CN105268021A - High-strength polypeptide hydrogel preparation method - Google Patents
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Abstract
The invention relates to a high-strength polypeptide hydrogel preparation method and belongs to the field of chemical preparation. The high-strength polypeptide hydrogel preparation method includes (1) selecting polypeptide molecules, which are provided with positive charges and capable of forming long fiber structures by self-assembly, to prepare an aqueous solution, mixing uniformly ultrasonically and standing so as to enable the aqueous solution to form the long fiber structures by self-assembly; (2) adding GO (graphene oxide) lamellas into the solution of the long fiber structures, mixing uniformly ultrasonically, regulating a pH (potential of hydrogen) value, standing and forming hybrid hydrogel after a system is aged and stabilizes. The high-strength polypeptide hydrogel preparation method has the advantages that the GO lamellas are introduced into the polypeptide hydrogel, and gelling capability of the system is improved, so that a high-strength polypeptide/GO hybrid hydrogel material is obtained; properties of mechanical strength, aperture size and the like of the polypeptide/GO hybrid hydrogel can be controlled by controlling the size, surface charge density and surface charge concentration of the GO lamellas.
Description
Technical field
The present invention relates to the preparation method of hydrogel, particularly relate to a kind of preparation method of high strength polypeptide hydrogel; Belong to chemical preparation field.
Background technology
Hydrogel take water as the gel rubber material of disperse medium, and the pore structure abundant due to it and tridimensional network can provide good permeability and mechanical support, therefore have a wide range of applications at numerous areas.In recent years, hydrogel material enjoys the concern of domestic and international scientist, has carried out large quantity research to it in biomedicine such as the application in organizational project, medicament transport etc.Common hydrogel can be obtained by the covalent cross-linking between the synthetic polymer of high molecular or between some micromolecule, but such crosslinking method needs to be undertaken by chemosynthesis.Because most chemical reagent is containing toxic, thus the hydrogel causing it to be formed does not have biological activity, yet non-degradable; Another kind method utilizes some micromolecule self can the characteristic of self assembly, forms network structure, further plastic under the physiological condition of gentleness by non-covalent interaction.Due to the method mild condition of the second synthesis, component units is optional controlled, and the glue therefore formed has the advantages such as good biocompatibility, reversibility, thus makes this method more and more cause the concern of people.
In the last few years, people were by the permutation and combination to amino acid residue, and the various not homotactic peptide molecule of design and synthesis, is regulated and controled by the molecular structure and property of exquisiteness, and a lot of peptide molecule self assembly can form stable hydrogel.Peptide molecule is as micromolecule species, and the formation of its hydrogel is generally that peptide molecule self assembly forms nanofiber, more three-dimensional cross-linkedly forms macroscopically hydrogel structure by fiber is further.This process is that this just makes peptide hydrogel have good Modulatory character and reversibility by various noncovalent interaction as electrostatic interaction, hydrogen bond, hydrophobic interaction, pi-pi accumulation etc. have driven.And because peptide molecule derives from organism at first, its basic structural unit is a-amino acid, has good biocompatibility, therefore can degrade in vivo, and metabolite is nontoxic.The hydrogel water content that self-assembling peptide is formed is very high, and the pore structure of this support and the similar of extracellular matrix, cell survival and movement can not only be made, somatomedin and nutrient etc. can also be made to spread discrepancy lentamente, be the desirable cell culture substrate that can control cell differentiation, and support the ideal material that cells grown is a complete tissue.The hydrogel material research formed based on self-assembling peptide in the last few years has become the study hotspot of Material Field gradually, and in nano material, the fields such as biomedicine and organizational project present wide application prospect.
