CN103301505B - Method for preparing bacterial cellulose three-dimensional exhibition microporous bracket - Google Patents
Method for preparing bacterial cellulose three-dimensional exhibition microporous bracket Download PDFInfo
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- CN103301505B CN103301505B CN201310224288.6A CN201310224288A CN103301505B CN 103301505 B CN103301505 B CN 103301505B CN 201310224288 A CN201310224288 A CN 201310224288A CN 103301505 B CN103301505 B CN 103301505B
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Abstract
The invention discloses a method for preparing a bacterial cellulose three-dimensional exhibition microporous bracket, and relates to the technical field of preparation and processing of bracket materials. The bacterial cellulose bracket is obtained by passivating, cutting and freeze-drying bacterial cellulose produced by fermentation of strains. By a computer tomography technology, a math model of a bacterial cellulose bracket three-dimensional structure is constructed; and the specific three-dimensional exhibition microporous bracket is designed by the math model. The math model for a required bacterial cellulose three-dimensional exhibition microporous structure is introduced into a carbon dioxide laser perforation machine and is processed under an environment of -5-10 DEG C; the processed bacterial cellulose bracket is cleaned by secondary distilled water; and then the bacterial cellulose three-dimensional exhibition microporous bracket is obtained by freeze-drying. According to the method, the technology is simple and convenient to operate; sizes of micropores and structures of three-dimensional exhibition holes of the bracket can be adjusted and controlled by methods, such as a method for controlling technical parameters; and the prepared three-dimensional exhibition microporous bracket can be applied to engineering field of construction of tissues, such as skins, bones, ribs and blood vessels.
Description
Technical field
The present invention relates to the processing technology of preparing field of biologic bracket material.Be particularly related to the preparation method of the three-dimensional display of a kind of Bacterial cellulose micropore bracket.
Background technology
Tissue engineering bracket material is one of organizational project three large key elements, and because tissue all has specific macro morphology, the timbering material that need to be used for organizational project reparation also possesses corresponding macro morphology.And because showing timbering material microcosmic (nanoscale, the micron order) structure of repairing for organizational project, large quantity research can affect growth and the protein expression of cell.Therefore,, in tissue engineering bracket material preparation process, in controlling macro morphology, also need to control microstructure.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 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 many, the highdensity displays of quantity hole 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.
The present invention adopts carbon dioxide laser punching technology, prepares the three-dimensional display of a kind of Bacterial cellulose micropore bracket.On the bacterial cellulose stent with macro morphology, prepare the Bacterial cellulose micropore bracket of specific three dimensional arrayed.Cell adhesion rate when this timbering material has increased cell culture, and improved the multiplication rate of cell.The present invention is prepared fast, easy to operate, can be by controlling pore size, shape and the three-dimensional structure that displays hole of the method accuracy controlling supports such as technological parameter.The mutual perforation degree that the three-dimensional display of the Bacterial cellulose micropore bracket of preparation has good mechanical strength, good structural stability, high porosity and hole is high, can be used as tissue engineering bracket material, be applied to tissue repair and the reconstructions such as skin, bone, cartilage, blood vessel.
Summary of the invention
The invention discloses the preparation method of the three-dimensional display of a kind of Bacterial cellulose micropore bracket.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 pore size of the method regulation and control supports such as technological parameter and the structure in three-dimensional display hole, and the three-dimensional display micropore bracket of preparation can be applicable to build the field of tissue engineering technology such as skin, bone, cartilage, blood vessel.
The invention discloses the preparation method of the three-dimensional display of a kind of Bacterial cellulose micropore bracket.Comprise,
(1) the purified processing of Bacterial cellulose, cutting, the lyophilization that are produced by strain fermentation obtain bacterial cellulose stent.
(2) adopt computed tomography to build the mathematical model of bacterial cellulose stent three dimensional structure.Taking mathematical model center of gravity as initial point, inoculating surfaces when bacterial cellulose stent inoculating cell is projected as XY plane downwards and sets up three-dimensional system of coordinate.Utilize the specific three-dimensional display microcellular structure of mathematical model design.
(3) by the three-dimensional display of required Bacterial cellulose microcellular structure mathematical model introducing carbon dioxide laser-beam drilling machine, under-5～10 DEG C of environment, process, bacterial cellulose stent after processing cleans through redistilled water, and lyophilization obtains the three-dimensional display of Bacterial cellulose micropore bracket.
As preferred technical scheme:
Wherein, the preparation method of the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, 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 the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, 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 cubical array microcellular structure of freeze-drying to keep obtaining after processing.
The preparation method of the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, described purified processing, cutting, lyophilization obtains its profile of bacterial cellulose stent and is: cylinder, square, cuboid, membranaceous or irregular body.The bacterial cellulose stent with certain profile can form in incubation, can be to be also processed to form through machine cuts.
The preparation method of the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, the three-dimensional display microcellular structure of described design has three processing planes, XY plane, XY plane, the YZ plane of the mathematical model of corresponding bacterial cellulose stent three dimensional structure respectively.Can realize by bacteria cellulose material on change laser-beam drilling machine the array perforation processing of different processing planes in the position of fixture.
