CN103691005A - Micro-nanofiber tissue engineering scaffold and preparation method thereof - Google Patents
Micro-nanofiber tissue engineering scaffold and preparation method thereof Download PDFInfo
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- CN103691005A CN103691005A CN201310720956.4A CN201310720956A CN103691005A CN 103691005 A CN103691005 A CN 103691005A CN 201310720956 A CN201310720956 A CN 201310720956A CN 103691005 A CN103691005 A CN 103691005A
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Abstract
The invention relates to the technical field of biological materials, in particular to a micro-nanofiber tissue engineering scaffold and a preparation method thereof. The micro-nanofiber tissue engineering scaffold comprises microfibers, bacterial cellulose nanofibers, micropores and bacterial cellulose micropores, wherein the microfibers are 5-500 microns in diameter, the bacterial cellulose nanofibers are 10-100 nm in diameter, the micropores are 200-500 microns in diameter, and the bacterial cellulose micropores are 10-100 nm in diameter. Compared with the prior art, the micro-nanofiber tissue engineering scaffold is high in bionic degree, stable in structure and proper in porosity and aperture, and can provide a proper external environment for cells; the preparation method is simple and easy to implement, low in cost, wide in raw material source, environment-friendly and pollution-free.
Description
Technical field
The present invention relates to technical field of biological materials, specifically a kind of micro--Na fibrous tissue engineering rack and preparation method thereof.
Background technology
No matter be organizational project, or regenerative medicine, the performance of timbering material has determined final success or failure.From geometrical property, cells in vivo epimatrix (being designated hereinafter simply as ECM) is three-dimensional fibrous structure, it not only comprises micron hole and fiber, there is hole and the fiber of nanoscale simultaneously, be that ECM has multiple dimensioned fiber (diameter is 50-500 nanometer) and pore structure, the structure of these nanoscales and micrometer structure synergism, and then control the behavior of cell and the function of tissue and organ.Therefore, from bionics angle, consider, must, simultaneously from micron and nano-level, from the geometry structure of the natural ECM of multiple dimensioned imitation, design and manufacture the fibrous biomimetic scaffolds with suitable geometrical property.Therefore, researcher was prepared the support (hereinafter to be referred as micro--Na fibrous framework) being comprised of micron and nanofiber before the several years simultaneously.
In recent years, Many researchers utilizes wet spinning and electrostatic spinning technique to prepare bionical fibrous framework.But wet spinning technology cannot be prepared nanoscale fiber, and brace aperture is large; Electrostatic spinning technique can be prepared the fiber of submicron order, but is not easy the Nano grade (being less than 100 nanometers) that reaches real, is more difficult to prepare micro--Na fibre structure, and preparation method is complicated, cost is high, support intensity is low etc.What lot of documents was reported is the supporting structure of similar stratiform, and micrometer fibers layer and nanofiber layer are alternately laminated.Therefore, prepare and a kind ofly have that cost is low, intensity is high, and have good biocompatibility micro--Na fibrous framework becomes the hot subject of domestic and international biomaterial circle.
Bacterial cellulose is a kind of fibrous natural nano-material being synthesized by acetobacter xylinum, intensity is large, Young's modulus can reach 15-30 GPa, purity is high, degree of crystallinity is high, and retentiveness is high, and biocompatibility is good, its diameter, just in time between 10-100 nanometer, is desirable nano fibrous tissue engineering timbering material.Therefore, Bacterial cellulose nanofiber can become the nanofiber constituent element in micro--Na fibrous framework.But, Bacterial cellulose nanofiber and micrometer fibers are not combined into so far to the relevant public technology of micro--Na fibrous framework.
Summary of the invention
For above-mentioned prior art, the invention provides a kind of bionical degree high, supporting structure is stable, has suitable porosity and pore size, can be cell provide suitable external environment micro--Na fibrous tissue engineering rack; Its preparation method is simple, with low cost, and raw material sources are extensive, and preparation process environmental protection is pollution-free.
