CN110746616A

Movatterモバイル変換

Info

Publication number: CN110746616A
Application number: CN201911025476.XA
Authority: CN
Inventors: 武伟; 张丹; 蒋锡群
Original assignee: Nanjing University
Current assignee: Nanjing University
Priority date: 2019-10-25
Filing date: 2019-10-25
Publication date: 2020-02-04

Abstract

The invention discloses a cellulose-based three-dimensional hydrogel containing phenylboronic acid, a preparation method and application thereof. The hydrogel is simple and rapid in preparation process, and has good biocompatibility, self-repairability and cell adhesion.

Description

Translated fromChinese

一种含苯硼酸的纤维素水凝胶及其制备方法和应用A kind of cellulose hydrogel containing phenylboronic acid and its preparation method and application

技术领域technical field

本发明属于高分子材料领域，具体涉及一类利用硼酸酯交联的含苯硼酸的纤维素水凝胶及其制备方法和应用。The invention belongs to the field of polymer materials, and in particular relates to a type of phenylboronic acid-containing cellulose hydrogel cross-linked by borate esters and a preparation method and application thereof.

技术背景technical background

纤维素具有理想的生物相容性、良好的细胞粘附性和较高的机械强度，近年来，作为细胞支架材料已经在骨组织工程、软骨组织工程、血管组织工程等领域被广泛研究。纤维素在自然资源中分布广泛，含量丰富，在各种植物中的含量几乎可以占到50％，因此植物纤维素来源广泛，且廉价易得。但是由植物纤维资源制备的纤维素不具备三维网络结构，无法直接用作细胞支架材料，大大限制了其应用。而如果将植物纤维素分子交联制备成生物相容的三维水凝胶材料，则可以作为支架材料用于组织工程领域，实现植物纤维素的高附加值应用，因此开发纤维素基三维水凝胶的高效制备方法可大大推动纤维素在组织工程领域的应用，对于林木剩余物的循环利用具有重要意义。此外，细胞粘附性是评价支架材料的重要指标之一，高的细胞粘附性有利于细胞沿着支架材料铺展和生长，因此提高纤维素三维水凝胶的细胞粘附性可显著改善其使用性能和应用潜力。Cellulose has ideal biocompatibility, good cell adhesion and high mechanical strength. In recent years, cellulose has been widely studied as a cell scaffold material in the fields of bone tissue engineering, cartilage tissue engineering, and vascular tissue engineering. Cellulose is widely distributed and abundant in natural resources, and the content in various plants can account for almost 50%. Therefore, plant cellulose has a wide range of sources and is cheap and easy to obtain. However, cellulose prepared from plant fiber resources does not have a three-dimensional network structure and cannot be directly used as a cell scaffold material, which greatly limits its application. However, if plant cellulose molecules are cross-linked to prepare biocompatible three-dimensional hydrogel materials, it can be used as a scaffold material in the field of tissue engineering to achieve high value-added applications of plant cellulose. Therefore, the development of cellulose-based three-dimensional hydrogel materials The efficient preparation method of glue can greatly promote the application of cellulose in the field of tissue engineering, which is of great significance for the recycling of forest residues. In addition, cell adhesion is one of the important indicators for evaluating scaffold materials. High cell adhesion is conducive to the spreading and growth of cells along the scaffold material. Therefore, improving the cell adhesion of cellulose three-dimensional hydrogels can significantly improve its cell adhesion. Use performance and application potential.

发明内容SUMMARY OF THE INVENTION

本发明专利提供一种以具有柔性二元或多元醇或顺式二醇的纤维素为主要高分子，以多苯硼酸化合物(如苯硼酸共价修饰的聚乙二醇、环糊精、聚氨基酸、蛋白质和多聚糖等)为交联剂的构建纤维素三维水凝胶的方法。该水凝胶制备过程简单快速,具有良好的生物相容性、可自修复性和细胞粘附性。The patent of the present invention provides a kind of cellulose with flexible dihydric or polyhydric alcohol or cis-diol as the main polymer, polyphenylboronic acid compound (such as polyethylene glycol, cyclodextrin, polyphenylboronic acid covalently modified with phenylboronic acid). amino acids, proteins and polysaccharides, etc.) as cross-linking agents for the construction of cellulose three-dimensional hydrogels. The preparation process of the hydrogel is simple and fast, and it has good biocompatibility, self-repairability and cell adhesion.