The preparation method of the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, described three-dimensional display microcellular structure: the micropore that is n × n array on the processing plane of every 5mm × 5mm, micro-pore diameter is 100～300 μ m, micropore spacing is not less than 200 μ m, and micropore and XY plane on inoculating surfaces is the angle of 45～80 °.In the cell and timbering material recombination process of organizational project, cell inoculation is to be dripped on timbering material and by the three-dimensional micropore of material and entered timbering material inside by cell suspension.In this course, cell enters the quantity of timbering material and time that cell contacts with timbering material all affects the adhesion rate of cell on timbering material.Enter the quantity of timbering material in order to increase cell, in the time that designing, need improve timbering material microcellular structure interpore mutual perforation degree, can be by regulating quantity and the position in three-dimensional display hole, the duct of three processing planes is intersected in duct crossing or three processing planes between two simultaneously.Meanwhile, the time contacting with timbering material in order to extend cell, the three-dimensional display microcellular structure of this patent makes micropore on inoculating surfaces and XY plane be certain angle.This display micropore design with certain angle of inclination can prevent that cell suspension passing hole channel from directly spilling timbering material, does not also affect cell suspension and enters timbering material inside.
The preparation method of the three-dimensional display of a kind of Bacterial cellulose as above micropore bracket, described three-dimensional display micropore perpendicular to cross section is radially: circle, square, triangle or polygon.
Compared with prior art, the invention has the beneficial effects as follows: adopt carbon dioxide laser punching technology, prepare the three-dimensional display of a kind of Bacterial cellulose micropore bracket.Preparation process can be controlled macro morphology and the microstructure of bacteria cellulose bracket material, and the arrangement of the cubical array of this Bacterial cellulose micropore bracket, is conducive to cell and enters timbering material inside, and improved cell adhesion rate.Can be by controlling pore size, shape and the three-dimensional structure that displays hole of the method accuracy controlling supports such as technological parameter.The mutual perforation degree that the three-dimensional display of the Bacterial cellulose micropore bracket of preparation has good mechanical strength, good structural stability, high porosity and hole is high, can be used as tissue engineering bracket material, be applied to tissue repair and the reconstructions such as skin, bone, cartilage, blood vessel.
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:
1, the Bacterial cellulose being obtained by acetobacter xylinum fermentation culture being immersed in to percentage by weight is, in 8% NaOH aqueous solution, at the temperature of 30 DEG C, to heat 6h, more repeatedly rinses to neutrality with redistilled water.Remove tropina and stick to the residual media on cellulose membrane.Machine cuts is processed into the square of 2cm × 2cm × 2cm.Under-40 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
2, adopt computed tomography to obtain the faultage image of bacterial cellulose stent, each faultage image is processed, machine simulation builds the mathematical model of bacterial cellulose stent three dimensional structure as calculated.Taking mathematical model center of gravity as initial point, inoculating surfaces when bacterial cellulose stent inoculating cell is projected as XY plane downwards and sets up three-dimensional system of coordinate.Utilize the three-dimensional display of mathematical model design microcellular structure.The three-dimensional display microcellular structure of design has three processing planes, XY plane, XY plane, the YZ plane of corresponding bacterial cellulose stent three dimensional structure mathematical model respectively.On the processing plane of every 5mm × 5mm, be the micropore of 15 × 15 arrays, micro-pore diameter is 100 μ m, and micropore spacing is not less than 200 μ m, and micropore and XY plane on inoculating surfaces is the angle of 45 °.Three-dimensional display micropore is circular perpendicular to cross section radially.
3, by the three-dimensional display of required Bacterial cellulose microcellular structure mathematical model introducing carbon dioxide laser-beam drilling machine, under-5 DEG C of environment, process, bacterial cellulose stent after processing cleans through redistilled water, and lyophilization obtains the three-dimensional display of Bacterial cellulose micropore bracket.
Embodiment 2:
It is in 4% 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.It is 1cm that machine cuts is processed into diameter, the cylinder of high 2cm.Under-30 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Adopt computed tomography to obtain the faultage image of bacterial cellulose stent, each faultage image is processed, machine simulation builds the mathematical model of bacterial cellulose stent three dimensional structure as calculated.Taking mathematical model center of gravity as initial point, inoculating surfaces when bacterial cellulose stent inoculating cell is projected as XY plane downwards and sets up three-dimensional system of coordinate.Utilize the three-dimensional display of mathematical model design microcellular structure.The three-dimensional display microcellular structure of design has three processing planes, XY plane, XY plane, the YZ plane of corresponding bacterial cellulose stent three dimensional structure mathematical model respectively.On the processing plane of every 5mm × 5mm, be the micropore of 9 × 9 arrays, micro-pore diameter is 300 μ m, and micropore spacing is not less than 200 μ m, and micropore and XY plane on inoculating surfaces is the angle of 60 °.Three-dimensional display micropore is square perpendicular to cross section radially.
