技术领域technical field
本发明涉及水凝胶的制备方法，尤其涉及一种高强度多肽水凝胶的制备方法；属于化学制备领域。The invention relates to a preparation method of a hydrogel, in particular to a preparation method of a high-strength polypeptide hydrogel; it belongs to the field of chemical preparation.
背景技术Background technique
水凝胶是以水为分散介质的凝胶材料，由于其丰富的孔结构和三维网状结构能够提供较好的渗透性和机械支撑，因此在诸多领域有着广泛的应用前景。近年来，水凝胶材料备受国内外科学家的关注，对其在生物医学如组织工程、药物运输等方面的应用进行了大量研究。普通的水凝胶可以通过高分子量的合成聚合物之间或者一些小分子之间的共价交联得到，但是，这样的交联方式需要通过化学合成来进行。由于大部分的化学试剂含有毒性，从而导致其形成的水凝胶没有生物活性，也不可降解；另一种方法是利用一些小分子自身可以自组装的特性，通过非共价的相互作用形成网状结构，在温和的生理条件下进一步成胶。由于第二种合成的方法条件温和，组成单元可选可控，因此形成的胶具有良好的生物相容性、可逆性等优势，从而使这种方法越来越多的引起人们的关注。Hydrogel is a gel material with water as the dispersion medium. Due to its rich pore structure and three-dimensional network structure that can provide better permeability and mechanical support, it has broad application prospects in many fields. In recent years, hydrogel materials have attracted the attention of scientists at home and abroad, and a lot of research has been done on their applications in biomedicine, such as tissue engineering and drug delivery. Ordinary hydrogels can be obtained by covalent cross-linking between high-molecular-weight synthetic polymers or between some small molecules, but such cross-linking methods need to be carried out by chemical synthesis. Since most of the chemical reagents are toxic, the hydrogel formed by them has no biological activity and is not degradable; another method is to use the self-assembly characteristics of some small molecules to form a network through non-covalent interactions. -like structure, further gelatinized under mild physiological conditions. Due to the mild conditions of the second synthesis method and the optional controllable constituent units, the formed glue has the advantages of good biocompatibility and reversibility, which makes this method attract more and more people's attention.
近些年来，人们通过对氨基酸残基的排列组合，设计合成各种不同序列的肽分子，通过精巧的分子结构和性质调控，很多肽分子可以自组装形成稳定的水凝胶。肽分子作为小分子物种，其水凝胶的形成一般是肽分子自组装形成纳米纤维，再由纤维进一步三维交联形成宏观上水凝胶结构。这一过程是通过各种非共价相互作用如静电作用、氢键、疏水作用、π-π堆积等驱动完成，这就使得肽水凝胶具有良好的可调控性和可逆性。而且，由于肽分子最初来源于生物体，其基本结构单元是α-氨基酸，有着较好的生物相容性，因此在体内可以降解，并且代谢产物无毒。肽自组装形成的水凝胶含水量非常高，并且这种支架的孔结构与细胞外间质的结构类似，不仅能使细胞存活和移动，还能使生长因子和营养素等缓慢地扩散出入，是可控制细胞分化的理想细胞培养基质，并且支撑细胞成长为一个完整组织的理想材料。近些年来基于肽自组装形成的水凝胶材料研究已经逐渐成为了材料领域的研究热点，在纳米材料，生物医学及组织工程等领域呈现出广阔的应用前景。In recent years, people have designed and synthesized peptide molecules of various sequences through the arrangement and combination of amino acid residues. Through exquisite molecular structure and property regulation, many peptide molecules can self-assemble to form stable hydrogels. Peptide molecules are small molecular species, and the formation of hydrogel is generally the self-assembly of peptide molecules to form nanofibers, and then the fibers are further three-dimensionally cross-linked to form a macroscopic hydrogel structure. This process is driven by various non-covalent interactions such as electrostatic interaction, hydrogen bond, hydrophobic interaction, π-π stacking, etc., which makes the peptide hydrogel have good controllability and reversibility. Moreover, since the peptide molecule is originally derived from organisms, its basic structural unit is α-amino acid, which has good biocompatibility, so it can be degraded in the body, and the metabolites are non-toxic. The hydrogel formed by peptide self-assembly has a very high water content, and the pore structure of this scaffold is similar to that of the extracellular matrix, which not only enables cells to survive and move, but also allows growth factors and nutrients to slowly diffuse in and out. It is an ideal cell culture substrate that can control cell differentiation and support the growth of cells into a complete tissue. In recent years, the research on hydrogel materials based on peptide self-assembly has gradually become a research hotspot in the field of materials, showing broad application prospects in the fields of nanomaterials, biomedicine, and tissue engineering.