本发明具体技术方案如下：The specific technical scheme of the present invention is as follows:

一种含苯硼酸的纤维素水凝胶的制备方法，包括如下步骤：A preparation method of a phenylboronic acid-containing cellulose hydrogel, comprising the steps:

(1)将具有柔性二元或多元醇或顺式二醇以及氨基的化合物在缩合剂存在下通过酰胺化反应与具有羧基的纤维素盐在水介质中共价偶联制得具有柔性二元或多元醇或顺式二醇的纤维素，所述具有柔性二元或多元醇以及氨基的化合物结构通式如下：(1) Covalently coupling a compound with a flexible dihydric or polyhydric alcohol or cis-diol and an amino group with a cellulose salt having a carboxyl group in an aqueous medium through an amidation reaction in the presence of a condensing agent to obtain a flexible dihydric or Cellulose of polyhydric alcohol or cis diol, the compound structure with flexible dihydric or polyhydric alcohol and amino group is as follows:

所述具有顺式二醇以及氨基的化合物结构通式如下：The general structural formula of the compound with cis-diol and amino group is as follows:

A代表饱和或不饱和的环，两个羟基为邻位或间位顺式取代，优选的，选自饱和五元环、饱和六元环或不饱和环状基团如环戊烯、环己烯、苯环、萘、蒽、喹啉等，上述结构中的

表示具有具有柔性二元或多元醇的化合物同时具有氨基取代基，二元醇基团与氨基之间可以通过任意数量或种类的连接基团连接，氨基取代基的位置为任意可取代的位置； A represents a saturated or unsaturated ring, the two hydroxyl groups are cis-substituted at ortho or meta positions, preferably, selected from saturated five-membered rings, saturated six-membered rings or unsaturated cyclic groups such as cyclopentene, cyclohexane alkene, benzene ring, naphthalene, anthracene, quinoline, etc., in the above structure

Indicates that the compound with flexible dihydric or polyhydric alcohol also has an amino substituent, the dihydric alcohol group and the amino group can be connected by any number or type of linking groups, and the position of the amino substituent is any substitutable position;

(2)以多苯硼酸化合物为交联剂，与具有柔性二元或多元醇或顺式二醇的纤维素在水介质中混合，即得含苯硼酸的纤维素水凝胶，所述多苯硼酸化合物为苯硼酸共价修饰的聚乙二醇、环糊精、聚氨基酸、蛋白质或多聚糖。(2) Using the polyphenylboronic acid compound as a cross-linking agent, and mixing it with cellulose with flexible dibasic or polyhydric alcohol or cis diol in an aqueous medium to obtain a phenylboronic acid-containing cellulose hydrogel, the polyphenylboronic acid-containing cellulose hydrogel is obtained. The phenylboronic acid compound is polyethylene glycol, cyclodextrin, polyamino acid, protein or polysaccharide covalently modified with phenylboronic acid.

优选的，所述步骤(1)具有羧基的纤维素盐选自羧甲基纤维素钠、羧乙基纤维素钠和羧甲基羟丙基纤维素钠中的一种或几种。Preferably, the cellulose salt having a carboxyl group in the step (1) is selected from one or more of sodium carboxymethyl cellulose, sodium carboxyethyl cellulose and sodium carboxymethyl hydroxypropyl cellulose.

优选的，于所述步骤(1)所述具有柔性二元或多元醇或顺式二醇以及氨基的化合物选自氨基葡萄糖或多巴胺。Preferably, in the step (1), the compound having a flexible dihydric or polyhydric alcohol or cis diol and an amino group is selected from glucosamine or dopamine.

优选的，步骤(1)所述的缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和/或N-羟基琥珀酰亚胺。Preferably, the condensing agent described in step (1) is selected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and/or N-hydroxysuccinimide.

优选的，步骤(1)所述的水介质为酸性或中性，优选pH值为4-7。Preferably, the aqueous medium described in step (1) is acidic or neutral, with a preferred pH value of 4-7.

优选的，步骤(2)所述水介质为中性或碱性，优选pH值为7-10。Preferably, the aqueous medium in step (2) is neutral or alkaline, preferably with a pH value of 7-10.

优选的，步骤(1)反应温度为10-50℃，反应时间为1-24小时。优选的反应温度为10-30℃，反应时间为5小时。Preferably, the reaction temperature in step (1) is 10-50° C., and the reaction time is 1-24 hours. The preferred reaction temperature is 10-30°C, and the reaction time is 5 hours.

优选的，步骤(2)所述具有柔性二元或多元醇或顺式二醇的纤维素的质量百分浓度为5％-20％，所述的多苯硼酸化合物的质量百分浓度为1％-20％。Preferably, the mass percentage concentration of the cellulose with flexible dihydric or polyhydric alcohol or cis diol in step (2) is 5%-20%, and the mass percentage concentration of the polyphenylboronic acid compound is 1 %-20%.

