CN102995145B - Integrated microfluidic spinning chip and method for preparing regenerated silk fibroin by using same - Google Patents
Integrated microfluidic spinning chip and method for preparing regenerated silk fibroin by using same Download PDFInfo
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- CN102995145B CN102995145B CN201210402427.5A CN201210402427A CN102995145B CN 102995145 B CN102995145 B CN 102995145B CN 201210402427 A CN201210402427 A CN 201210402427A CN 102995145 B CN102995145 B CN 102995145B
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Abstract
The invention belongs to the technical field of microfluidic spinning, relating to an integrated microfluidic spinning chip and a method for preparing regenerated silk fibroin by using the same. The integrated microfluidic spinning chip is of a three-layer structure, wherein the upper layer and the lower layer are provided with polydimethylsiloxane membranes of grooves, and the grooves are adhered with a middle dialysis membrane to form an upper microfluidic channel and a lower microfluidic channel. The depths of the two microfluidic channels are the same, and the widths gradually become small. The method for preparing the regenerated silk fibroin disclosed by the invention comprises the following steps: injecting spinning liquid in the upper channel and the lower channel at room temperature; injecting the mixed liquid of water-absorbent resin and ion adjusting liquid in the lower channel; and after the spinning liquid is adjusted via ions, cut, stretched and concentrated, extruding from an outlet, then solidifying in air as threads, and then winding the threads on rolls. The microfluidic chip disclosed by the invention can be used for simulating complex flow fields during the biological spinning process, and realizing the operations of the spanning liquid, such as cutting, stretching, component adjusting and concentrating. Compared with the spinning technology using a dry method, the method disclosed by the invention can be used for preparing good regenerated silk fibroin.
Description
Technical field
The invention belongs to bionics, microfluid spinning field, relate to a kind of integrated microfluidic spinning chip and prepare the method for regenerated silk fibroin fiber; Particularly relate to and a kind ofly in same chip, realize the integrated microfluid dry method spinning regenerated silk fibroin fiber process of the several functions such as shear tension to spinning solution, composition regulation, concentrated and spinning.
Background technology
The organism such as spider or silkworm can be solvent with water at normal temperatures and pressures, the azelon of obtained excellent in mechanical performance, and whole process power consumption is low, and does not have the use of organic solvent, environmentally friendly.So bionical spinning research is produced significant to industrial fiber.
Although the comprehensive mechanical property of spider dragline silk is best, because the silk of spider and generation thereof is of a great variety, be difficult to a large amount of acquisition, bring very large restriction to the bionics fiber of spider silk.Compared to spider silk be difficult to obtain, silk before several thousand just use by people, and its mechanical property is close with spider dragline silk with amino acid sequence; And the just medium research of Shao shows, the spinning technique optimizing silk obviously can improve its mechanical property, so, people then bionics fiber is carried out to silkworm.
Research shows, organism is in spinning process, and the metal ion content of spinning solution changes, pH reduces, concentration increases gradually; The tensile shear effect that spinning solution is subject to when flowing through some special area of biological spinner, makes the conformation of animal silk-fibroin be changed to water-fast beta sheet by random coil and/or alpha-helix conformation.So the spinning process of organism is a unified coordination process being integrated with the several functions such as composition regulation, tensile shear, pH reduction and protein concentration.And with fibroin be at present the artificial spinning research of raw material more be wet spinning, dry-spinning and Static Spinning.Although these spinning process technical maturities, because device is simple, bionical degree is lower, and comparatively natural silk is poor for the azelon mechanical property prepared.Micro-fluidic technologies, due to its special character, as solution presentation layer properties of flow, channels designs etc. controlled flexibly in the channel, and causes the extensive concern of people.Micro-fluidic technologies is applied in spinning field, the unapproachable condition of traditional spinning technique can be realized.Such as, patent WO2007/141131 (A1) and US2010029553 (A1) utilizes laminar flow diffusion principle, devises a kind of micro-fluid chip, achieves the adjustment of pH to regenerated silk fibroin solution and concentration of metal ions.Number of patent application is the patent of 201019063008.1 and 201010581835.2, is combined by micro-fluid chip with Static Spinning technique, has prepared regenerated silk fibroin nanofiber.Application number is the patent of 201110007230.7, by the spinning process that micro-fluid chip combines with wet spinning, dry-spinning and Static Spinning, is applied in regenerated silk fibroin solution system further, prepares the regeneration animal silk of excellent performance.The patent No. is the patent of ZL201110007232.6, devise the micro-fluid chip with shear tension function, be combined with dry-spinning, be applied in regenerated silk fibroin solution system, achieve the shear tension to spinning solution, but compared with the multiple effect of biological spinning process, function singleness, bionical degree are low.
