CN103272265A - Bacterial cellulose three-dimensional microporous scaffold preparation method - Google Patents
Bacterial cellulose three-dimensional microporous scaffold preparation method Download PDFInfo
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- CN103272265A CN103272265A CN2013102242890A CN201310224289A CN103272265A CN 103272265 A CN103272265 A CN 103272265A CN 2013102242890 A CN2013102242890 A CN 2013102242890A CN 201310224289 A CN201310224289 A CN 201310224289A CN 103272265 A CN103272265 A CN 103272265A
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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 use principle and the technology of 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 the normal tissue cell with cultured and amplified in vitro, be adsorbed on the good and porous three-dimensional biomaterial scaffolds that can be absorbed by body of biocompatibility and form active complex, then with cell-biomaterial composites implanting to human body tissue, the disease of organ is decreased the position, cell forms in gradually by the process of body degraded and absorbed new for form and function aspects and impaired organ at biomaterial, organize consistent substitute, thereby reach the purpose of repairing wound and rebuilding function.
Tissue engineering bracket material is one of organizational project three big key elements, and desirable tissue engineering bracket material should have the 3-D solid structure of the favorable tissue compatibility, biodegradability, degraded avirulence, excellent mechanical intensity and 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.Though the tissue engineering bracket that these methods can be succeedd, but resulting tissue engineering bracket lacks, and the mutual perforation degree of mechanical strength, hole is low, the poor controllability of porosity and pore size distribution, varying aperture at random, thereby influence the vascularization of growing into, organizing of cell, the transmission of nutrient and the discharging of metabolite.
Laser has that monochromaticity is good, energy density is high, controlled series of advantages such as good of spatial control and time, and is all very big to the degree of freedom of material, shape, size and the processing environment of processing object.Laser boring is the laser processing technology that reaches practicability the earliest, has been widely used in the processing of metal material, inorganic material and macromolecular material.Laser drilling has: untouchable; Punching speed is fast, efficient is high; The aperture is small, adjustable; Be fit to characteristics such as quantity is many, highdensity capillary processing.It is tens of to hundreds of microns that the laser micropore aperture can reach, and the organizational project three-dimensional micropore support that adopts this technology to prepare can be conducive to cell and enter internal stent, and 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 prepares fast, and is easy to operate, can be by the pore size of method accuracy controlling supports such as control technological parameter and the structure of three-dimensional micropore.The Bacterial cellulose three-dimensional micropore support of preparation has the mutual perforation degree height of good mechanical intensity, good structural stability, high porosity and hole, 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.The processing technology of preparing field that relates to a kind of timbering material.Technology of the present invention is simple, and is easy to operate, can be by the pore size of method regulation and control supports such as control technological parameter and the structure of three-dimensional micropore, 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 that by the purified processing of Bacterial cellulose that strain fermentation produces, lyophilization obtains the Bacterial cellulose support.Under-5～10 ℃ of environment, adopt laser drilling that it is processed, the Bacterial cellulose support after the processing cleans through redistilled water, and lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 100～500 μ m, and the micropore spacing is 0.8～2.5mm.
As optimized technical scheme:
Wherein, the preparation method of aforesaid a kind of Bacterial cellulose three-dimensional micropore support, the described bacterial strain that can ferment the generation Bacterial cellulose are one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter or the azotobacter.Described purification treating method can be, tunning heats 3～6h under 30～100 ℃ temperature in percentage by weight is 1～8% NaOH aqueous solution.Water washes repeatedly to neutrality again.To remove tropina and the residual media that sticks on the cellulose membrane.
The preparation method of aforesaid a kind of Bacterial cellulose three-dimensional micropore support, described lyophilization temperature is: under-40～-10 ℃ of conditions.Bacterial cellulose is to be made of the cellulose nano-fibrous of three-dimensional manometer network structure, has only the freeze-drying of employing could keep its original three-dimensional net structure in dry run.In like manner, the Bacterial cellulose support after the processing also needs to adopt the three-dimensional micropore structure of freeze-drying to keep obtaining after the processing.
The preparation method of aforesaid a kind of Bacterial cellulose three-dimensional micropore support, the used laser instrument of described laser drilling is: carbon dioxide laser machine, excimer laser or YAG laser instrument.
The preparation method of aforesaid a kind of Bacterial cellulose three-dimensional micropore support, described laser drilling are 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 axle) of Bacterial cellulose support respectively.Can realize the capillary processing of different change in coordinate axis direction by bacteria cellulose material on the change laser-beam drilling machine in the position of anchor clamps.That laser boring has is untouchable, the small advantage such as adjustable of the fast efficient height of punching speed, aperture, and this processing method is fit to many, the highdensity capillary processings of quantity.In order to improve interpore mutual perforation degree, can the duct of three directions be intersected in ducts crossing or three directions in twos simultaneously by regulating the structure of three-dimensional micropore.The Bacterial cellulose three-dimensional micropore support of preparation can be conducive to cell and enter internal stent, and nutritional labelings such as convenient egg white matter are passed through, and are convenient to microvascular reconstruction.In addition, process under-5～10 ℃ of environment, purpose is to reduce the heat of Laser Processing generation to the influence of material.
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 prepares fast, and is easy to operate, can be by the pore size of method accuracy controlling supports such as control technological parameter and the structure of three-dimension hole.The Bacterial cellulose three-dimensional micropore support of preparation has the mutual perforation degree height of good mechanical intensity, good structural stability, high porosity and hole, can be applicable to field of tissue engineering technology.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1:
To be immersed in percentage by weight by the Bacterial cellulose that the acetobacter xylinum fermentation culture obtains and be in 1% the NaOH aqueous solution, and heat 3h under 100 ℃ temperature, the reuse redistilled water washes repeatedly to neutrality.Remove tropina and the residual media that sticks on the cellulose membrane.Under-10 ℃ of conditions, lyophilization obtains the Bacterial cellulose support.