But also just because of the formation of peptide molecule hydrogel be generally peptide molecule first self assembly form nanofiber, then by the further three-dimensional cross-linked hydrogel structure formed macroscopically of fiber.This process is formed based on noncovalent interaction, and gel-formation power is limited, and its interfibrous cross-link intensity is often lower, and cause its gel strength limited, self-supporting poor performance, is easily destroyed; And it is often less and be difficult to Effective Regulation that its fiber is cross-linked formed three-dimensional net structure aperture, which greatly limits its application in fields such as tissue engineering materials.
Summary of the invention
The object of this invention is to provide a kind of preparation method of high strength polypeptide hydrogel, for the preparation of hydrogel, and the hydrogel mechanical strength prepared is high, and simultaneously the mechanical strength of hydrogel and aperture size are adjustable.
Technical scheme of the present invention is:
A preparation method for high strength polypeptide hydrogel, comprises the following steps:
(1) select self assembly to form long fibre structure and positively charged peptide molecule, prepare the aqueous solution of described peptide molecule, concentration is 2mmol/L-10mmol/L; Ultrasonic mixing, leaves standstill, makes its self assembly be long fibre structure;
(2) in the long fibre structure-solution of step (1) gained, graphene oxide (GO) lamella is added, the concentration of GO lamella is made to be 0.02 ~ 1.0mg/mL, ultrasonic mixing, regulate the pH value of gained polypeptide/GO lamella mixed solution between 8-11, leave standstill, hybridized hydrogel is formed, namely described high strength polypeptide hydrogel after system ripening is stable.
In described step (1), described peptide molecule contains the positively charged amino acid residue of more than 1, preferred I
3k or K
3a
3i
3g
3v
3, especially preferably K
3a
3i
3g
3v
3, the two molecular structural formula is respectively:
I
3k molecular structural formula is:
K
3a
3i
3g
3v
3molecular structural formula is:
In described step (1), the preferred 3-5mmol/L of described peptide molecule concentration.
In described step (1), ultrasonic object is to solution mixing, as long as can reach the object of mixing; Preferably ultrasonic condition is: ultrasonic power is: 90-110W, and ultrasonic time is: 5-15min.
In described step (2), described GO lamella dimensional parameters is: length is 100-1500nm, and width is 100-1500nm, and thickness is 1-2nm; Preferred length is 300 ± 200nm, and width is 300 ± 200nm.
In described step (2), ultrasonic object is to solution mixing, as long as can reach the object of mixing; Preferably ultrasonic condition is: ultrasonic power is: 90-110W, and ultrasonic time is: 5-15min.
In described step (1), place 1-7 days under general room temperature, peptide molecule can self assembly be just long fibre structure; Whether long fibre structure is formed can be detected by transmission electron microscope (TEM).
In described step (2), regulate the pH value of gained polypeptide/GO lamella mixed solution can reduce peptide molecule electric charge, be conducive to plastic.Whether system ripening completes, and can be judged by the disappearance of solution mobility, if ripening completes, the bottle holding solution is inverted solution can not be flowed down; General leaving standstill just can complete for 5-8 hour in ripening.
GO lamella is graphite through strong acid oxidation preparation, and its length and width dimensions are difficult to control to very accurate scope, normally used size range centered by a certain size point, in left and right diffusion tendency; Such as length is that the GO lamella of 300 ± 200nm refers to that length concentrates near 300nm, and the lamella spread to the left and right in the size range of 200nm can accept (namely admissible size range is 100-500nm).The size of GO lamella is also normal to be selected according to carrying out adjustment to the demand of hydrogel aperture size size in reality use.
The thin slice of GO lamella to be thickness be 1-2nm, its lateral dimension can be tens nanometers to tens micrometer ranges, and its architectural feature makes it have polymer, colloid, thin film and amphiphilic characteristic, is a kind of excellent soft material with carbon element.GO lamella is generally oxidized through strong acid by graphite and obtains, and there are carboxyl, phenolic hydroxyl group and the charged group of epoxide group isopolarity in its surface and edge, have good dispersibility, and be easy to and other matter interaction in water.