Although, current micro-fluidic technologies has certain development in bionical spinning field, but compared with biological spinning process, microfluid spinning chip is too simple, function singleness still, in same chip, not yet realize the several functions such as the shear tension to spinning solution, composition regulation, concentrated and spinning, also have very large gap with real bionical spinning.
Summary of the invention
The present invention with the biological spinning process of spider or silkworm for starting point, devise complicated microfluidic channel, the complex flowfield of spinning solution in simulation organism, and adopt multilayer micro-fluidic technologies and dry-spinning process, achieve the multifunctional unit of chip, bionical degree improves greatly; Under room temperature with the regenerated silk fibroin water solution of 5 ~ 50% for spinning solution, through the ion regulation of micro-fluid chip, shear tension and protein concentration, obtain the regeneration animal wire protein fiber of excellent performance.
The object of the present invention is to provide and a kind ofly simulate the microfluid spinning chip of biological spinning process and the spinning process for regenerated silk fibroin solution thereof.
A kind of integrated microfluidic spinning chip of the present invention, described integrated microfluidic spinning chip is a kind of three-decker, comprises upper and lower two-layer dimethyl silicone polymer (PDMS) diaphragm and the dialysis membrane between upper and lower two-layer PDMS diaphragm; Described upper and lower two-layer PDMS diaphragm there is corresponding groove, fitting and form upper strata microfluidic channel in the groove of upper strata PDMS diaphragm and a face of described dialysis membrane, fits and form lower floor's microfluidic channel in the groove of lower floor PDMS diaphragm and another face of described dialysis membrane; The slot opening of upper strata PDMS diaphragm is relative with the slot opening of lower floor PDMS diaphragm.
Described upper strata microfluidic channel, is divided into successively according to function and regulates section, stretch section and shearing section; The width of described upper strata microfluidic channel changes from the inlet to the outlet, and wherein, regulate the width of section and shearing section constant, the width of stretch section diminishes gradually; The adjustment section of described lower floor's microfluidic channel, stretch section and the horizontal cross-sectional shape of shearing section and the horizontal cross-sectional shape of upper strata microfluidic channel are mirror; The deep equality of described upper and lower two-layer microfluidic channel whole process; During spinning, solution is all flow to from the adjustment section of passage, shears section and flows out.
As preferred technical scheme:
A kind of integrated microfluidic spinning chip as above, described microfluidic channel is vertical or horizontal snakelike arrangement on PDMS diaphragm; The snakelike arrangement of described longitudinal direction refers to that the longitudinally vertical aligning part of passage is parallel to the minor face of PDMS diaphragm; The snakelike arrangement of described transverse direction refers to that the horizontal part of passage is parallel to the long limit of PDMS diaphragm;
A kind of integrated microfluidic spinning chip as above, the passage of the snakelike arrangement of described longitudinal direction, it regulates section and shears section and has 4 ~ 6 bendings, the vertical portion length of each serpentine channel is 7 ~ 14mm, connected by semi arch between each Vertical Channel, half diameter of a circle is 3 ~ 10mm, and the outlet of shearing section is positioned at the center of PDMS diaphragm minor face; The stretch section of described microfluidic channel, the line of symmetry of its passage and the long limit angle of PDMS diaphragm are 20 ~ 60 °, and the length of described stretch section is 15 ~ 30mm; The two sides of the described stretch section passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line.
A kind of integrated microfluidic spinning chip as above, the passage of the snakelike arrangement of described transverse direction, it regulates section and shears section and has 1 ~ 3 bending, the horizontal component length of each serpentine channel is 50 ~ 80mm, connected by semi arch between each horizontal channel, half diameter of a circle is 4 ~ 12mm, and the center 5 ~ 8mm of PDMS diaphragm minor face is departed from the exit position of shearing section; The stretch section of described microfluidic channel, is positioned at the horizontal component of passage, parallel with the long limit of PDMS diaphragm; The length of described stretch section is 15 ~ 30mm; The two sides of the described stretch section passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line.
A kind of integrated microfluidic spinning chip as above, described adjustment section is the widest one section of passage of width, and width is 1 ~ 2.5mm, and length is 60 ~ 100mm; Described shearing section is the narrowest one section of passage of width, and width is 100 ~ 500 μm, and length is 20 ~ 150mm.
A kind of integrated microfluidic spinning chip as above, the shape of cross section of described groove is rectangle; Described adjustment section, between stretch section and shearing section by circular arc or directly connect.
