CN105624832A - Method for preparing calcium alginate microfibers having bat structure based on micro-fluidic technology - Google Patents
Method for preparing calcium alginate microfibers having bat structure based on micro-fluidic technology Download PDFInfo
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Abstract
The present invention provides a method for preparing calcium alginate microfibers having a bat structure based on a micro-fluidic technology. According to the method, a liquid droplet micro-fluidic device is used to produce a lot of oil phase liquid droplets having uniform size, and the oil phase liquid droplets are aligned in a continuous phase sodium alginate solution in a liner manner; and the sodium alginate solution containing the liquid droplets is continuously introduced into an iso propanol solution through a chip outlet, the sodium alginate solute is precipitated to form sodium alginate fibers, the sodium alginate fibers are immersed in calcium chloride, and through an ion chelation effect, the bat structure calcium alginate microfibers with the uniform size oil phase liquid droplets inside are formed. According to the present invention, with the liquid droplet micro-fluidic device, the high control of the alignment, the distance and the size of the ball type structure inside the calcium alginate microfibers can be achieved, the high control of the diameter of the bat type structure of the calcium alginate microfibers can be achieved, and the high maintenance property and the swelling resistance of the calcium alginate microfibers having the bat structure can be achieved.
Description
Technical field
The invention belongs to the crossing domain of new material technology and microflow control technique, a kind of method being specifically related to calcium alginate microfibre based on microflow control technique preparation tool bat structure.
Background technology
The present invention is with microflow control technique for relying on. Micro-fluidic chip is a kind of at minim channel or the system of manipulation micro volume fluid flowing in building, and wherein the yardstick of passage and component is tens to hundreds of micron. Each operating unit of micro-fluidic chip is connected each other by the flowing of fluid in microchannel network, and fluid flows in a certain way in this microchannel network, and micro-fluidic chip just can realize specific function on the whole. Microfluidic chip technology is widely used in biology, chemistry, the sample preparation of medical analysis process, reaction, separation, detection etc. In recent years along with the development of material science, micro-nano process technology, microfluidic chip technology also achieves breakthrough at micron order Material Field. Highgrade integration and homogeneity due to micro-fluidic chip so that microsize, pattern are various, the preparation of height homogeneous material structure is achieved, and possess very big development prospect at biomedical and Material Field.
Fiber refers to the material being made up of continuous or discrete filament, has elastic modelling quantity big, and plastic deformation is little, intensity high. The multilevel hierarchy (blood vessel, muscle fiber, nerve tract, hepatic cords etc.) of human body is all made up of fibre structure, and therefore fiber plays an important role maintaining organizational aspects. The preparation method of current fibrous material is many based on spinning with microflow control technique. Spinning can prepare submicron order, even nano-scale fiber, the single-size solid fibers silk mostly being one-component that polymer solution extrudes through spinning head pore, forms variously-shaped fibrous material by regulating receiver path, and the fibrous material formed is to be piled up molding by many cellosilks; Microflow control technique one-time formed can prepare difform Sub-micro Fibers, can prepare the fibre structure of porous, hollow and complexity, it may be achieved the preparation of one pack system or multi-component fiber. Microflow control technique, with its Highgrade integration, small but excellent thin platform advantage, is occupied an leading position in the research of fibrous material.
Sodium alginate is similar to the glycosaminoglycans in extracellular matrix, possesses good biocompatibility. Sodium alginate is easily combined with some bivalent cations, forms gel, sodium alginate and Ca2+Chelating, it is possible to form stable calcium alginate gel. Due to the collosol and gel character that sodium alginate is gentle so that microflow control technique is prepared calcium alginate fibre and is possibly realized. At present, the preparation of different size calcium alginate fibre can be realized by microflow control technique, it may be achieved the preparation of hollow calcium alginate fibre, can as the carrier of release or embedding medicinal, albumen and cell, micro structure in analogue body. Microflow control technique combines with biomaterial, it may be achieved the preparation of various micro structures, has very big development prospect in biomedical and field of tissue engineering technology.