Graphene oxide (GO) lamella is incorporated in polypeptide hydrogel by the present invention, space three-dimensional network structure is formed by the interaction of GO lamella and self-assembling polypeptide fiber, realization is to effective combination of hydrone and fix, form hydrogel structure, substantially increase the gel-formation power of system, obtain the peptide/GO hybridized hydrogel material of high strength.GO lamella has excellent mechanical property, its promoter as gel formation and cross-linking agent, greatly can improve the mechanical strength of hybridized hydrogel, and by can the mechanical strength of Effective Regulation hybridized hydrogel material and pore size to the adjustment of GO lamella size and surface charge density.
The present invention, by regulating size and the concentration of GO lamella, realizes the regulation and control to hybridized hydrogel mechanical strength, aperture size.
In general, in suitable GO lamella concentration range, GO lamella concentration is higher, and (G ") is larger, and the hydrogel mechanical property obtained is better for storage modulus (G') and Loss modulus; Described debita spissitudo scope refers to that adding of GO lamella can not cause the precipitation of system to produce, and GO lamella addition is too large, can produce precipitation, system phase-splitting, cannot form homogeneous hydrogel.
Different size GO lamella affects the mechanical property of hydrogel equally, when GO lamella addition is identical, along with GO lamella size increases, system mechanical properties decrease, in addition oversize GO lamella is difficult to homogeneous dispersion in the solution, causes the uniformity of hydrogel to decline.Different size GO lamella add the pore size that membership affects hydrogel.
Compared with prior art, beneficial effect of the present invention is:
(1) GO lamella is incorporated in polypeptid solution, by the interaction of GO lamella and self-assembling polypeptide fiber, the gel-formation power of system can be improved, obtain the polypeptide/GO hybridized hydrogel material of high strength;
(2) can control to regulate and control the character such as the mechanical strength of polypeptide/GO hybridized hydrogel and aperture size by the size of GO lamella and concentration.
Accompanying drawing explanation
Fig. 1 polypeptide/GO hybridized hydrogel preparation flow schematic diagram,
The rheometry result of Fig. 2 embodiment 3 hybridized hydrogel,
Transmission electron microscope (staining counter) picture of Fig. 3 embodiment 3 hybridized hydrogel.
Detailed description of the invention
Be explained in detail the present invention below in conjunction with embodiment and accompanying drawing, wherein Fig. 1 is polypeptide of the present invention/GO hybridized hydrogel preparation flow schematic diagram, presents preparation flow of the present invention briefly in figure.
Embodiment 1
(1) I is prepared
3k peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Leave standstill 3 days under room temperature, make its self assembly be long fibre structure, now system can not form hydrogel;
(2) in the above-mentioned solution containing polypeptide long fibre structure, adding length is 300 ± 200nm, width is 300 ± 200nm, thickness is the GO lamella of 1-2nm, GO lamella concentration is made to be 0.1mg/mL (namely every mL solution adds 0.1mgGO lamella), ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 12min; Employing concentration is that the NaOH solution of 0.5mol/L regulates mixed solution pH value to be 10.5, and room temperature leaves standstill 6 hours, forms hybridized hydrogel.
Embodiment 1 gained hydrogel storage modulus (G') can reach 4000Pa, and (G ") can reach 500Pa to Loss modulus, and gel aperture size is less than 400nm; The hybridized hydrogel of self-supporting can be formed by this enforcement preparation method.
Embodiment 2
(1) K is prepared
3a
3i
3g
3v
3peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Leave standstill 3 days under room temperature, make its self assembly be long fibre structure;
(2) in above-mentioned long fibre structure-solution, adding length is 300 ± 200nm, and width is 300 ± 200nm, and thickness is the GO lamella of 1-2nm, and make GO lamella concentration be 0.02mg/mL, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Employing concentration is that the NaOH solution of 0.5mol/L regulates mixed solution pH value to be 10.0, and room temperature leaves standstill 6 hours, forms hybridized hydrogel.