然而也正是由于肽分子水凝胶的形成一般是肽分子首先自组装形成纳米纤维，再由纤维进一步三维交联形成宏观上的水凝胶结构。这一过程是基于非共价相互作用形成，成胶能力有限，且其纤维间的交联强度往往较低，导致其凝胶强度有限，自支撑性能差，容易被破坏；而且，其纤维交联所形成的三维网络结构孔径往往较小且难于有效调控，这大大限制了其在组织工程材料等领域的应用。However, it is precisely because of the formation of peptide molecular hydrogel that the peptide molecules first self-assemble to form nanofibers, and then the fibers are further three-dimensionally cross-linked to form a macroscopic hydrogel structure. This process is based on the formation of non-covalent interactions, the ability to form gels is limited, and the cross-linking strength between fibers is often low, resulting in limited gel strength, poor self-supporting performance, and easy to be destroyed; moreover, its fiber cross-linking The pore size of the three-dimensional network structure formed by the association is often small and difficult to effectively control, which greatly limits its application in tissue engineering materials and other fields.
发明内容Contents of the invention
本发明的目的是提供一种高强度多肽水凝胶的制备方法，用于制备水凝胶，且制备出的水凝胶力学强度高，同时水凝胶的力学强度和孔径尺寸是可调的。The purpose of the present invention is to provide a method for preparing high-strength polypeptide hydrogel, which is used to prepare hydrogel, and the prepared hydrogel has high mechanical strength, and the mechanical strength and pore size of the hydrogel are adjustable at the same time .
本发明的技术方案为：Technical scheme of the present invention is:
一种高强度多肽水凝胶的制备方法，包括以下步骤：A method for preparing a high-strength polypeptide hydrogel, comprising the following steps:
(1)选择能够自组装形成长纤维结构且带正电荷的多肽分子，配制所述多肽分子的水溶液，浓度为2mmol/L-10mmol/L；超声混匀，静置，使其自组装为长纤维结构；(1) Select a positively charged polypeptide molecule capable of self-assembling to form a long fiber structure, prepare an aqueous solution of the polypeptide molecule at a concentration of 2mmol/L-10mmol/L; fiber structure;
(2)向步骤(1)所得的长纤维结构溶液中加入氧化石墨烯(GO)片层，使GO片层的浓度为0.02～1.0mg/mL，超声混匀，调节所得多肽/GO片层混合溶液的pH值在8-11之间，静置，待体系熟化稳定后形成杂化水凝胶，即所述的高强度多肽水凝胶。(2) Add graphene oxide (GO) sheets to the long fiber structure solution obtained in step (1), so that the concentration of the GO sheets is 0.02 to 1.0 mg/mL, and ultrasonically mix to adjust the obtained polypeptide/GO sheets The pH value of the mixed solution is between 8-11, and it is left to stand until the system matures and stabilizes to form a hybrid hydrogel, that is, the high-strength polypeptide hydrogel.
所述步骤(1)中，所述多肽分子含有1个以上的带正电荷的氨基酸残基，优选I3K或K3A3I3G3V3，尤其优选K3A3I3G3V3，二者分子结构式分别为：In the step (1), the polypeptide molecule contains more than one positively charged amino acid residue, preferably I3 K or K3 A3 I3 G3 V3 , especially preferably K3 A3 I3 G3 V3 , the molecular structural formulas of the two are:
I3K分子结构式为：The molecular structural formula of I3 K is:
K3A3I3G3V3分子结构式为：The molecular structural formula of K3 A3 I3 G3 V3 is:
所述步骤(1)中，所述多肽分子浓度优选3-5mmol/L。In the step (1), the molecular concentration of the polypeptide is preferably 3-5 mmol/L.
所述步骤(1)中，超声的目的是对溶液混匀，只要能达到混匀的目的即可；优选超声的条件为：超声功率为：90-110W，超声时间为：5-15min。In the step (1), the purpose of ultrasound is to mix the solution, as long as the purpose of mixing can be achieved; the preferred conditions of ultrasound are: ultrasonic power: 90-110W, ultrasonic time: 5-15min.