The present invention is achieved through the following technical solutions:
Micro--Na fibrous tissue engineering rack, comprises micrometer fibers, Bacterial cellulose nanofiber, micron hole and Bacterial cellulose nanoaperture; The diameter of described micrometer fibers is 5-500 micron, and the diameter of described Bacterial cellulose nanofiber is 10-100 nanometer; The diameter of described micron hole is 100-500 micron, and the diameter of described Bacterial cellulose nanoaperture is 10-100 nanometer.
The material of described micrometer fibers is the degradation material that biocompatibility is good, wherein a kind of in cellulose, collagen, gelatin, polylactic acid, chitosan, sodium alginate.
A preparation method for micro--Na fibrous tissue engineering rack, is characterized in that comprising the steps:
(1) prepare bacterial fermentation culture medium: take deionized water as solvent, the mass fraction of each solute is respectively: glucose 2.5%, yeast powder 0.75%, peptone 1.0%, Na2HPO41.0%, in beaker, stirring at room to solute dissolves completely, by dripping acetic acid adjusting medium pH, is 4-5;
(2) autoclaving: previously prepared good micrometer fibers support is put into culture medium prepared by step (1), then, to 115 ℃ of autoclavings of culture medium 30 minutes, take out culture medium and be cooled to room temperature under ultra violet lamp;
(3) inoculated bacteria: the bacteria culture of a certain amount of three ages in days of inoculation of medium after step (2) sterilizing;
(4) the preparation of micro--Na fibrous framework: the postvaccinal culture medium of step (3) is put into shaking table and implement the dynamic and stalic state alternate culture method, be first dynamic cultivation 30-60 minute, then static culture 60-120 minute, so repeatedly, during dynamic cultivation, the rotating speed of shaking table is 140-500rpm, and total incubation time is 24-168 hour; Culture medium temperature is 30-35 ℃, and medium pH is 4-5; Then take out support, first with 0.1 mol/L sodium hydroxide, boil and clean 20 minutes, then with washed with de-ionized water to solution, be neutral, by support liquid nitrogen freezing dry, obtain micro--Na fibrous tissue engineering rack.
The micron hole of described micrometer fibers support is greater than 300 microns, and described micrometer fibers support is made by wet spinning or electrospinning process.
The beneficial effect that the present invention brings is:
This preparation method is simple, with low cost, and raw material sources are extensive; Preparation process environmental protection, without any pollution, prepared micro--the bionical degree of Na fibrous tissue engineering rack is high, supporting structure is stable, has suitable porosity and pore size, can be cell suitable external environment is provided.The present invention combines wet spinning and electrostatic spinning technique with bacterial fibers nano-cellulose culture technique, first adopt traditional wet spinning or electrostatic spinning technique to prepare micrometer fibers support, then this micron is placed on and in the culture medium of Bacterial cellulose, carries out original position and cultivate altogether, the method of cultivating by the dynamic and stalic state, make the nanofiber of Bacterial cellulose through between micrometer fibers, prepare intercrossed network type micro--Na fibrous tissue engineering rack.Intercrossed network type prepared by the present invention is micro--and Na fibrous tissue engineering rack can overcome the defect of existing micro--Na fibrous tissue engineering rack, be conducive to adhesion, propagation and the differentiation of cell, for Growth of Cells provides suitable external environment, and preparation method is simple, for green manufacturing process, process controllability is strong.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the present invention is described in further detail.
Fig. 1 is the SEM photo of cellulose microfibers-Bacterial cellulose nano fiber scaffold;
Fig. 2 is the SEM photo of gelatin microfibre-Bacterial cellulose nano fiber scaffold.
