CN103272265B - Bacterial cellulose three-dimensional microporous scaffold preparation method - Google Patents
Bacterial cellulose three-dimensional microporous scaffold preparation method Download PDFInfo
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- CN103272265B CN103272265B CN201310224289.0A CN201310224289A CN103272265B CN 103272265 B CN103272265 B CN 103272265B CN 201310224289 A CN201310224289 A CN 201310224289A CN 103272265 B CN103272265 B CN 103272265B
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- 229920002749 Bacterial cellulose Polymers 0.000 title claims abstract description 70
- 239000005016 bacterial cellulose Substances 0.000 title claims abstract description 70
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 238000012545 processing Methods 0.000 claims abstract description 36
- 238000004108 freeze drying Methods 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000005553 drilling Methods 0.000 claims abstract description 11
- 238000000855 fermentation Methods 0.000 claims abstract description 8
- 230000004151 fermentation Effects 0.000 claims abstract description 8
- 230000001580 bacterial effect Effects 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 13
- 239000011148 porous material Substances 0.000 claims description 11
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 8
- 239000001569 carbon dioxide Substances 0.000 claims description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 3
- 241000590020 Achromobacter Species 0.000 claims description 3
- 241001136169 Komagataeibacter xylinus Species 0.000 claims description 3
- 241000589180 Rhizobium Species 0.000 claims description 3
- 241000190932 Rhodopseudomonas Species 0.000 claims description 3
- 241000192023 Sarcina Species 0.000 claims description 3
- 241000588986 Alcaligenes Species 0.000 claims description 2
- 241000589151 Azotobacter Species 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 14
- 238000005516 engineering process Methods 0.000 abstract description 11
- 230000001276 controlling effect Effects 0.000 abstract description 6
- 238000000746 purification Methods 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract description 2
- 238000005406 washing Methods 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- 229920002678 cellulose Polymers 0.000 description 8
- 239000001913 cellulose Substances 0.000 description 8
- 239000007864 aqueous solution Substances 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 239000012620 biological material Substances 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 235000014103 egg white Nutrition 0.000 description 2
- 210000000969 egg white Anatomy 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 238000004080 punching Methods 0.000 description 2
- 230000017423 tissue regeneration Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 230000000680 avirulence Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000010041 electrostatic spinning Methods 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000005325 percolation Methods 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a bacterial cellulose three-dimensional microporous scaffold preparation method which relates to the technical field of scaffold material preparation processing. The preparation method provided by the invention comprises the following steps: carrying out purification processing on bacterial cellulose generated by bacterial strain fermentation and carrying out freeze drying to obtain bacterial cellulose scaffold; and processing the bacterial cellulose scaffold in the environment of minus 5-10 DEG C by the adoption of a laser drilling technology, washing the processed bacterial cellulose scaffold by the use of redistilled water, and carrying out freeze drying to obtain the bacterial cellulose three-dimensional microporous scaffold, wherein the diameter of the micropores is 100-500 microns, and micropore spacing is 0.8-2.5mm. The preparation method provided by the invention requires a simple technology and is convenient to operate. Micropore size and three-dimensional micropore structure of the scaffold can be regulated and controlled by means of controlling technological parameters and the like. The prepared bacterial cellulose three-dimensional microporous scaffold can be applied in the field of tissue engineering.
Description
Technical field
The present invention relates to the processing technology of preparing field of biologic bracket material.Be particularly related to a kind of preparation method of Bacterial cellulose three-dimensional micropore support.
Background technology
Organizational project refers to principle and the technology of application life sciences and engineering, design, set up, safeguard the growth of human body cell and tissue, and recover the function of impaired tissue or organ, its core is exactly by the normal tissue cell of cultured and amplified in vitro, be adsorbed on the good porous three-dimensional biomaterial scaffolds that also can be absorbed by body of biocompatibility and form active complex, then by cell-biomaterial composites implanting to human body tissue, the disease damage position of organ, cell forms in gradually by the process of body degraded and absorbed new for aspect morphology and function and damaged organ at biomaterial, organize consistent substitute, thereby reach the object of repairing wound and Reconstruction of The Function.
Tissue engineering bracket material is one of organizational project three large key elements, and desirable tissue engineering bracket material should have good histocompatibility, biodegradability, degraded avirulence, good mechanical strength and the 3-D solid structure of high porosity.Wherein the preparation method of timbering material is often depended in the high porosity of tissue engineering bracket material, high-specific surface area and suitable aperture.At present the preparation method of tissue engineering bracket material mainly contains: method of electrostatic spinning, phase separation method, gas foaming are sent out, solution casting method, percolation etc.Although the tissue engineering bracket that these methods can succeed, but the tissue engineering bracket obtaining lacks, and the mutual perforation degree of mechanical strength, hole is low, the poor controllability of porosity and pore size distribution, varying aperture is random, thereby affects the vascularization of growing into, organizing, the transmission of nutrient and the discharge of metabolite of cell.