所述步骤(2)中，所述GO片层尺寸参数为：长度为100-1500nm，宽度为100-1500nm，厚度为1-2nm；优选长度为300±200nm，宽度为300±200nm。In the step (2), the size parameters of the GO sheet are: length 100-1500nm, width 100-1500nm, thickness 1-2nm; preferably length 300±200nm, width 300±200nm.
所述步骤(2)中，超声的目的是对溶液混匀，只要能达到混匀的目的即可；优选超声的条件为：超声功率为：90-110W，超声时间为：5-15min。In the step (2), the purpose of ultrasound is to mix the solution, as long as the purpose of mixing can be achieved; the preferred conditions of ultrasound are: ultrasonic power: 90-110W, ultrasonic time: 5-15min.
所述步骤(1)中，一般常温下放置1-7天，多肽分子便可自组装为长纤维结构；长纤维结构是否形成可以通过透射电镜(TEM)进行检测。In the step (1), the polypeptide molecules can be self-assembled into a long fiber structure after standing at room temperature for 1-7 days; whether the long fiber structure is formed can be detected by transmission electron microscope (TEM).
所述步骤(2)中，调节所得多肽/GO片层混合溶液的pH值可以降低多肽分子电荷，有利于成胶。体系熟化是否完成则可以通过溶液流动性的消失来判断，若熟化完成，将盛放溶液的小瓶倒置溶液不会流下；一般静置5-8小时便可熟化完成。In the step (2), adjusting the pH value of the obtained polypeptide/GO sheet mixed solution can reduce the molecular charge of the polypeptide, which is conducive to gelation. Whether the aging of the system is completed can be judged by the disappearance of the fluidity of the solution. If the aging is completed, the solution will not flow down when the vial containing the solution is turned upside down; generally, the aging can be completed after standing for 5-8 hours.
GO片层是石墨经强酸氧化制备，其长度和宽度尺寸难以控制到很精确的范围，通常使用的尺寸范围以某一尺寸点为中心，呈左右扩散趋势；例如长度为300±200nm的GO片层是指长度集中在300nm附近，并向左右扩散200nm的尺寸范围内的片层都可以接受(即可接纳的尺寸范围为100-500nm)。GO片层的尺寸也常根据实际使用中对水凝胶孔径尺寸大小的需求进行调节选择。GO flakes are prepared by oxidation of graphite with strong acid, and its length and width are difficult to control to a very precise range. The commonly used size range is centered on a certain size point, showing a tendency to diffuse left and right; for example, a GO flake with a length of 300±200nm A layer refers to a sheet whose length is concentrated around 300nm and diffuses to the left and right within a size range of 200nm is acceptable (that is, the acceptable size range is 100-500nm). The size of GO sheet is also often adjusted and selected according to the demand for hydrogel pore size in actual use.
GO片层是厚度为1-2nm的薄片，其横向尺寸可为几十纳米到十几微米范围，其结构特征使其具有聚合物、胶体、薄膜以及两性分子的特性，是一种优良的软性碳材料。GO片层一般由石墨经强酸氧化而得，其表面和边缘有羧基、酚羟基和环氧基团等极性带电集团，在水中具有很好的分散性，且易于与其它物质相互作用。GO sheet is a thin sheet with a thickness of 1-2nm, and its lateral size can range from tens of nanometers to more than ten microns. Its structural characteristics make it have the characteristics of polymers, colloids, films and amphiphilic molecules. It is an excellent soft permanent carbon material. GO sheets are generally obtained by oxidation of graphite with strong acid. There are polar charged groups such as carboxyl groups, phenolic hydroxyl groups, and epoxy groups on the surface and edges. They have good dispersion in water and are easy to interact with other substances.
本发明将氧化石墨烯(GO)片层引入到多肽水凝胶中，通过GO片层与多肽自组装纤维的相互作用形成空间三维网络结构，实现对水分子的有效结合和固定，形成水凝胶结构，大大提高了体系的成胶能力，得到高强度的肽/GO杂化水凝胶材料。GO片层具有优良的力学性能，其作为凝胶形成的促进剂和交联剂，可以大大提高杂化水凝胶的力学强度，而且通过对GO片层尺寸和表面电荷密度的调节可以有效调控杂化水凝胶材料的力学强度和孔径大小。In the present invention, graphene oxide (GO) sheets are introduced into the polypeptide hydrogel, and a spatial three-dimensional network structure is formed through the interaction between GO sheets and polypeptide self-assembled fibers, so as to realize the effective combination and fixation of water molecules and form hydrogels. The gel structure greatly improves the gel-forming ability of the system, and a high-strength peptide/GO hybrid hydrogel material is obtained. GO sheets have excellent mechanical properties. As accelerators and crosslinkers for gel formation, they can greatly improve the mechanical strength of hybrid hydrogels, and can effectively control the size and surface charge density of GO sheets. Mechanical strength and pore size of hybrid hydrogel materials.