A kind of integrated microfluidic spinning chip as above, described dialysis membrane is molecular cut off is 5000 ~ 30000 daltonian regenerated cellulose dialysis membranes; The upper and lower two-layer PDMS diaphragm of described integrated microfluidic spinning chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by adhesive package laminating; Described adhesive is one in silica gel, epobond epoxyn or double faced adhesive tape or combination.
A kind of integrated microfluidic spinning chip as above prepares the method for regenerated silk fibroin fiber, take mass percent as the regenerated silk fibroin water solution of 5% ~ 50% be spinning solution, under room temperature, upper strata microfluidic channel injects spinning solution, and lower floor's microfluidic channel injects the mixed liquor of water-absorbing resin and ion-select electrode liquid; The tensile shear of the passage that spinning solution diminishes gradually through width and the concentrated of lower channel solution and ion-select electrode effect, silk is solidified in atmosphere and the top roll that reels after outlet is extruded, obtained regenerated silk fibroin fiber, the fracture strength of obtained regenerated silk fibroin fiber is 20 ~ 100MPa, and elongation at break is 1 ~ 10%; In outlet, the solution of lower channel is collected simultaneously.
A kind of method as above, described ion-select electrode liquid is CaCl
2the aqueous solution; Described water-absorbing resin is molecular weight is 10000 ~ 40000 daltonian polyethylene glycol, wherein Ca in mixed liquor
2+concentration be 80 ~ 120g/L, Polyethylene glycol is 70 ~ 600g/L.
A kind of method as above, the air gap exporting to winding up roller of described micro-fluid chip upper strata microfluidic channel is 5 ~ 30cm, and winding speed is 2 ~ 10cm/s.
A kind of method as above, described spinning solution, ion-select electrode liquid and water-absorbing resin inject by micro-injection pump.
A kind of method as above, the diameter of described regenerated silk fibroin fiber is 7 ~ 30 μm; Described regenerated silk fibroin fiber is long filament or staple fibre.
The operating procedure of said method is as follows:
(1) preparation method of the micro-fluid chip of the present invention's proposition, with slide (75 × 25mm) for substrate, carries out uv-exposure by SU-8 photoresist coating under the mask being printed on microchannel pattern.Just can obtain the formpiston of microchannel pattern after development, then at this mould top casting PDMS prepolymer and curing agent thereof, after solidification, just can obtain the PDMS diaphragm being printed on microchannel pattern.Finally, bilevel PDMS diaphragm and dialysis membrane are fitted by adhesive package after Cement Composite Treated by Plasma.
(2) silk cocoon is used 0.5%(mass volume ratio) Na
2cO
3after coming unstuck, be dissolved in 9.0mol/L BrLi solution, then spin dialysis, and carry out preliminary concentrated after, obtain final spinning solution;
(3) by polyethylene glycol and CaCl
22H
2o is dissolved in a certain amount of deionized water in required ratio, obtains the concentrate of certain concentration and the mixed liquor of ion-select electrode liquid;
(4) with low concentration regenerated silk fibroin water solution for spinning solution, at room temperature, spinning solution is injected the upper channel of micro-fluid chip, lower channel then injects the mixed liquor of water-absorbing resin and ion-select electrode liquid, extrude through outlet after spinning solution flows through the passage of chip, be solidified into silk in atmosphere and the top roll that reels.
Beneficial effect
(1) micro-fluid chip prepared by, according to the style characteristic of spider or silkworm body of gland, simulate the complex flowfield of spinning solution in organism, achieve the composition regulation to spinning solution, shear tension and concentrated, achieving multifunctional unit, is a kind of three-dimensional multi-functional integrated microfluidic chip;
(2) can directly with low concentration regenerated silk fibroin water solution for spinning solution, do not need the process such as further concentrated, the pH in early stage and the adjustment of metal ion, and avoid the use of organic solvent, effectively simplify operating procedure, reduce production cost;
(3) micro-fluidic technologies is combined with dry-spinning process, improve spinning technique, improve spinning effect, further increase bionical spinning degree.