The specific embodiment
Below by the specific embodiment, content of the present invention is described further, but these embodiment do not limit the scope of the invention:
Embodiment mono-:
As of the present invention micro--embodiment of Na fibrous tissue engineering rack, comprise micrometer fibers, Bacterial cellulose nanofiber, micron hole and Bacterial cellulose nanoaperture; The diameter of described micrometer fibers is 5 microns, and the diameter of described Bacterial cellulose nanofiber is 10 nanometers; The diameter of described micron hole is 100 microns, and the diameter of described Bacterial cellulose nanoaperture is 10 nanometers.
The cellulose of take is prepared the preparation method of micro--Na fibrous tissue engineering rack as microfibre support, cellulose microfibers support is cut into the square sheets that is of a size of 20 * 20 * 0.5mm, preparation acetobacter xylinum culture medium, take deionized water as solvent, the mass fraction of each solute is respectively: 2.5% glucose, 0.75% yeast powder, 1.0% peptone, 1.0%Na2HPO4, in beaker, stirring at room to solute dissolves completely, by dripping acetic acid, regulates medium pH=4; Cellulose microfibers support is put into culture medium, and sealing, then to 115 ℃ of autoclaving 30 min of culture medium, takes out culture medium and be cooled to room temperature under ultra violet lamp; The acetobacter xylinum strain of inoculation of medium 40 ml tri-ages in days after sterilizing; Postvaccinal culture medium is put into shaking table, under 30 ℃, 160 rpm rotating speeds, carry out static-dynamic state alternate culture: dynamic 30 minutes → static 60 minutes → dynamic 30 minutes → static 60 minutes, cultivate altogether after 30 hours and take out support, first with 0.1 mol/L sodium hydroxide, boil and clean 20 min, then with washed with de-ionized water to solution, be neutral, by support liquid nitrogen freezing dry, obtain micro--Na fibrous framework.
Fig. 1 be embodiment 1 take cellulose as microfibre support prepare micro--the SEM photo of Na fibrous framework.As can be seen from the figure, on cellulose microfibers support, deposited nanoscale Bacterial cellulose, and between micro--Na fiber, combination is good.
Embodiment bis-:
As of the present invention micro--embodiment of Na fibrous tissue engineering rack, be with the difference of embodiment mono-, in the present embodiment, the diameter of described micrometer fibers is 500 microns, the diameter of described Bacterial cellulose nanofiber is 100 nanometers; The diameter of described micron hole is 500 microns, and the diameter of described Bacterial cellulose nanoaperture is 100 nanometers.
The gelatin of take is prepared the preparation method of micro--Na fibrous tissue engineering rack as microfibre support, gelatin microfibre support is cut into the square sheets that is of a size of 20 * 20 * 2mm, preparation acetobacter xylinum culture medium (be take deionized water as solvent, the mass fraction of each solute is respectively: 2.5% glucose, 0.75% yeast powder, 1.0% peptone, 1.0%Na2HPO4), in beaker, stirring at room to solute dissolves completely, by dripping acetic acid, regulates medium pH=5; Cellulose microfibers support is put into culture medium, and sealing, then to 115 ℃ of autoclaving 30 min of culture medium, takes out culture medium and be cooled to room temperature under ultra violet lamp; The acetobacter xylinum strain of inoculation of medium 40 ml tri-ages in days after sterilizing; Postvaccinal culture medium is put into shaking table, under 30 ℃, 400 rpm rotating speeds, carry out static-dynamic state alternate culture: dynamic 60 minutes → static 60 minutes → dynamic 60 minutes → static 60 minutes, cultivate altogether after 48 hours and take out support, first with 0.1 mol/L sodium hydroxide, boil and clean 20 min, then with washed with de-ionized water to solution, be neutral, by support liquid nitrogen freezing dry, obtain micro--Na fibrous framework.
Fig. 2 be embodiment 2 take gelatin as microfibre support prepare micro--the SEM photo of Na fibrous framework.As can be seen from the figure, on gelatin microfibre support, deposited nanoscale Bacterial cellulose, and between micro--Na fiber, combination is good.