本发明可通过调节GO片层的尺寸及浓度，实现对杂化水凝胶力学强度、孔径尺寸的调控。The invention can adjust the mechanical strength and pore size of the hybrid hydrogel by adjusting the size and concentration of the GO sheets.
一般来讲，在适当的GO片层浓度范围内，GO片层浓度越高，储能模量(G')和耗能模量(G")越大，得到的水凝胶力学性能越好；所述适当浓度范围是指GO片层的加入不能引起体系的沉淀产生，GO片层加入量太大，会产生沉淀，体系分相，无法形成均一的水凝胶。Generally speaking, within the appropriate range of GO sheet concentration, the higher the GO sheet concentration, the larger the storage modulus (G') and loss modulus (G"), and the better the mechanical properties of the obtained hydrogel. The appropriate concentration range means that the addition of GO sheets cannot cause precipitation of the system. If the amount of GO sheets added is too large, precipitation will occur, the system will separate phases, and a uniform hydrogel cannot be formed.
不同尺寸GO片层同样影响水凝胶的力学性能，在GO片层加入量相同的情况下，随着GO片层尺寸增加，体系力学性能下降，此外尺寸过大的GO片层在溶液中难于均一分散，导致水凝胶的均匀性下降。不同尺寸GO片层的加入会影响水凝胶的孔径大小。Different sizes of GO sheets also affect the mechanical properties of the hydrogel. When the amount of GO sheets added is the same, as the size of GO sheets increases, the mechanical properties of the system decrease. In addition, GO sheets with too large sizes are difficult to dissolve in solution. Uniform dispersion, resulting in a decrease in the homogeneity of the hydrogel. The addition of GO sheets with different sizes will affect the pore size of the hydrogel.
与现有技术相比，本发明的有益效果为：Compared with prior art, the beneficial effect of the present invention is:
(1)将GO片层引入到多肽水溶液中，通过GO片层与多肽自组装纤维的相互作用，可以提高体系的成胶能力，得到高强度的多肽/GO杂化水凝胶材料；(1) The GO sheet is introduced into the polypeptide aqueous solution, and the gelation ability of the system can be improved through the interaction between the GO sheet and the polypeptide self-assembled fiber, and a high-strength polypeptide/GO hybrid hydrogel material can be obtained;
(2)可以通过GO片层的尺寸及浓度控制来调控多肽/GO杂化水凝胶的力学强度和孔径尺寸等性质。(2) The mechanical strength and pore size of the polypeptide/GO hybrid hydrogel can be regulated by controlling the size and concentration of the GO sheet.
附图说明Description of drawings
图1多肽/GO杂化水凝胶制备流程示意图，Figure 1 Schematic diagram of the preparation process of polypeptide/GO hybrid hydrogel,
图2实施例3杂化水凝胶的流变测定结果，The rheological measurement result of Fig. 2 embodiment 3 hybrid hydrogels,
图3实施例3杂化水凝胶的透射电镜(复染法)图片。Figure 3 is a transmission electron microscope (counter-staining) picture of the hybrid hydrogel of Example 3.
具体实施方式detailed description
以下结合实施例和附图对本发明进行详细的阐述，其中图1是本发明的多肽/GO杂化水凝胶制备流程示意图，图中简要地呈现了本发明的制备流程。The present invention will be described in detail below with reference to the examples and accompanying drawings, wherein Figure 1 is a schematic diagram of the preparation process of the polypeptide/GO hybrid hydrogel of the present invention, which briefly presents the preparation process of the present invention.