Under 10 ℃ of environment, adopt the carbon dioxide laser machine that the Bacterial cellulose support is carried out laser boring processing.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively.Bacterial cellulose support after the processing cleans through redistilled water.Under-40 ℃ of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 100 μ m, and the micropore spacing is 0.8mm.
Embodiment 2:
To be immersed in percentage by weight by the Bacterial cellulose that the rhizobium fermentation culture obtains and be in 3% the NaOH aqueous solution, and heat 4h under 80 ℃ temperature, the reuse redistilled water washes repeatedly to neutrality.Remove tropina and the residual media that sticks on the cellulose membrane.Under-20 ℃ of conditions, lyophilization obtains the Bacterial cellulose support.
Under 8 ℃ of environment, adopt the carbon dioxide laser machine that the Bacterial cellulose support is carried out laser boring processing.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively.Bacterial cellulose support after the processing cleans through redistilled water.Under-30 ℃ of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 150 μ m, and the micropore spacing is 1.0mm.
Embodiment 3:
To be immersed in percentage by weight by the Bacterial cellulose that the Sarcina fermentation culture obtains and be in 5% the NaOH aqueous solution, and heat 5h under 60 ℃ temperature, the reuse redistilled water washes repeatedly to neutrality.Remove tropina and the residual media that sticks on the cellulose membrane.Under-30 ℃ of conditions, lyophilization obtains the Bacterial cellulose support.
Under 4 ℃ of environment, adopt excimer laser that the Bacterial cellulose support is carried out laser boring processing.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively.Bacterial cellulose support after the processing cleans through redistilled water.Under-20 ℃ of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter be 250 μ m(100,150), the micropore spacing is 1.5mm.
Embodiment 4:
To be immersed in percentage by weight by the Bacterial cellulose that the Rhodopseudomonas fermentation culture obtains and be in 7% the NaOH aqueous solution, and heat 6h under 40 ℃ temperature, the reuse redistilled water washes repeatedly to neutrality.Remove tropina and the residual media that sticks on the cellulose membrane.Under-40 ℃ of conditions, lyophilization obtains the Bacterial cellulose support.
Under 0 ℃ of environment, adopt the YAG laser instrument that the Bacterial cellulose support is carried out laser boring processing.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively.Bacterial cellulose support after the processing cleans through redistilled water.Under-10 ℃ of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 500 μ m, and the micropore spacing is 2.5mm.
Embodiment 5:
To be immersed in percentage by weight by the Bacterial cellulose that the achromobacter fermentation culture obtains and be in 8% the NaOH aqueous solution, and heat 3h under 30 ℃ temperature, the reuse redistilled water washes repeatedly to neutrality.Remove tropina and the residual media that sticks on the cellulose membrane.Under-40 ℃ of conditions, lyophilization obtains the Bacterial cellulose support.
Under-5 ℃ of environment, adopt excimer laser that the Bacterial cellulose support is carried out laser boring processing.Capillary processing is carried out along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively.Bacterial cellulose support after the processing cleans through redistilled water.Under-40 ℃ of conditions, lyophilization obtains Bacterial cellulose three-dimensional micropore support.Wherein micro-pore diameter is 350 μ m, and the micropore spacing is-2.0mm.
Claims (5)
1. the preparation method of a Bacterial cellulose three-dimensional micropore support, it is characterized in that: by the purified processing of Bacterial cellulose of strain fermentation generation, lyophilization obtains the Bacterial cellulose support, under-5～10 ℃ of environment, adopt laser drilling that it is processed, the Bacterial cellulose support after the processing cleans through redistilled water, and lyophilization obtains Bacterial cellulose three-dimensional micropore support, wherein micro-pore diameter is 100～500 μ m, and the micropore spacing is 0.8～2.5mm.
2. the preparation method of Bacterial cellulose three-dimensional micropore support as claimed in claim 1 is characterized in that: the described bacterial strain that can fermentation produces Bacterial cellulose is one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, Aerobacter or the azotobacter.
3. the preparation method of as claimed in claim a kind of Bacterial cellulose three-dimensional micropore support, it is characterized in that: described lyophilization temperature is: under-40～-10 ℃ of conditions.
4. the preparation method of Bacterial cellulose three-dimensional micropore support as claimed in claim 1, it is characterized in that: the used laser instrument of described laser drilling 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 is characterized in that: described laser drilling carries out capillary processing along the 3 d space coordinate direction of principal axis (X-axis, Y-axis and Z axle) of Bacterial cellulose support respectively for adopting laser instrument.
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Cited By (2)
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| CN103861154A (en) * | 2014-03-31 | 2014-06-18 | 宁夏医科大学 | Novel dual-layer composite 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 |
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| CN101288778A (en) * | 2008-06-18 | 2008-10-22 | 天津大学 | Preparation method of porous bacteria cellulose sponges |
| 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 |
| US20120135086A1 (en) * | 2002-11-15 | 2012-05-31 | University Of Tennessee Research Foundation | Method of tissue repair using a composite material |
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Patent Citations (4)
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| US20120135086A1 (en) * | 2002-11-15 | 2012-05-31 | University Of Tennessee Research Foundation | Method of tissue repair using a composite material |
| 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 bacteria cellulose sponges |
| 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 |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103861154A (en) * | 2014-03-31 | 2014-06-18 | 宁夏医科大学 | Novel dual-layer composite bone tissue engineering scaffold and preparation method thereof |
| 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 |
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