Laser has that monochromaticity is good, energy density is high, spatial control and the controlled series of advantages such as good of time, and the degree of freedom of material, shape, size and processing environment to processing object is all very large.Laser boring is the laser processing technology that reaches the earliest practical, has been widely used in the processing of metal material, inorganic material and macromolecular material.Laser drilling has: untouchable; Punching speed is fast, efficiency is high; Aperture is small, adjustable; Be applicable to the features such as quantity is many, highdensity capillary processing.It is tens of to hundreds of micron that laser micropore aperture can reach, and adopts organizational project three-dimensional micropore support prepared by this technology can be conducive to cell and enter internal stent, and the nutritional labelings such as convenient egg white matter are passed through, and are convenient to microvascular reconstruction.
For this reason, the present invention adopts laser drilling, prepares a kind of Bacterial cellulose three-dimensional micropore support.This support can be used as tissue engineering bracket material, is applied to tissue repair and reconstruction.The present invention is prepared fast, easy to operate, can be by controlling the pore size of the method accuracy controlling supports such as technological parameter and the structure of three-dimensional micropore.The mutual perforation degree that the Bacterial cellulose three-dimensional micropore support of preparation has good mechanical strength, good structural stability, high porosity and hole is high, can be applicable to field of tissue engineering technology.
Summary of the invention
The invention discloses a kind of preparation method of Bacterial cellulose three-dimensional micropore support.Relate to a kind of processing technology of preparing field of timbering material.Technique of the present invention is simple, easy to operate, can be by controlling the method regulation and control pore size of support and the structures of three-dimensional micropore such as technological parameter, and the three-dimensional micropore support of preparation can be applicable to field of tissue engineering technology.
The invention discloses a kind of preparation method of Bacterial cellulose three-dimensional micropore support.Comprise, the purified processing of Bacterial cellulose being produced by strain fermentation, lyophilization obtains bacterial cellulose stent.Under-5～10 DEG C of environment, adopt laser drilling to process it, the bacterial cellulose stent after processing cleans through redistilled water, and lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 100～500 μ m, and micropore spacing is 0.8～2.5mm.
As preferred technical scheme:
Wherein, the preparation method of a kind of Bacterial cellulose three-dimensional micropore support as above, the bacterial strain that described fermenting produces Bacterial cellulose is one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter or azotobacter.Described purification treating method can be, in the NaOH aqueous solution that tunning is 1～8% at percentage by weight, at the temperature of 30～100 DEG C, to heat 3～6h.Water rinses repeatedly to neutrality again.To remove tropina and to stick to the residual media on cellulose membrane.
The preparation method of a kind of Bacterial cellulose three-dimensional micropore support as above, described lyophilization temperature is: under-40～-10 DEG C of conditions.Bacterial cellulose is to be made up of the cellulose nano-fibrous of three-dimensional manometer network structure, only has the freeze-drying of employing could in dry run, keep its original three-dimensional net structure.In like manner, the bacterial cellulose stent after processing also needs to adopt the three-dimensional micropore structure of freeze-drying to keep obtaining after processing.
The preparation method of a kind of Bacterial cellulose three-dimensional micropore support as above, described laser drilling laser instrument used is: carbon dioxide laser machine, excimer laser or YAG laser instrument.
The preparation method of a kind of Bacterial cellulose three-dimensional micropore support as above, described laser drilling is to adopt laser instrument to carry out capillary processing along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Can realize by bacteria cellulose material on change laser-beam drilling machine the capillary processing of different change in coordinate axis direction in the position of fixture.That laser boring has is untouchable, the fast efficiency of punching speed is high, the small advantage such as adjustable in aperture, and this processing method is applicable to many, the highdensity capillary processings of quantity.In order to improve interpore mutual perforation degree, can be by regulating the structure of three-dimensional micropore, the duct of three directions is intersected in duct crossing or three directions between two simultaneously.The Bacterial cellulose three-dimensional micropore support of preparation can be conducive to cell and enter internal stent, and the nutritional labelings such as convenient egg white matter are passed through, and are convenient to microvascular reconstruction.In addition, under-5～10 DEG C of environment, process, object is to reduce the impact of heat on material that Laser Processing produces.
Compared with prior art, the invention has the beneficial effects as follows: adopt laser drilling, prepare a kind of Bacterial cellulose three-dimensional micropore support.This support can be used as tissue engineering bracket material, is applied to tissue repair and reconstruction.The present invention is prepared fast, easy to operate, can be by controlling the pore size of the method accuracy controlling supports such as technological parameter and the structure of three-dimension hole.The mutual perforation degree that the Bacterial cellulose three-dimensional micropore support of preparation has good mechanical strength, good structural stability, high porosity and hole is high, can be applicable to field of tissue engineering technology.
Detailed description of the invention
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read the content of the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1:
It is in 1% NaOH aqueous solution that the Bacterial cellulose being obtained by acetobacter xylinum fermentation culture is immersed in to percentage by weight, at the temperature of 100 DEG C, heats 3h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Under-10 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Under 10 DEG C of environment, adopt carbon dioxide laser machine to carry out laser boring processing to bacterial cellulose stent.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Bacterial cellulose stent after processing cleans through redistilled water.Under-40 DEG C of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 100 μ m, and micropore spacing is 0.8mm.