实施例1Example 1
(1)配制I3K多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温下静置3天，使其自组装为长纤维结构，此时体系不能形成水凝胶；(1) Prepare a 4.0mmol/L aqueous solution of I3 K polypeptide molecules, mix them evenly by ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand for 3 days at room temperature to make it self-assemble into a long fiber structure, and the system cannot form at this time Hydrogels;
(2)在上述含有多肽长纤维结构的溶液中加入长度为300±200nm，宽度为300±200nm，厚度为1-2nm的GO片层，使GO片层浓度为0.1mg/mL(即每mL溶液添加0.1mgGO片层)，超声混匀，超声功率为100W，超声时间为12min；采用浓度为0.5mol/L的NaOH溶液调节混合溶液pH值为10.5，常温静置6小时，形成杂化水凝胶。(2) Add GO sheets with a length of 300±200nm, a width of 300±200nm, and a thickness of 1-2nm in the solution containing the long polypeptide fiber structure, so that the concentration of the GO sheet is 0.1mg/mL (that is, every mL Add 0.1 mg GO flakes to the solution), and ultrasonically mix, the ultrasonic power is 100W, and the ultrasonic time is 12min; the pH value of the mixed solution is adjusted to 10.5 by using NaOH solution with a concentration of 0.5mol/L, and it is allowed to stand at room temperature for 6 hours to form hybrid water gel.
实施例1所得水凝胶储能模量(G')可达4000Pa，耗能模量(G")可达500Pa，凝胶孔径尺寸小于400nm；通过本实施制备方法可以形成自支撑的杂化水凝胶。The storage modulus (G') of the hydrogel obtained in Example 1 can reach 4000Pa, the energy dissipation modulus (G") can reach 500Pa, and the gel pore size is less than 400nm; the self-supporting hybrid Hydrogels.
实施例2Example 2
(1)配制K3A3I3G3V3多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温下静置3天，使其自组装为长纤维结构；(1) Prepare 4.0mmol/L aqueous solution of K3 A3 I3 G3 V3 polypeptide molecules, and mix them by ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand at room temperature for 3 days to make it self-assemble into long fibers structure;
(2)在上述长纤维结构溶液中加入长度为300±200nm，宽度为300±200nm，厚度为1-2nm的GO片层，使GO片层浓度为0.02mg/mL，超声混匀，超声功率为100W，超声时间为10min；采用浓度为0.5mol/L的NaOH溶液调节混合溶液pH值为10.0，常温静置6小时，形成杂化水凝胶。(2) Add GO sheets with a length of 300±200nm, a width of 300±200nm, and a thickness of 1-2nm to the above-mentioned long fiber structure solution, so that the concentration of the GO sheet is 0.02mg/mL, ultrasonically mix, and the ultrasonic power 100W, ultrasonic time is 10min; use 0.5mol/L NaOH solution to adjust the pH of the mixed solution to 10.0, and let it stand at room temperature for 6 hours to form a hybrid hydrogel.
实施例2所得水凝胶储能模量(G')可达5000Pa，耗能模量(G")可达600Pa，凝胶孔径尺寸小于500nm；通过本实施制备方法可以形成自支撑的杂化水凝胶。The storage modulus (G') of the hydrogel obtained in Example 2 can reach 5000Pa, the energy dissipation modulus (G") can reach 600Pa, and the gel pore size is less than 500nm; the self-supporting hybrid Hydrogels.
实施例3Example 3
(1)配制K3A3I3G3V3多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温静置3天，使其自组装为长纤维结构；(1) Prepare 4.0mmol/L aqueous solution of K3 A3 I3 G3 V3 polypeptide molecule, mix it with ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand at room temperature for 3 days to make it self-assemble into a long fiber structure ;
(2)在上述长纤维结构溶液中加入长度为300±200nm，宽度为300±200nm，厚度为1-2nm的GO片层，使GO片层浓度为0.1mg/mL，超声混匀，超声功率为100W，超声时间为10min；采用浓度为0.5mol/L的NaOH溶液调节混合溶液pH值为10.0，常温静置6小时，形成杂化水凝胶。(2) Add GO sheets with a length of 300±200nm, a width of 300±200nm, and a thickness of 1-2nm to the above-mentioned long fiber structure solution, so that the concentration of the GO sheet is 0.1mg/mL, ultrasonically mix, and the ultrasonic power 100W, ultrasonic time is 10min; use 0.5mol/L NaOH solution to adjust the pH of the mixed solution to 10.0, and let it stand at room temperature for 6 hours to form a hybrid hydrogel.