Accompanying drawing explanation
Fig. 1 is micro-fluid chip spinning technique figure
Fig. 2 is micro-fluid chip cross sectional representation
Fig. 3 is the upper strata microfluidic chip channel structural representation of laterally snakelike arrangement
Fig. 4 is lower floor's microfluidic chip channel structural representation of laterally snakelike arrangement
Fig. 5 is the upper strata microfluidic chip channel structural representation of longitudinally snakelike arrangement
Fig. 6 is lower floor's microfluidic chip channel structural representation of longitudinally snakelike arrangement
1. the adjustment section of passage
2. the stretch section of passage
3. the shearing section of passage
4. upper strata PDMS diaphragm
5. dialysis membrane
6. lower floor PDMS diaphragm
The mixed solution of the 7.PEG aqueous solution and ion-select electrode liquid
8. regenerated silk fibroin solution
9. micro-injection pump
10. winding up roller
11. solution collection devices
12. upper strata microfluidic channel
13. lower floor's microfluidic channel
Detailed description of the invention
Below in conjunction with detailed description of the invention, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
A kind of integrated microfluidic spinning chip of the present invention, described integrated microfluidic spinning chip is a kind of three-decker, comprises upper and lower two-layer PDMS diaphragm 4,6 and the dialysis membrane 5 between upper and lower two-layer PDMS diaphragm 4,6; Described upper and lower two-layer PDMS diaphragm there is corresponding groove; The groove of upper strata PDMS diaphragm 4 and described dialysis membrane 5 are fitted and are formed upper strata microfluidic channel 12, and the groove of lower floor PDMS diaphragm 6 and described dialysis membrane 5 are fitted and formed lower floor's microfluidic channel 13; The slot opening of upper strata PDMS diaphragm 4 is relative with the slot opening of lower floor PDMS diaphragm 6.
Described upper strata microfluidic channel 12, is divided into successively according to function and regulates section 1, stretch section 2 and shear section 3.The width of described upper strata microfluidic channel 12 changes from the inlet to the outlet, and wherein, regulate the width of section 1 and shearing section 3 constant, the width of stretch section 2 diminishes gradually; The adjustment section 1 of described lower floor's microfluidic channel 13, stretch section 2 and shear the horizontal cross-sectional shape of section 3 and the horizontal cross-sectional shape of upper strata microfluidic channel 12 is mirror; The deep equality of described upper and lower two-layer microfluidic channel 12,13 whole process; During spinning, solution 7,8 is all flow to by the adjustment section 1 of micro-injection pump 9 from passage, shears section 3 and flows out.
A kind of integrated microfluidic spinning chip as above, described microfluidic channel is vertical or horizontal snakelike arrangement on PDMS diaphragm; The snakelike arrangement of transverse direction described in Fig. 3, Fig. 4 refers to that the horizontal part of passage is parallel to the long limit of PDMS diaphragm; The snakelike arrangement of longitudinal direction described in Fig. 5, Fig. 6 refers to that the longitudinally vertical aligning part of passage is parallel to the minor face of PDMS diaphragm.
A kind of integrated microfluidic spinning chip as above, the passage of the snakelike arrangement of the transverse direction described in Fig. 3, Fig. 4, it regulates section 1 and shears section 3 and has 1 ~ 3 bending, the horizontal component length of each serpentine channel is 50 ~ 80mm, connected by semi arch between each horizontal channel, half diameter of a circle is 4 ~ 12mm, and the center 5 ~ 8mm of PDMS diaphragm minor face is departed from the exit position of shearing section 3; The stretch section 2 of described microfluidic channel, is positioned at the horizontal component of passage, parallel with the long limit of PDMS diaphragm; The length of described stretch section 2 is 15 ~ 30mm; The two sides of the described stretch section 2 passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line.
A kind of integrated microfluidic spinning chip as above, the passage of the snakelike arrangement of the longitudinal direction described in Fig. 5, Fig. 6, it regulates section 1 and shears section 3 and has 4 ~ 6 bendings, the vertical portion length of each serpentine channel is 7 ~ 14mm, connected by semi arch between each Vertical Channel, half diameter of a circle is 3 ~ 10mm, and the outlet of shearing section 3 is positioned at the center of PDMS diaphragm minor face; The stretch section 2 of described microfluidic channel, the line of symmetry of its passage and the long limit angle of PDMS diaphragm are 20 ~ 60 °, and the length of described stretch section 2 is 15 ~ 30mm; The two sides of the described stretch section 2 passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line.
A kind of integrated microfluidic spinning chip as above, described adjustment section 1 is the widest one section of passage of width, and width is 1 ~ 2.5mm, and length is 60 ~ 100mm.
A kind of integrated microfluidic spinning chip as above, described shearing section 3 is the narrowest one section of passage of width, and width is 100 ~ 500 μm, and length is 20 ~ 150mm.
A kind of integrated microfluidic spinning chip as above, the shape of cross section of described upper strata PDMS diaphragm 4 and the groove of lower floor PDMS diaphragm 6 is rectangle; Described adjustment section 1, stretch section 2 and shear between section 3 and be by circular arc or directly connect.