优选的，步骤(2)反应温度为10-50℃，反应时间为10min到1小时，反应条件为静置，进一步优选的反应温度为10-30℃，反应时间为10min。Preferably, in step (2), the reaction temperature is 10-50°C, the reaction time is 10min to 1 hour, and the reaction condition is standing, and a further preferred reaction temperature is 10-30°C, and the reaction time is 10min.

本发明所述步骤(2)所述多苯硼酸化合物通过具有两个或两个以上官能团的化合物与带有氨基或羧基的苯硼酸或苯硼酸衍生物偶联制得，所述的具有两个或两个以上官能团的化合物为具有羧基、氨基、羟基、卤素、炔基和叠氮中的一种或几种的聚乙二醇、环糊精、聚氨基酸、蛋白质或多聚糖，优选二氨基聚乙二醇、二羟基聚乙二醇、α-、β-、γ-环糊精、聚氨基酸、蛋白质、多聚糖中的一种或几种。The polyphenylboronic acid compound in the step (2) of the present invention is prepared by coupling a compound with two or more functional groups with phenylboronic acid or a phenylboronic acid derivative with an amino group or a carboxyl group. Or the compound with two or more functional groups is polyethylene glycol, cyclodextrin, polyamino acid, protein or polysaccharide with one or more of carboxyl, amino, hydroxyl, halogen, alkynyl and azide, preferably two. One or more of amino polyethylene glycol, dihydroxy polyethylene glycol, α-, β-, γ-cyclodextrin, polyamino acid, protein, and polysaccharide.

本发明另一目的在于提供一种含苯硼酸的纤维素水凝胶，采用本发明所述方法制备而成。Another object of the present invention is to provide a phenylboronic acid-containing cellulose hydrogel prepared by the method of the present invention.

本发明所述的含苯硼酸的纤维素水凝胶可以用于制备组织工程材料。The phenylboronic acid-containing cellulose hydrogel of the present invention can be used to prepare tissue engineering materials.

本发明优点：Advantages of the present invention:

1)本发明所述制备方法中苯硼酸基团与柔性二醇或顺式二醇基团反应生成环状硼酸酯，该反应可以在水介质中进行、快速可逆、而且不需要任何催化剂和助剂，因此利用该反应可以通过简单混合的方式快捷地制备纤维素基三维水凝胶。2)苯硼酸基团与二醇基团之间的反应具有快速可逆的特点，生成的硼酸酯属于一种典型的动态共价键，因而利用该交联反应制备的水凝胶具有自修复的功能。3)该方法制备的水凝胶骨架上还有大量的苯硼酸基团，这些苯硼酸基团可以与细胞表面的唾液酸残基发生反应生成硼酸酯，因而可以显著提升水凝胶材料的细胞粘附性。4)水凝胶所用材料均具有良好的生物相容性，在组织工程领域具有巨大的应用前景。1) In the preparation method of the present invention, a phenylboronic acid group reacts with a flexible diol or a cis diol group to generate a cyclic boronic acid ester, and the reaction can be carried out in an aqueous medium, is rapidly reversible, and does not require any catalyst and Therefore, cellulose-based three-dimensional hydrogels can be quickly prepared by simple mixing by using this reaction. 2) The reaction between the phenylboronic acid group and the diol group is fast and reversible, and the resulting boronic acid ester belongs to a typical dynamic covalent bond, so the hydrogel prepared by this cross-linking reaction has self-healing properties. function. 3) There are also a large number of phenylboronic acid groups on the hydrogel skeleton prepared by this method, and these phenylboronic acid groups can react with sialic acid residues on the cell surface to form boronic acid esters, which can significantly improve the performance of the hydrogel material. cell adhesion. 4) The materials used in the hydrogel have good biocompatibility and have great application prospects in the field of tissue engineering.

附图说明Description of drawings

图1为氨基葡萄糖共价修饰纤维素的核磁共振氢谱。Figure 1 shows the H NMR spectrum of covalently modified cellulose with glucosamine.

图2为苯硼酸共价修饰聚谷氨酸的核磁共振氢谱。Figure 2 is the H NMR spectrum of phenylboronic acid covalently modified polyglutamic acid.

图3为实施例10制备的纤维素水凝胶的扫描电镜照片及实物照片。3 is a scanning electron microscope photo and a real photo of the cellulose hydrogel prepared in Example 10.