Embodiment tri-:
As of the present invention micro--embodiment of Na fibrous tissue engineering rack, be with the difference of embodiment mono-, in the present embodiment, the diameter of described micrometer fibers is 250 microns, the diameter of described Bacterial cellulose nanofiber is 50 nanometers; The diameter of described micron hole is 300 microns, and the diameter of described Bacterial cellulose nanoaperture is 60 nanometers.
(1) step in the preparation method of micro--Na fibrous tissue engineering rack prepares bacterial fermentation culture medium: take deionized water as solvent, the mass fraction of each solute is respectively: glucose 2.5%, yeast powder 0.75%, peptone 1.0%, Na2HPO41.0%, in beaker, stirring at room to solute dissolves completely, by dripping acetic acid adjusting medium pH, is 4.5.
Claims (4)
1. micro--Na fibrous tissue engineering rack, is characterized in that comprising micrometer fibers, Bacterial cellulose nanofiber, micron hole and Bacterial cellulose nanoaperture; The diameter of described micrometer fibers is 5-500 micron, and the diameter of described Bacterial cellulose nanofiber is 10-100 nanometer; The diameter of described micron hole is 100-500 micron, and the diameter of described Bacterial cellulose nanoaperture is 10-100 nanometer.
As claimed in claim 1 micro--Na fibrous tissue engineering rack, the material that it is characterized in that described micrometer fibers is the degradation material that biocompatibility is good, wherein a kind of in cellulose, collagen, gelatin, polylactic acid, chitosan, sodium alginate.
3. a preparation method for micro--Na fibrous tissue engineering rack, is characterized in that comprising the steps:
(1) prepare bacterial fermentation culture medium: take deionized water as solvent, the mass fraction of each solute is respectively: glucose 2.5%, yeast powder 0.75%, peptone 1.0%, Na2HPO41.0%, in beaker, stirring at room to solute dissolves completely, by dripping acetic acid adjusting medium pH, is 4-5;
(2) autoclaving: previously prepared good micrometer fibers support is put into culture medium prepared by step (1), then to 115 ℃ of autoclavings of culture medium 30 minutes, take out culture medium and be cooled to room temperature under ultra violet lamp;
(3) inoculated bacteria: the bacteria culture of a certain amount of three ages in days of inoculation of medium after step (2) sterilizing;
(4) preparation of micro--Na fibrous framework: the postvaccinal culture medium of step (3) is put into shaking table and implement the dynamic and stalic state alternate culture method, be first dynamic cultivation 30-60 minute, then static culture 60-120 minute, so repeatedly, during dynamic cultivation, the rotating speed of shaking table is 140-500rpm, and total incubation time is 24-168 hour; Culture medium temperature is 30-35 ℃, and medium pH is 4-5; Then take out support, first with 0.1 mol/L sodium hydroxide, boil and clean 20 minutes, then with washed with de-ionized water to solution, be neutral, by support liquid nitrogen freezing dry, obtain micro--Na fibrous tissue engineering rack.
As claimed in claim 3 micro--preparation method of Na fibrous tissue engineering rack, it is characterized in that a micron hole for described micrometer fibers support is greater than 300 microns, described micrometer fibers support is made by wet spinning or electrospinning process.