Embodiment 2 gained hydrogel storage modulus (G') can reach 5000Pa, and (G ") can reach 600Pa to Loss modulus, and gel aperture size is less than 500nm; The hybridized hydrogel of self-supporting can be formed by this enforcement preparation method.
Embodiment 3
(1) K is prepared
3a
3i
3g
3v
3peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Room temperature leaves standstill 3 days, makes its self assembly be long fibre structure;
(2) in above-mentioned long fibre structure-solution, adding length is 300 ± 200nm, and width is 300 ± 200nm, and thickness is the GO lamella of 1-2nm, and make GO lamella concentration be 0.1mg/mL, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Employing concentration is that the NaOH solution of 0.5mol/L regulates mixed solution pH value to be 10.0, and room temperature leaves standstill 6 hours, forms hybridized hydrogel.
Embodiment 3 gained hydrogel storage modulus (G') can reach 6000Pa, and (G ") can reach 1000Pa to Loss modulus, and gel aperture size is less than 500nm, and Fig. 3 is the rheometry result of the present embodiment hybridized hydrogel; The hybridized hydrogel of self-supporting can be formed by this enforcement preparation method.
Fig. 3 is transmission electron microscope (staining counter) picture of the present embodiment gained hybridized hydrogel; Result shows, K
3a
3i
3g
3v
3define a large amount of long fibre structures, these fibers by with GO lamella (darker regions in picture) be cross-linked to form network structure.It is pointed out that because GO lamellar spacing is minimum and with shape in peptide fiber mechanism, larger change occurs, be difficult to by its border of the clear resolution of transmission electron microscope.
Embodiment 4
(1) K is prepared
3a
3i
3g
3v
3peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Room temperature leaves standstill 3 days, makes its self assembly be long fibre structure;
(2) in above-mentioned long fibre structure-solution, adding length is 300 ± 200nm, and width is 300 ± 200nm, and thickness is the GO lamella of 1-2nm, and make the concentration of GO lamella be 1.0mg/mL, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Employing concentration is that the NaOH solution of 0.5mol/L regulates mixed solution pH value to be 10.0, places 6 hours, forms hybridized hydrogel.
Embodiment 4 gained hydrogel storage modulus (G') can reach 17000Pa, and (G ") can reach 2500Pa to Loss modulus, and gel aperture size is less than 450nm; The hybridized hydrogel of self-supporting can be formed by this enforcement preparation method.
Comparative example 2,3,4, at K
3a
3i
3g
3v
3under the condition that solution concentration is identical with pH value, the addition of GO lamella is larger, and the mechanical strength of gained hydrogel is larger, and aperture size then reduces to some extent.
Embodiment 5
(1) K is prepared
3a
3i
3g
3v
3peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Room temperature leaves standstill 3 days, makes its self assembly be long fibre structure, obtains long fibre structure-solution;
(2) in above-mentioned long fibre structure-solution, adding length is 1000 ± 500nm, and width is 1000 ± 500nm, and thickness is the GO lamella of 1-2nm, and make the concentration of GO lamella be 1.0mg/mL, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Employing concentration is that the NaOH solution of 0.5mol/L regulates mixed solution pH value to be 10.0, places 6 hours, forms hybridized hydrogel.
Embodiment 5 gained hydrogel storage modulus (G') can reach 13000Pa, and (G ") can reach 1500Pa to Loss modulus, and gel aperture size is less than 900nm; The hybridized hydrogel of self-supporting can be formed by this enforcement preparation method.
Comparative example 4 and 5, at K
3a
3i
3g
3v
3under solution concentration, solution ph and GO lamella add condition identical in quality, GO lamella size becomes large, and the aperture size of gained hydrogel increases, and mechanical strength reduces to some extent.