具体实施方式Detailed ways

以下通过实施例说明本发明的具体步骤，但不受实施例限制。The specific steps of the present invention are illustrated by the following examples, but are not limited by the examples.

在本发明中使用的术语，除非另有说明，一般具有本领域普通技术人员通常理解的含义。Terms used in the present invention generally have the meanings commonly understood by those of ordinary skill in the art unless otherwise specified.

下面结合具体实施例并参照数据进一步详细描述本发明，应理解，这些实施例只是为了举例说明本发明，而非以任何方式限制本发明的范围。The present invention will be described in further detail below in conjunction with specific embodiments and with reference to data. It should be understood that these embodiments are only for illustrating the present invention, rather than limiting the scope of the present invention in any way.

在以下实施例中，未详细描述的各种过程和方法是本领域中公知的常规方法。In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

本发明以羧甲基纤维素和氨基葡萄糖或多巴胺偶联制备带顺式二醇的纤维素，以苯硼酸修饰的二氨基聚乙二醇、聚谷氨酸、聚赖氨酸为多苯硼酸交联剂为例进一步详细说明本发明，但应注意本发明的范围并不受这些实施例的任何限制。In the present invention, cellulose with cis-diol is prepared by coupling carboxymethyl cellulose and glucosamine or dopamine, and diamino polyethylene glycol, polyglutamic acid and polylysine modified with phenylboronic acid are used as polyphenylboronic acid. The present invention is further described in detail by taking a crosslinking agent as an example, but it should be noted that the scope of the present invention is not limited in any way by these examples.

实施例1氨基葡萄糖修饰的纤维素的制备Example 1 Preparation of glucosamine-modified cellulose

将2g的羧甲基纤维素，溶于200mL去离子水中，加入10.8g(10mol)的EDC·HCL，6.4g(10mol)的NHS，6.04g(10mol)的氨基葡萄糖,室温下搅拌8小时，将反应混合物在截留分子量为14000Da的透析袋中透析2天，冷冻干燥后得产物，核磁共振氢谱如图1所示。显示氨基葡萄糖被共价连接到纤维素上，其取代度可达30％。Dissolve 2g of carboxymethyl cellulose in 200mL of deionized water, add 10.8g (10mol) of EDC·HCl, 6.4g (10mol) of NHS, 6.04g (10mol) of glucosamine, stir at room temperature for 8 hours, The reaction mixture was dialyzed in a dialysis bag with a molecular weight cut-off of 14,000 Da for 2 days, and the product was obtained after freeze-drying. Glucosamine was shown to be covalently attached to cellulose with a degree of substitution of up to 30%.

实施例2多巴胺修饰的纤维素的制备Example 2 Preparation of dopamine-modified cellulose

将2g的羧甲基纤维素，溶于200mL去离子水中，加入10.8g(10mol)的EDC·HCL，6.4g(10mol)的NHS，1.53g(10mol)的多巴胺,室温氮气气氛下搅拌8小时，将反应混合物在截留分子量为14000Da的透析袋中透析2天，冷冻干燥后得产物。Dissolve 2 g of carboxymethyl cellulose in 200 mL of deionized water, add 10.8 g (10 mol) of EDC·HCl, 6.4 g (10 mol) of NHS, 1.53 g (10 mol) of dopamine, and stir for 8 hours at room temperature under a nitrogen atmosphere. , the reaction mixture was dialyzed in a dialysis bag with a molecular weight cut-off of 14000 Da for 2 days, and the product was obtained after freeze-drying.

实施例3苯硼酸修饰的聚乙二醇的制备The preparation of embodiment 3 phenylboronic acid-modified polyethylene glycol

将2g(2mmol)二氨基聚乙二醇和0.5mL三乙胺溶解在10mL DMF中，将1.53g EDC·HCL(8mmol)，0.92g NHS(0.021mol)和3-氨基苯硼酸1.1g(8mmol)加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后用柱色谱分离纯化并真空干燥后得产物。2 g (2 mmol) of diaminopolyethylene glycol and 0.5 mL of triethylamine were dissolved in 10 mL of DMF, 1.53 g of EDC·HCl (8 mmol), 0.92 g of NHS (0.021 mol) and 1.1 g (8 mmol) of 3-aminophenylboronic acid were dissolved It was added to the above solution and stirred at room temperature for 8 hours. The reaction mixture was concentrated, purified by column chromatography and dried in vacuo to give the product.