A kind of integrated microfluidic spinning chip as above, described dialysis membrane 5 is molecular cut offs is 5000 ~ 30000 daltonian regenerated cellulose dialysis membranes.
A kind of integrated microfluidic spinning chip as above, the upper strata PDMS diaphragm 4 of described integrated microfluidic spinning chip, lower floor PDMS diaphragm 6 and the dialysis membrane 5 between upper and lower two-layer PDMS diaphragm 4,6 are formed by adhesive package laminating; Described adhesive is one in silica gel, epobond epoxyn or double faced adhesive tape or combination.
A kind of integrated microfluidic spinning chip as above prepares the method for regenerated silk fibroin fiber, take mass percent as the regenerated silk fibroin water solution of 5% ~ 50% be spinning solution 8, under room temperature, upper strata microfluidic channel 12 injects spinning solution 8, and lower floor's microfluidic channel 13 injects the mixed liquor 7 of the water-absorbing resin aqueous solution and ion-select electrode liquid; The tensile shear of the stretch section 2 that spinning solution diminishes gradually through width and the concentrated of lower channel solution 7 and ion-select electrode effect, silk is solidified in atmosphere and the top roll 11 that reels after outlet is extruded, obtained regenerated silk fibroin fiber, the fracture strength of obtained regenerated silk fibroin fiber is 20 ~ 100MPa, and elongation at break is 1 ~ 10%; The solution 7 simultaneously flowed out at outlet gathering-device 11 pairs of lower channel 13 is collected.
A kind of method as above, the ion-select electrode liquid of described lower channel 13 and the ion-select electrode liquid of water-absorbing resin mixed aqueous solution 7 are CaCl
2, water-absorbing resin is molecular weight is 10000 ~ 40000 daltonian polyethylene glycol, wherein Ca in mixed liquor
2+concentration be 80 ~ 120g/L, Polyethylene glycol is 70 ~ 600g/L.
A kind of method as above, the air gap exporting to winding up roller 10 of described micro-fluid chip upper strata microfluidic channel 12 is 5 ~ 30cm, and winding up roller 10 winding speed is 2 ~ 10cm/s.
A kind of method as above, the mixed liquor 7 of described spinning solution 8, ion-select electrode liquid and water-absorbing resin injects by micro-injection pump 9.
A kind of method as above, the diameter of described regenerated silk fibroin fiber is 7 ~ 30 μm; Described regenerated silk fibroin fiber is long filament or staple fibre.
Embodiment 1
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The omnidistance deep equality of passage is 50 μm.The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.Passage is longitudinally snakelike arrangement on diaphragm: regulate section serpentine channel to have 3 bendings, width is 2mm, the long 7mm of vertical portion of its passage, and half diameter of a circle of sweep is 4mm; Shear section and have 3 bendings, width is 150 μm, the long 14mm of vertical portion of serpentine path, and half diameter of a circle of sweep is 3mm.The stretch section length of microfluidic channel is 15mm, is 20 ° with the long limit angle of the PDMS diaphragm of chip, and its two sides trace crossing with horizontal plane is first order exponential function curve, and exponential function formula is: R (x)=e
a*ln (x).The circular arc radian connecting each section of passage is 6.The mid point 8mm of PDMS diaphragm minor face is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by epoxy resin and silica adhesive encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 5% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 80g/L, and water-absorbent resin concentration is 600g/L, and molecular weight is polyethylene glycol (PEG-10000) aqueous solution of 10000; The molecular cut off of dialysis membrane is 5000, and spinning solution injects upper channel with the flow velocity of 5 μ L/min, and the PEG-10000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 10 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 5cm from the air-gap exporting to winding up roller, winding speed is 2cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 7 μm, and the intensity of described regenerated silk fibroin fiber is 20MPa, and elongation at break is 7%.Described regenerated silk fibroin fiber is staple fibre.