Embodiment 6 (not adding GO lamella)
(1) K is prepared
3a
3i
3g
3v
3peptide molecule 4.0mmol/L aqueous solution, ultrasonic mixing, ultrasonic power is 100W, and ultrasonic time is 10min; Room temperature leaves standstill 3 days, makes its self assembly be long fibre structure;
(2) carry out ultrasonic mixing to above-mentioned long fibre structure-solution, ultrasonic power is 100W, and ultrasonic time is 10min; Employing concentration is that the NaOH solution of 0.5mol/L regulates solution ph to be 10.0, places 6 hours, obtains hydrogel reference substance 1.
Embodiment 6 gained hydrogel storage modulus (G') can reach 300Pa, and (G ") can reach 50Pa to Loss modulus, and gel aperture size is less than 400nm.
Embodiment 6 and the embodiment of the present invention 2,3,4,5 compare, at K
3a
3i
3g
3v
3under the condition that solution concentration is identical with pH value, the hydrogel mechanical strength being added with GO lamella improves greatly, and aperture size obtains Effective Regulation.
With I in various embodiments of the present invention
3k peptide molecule or K
3a
3i
3g
3v
3peptide molecule is that hybridized hydrogel prepared by raw material, it should be noted that when not adding GO lamella, I
3k aqueous solution cannot form hydrogel under 4.0mmol/L concentration, K
3a
3i
3g
3v
3aqueous solution can form hydrogel under 4.0mmol/L concentration, but mechanical property is poor.
Above-described embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under the prerequisite not departing from the invention spirit; the various distortion that those of ordinary skill in the art make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determines.
Claims (10)
1. a preparation method for high strength polypeptide hydrogel, is characterized in that: comprise the following steps:
(1) select self assembly to form long fibre structure and positively charged peptide molecule, prepare the aqueous solution of described peptide molecule, concentration is 2mmol/L-10mmol/L; Ultrasonic mixing, leaves standstill, makes its self assembly be long fibre structure;
(2) in the long fibre structure-solution of step (1) gained, graphene oxide (GO) lamella is added, the concentration of GO lamella is made to be 0.02 ~ 1.0mg/mL, ultrasonic mixing, regulate the pH value of gained polypeptide/GO lamella mixed solution between 8-11, leave standstill, hybridized hydrogel is formed, namely described high strength polypeptide hydrogel after system ripening is stable.
2. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (1), and described peptide molecule contains the positively charged amino acid residue of more than 1.
3. the preparation method of high strength polypeptide hydrogel according to claim 2, is characterized in that: described peptide molecule chooses I
3k or K
3a
3i
3g
3v
3.
4. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (1), and described peptide molecule concentration chooses 3-5mmol/L.
5. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (1), ultrasonic condition is: ultrasonic power is: 90-110W, and ultrasonic time is: 5-15min.
6. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (1), after ultrasonic mixing, leaves standstill 1-7 days under room temperature.
7. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (2), and GO lamella dimensional parameters is: length is 100-1500nm, and width is 100-1500nm, and thickness is 1-2nm.
8. the preparation method of high strength polypeptide hydrogel according to claim 7, is characterized in that: described graphene oxide lamella dimensional parameters is: length is 300 ± 200nm, and width is 300 ± 200nm.
9. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (2), ultrasonic condition is: ultrasonic power is: 90-110W, and ultrasonic time is: 5-15min.
10. the preparation method of high strength polypeptide hydrogel according to claim 1, is characterized in that: in described step (2), leaves standstill after 5-8 hour, and system reaches ripening and stablizes.
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| CN105801914A (en) * | 2016-03-17 | 2016-07-27 | 四川大学 | Functional graphene composite material modified with polypeptide and preparing method thereof |
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| CN108503860A (en) * | 2018-04-24 | 2018-09-07 | 中国石油大学(华东) | The preparation method and application of the reversible hydrogel of temperature-responsive |
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| CN110010372A (en) * | 2019-03-06 | 2019-07-12 | 常州沃兰特电子有限公司 | A kind of preparation method of high mating type electrode for super capacitor material |
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