Embodiment 2:
It is in 3% NaOH aqueous solution that the Bacterial cellulose being obtained by rhizobium fermentation culture is immersed in to percentage by weight, at the temperature of 80 DEG C, heats 4h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Under-20 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Under 8 DEG C of environment, adopt carbon dioxide laser machine to carry out laser boring processing to bacterial cellulose stent.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Bacterial cellulose stent after processing cleans through redistilled water.Under-30 DEG C of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 150 μ m, and micropore spacing is 1.0mm.
Embodiment 3:
It is in 5% NaOH aqueous solution that the Bacterial cellulose being obtained by Sarcina fermentation culture is immersed in to percentage by weight, at the temperature of 60 DEG C, heats 5h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Under-30 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Under 4 DEG C of environment, adopt excimer laser to carry out laser boring processing to bacterial cellulose stent.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Bacterial cellulose stent after processing cleans through redistilled water.Under-20 DEG C of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter be 250 μ m(100,150), micropore spacing is 1.5mm.
Embodiment 4:
It is in 7% NaOH aqueous solution that the Bacterial cellulose being obtained by Rhodopseudomonas fermentation culture is immersed in to percentage by weight, at the temperature of 40 DEG C, heats 6h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Under-40 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Under 0 DEG C of environment, adopt YAG laser instrument to carry out laser boring processing to bacterial cellulose stent.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Bacterial cellulose stent after processing cleans through redistilled water.Under-10 DEG C of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 500 μ m, and micropore spacing is 2.5mm.
Embodiment 5:
It is in 8% NaOH aqueous solution that the Bacterial cellulose being obtained by achromobacter fermentation culture is immersed in to percentage by weight, at the temperature of 30 DEG C, heats 3h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Under-40 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Under-5 DEG C of environment, adopt excimer laser to carry out laser boring processing to bacterial cellulose stent.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axis) of bacterial cellulose stent respectively.Bacterial cellulose stent after processing cleans through redistilled water.Under-40 DEG C of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 350 μ m, and micropore spacing is-2.0mm.
Claims (5)
1. the preparation method of a Bacterial cellulose three-dimensional micropore support, it is characterized in that: the purified processing of Bacterial cellulose being produced by strain fermentation, lyophilization obtains bacterial cellulose stent, under-5～10 DEG C of environment, adopt laser drilling to process it, bacterial cellulose stent after processing cleans through redistilled water, lyophilization obtains Bacterial cellulose three-dimensional micropore support, wherein micro-pore diameter is 100～500 μ m, micropore spacing is 0.8～2.5mm, and described lyophilization temperature is: under-40～-10 DEG C of conditions.
2. the preparation method of Bacterial cellulose three-dimensional micropore support as claimed in claim 1, is characterized in that: the bacterial strain that described fermenting produces Bacterial cellulose is one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter or azotobacter.
3. the preparation method of as claimed in claim a kind of Bacterial cellulose three-dimensional micropore support, is characterized in that: described lyophilization temperature is: under-40～-10 DEG C of conditions.
4. the preparation method of Bacterial cellulose three-dimensional micropore support as claimed in claim 1, is characterized in that: described laser drilling laser instrument used is: carbon dioxide laser machine, excimer laser or YAG laser instrument.
5. the preparation method of Bacterial cellulose three-dimensional micropore support as claimed in claim 1, it is characterized in that: described laser drilling is to adopt laser instrument respectively along the 3 d space coordinate direction of principal axis of bacterial cellulose stent, be X-axis, Y-axis and Z axis, carry out capillary processing.
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| CN103861154B (en) * | 2014-03-31 | 2016-02-24 | 宁夏医科大学 | A kind of two-layer compound bone tissue engineering scaffold and preparation method thereof |
| CN108126248A (en) * | 2018-03-05 | 2018-06-08 | 涂青山 | A kind of preparation method of porous bacteria cellulose membrane material |
| WO2025177106A1 (en) | 2024-02-21 | 2025-08-28 | Universidade Do Minho | Bacterial nanocellulose material, methods of production, and uses thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101288778A (en) * | 2008-06-18 | 2008-10-22 | 天津大学 | Preparation method of porous bacterial cellulose sponge |
| CN101455862A (en) * | 2007-12-12 | 2009-06-17 | 中国科学院金属研究所 | Preparation method of polyporous material for biological medicine tissue engineering scaffold |
| CN101584882A (en) * | 2009-06-10 | 2009-11-25 | 海南椰国食品有限公司 | Vascular stent material of tissue engineering and manufacturing method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101455862A (en) * | 2007-12-12 | 2009-06-17 | 中国科学院金属研究所 | Preparation method of polyporous material for biological medicine tissue engineering scaffold |
| CN101288778A (en) * | 2008-06-18 | 2008-10-22 | 天津大学 | Preparation method of porous bacterial cellulose sponge |
| CN101584882A (en) * | 2009-06-10 | 2009-11-25 | 海南椰国食品有限公司 | Vascular stent material of tissue engineering and manufacturing method thereof |
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