crystal mass ratio are 1: 3-5.
5, method according to claim 1 is characterized in that: the concentration of described Bacterial cellulose and ice crystal homodisperse system is 0.1～1%.
6, method according to claim 1 is characterized in that: described phase disengagement method is lyophilization, and freeze-drying time is 12～36h.
7, a kind of preparation method of porous bacteria cellulose sponges is characterized in that comprising the steps:
1) utilizes acetobacter xylinum as bioreactor, prepare the bacteria cellulose aquagel film by fermentation;
2) above-mentioned bacteria cellulose aquagel film is fully mixed with ice crystal;
3) under-20～-50 ℃, carry out pre-freeze and make the homodisperse system, utilize freeze-drying to prepare porous bacteria cellulose sponges at last.
8, method according to claim 7, the cryodesiccated time is 12～48h.
9, the porous bacteria cellulose sponges product that obtains of each described method of claim 1-8.
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| CN102210888A (en) * | 2010-04-08 | 2011-10-12 | 上海交通大学医学院附属第九人民医院 | Agarose-enhanced three-dimensional nanometer porous bacterial cellulose stent and application thereof |
| CN102276876A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Bacterial cellulose porous foam material and preparation method thereof |
| CN102274549A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Method for preparing bacteria cellulose bracket material and product thereof |
| CN102600507A (en) * | 2012-03-08 | 2012-07-25 | 天津大学 | Cell culture support frame material and preparation method of cell culture support frame material |
| CN102764451A (en) * | 2012-06-08 | 2012-11-07 | 钟春燕 | Preparation method and application of porous biological cellulose gel material |
| CN102791300A (en) * | 2010-03-19 | 2012-11-21 | 耶拿市弗里德里希·席勒大学 | Multi-phase bacterially-synthesized-nanocellulose biomaterials and method for producing same |
| CN101569764B (en) * | 2009-06-10 | 2012-12-05 | 天津大学 | Bacteria cellulose aquagel with undirectional hole array and preparation method |
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| CN103536954A (en) * | 2013-10-22 | 2014-01-29 | 山东颐诺生物科技有限公司 | Crosslinked alginate-bacterial cellulose sponge and preparation method thereof |
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| CN109384953A (en) * | 2018-09-26 | 2019-02-26 | 德清舒华泡沫座椅有限公司 | A kind of bacteria cellulose sponges of conductive modified |
| CN109841899A (en) * | 2017-11-28 | 2019-06-04 | 中国科学院上海硅酸盐研究所 | A kind of preparation method of the three-dimensional network gel-form solid polymer electrolyte film based on bacteria cellulose |
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2008
- 2008-06-18 CN CNA2008100535523A patent/CN101288778A/en active Pending
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