By in the three-dimensional display of required Bacterial cellulose microcellular structure mathematical model introducing carbon dioxide laser-beam drilling machine, under 0 DEG C of environment, process, bacterial cellulose stent after processing cleans through redistilled water, and lyophilization obtains the three-dimensional display of Bacterial cellulose micropore bracket.
Embodiment 3:
It is in 1～8% NaOH aqueous solution that the Bacterial cellulose being obtained by Alcaligenes and azotobacter 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.Machine cuts is processed into the film material of 5cm × 5cm × 1cm.Under--10 DEG C of conditions, lyophilization obtains bacterial cellulose stent.
Adopt computed tomography to obtain the faultage image of bacterial cellulose stent, each faultage image is processed, machine simulation builds the mathematical model of bacterial cellulose stent three dimensional structure as calculated.Taking mathematical model center of gravity as initial point, inoculating surfaces when bacterial cellulose stent inoculating cell is projected as XY plane downwards and sets up three-dimensional system of coordinate.Utilize the three-dimensional display of mathematical model design microcellular structure.The three-dimensional display microcellular structure of design has three processing planes, XY plane, XY plane, the YZ plane of corresponding bacterial cellulose stent three dimensional structure mathematical model respectively.On the processing plane of every 5mm × 5mm, be the micropore of 6 × 6 arrays, micro-pore diameter is 200 μ m, and micropore spacing is not less than 200 μ m, and micropore and XY plane on inoculating surfaces is the angle of 80 °.Three-dimensional display micropore is equilateral triangle perpendicular to cross section radially.
By in the three-dimensional display of required Bacterial cellulose microcellular structure mathematical model introducing carbon dioxide laser-beam drilling machine, under 10 DEG C of environment, process, bacterial cellulose stent after processing cleans through redistilled water, and lyophilization obtains the three-dimensional display of Bacterial cellulose micropore bracket.
Claims (7)
1. a preparation method for the three-dimensional display of Bacterial cellulose micropore bracket, is characterized in that:
(1) the purified processing of Bacterial cellulose, cutting, the lyophilization that are produced by strain fermentation obtain bacterial cellulose stent;
(2) adopt computed tomography to build the mathematical model of bacterial cellulose stent three dimensional structure, taking mathematical model center of gravity as initial point, inoculating surfaces when bacterial cellulose stent inoculating cell is projected as XY plane downwards and sets up three-dimensional system of coordinate, utilizes the specific three-dimensional display microcellular structure of mathematical model design;
(3) by the three-dimensional display of required Bacterial cellulose microcellular structure mathematical model introducing carbon dioxide laser-beam drilling machine, under-5～10 DEG C of environment, process, bacterial cellulose stent after processing cleans through redistilled water, lyophilization obtains the three-dimensional display of Bacterial cellulose micropore bracket, and described lyophilization temperature is: under-40～-10 DEG C of conditions.
2. the preparation method of the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 1 micropore bracket, 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 the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 1 micropore bracket, is characterized in that: described purified processing, cutting, lyophilization obtains its profile of bacterial cellulose stent and be: cylinder, square, cuboid or irregular body.
4. the preparation method of the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 1 micropore bracket, it is characterized in that: the three-dimensional display microcellular structure of design has three processing planes, XY plane, XY plane, the YZ plane of the mathematical model of corresponding bacterial cellulose stent three dimensional structure respectively.
5. the preparation method of the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 1 micropore bracket, it is characterized in that: three-dimensional display microcellular structure: the micropore that is n × n array on the processing plane of every 5mm × 5mm, micro-pore diameter is 100～300 μ m, micropore spacing is not less than 200 μ m, and micropore and XY plane on inoculating surfaces is the angle of 45～80 °.
6. the preparation method of the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 1 micropore bracket, is characterized in that: three-dimensional display micropore perpendicular to cross section is radially: circle or polygon.
7. the preparation method of the three-dimensional display of a kind of Bacterial cellulose as claimed in claim 6 micropore bracket, is characterized in that: described polygon is triangle or square.
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| CN103691005B (en) * | 2013-12-24 | 2015-10-28 | 华东交通大学 | A kind of micro--Na fibrous tissue engineering rack and preparation method thereof |
| CN104958782B (en) * | 2015-06-08 | 2018-01-02 | 东华大学 | A kind of bacterial cellulose porous foam material and preparation method thereof |
| CN111001951B (en) * | 2019-12-26 | 2022-02-18 | 上海百心安生物技术股份有限公司 | Vascular stent structure beneficial to wall adhesion and processing device and method thereof |
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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 |
| CN102078642A (en) * | 2011-01-19 | 2011-06-01 | 北京大学第三医院 | Articular cartilage restoration and regeneration stent and preparation method thereof |
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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 bacteria cellulose sponges |
| CN102078642A (en) * | 2011-01-19 | 2011-06-01 | 北京大学第三医院 | Articular cartilage restoration and regeneration stent and preparation method thereof |
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