Summary of the invention
A kind of method that it is an object of the invention to provide calcium alginate microfibre based on microflow control technique preparation tool bat structure, preparation process of the present invention is stable, and product morphology is homogeneous controlled, simple to operate.
The present invention adopts drop micro fluidic device to produce uniform oil phase drop, and in linear array in continuous phase sodium alginate soln. It is passed into forgiving the sodium alginate soln that there the is oil phase drop liquid spout by micro fluidic device continuously in isopropanol, the polarity (3.9) of isopropanol is more weak than water (9.0), at low polar solvent ionic polymer owing to the captivation effect of ion dipole is susceptible to assemble, therefore, sodium alginate soln passes in aqueous isopropanol, occurs dehydration to assemble, and sodium alginate solute precipitates out and solidifies, form sodium alginate solid fiber, and present the bat form of series connection; The sodium alginate fiber of solidification is immersed in high calcium chloride concentration solution and makes sodium alginate generate stable calcium alginate fibre with calcium ion chelating. Oil phase drop of the present invention is water insoluble also liquid insoluble in isopropanol, and after sodium alginate soln is passed in isopropanol, oil phase drop is fixed in sodium alginate fiber, and is linear array along fiber. Drop micro fluidic device of the present invention can generate oil phase drop, by regulating droplet distribution phase and continuous phase flow velocity, controls the spacing between oil phase drop, makes between each drop interval fixed range or makes each drop closely adjacent; Droplet distribution phase flow rates is 0.5ul/min-2ul/min, and continuous phase flow rates is 3ul/min-10ul/min; The oil phase dispersion phase unit of drop micro fluidic device can suitably increase and decrease, to increase or to reduce oil phase dispersion phase kind.
The formation of the bat form of series connection of the present invention, it is owing to the oil phase liquid-drop diameter in sodium alginate soln is slightly less than the diameter of the sodium alginate soln liquid stream by the ejection of micro fluidic device liquid spout, after sodium alginate soln is passed in isopropanol by spout, precipitate out the sodium alginate fiber solidified and there occurs violent contraction relative to sodium alginate soln liquid stream in diametric(al), the sodium alginate soln of variable concentrations, shrinkage degree is different, and concentration is more little, shrinks more violent; At the fiber position not having oil phase drop occupy-place, defining thin " rod " form, at the fiber position having oil phase drop occupy-place, define thick " ball " form, " ball " " rod " spacing structure arranges, and forms the sodium alginate fiber with bat structure. The sodium alginate bat fiber of formation is immersed in calcium chloride solution by the present invention, fiber is made to be changed into calcium alginate fibre, fibre morphology structure can be made more stable, it is prevented that when in fiber submerged in water or saline solution, occur fiber swelling set even to become solution.
Accompanying drawing explanation
The micro-fluidic single unit system schematic diagram of Fig. 1 drop.
Fig. 2 drop micro-fluidic chip schematic diagram.
Wherein 1 micro-fluidic chip, 2 delivery hoses, 3 sodium alginate soln liquid spouts, 4 oil are as drop, and 5 aqueous isopropanols, 11 sodium alginate soln entrances, 12 oil phase entrances, 13 chips export, 14 oil phase drop formation places.
Fig. 3 generates the principle schematic of tool bat structure calcium alginate microfibre.
Detailed description of the invention:
Embodiment 1
Include the preparation of the calcium alginate microfibre of the tool bat structure of single-size drop.