Embodiment 2
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.The deep equality of microfluidic channel whole process is 100 μm.Microfluidic channel is longitudinally snakelike arrangement on diaphragm: regulate section serpentine channel to have 2 bendings, width is equal is 2.5mm, the long 8mm of vertical portion of its passage, and half diameter of a circle of sweep is 10mm; Shear section and have 2 bendings, width is 200 μm, the long 10mm of vertical portion of serpentine path, and half diameter of a circle of sweep is 4mm.Stretch section length is 30mm, is 60 ° with the long limit angle of chip, and its two sides trace crossing with horizontal plane is bi-exponential function curve, and exponential function formula is R (x)=ae
bx+ ce
dx.The circular arc radian connecting each section of passage is 6.The mid point 8mm of PDMS diaphragm minor face is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by adhered by double sided plaster agent encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 40% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 95g/L, and water-absorbent resin concentration is 70g/L, and molecular weight is polyethylene glycol (PEG-20000) aqueous solution of 20000; The molecular cut off of dialysis membrane is 14000, and spinning solution injects upper channel with the flow velocity of 3 μ L/min, and the PEG-20000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 7 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 12cm from the air-gap exporting to winding up roller, winding speed is 6cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 20 μm, and the intensity of described regenerated silk fibroin fiber is 50MPa, and elongation at break is 8%.Described regenerated silk fibroin fiber is long filament.
Embodiment 3
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.The deep equality of microfluidic channel whole process is 85 μm.Microfluidic channel is longitudinally snakelike arrangement on diaphragm: regulate section serpentine channel to have 3 bendings, width is equal is 2.5mm, the long 7mm of vertical portion of its passage, and half diameter of a circle of sweep is 9mm; Shear section and have 1 bending, width is 300 μm, the long 10mm of vertical portion of serpentine path, and half diameter of a circle of sweep is 4mm.Stretch section length is 25mm, is 45 ° with the long limit angle of chip, and its two sides trace crossing with horizontal plane is straight line, and the function formula of straight line is R (x)=ax+b.The circular arc radian connecting each section of passage is 5.The mid point 6mm of PDMS diaphragm minor face is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by silica adhesive encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 35% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 90g/L, and water-absorbent resin concentration is 200g/L, and component is polyethylene glycol (PEG-20000) aqueous solution of 20000; The molecular cut off of dialysis membrane is 10000, and spinning solution injects upper channel with the flow velocity of 10 μ L/min, and the PEG-20000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 5 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 10cm from the air-gap exporting to winding up roller, winding speed is 5cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 18 μm, and the intensity of described regenerated silk fibroin fiber is 42MPa, and elongation at break is 10%.Described regenerated silk fibroin fiber is staple fibre.
Embodiment 4
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.The deep equality of microfluidic channel whole process is 80 μm.Microfluidic channel is laterally snakelike arrangement on diaphragm: whole serpentine channel only has 1 bending, and adjusting segment width equal is 2.0mm, the long 80mm of horizontal component of its passage, and half diameter of a circle of sweep is 5mm; Shearing section width is 100 μm, and length is 20mm, and half diameter of a circle of sweep is 12mm.The stretch section of microfluidic channel is horizontal, and its two sides trace crossing with horizontal plane is bi-exponential function curve and about the central axis symmetry of passage, exponential function formula is: R (x)=ae
bx+ ce
dx; Stretching segment length 20mm, the mid point 5mm of PDMS diaphragm minor face is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by silica adhesive encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 35% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 108g/L, and water-absorbent resin concentration is 300g/L, and molecular weight is polyethylene glycol (PEG-30000) aqueous solution of 30000; The molecular cut off of dialysis membrane is 14000, and spinning solution injects upper channel with the flow velocity of 15 μ L/min, and the PEG-30000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 15 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 16cm from the air-gap exporting to winding up roller, winding speed is 10cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 20 μm, and the intensity of described regenerated silk fibroin fiber is 100MPa, and elongation at break is 1%.Described regenerated silk fibroin fiber is long filament.
Embodiment 5
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.The deep equality of described microfluidic channel whole process is 80 μm, microfluidic channel is laterally snakelike arrangement on diaphragm: regulate section serpentine channel to have 1 bending, width is equal is 1.0mm, the long 60mm of horizontal component of its passage, and half diameter of a circle of sweep is 6mm; Shear section and have 2 bendings, width is 500 μm, the long 55mm of horizontal component of serpentine path, and half diameter of a circle of sweep is 4mm.The stretch section of microfluidic channel is horizontal, and its two sides trace crossing with horizontal plane is hyperbolic function curve, and hyperbolic function formula is: R (x)=1/2 (e
x-e
-x), the length of stretch section is 15mm; PDMS diaphragm minor face mid point 8mm is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by epobond epoxyn encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 50% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 120g/L, and molecular weight is the concentration of polyethylene glycol (PEG-40000) water-absorbing resin of 40000 is 100g/L; The molecular cut off of dialysis membrane is 30000, and spinning solution injects upper channel with the flow velocity of 5 μ L/min, and the PEG-40000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 10 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 30cm from the air-gap exporting to winding up roller, winding speed is 6cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 30 μm, and the intensity of described regenerated silk fibroin fiber is 60MPa, and elongation at break is 8%.Described regenerated silk fibroin fiber is long filament.