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| CN105536075A (en) * | 2016-02-27 | 2016-05-04 | 青岛大学 | Nano-micron fiber, chitosan and polylactic acid composite stent and making method thereof |
| CN105582573A (en) * | 2016-02-27 | 2016-05-18 | 青岛大学 | Nano-micron multi-scale chitosan three-dimensional scaffold and preparation method thereof |
| CN105641744A (en) * | 2016-02-27 | 2016-06-08 | 青岛大学 | Nano-micro multi-scale chitosan and polylactic acid composite scaffold and preparation method thereof |
| CN105664246A (en) * | 2016-02-27 | 2016-06-15 | 青岛大学 | Nano-micron multiscale tissue engineering composite three-dimension bracket and preparation method thereof |
| CN105664243A (en) * | 2016-02-27 | 2016-06-15 | 青岛大学 | Three-dimensional stent with polylactic acid nano-mircon fiber structure and preparation method thereof |
| CN107177048A (en) * | 2017-05-12 | 2017-09-19 | 陕西科技大学 | A kind of bacteria cellulose polymeric lactic acid compound film and preparation method thereof and load medicine gauze and preparation method based on the composite membrane |
| CN107569718A (en) * | 2017-08-30 | 2018-01-12 | 哈尔滨工业大学 | A kind of preparation method of 3D printing PLA and bacteria cellulose compound support frame material |
| CN108273130A (en) * | 2018-02-27 | 2018-07-13 | 华东交通大学 | A kind of three-D micro-nano fibrous composite scaffold and preparation method |
| CN108601860A (en) * | 2016-02-12 | 2018-09-28 | 金珂生物医疗公司 | Chitosan ultrafine fiber system |
| CN112746350A (en) * | 2020-12-17 | 2021-05-04 | 嘉兴学院 | Composite fiber with nanofiber surface modified and preparation method thereof |
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| CN105311683B (en) * | 2015-11-16 | 2019-01-04 | 清华大学 | A kind of network containing internal channel and the bionical tissue engineering bracket of directional pore structure and the preparation method and application thereof |
| CN105311683A (en) * | 2015-11-16 | 2016-02-10 | 清华大学 | Bionic tissue engineering scaffold containing inner channel network and oriented pore structure as well as preparation method and application of bionic tissue engineering scaffold |
| CN108601860A (en) * | 2016-02-12 | 2018-09-28 | 金珂生物医疗公司 | Chitosan ultrafine fiber system |
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| CN105641744A (en) * | 2016-02-27 | 2016-06-08 | 青岛大学 | Nano-micro multi-scale chitosan and polylactic acid composite scaffold and preparation method thereof |
| CN105664243A (en) * | 2016-02-27 | 2016-06-15 | 青岛大学 | Three-dimensional stent with polylactic acid nano-mircon fiber structure and preparation method thereof |
| CN105664246A (en) * | 2016-02-27 | 2016-06-15 | 青岛大学 | Nano-micron multiscale tissue engineering composite three-dimension bracket and preparation method thereof |
| CN105582573B (en) * | 2016-02-27 | 2018-10-26 | 青岛大学 | One kind receives multiple dimensioned chitosan three-dimensional holder of micron and preparation method thereof |
| CN105582573A (en) * | 2016-02-27 | 2016-05-18 | 青岛大学 | Nano-micron multi-scale chitosan three-dimensional scaffold and preparation method thereof |
| CN105536075A (en) * | 2016-02-27 | 2016-05-04 | 青岛大学 | Nano-micron fiber, chitosan and polylactic acid composite stent and making method thereof |
| CN107177048A (en) * | 2017-05-12 | 2017-09-19 | 陕西科技大学 | A kind of bacteria cellulose polymeric lactic acid compound film and preparation method thereof and load medicine gauze and preparation method based on the composite membrane |
| CN107177048B (en) * | 2017-05-12 | 2023-04-07 | 陕西科技大学 | Bacterial cellulose-polylactic acid composite membrane and preparation method thereof, and drug-loaded gauze based on composite membrane and preparation method thereof |
| CN107569718A (en) * | 2017-08-30 | 2018-01-12 | 哈尔滨工业大学 | A kind of preparation method of 3D printing PLA and bacteria cellulose compound support frame material |
| CN108273130A (en) * | 2018-02-27 | 2018-07-13 | 华东交通大学 | A kind of three-D micro-nano fibrous composite scaffold and preparation method |
| CN112746350A (en) * | 2020-12-17 | 2021-05-04 | 嘉兴学院 | Composite fiber with nanofiber surface modified and preparation method thereof |
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