Mineral oil passes into chip liquid drop dispersion phase entrance, droplet distribution phase and continuous phase flow velocity be respectively set to:
A. the flow velocity that droplet distribution phase entrance is corresponding is 0.5ul/min, and the flow velocity of continuous phase and cross folder circulation road sodium alginate soln is 3ul/min;
B. the flow velocity that droplet distribution phase entrance is corresponding is 1ul/min, and the flow velocity of continuous phase and cross folder circulation road sodium alginate soln is 6ul/min;
C. the flow velocity that droplet distribution phase entrance is corresponding is 2ul/min, and the flow velocity of continuous phase and cross folder circulation road sodium alginate soln is 10ul/min;
Test the sodium alginate soln of working concentration respectively 1% (w/w), 2% (w/w), 4% (w/w), chip outlet is kept vertical with the receiving flask bottleneck containing aqueous isopropanol, chip exit is immersed under aqueous isopropanol liquid level, and outlet is wide, height is all 100um. By syringe pump pushing syringe, mineral oil is made to pass through T-shaped passage, form the drop of size uniformity, by regulating cross clevis circulation road sodium alginate soln continuous phase flow velocity, control spacing between drop, it is passed into forgiving the sodium alginate soln that there the is mineral oil liquid spout by micro fluidic device continuously in isopropanol, sodium alginate generation dehydration clustering phenomena, form sodium alginate fiber, fiber is immersed in 4% (w/w) calcium chloride, through ion sequestration, form the bat structure calcium alginate microfibre of the internal mineral oil drop comprising single-size. This experiment, by orthogonal test, finally prepares 9 groups of calcium alginate microfibres.
Embodiment 2
The cell of tool bat structure calcium alginate microfibre is cultivated and assay
Utilizing laboratory designed, designed the 9 groups of tool bat structure calcium alginate microfibres prepared, configuration is as shown in Figure 3. 9 groups of bat structure calcium alginate microfibres are carried out Gradient elution using ethanol (20%, 40%, 60%, 80%, 100%) respectively, it is soaked in 75% ethanol, ultraviolet sterilization 2h, containing 10% dual anti-PBS solution soaked overnight, after adopting NTx to carry out finishing, NSCs cell inoculated by bat structure calcium alginate microfibre, and inoculum density is 1 �� 106Liquid is changed after individual/mL, 24h cell attachment, after cell is cultivated 3 days, the relevant dyeing of the aspects such as cell life or death, apoptosis, cell proliferation, differentiation detection, ImagePro software analysis.
Claims (5)
1. the method based on the calcium alginate microfibre of microflow control technique preparation tool bat structure, it is characterised in that:
(1) drop micro fluidic device is utilized to produce uniform oil phase drop, and in linear array in continuous phase sodium alginate soln;
(2) it is passed into forgiving the sodium alginate soln that there the is oil phase drop liquid spout by micro fluidic device continuously in isopropanol, make sodium alginate soln dehydration, sodium alginate solute precipitates out and solidifies, and forms sodium alginate solid fiber, and presents the bat form of series connection;
(3) the sodium alginate fiber of solidification is immersed in high calcium chloride concentration solution and makes sodium alginate generate stable calcium alginate fibre with calcium ion chelating; The concentration of described sodium alginate soln is 1%-4% (w/w); The concentration of described calcium chloride solution is 4% (w/w).
2. based on the method for the calcium alginate microfibre of microflow control technique preparation tool bat structure described in claim 1, it is characterized in that: described oil phase drop is water insoluble also hydrophobic liquid (fluorinated oil insoluble in isopropanol, FC-40), after sodium alginate soln is passed in isopropanol, oil phase drop is fixed in sodium alginate fiber, and is linear array along fiber.
3. based on the method for the calcium alginate microfibre of microflow control technique preparation tool bat structure described in claim 1, it is characterized in that: described drop micro fluidic device can generate oil phase drop, by regulating droplet distribution phase and continuous phase flow velocity, control spacing between oil phase drop, make interval between each drop fix or make each drop closely adjacent; Droplet distribution phase flow rates is 0.5ul/min-2ul/min, and continuous phase flow rates is 3ul/min-10ul/min.