Embodiment 6
Micro-fluid chip of the present invention is a kind of three-decker, comprises the PDMS diaphragm of levels and middle dialysis rete, and fluted on PDMS diaphragm, reeded of upper and lower diaphragm and dialysis membrane are fitted and formed upper and lower two microfluidic channel; The horizontal cross-sectional shape of lower floor's microfluidic channel and the horizontal cross-sectional shape of upper channel are mirror.The deep equality of described microfluidic channel whole process is 100 μm, microfluidic channel is laterally snakelike arrangement on diaphragm: regulate section serpentine channel to have 1 bending, width is equal is 1.5mm, the long 50mm of horizontal component of its passage, and half diameter of a circle of sweep is 6mm; Shear section and have 2 bendings, width is 350 μm, the long 66mm of horizontal component of serpentine path, and half diameter of a circle of sweep is 4mm.The stretch section of microfluidic channel is horizontal, and its two sides trace crossing with horizontal plane is straight line, and the formula of straight line is: R (x)=ax+b, and the length of stretch section is 30mm; PDMS diaphragm minor face mid point 6mm is departed from the exit position of lower channel.The upper and lower two-layer PDMS diaphragm of chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by epoxy resin and double faced adhesive tape encapsulation laminating.
The preparation method of a kind of regenerated silk fibroin fiber of the present invention, at room temperature, the regenerated silk fibroin water solution with 40% is spinning solution, Ca in ion-select electrode liquid and water-absorbing resin mixed liquor
2+concentration is 105g/L, and molecular weight is the concentration of polyethylene glycol (PEG-30000) water-absorbing resin of 30000 is 200g/L; The molecular cut off of dialysis membrane is 20000, and spinning solution injects upper channel with the flow velocity of 6 μ L/min, and the PEG-30000 aqueous solution and ion-select electrode liquid mixed liquor inject lower channel with the speed of 10 μ L/min; Spinning solution is solidified into silk and the top roll that reels after being extruded by the outlet of microchannel in atmosphere; Be 20cm from the air-gap exporting to winding up roller, winding speed is 6cm/s.
Regenerated silk fibroin fibre diameter obtained by the present invention is 28 μm, and the intensity of described regenerated silk fibroin fiber is 70MPa, and elongation at break is 5%.Described regenerated silk fibroin fiber is staple fibre.
Claims (9)
1. an integrated microfluidic spinning chip, is characterized in that: described integrated microfluidic spinning chip is a kind of three-decker, comprises upper and lower two-layer PDMS diaphragm and the dialysis membrane between upper and lower two-layer PDMS diaphragm; Described upper and lower two-layer PDMS diaphragm there is corresponding groove; Fit and form upper strata microfluidic channel in the groove of upper strata PDMS diaphragm and a face of described dialysis membrane, the groove of lower floor PDMS diaphragm and the another side of described dialysis membrane are fitted and formed lower floor's microfluidic channel; The slot opening of upper strata PDMS diaphragm is relative with the slot opening of lower floor PDMS diaphragm;
Described upper strata microfluidic channel, is divided into successively according to function and regulates section, stretch section and shearing section; The width of described upper strata microfluidic channel changes from the inlet to the outlet, and wherein, regulate the width of section and shearing section constant, the width of stretch section diminishes gradually;
The adjustment section of described lower floor's microfluidic channel, stretch section and the horizontal cross-sectional shape of shearing section and the horizontal cross-sectional shape of upper strata microfluidic channel are mirror;
The deep equality of described upper and lower two-layer microfluidic channel whole process; During spinning, solution is all flow to from the adjustment section of passage, shears section and flows out;
Described microfluidic channel is vertical or horizontal snakelike arrangement on PDMS diaphragm; The snakelike arrangement of described longitudinal direction refers to that the longitudinally vertical aligning part of passage is parallel to the minor face of PDMS diaphragm; The snakelike arrangement of described transverse direction refers to that the horizontal part of passage is parallel to the long limit of PDMS diaphragm;
The passage of the snakelike arrangement of described longitudinal direction, it regulates section and shears section and has 4 ~ 6 bendings, and the vertical portion length of each serpentine channel is 7 ~ 14mm, is connected between each Vertical Channel by semi arch, half diameter of a circle is 3 ~ 10mm, and the outlet of shearing section is positioned at the center of PDMS diaphragm minor face; The stretch section of described microfluidic channel, the line of symmetry of its passage and the long limit angle of PDMS diaphragm are 20 ~ 60 °, and the length of described stretch section is 15 ~ 30mm; The two sides of the described stretch section passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line;
The passage of the snakelike arrangement of described transverse direction, it regulates section and shears section and has 1 ~ 3 bending, the horizontal component length of each serpentine channel is 50 ~ 80mm, connected by semi arch between each horizontal channel, half diameter of a circle is 4 ~ 12mm, and the center 5 ~ 8mm of PDMS diaphragm minor face is departed from the exit position of shearing section; The stretch section of described microfluidic channel, is positioned at the horizontal component of passage, parallel with the long limit of PDMS diaphragm; The length of described stretch section is 15 ~ 30mm; The two sides of the described stretch section passage trace crossing with horizontal plane is the one in exponential function, hyperbolic function or straight line.