实施例3所得水凝胶储能模量(G')可达6000Pa，耗能模量(G")可达1000Pa，凝胶孔径尺寸小于500nm，图3是本实施例杂化水凝胶的流变测定结果；通过本实施制备方法可以形成自支撑的杂化水凝胶。The storage modulus (G') of the hydrogel obtained in Example 3 can reach 6000Pa, the loss modulus (G") can reach 1000Pa, and the gel pore size is less than 500nm. Figure 3 is the hybrid hydrogel of this embodiment. Rheological measurement results; Self-supporting hybrid hydrogels can be formed through the implementation of the preparation method.
图3为本实施例所得杂化水凝胶的透射电镜(复染法)图片；结果显示，K3A3I3G3V3形成了大量的长纤维结构，这些纤维通过与GO片层(图片中的深色区域)的交联形成网络结构。需要指出的是，由于GO片层厚度极小且与肽纤维作用过程中形状发生较大变化，难于通过透射电镜清晰分辨其边界。Figure 3 is a transmission electron microscope (counterstaining) picture of the hybrid hydrogel obtained in this example; the results show that K3 A3 I3 G3 V3 forms a large number of long fiber structures, and these fibers pass through the GO sheet The cross-links (dark areas in the picture) form a network structure. It should be pointed out that due to the extremely small thickness of GO sheets and the large change in shape during the interaction with peptide fibers, it is difficult to clearly distinguish their boundaries by transmission electron microscopy.
实施例4Example 4
(1)配制K3A3I3G3V3多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温静置3天，使其自组装为长纤维结构；(1) Prepare 4.0mmol/L aqueous solution of K3 A3 I3 G3 V3 polypeptide molecule, mix it with ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand at room temperature for 3 days to make it self-assemble into a long fiber structure ;
(2)在上述长纤维结构溶液中加入长度为300±200nm，宽度为300±200nm，厚度为1-2nm的GO片层，使GO片层的浓度为1.0mg/mL，超声混匀，超声功率为100W，超声时间为10min；采用浓度为0.5mol/L的NaOH溶液调节混合溶液pH值为10.0，放置6小时，形成杂化水凝胶。(2) Add GO sheets with a length of 300±200nm, a width of 300±200nm, and a thickness of 1-2nm to the above-mentioned long fiber structure solution, so that the concentration of the GO sheet is 1.0mg/mL, ultrasonically mix, and ultrasonically The power is 100W, and the ultrasonic time is 10min; the pH value of the mixed solution is adjusted to 10.0 by using NaOH solution with a concentration of 0.5mol/L, and left for 6 hours to form a hybrid hydrogel.
实施例4所得水凝胶储能模量(G')可达17000Pa，耗能模量(G")可达2500Pa，凝胶孔径尺寸小于450nm；通过本实施制备方法可以形成自支撑的杂化水凝胶。The storage modulus (G') of the hydrogel obtained in Example 4 can reach 17000Pa, the energy dissipation modulus (G") can reach 2500Pa, and the gel pore size is less than 450nm; the self-supporting hybrid Hydrogels.
对比实施例2，3，4，在K3A3I3G3V3溶液浓度和pH值相同的条件下，GO片层的添加量越大，所得水凝胶的力学强度越大，孔径尺寸则有所减小。Comparative examples 2, 3, and 4, under the same K3 A3 I3 G3 V3 solution concentration and pH value, the greater the amount of GO sheets added, the greater the mechanical strength of the obtained hydrogel, and the pore diameter The size is reduced.
实施例5Example 5
(1)配制K3A3I3G3V3多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温静置3天，使其自组装为长纤维结构，得到长纤维结构溶液；(1) Prepare 4.0mmol/L aqueous solution of K3 A3 I3 G3 V3 polypeptide molecule, mix it with ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand at room temperature for 3 days to make it self-assemble into a long fiber structure , to obtain a long fiber structure solution;
(2)在上述长纤维结构溶液中加入长度为1000±500nm，宽度为1000±500nm，厚度为1-2nm的GO片层，使GO片层的浓度为1.0mg/mL，超声混匀，超声功率为100W，超声时间为10min；采用浓度为0.5mol/L的NaOH溶液调节混合溶液pH值为10.0，放置6小时，形成杂化水凝胶。(2) Add GO sheets with a length of 1000±500nm, a width of 1000±500nm, and a thickness of 1-2nm to the above-mentioned long fiber structure solution, so that the concentration of the GO sheet is 1.0mg/mL, ultrasonically mix, and ultrasonically The power is 100W, and the ultrasonic time is 10min; the pH value of the mixed solution is adjusted to 10.0 by using NaOH solution with a concentration of 0.5mol/L, and left for 6 hours to form a hybrid hydrogel.