4. in accordance with the method for claim 1, it is characterized in that: the formation of the bat form of described series connection, it is owing to the oil phase liquid-drop diameter in sodium alginate soln is slightly less than the diameter of the sodium alginate soln liquid stream by the ejection of micro fluidic device liquid spout, after sodium alginate soln is passed in isopropanol by spout, precipitates out the sodium alginate fiber solidified and there occurs violent contraction relative to sodium alginate soln liquid stream in diametric(al); The sodium alginate soln of variable concentrations, shrinkage degree is different, and concentration is more little, shrinks more violent; At the fiber position not having oil phase drop occupy-place, defining thin " rod " form, at the fiber position having oil phase drop occupy-place, define thick " ball " form, " ball " " rod " spacing structure arranges, and forms the sodium alginate fiber with bat structure.
5. in accordance with the method for claim 1, it is characterized in that: the sodium alginate bat fiber of formation is immersed in calcium chloride solution, fiber is made to be changed into calcium alginate fibre, fibre morphology structure can be made more stable, prevent when in fiber submerged in water or saline solution, occur fiber swelling set even to become solution.
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| WO2017219305A1 (en) * | 2016-06-23 | 2017-12-28 | The University Of Hong Kong | Cavity-microfibers, methods and apparatuses for the fabrication of microfibers, threads, and filaments |
| CN108071007A (en) * | 2017-11-08 | 2018-05-25 | 韩金玲 | It is a kind of to prepare nano silver wire and graphene-based calcium alginate compounded conductive fiber method |
| CN108149334A (en) * | 2016-12-05 | 2018-06-12 | 中国科学院大连化学物理研究所 | The method and special chip of complicated form microfibre are prepared based on micro-fluidic chip |
| CN108360088A (en) * | 2018-02-28 | 2018-08-03 | 清华大学深圳研究生院 | The method and apparatus for preparing calcium alginate fibre |
| CN109652359A (en) * | 2017-10-12 | 2019-04-19 | 中国科学院大连化学物理研究所 | A kind of preparation method of the cell 3D culture hydrogel microsphere based on aqueous two-phase drop |
| CN111250009A (en) * | 2018-12-03 | 2020-06-09 | 成都市银隆新能源有限公司 | Method for preparing lithium ion battery material by using microfluidic technology |
| CN112513245A (en) * | 2018-08-10 | 2021-03-16 | 持田制药株式会社 | Alginic acid hollow microfiber |
| CN113355754A (en) * | 2021-07-07 | 2021-09-07 | 中国计量大学 | Method for preparing multiphase fiber based on microfluidic-electrostatic spinning technology |
| CN114981006B (en) * | 2020-02-03 | 2024-03-29 | 创技公司株式会社 | Microparticle production system including carrier liquid and control method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108149334A (en) * | 2016-12-05 | 2018-06-12 | 中国科学院大连化学物理研究所 | The method and special chip of complicated form microfibre are prepared based on micro-fluidic chip |
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| CN108071007A (en) * | 2017-11-08 | 2018-05-25 | 韩金玲 | It is a kind of to prepare nano silver wire and graphene-based calcium alginate compounded conductive fiber method |
| CN108360088A (en) * | 2018-02-28 | 2018-08-03 | 清华大学深圳研究生院 | The method and apparatus for preparing calcium alginate fibre |
| CN112513245A (en) * | 2018-08-10 | 2021-03-16 | 持田制药株式会社 | Alginic acid hollow microfiber |
| CN111250009A (en) * | 2018-12-03 | 2020-06-09 | 成都市银隆新能源有限公司 | Method for preparing lithium ion battery material by using microfluidic technology |
| CN114981006B (en) * | 2020-02-03 | 2024-03-29 | 创技公司株式会社 | Microparticle production system including carrier liquid and control method thereof |
| CN113355754A (en) * | 2021-07-07 | 2021-09-07 | 中国计量大学 | Method for preparing multiphase fiber based on microfluidic-electrostatic spinning technology |
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