2. a kind of integrated microfluidic spinning chip according to claim 1, is characterized in that, described adjustment section is the widest one section of passage of width, and width is 1 ~ 2.5mm, and length is 60 ~ 100mm; Described shearing section is the narrowest one section of passage of width, and width is 100 ~ 500 μm, and length is 20 ~ 150mm.
3. a kind of integrated microfluidic spinning chip according to claim 1, is characterized in that, the shape of cross section of described groove is rectangle, and the degree of depth of groove is 50 ~ 100 μm; Described adjustment section, between stretch section and shearing section by circular arc or directly connect.
4. a kind of integrated microfluidic spinning chip according to claim 1, is characterized in that, described dialysis membrane is molecular cut off is 5000 ~ 30000 daltonian regenerated cellulose dialysis membranes; The upper and lower two-layer PDMS diaphragm of described integrated microfluidic spinning chip and the dialysis membrane between upper and lower two-layer PDMS diaphragm are formed by adhesive package laminating; Described adhesive is one in silica gel, epobond epoxyn or double faced adhesive tape or combination.
5. any one integrated microfluidic spinning chip according to claims 1 to 4 prepares the method for regenerated silk fibroin fiber, it is characterized in that: take mass percent as the regenerated silk fibroin water solution of 5% ~ 50% be spinning solution, under room temperature, upper strata microfluidic channel injects spinning solution, and lower floor's microfluidic channel injects the mixed liquor of the water-absorbing resin aqueous solution and ion-select electrode liquid; The tensile shear of the passage that spinning solution diminishes gradually through width and the concentrated of lower channel solution and ion-select electrode effect, silk is solidified in atmosphere and the top roll that reels after outlet is extruded, obtained regenerated silk fibroin fiber, the fracture strength of obtained regenerated silk fibroin fiber is 20 ~ 100MPa, and elongation at break is 1 ~ 10%; In outlet, the solution of lower channel is collected simultaneously.
6. method according to claim 5, is characterized in that, described ion-select electrode liquid is CaCl
2the aqueous solution; Described water-absorbing resin is molecular weight is 10000 ~ 40000 daltonian polyethylene glycol, wherein Ca in mixed liquor
2+concentration be 80 ~ 120g/L, Polyethylene glycol is 70 ~ 600g/L.
7. method according to claim 5, is characterized in that, the air gap exporting to winding up roller of described micro-fluid chip upper strata microfluidic channel is 5 ~ 30cm, and winding speed is 2 ~ 10cm/s.
8. method according to claim 5, is characterized in that, described spinning solution, ion-select electrode liquid and water-absorbing resin inject by micro-injection pump.
9. method according to claim 5, is characterized in that, the diameter of described regenerated silk fibroin fiber is 7 ~ 30 μm; Described regenerated silk fibroin fiber is long filament or staple fibre.
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CN104947320B (en) * | 2015-05-28 | 2017-11-14 | 清华大学深圳研究生院 | A kind of bio-medical energy-saving and temperature-regulating tunica fibrosa and preparation method thereof |
CN106245134B (en) * | 2016-06-29 | 2018-05-25 | 东华大学 | A kind of spinning process that micro-fluid chip is used for recombinant spider silk proteins |
CN107597216A (en) * | 2017-03-17 | 2018-01-19 | 李芳芳 | A kind of method that excretion body is separated using micro-fluid chip |
CN110359103B (en) * | 2019-07-10 | 2020-09-08 | 西安交通大学 | Method for preparing polymer micro-nanofiber based on stepping stretching method |
CN113413936B (en) * | 2021-08-24 | 2021-11-30 | 北京寻因生物科技有限公司 | Flow regulation structure and microfluid chip |
CN115487360B (en) * | 2022-09-29 | 2023-12-22 | 南京工业大学 | Preparation method of sericin-based skin tissue scaffold |
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