实施例5所得水凝胶储能模量(G')可达13000Pa，耗能模量(G")可达1500Pa，凝胶孔径尺寸小于900nm；通过本实施制备方法可以形成自支撑的杂化水凝胶。The storage modulus (G') of the hydrogel obtained in Example 5 can reach 13000Pa, the energy dissipation modulus (G") can reach 1500Pa, and the gel pore size is less than 900nm; the preparation method of this implementation can form a self-supporting hybrid Hydrogels.
对比实施例4和5，在K3A3I3G3V3溶液浓度、溶液pH值和GO片层添加质量相同的条件下，GO片层尺寸变大，所得水凝胶的孔径尺寸增加，而力学强度有所减小。Comparing Examples 4 and 5, under the same conditions of K3 A3 I3 G3 V3 solution concentration, solution pH value and GO sheet added mass, the GO sheet size becomes larger, and the pore size of the obtained hydrogel increases , while the mechanical strength decreases.
实施例6(不添加GO片层)Embodiment 6 (do not add GO sheet)
(1)配制K3A3I3G3V3多肽分子4.0mmol/L水溶液，超声混匀，超声功率为100W，超声时间为10min；常温静置3天，使其自组装为长纤维结构；(1) Prepare 4.0mmol/L aqueous solution of K3 A3 I3 G3 V3 polypeptide molecule, mix it with ultrasonic, the ultrasonic power is 100W, and the ultrasonic time is 10min; stand at room temperature for 3 days to make it self-assemble into a long fiber structure ;
(2)对上述长纤维结构溶液进行超声混匀，超声功率为100W，超声时间为10min；采用浓度为0.5mol/L的NaOH溶液调节溶液pH值为10.0，放置6小时，得到水凝胶对照品1。(2) Ultrasonic mixing is carried out on the above-mentioned long fiber structure solution, the ultrasonic power is 100W, and the ultrasonic time is 10min; the pH value of the solution is adjusted to 10.0 by using NaOH solution with a concentration of 0.5mol/L, and left for 6 hours to obtain the hydrogel control Product 1.
实施例6所得水凝胶储能模量(G')可达300Pa，耗能模量(G")可达50Pa，凝胶孔径尺寸小于400nm。The storage modulus (G') of the hydrogel obtained in Example 6 can reach 300Pa, the dissipation modulus (G") can reach 50Pa, and the gel pore size is less than 400nm.
实施例6与本发明实施例2，3，4，5相比较，在K3A3I3G3V3溶液浓度和pH值相同的条件下，添加有GO片层的水凝胶力学强度大大提高，孔径尺寸得到有效调控。Example 6 Compared with Examples 2, 3, 4, and 5 of the present invention, the mechanical strength of the hydrogel with GO sheets added under the same K3 A3 I3 G3 V3 solution concentration and pH value Greatly improved, the pore size is effectively regulated.
本发明各实施例中以I3K多肽分子或K3A3I3G3V3多肽分子为原料制备杂化水凝胶，值得注意的是，在不添加GO片层的情况下，I3K水溶液在4.0mmol/L浓度下无法形成水凝胶，K3A3I3G3V3水溶液在4.0mmol/L浓度下可以形成水凝胶，但是力学性能较差。In each embodiment of the present invention, I3 K polypeptide molecules or K3 A3 I3 G3 V3 polypeptide molecules are used as raw materials to prepare hybrid hydrogels. It is worth noting that, without adding GO sheets, I The3 K aqueous solution cannot form a hydrogel at the concentration of 4.0mmol/L, and the K3 A3 I3 G3 V3 aqueous solution can form a hydrogel at the concentration of 4.0mmol/L, but the mechanical properties are poor.
以上所述的实施例仅仅是对本发明的优选实施方式进行描述，并非对本发明的范围进行限定，在不脱离本发明创造精神的前提下，本领域普通技术人员对本发明的技术方案作出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。The above-mentioned embodiments are only descriptions of preferred implementations of the present invention, and are not intended to limit the scope of the present invention. Variations and improvements should fall within the scope of protection defined by the claims of the present invention.
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| CN201510697529.8ACN105268021B (en) | 2015-10-23 | 2015-10-23 | A kind of preparation method of high intensity polypeptide hydrogel |
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| CN201510697529.8ACN105268021B (en) | 2015-10-23 | 2015-10-23 | A kind of preparation method of high intensity polypeptide hydrogel |
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