实施例4苯硼酸修饰的β-环糊精的制备Example 4 Preparation of phenylboronic acid-modified β-cyclodextrin

将2gβ-环糊精和1mL三乙胺溶解在15mL DMF中，将3.811g DCC(0.019mol)和4-羧基苯硼酸3.15g(0.019mol)加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后在丙酮中沉淀洗涤6次，真空干燥后得产物。2 g of β-cyclodextrin and 1 mL of triethylamine were dissolved in 15 mL of DMF, 3.811 g of DCC (0.019 mol) and 3.15 g (0.019 mol) of 4-carboxyphenylboronic acid were added to the above solution, and stirred at room temperature for 8 hours. The reaction mixture was concentrated, precipitated in acetone, washed 6 times, and dried in vacuo to obtain the product.

实施例5苯硼酸修饰的聚谷氨酸的制备Example 5 Preparation of phenylboronic acid-modified polyglutamic acid

将2g聚谷氨酸(分子量为5000)溶解在20mL去离子水中，将4.01g EDC·HCL(0.021mol)，2.4g NHS(0.021mol)和3-氨基苯硼酸2.88g(0.021mol)加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后在丙酮中沉淀洗涤6次，真空干燥后得产物，核磁共振氢谱如图2所示。显示苯硼酸被共价连接到聚谷氨酸上，其取代度可达25％。Dissolve 2g polyglutamic acid (molecular weight 5000) in 20mL deionized water, add 4.01g EDC·HCl (0.021mol), 2.4g NHS (0.021mol) and 2.88g (0.021mol) of 3-aminophenylboronic acid to the solution. The above solution was stirred at room temperature for 8 hours. The reaction mixture was concentrated, precipitated and washed in acetone for 6 times, and the product was obtained after vacuum drying. Phenylboronic acid was shown to be covalently attached to polyglutamic acid with a degree of substitution of up to 25%.

实施例6苯硼酸修饰的聚赖氨酸的制备Example 6 Preparation of phenylboronic acid-modified polylysine

将2g聚赖氨酸(分子量为5000)溶解在20mL去离子水中，将4.01g EDC·HCL(0.021mol)，2.4g NHS(0.021mol)和4-羧基苯硼酸3.49g(0.021mol)加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后在丙酮中沉淀洗涤6次，真空干燥后得产物。2g polylysine (molecular weight 5000) was dissolved in 20mL deionized water, 4.01g EDC·HCl (0.021mol), 2.4g NHS (0.021mol) and 3.49g (0.021mol) of 4-carboxyphenylboronic acid were added to the solution. The above solution was stirred at room temperature for 8 hours. The reaction mixture was concentrated, precipitated in acetone, washed 6 times, and dried in vacuo to obtain the product.

实施例7苯硼酸修饰的牛血清白蛋白(BSA)的制备Example 7 Preparation of bovine serum albumin (BSA) modified with phenylboronic acid

将0.58g(0.003mol)EDC·HCL，0.35g(0.003mol)NHS和0.5g(0.003mol)4-羧基苯硼酸溶解在2mL DMF中，室温搅拌1小时。将2g BSA溶解在20mL去离子水中，并加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后透析5天，冷冻干燥后得产物。0.58 g (0.003 mol) of EDC·HCl, 0.35 g (0.003 mol) of NHS and 0.5 g (0.003 mol) of 4-carboxyphenylboronic acid were dissolved in 2 mL of DMF, and stirred at room temperature for 1 hour. 2 g of BSA was dissolved in 20 mL of deionized water, added to the above solution, and stirred at room temperature for 8 hours. The reaction mixture was concentrated, dialyzed for 5 days, and freeze-dried to obtain the product.

实施例8苯硼酸修饰的壳聚糖的制备Example 8 Preparation of phenylboronic acid-modified chitosan

将0.58g(0.003mol)EDC·HCL，0.35g(0.003mol)NHS和0.5g(0.003mol)4-羧基苯硼酸溶解在2mL DMF中，室温搅拌1小时。将1g壳聚糖溶解在20mL去离子水中，并加入到上述溶液中，室温下搅拌8小时。将反应混合物浓缩后在丙酮中沉淀洗涤6次，真空干燥后得产物。0.58 g (0.003 mol) of EDC·HCl, 0.35 g (0.003 mol) of NHS and 0.5 g (0.003 mol) of 4-carboxyphenylboronic acid were dissolved in 2 mL of DMF, and stirred at room temperature for 1 hour. 1 g of chitosan was dissolved in 20 mL of deionized water, added to the above solution, and stirred at room temperature for 8 hours. The reaction mixture was concentrated, precipitated in acetone, washed 6 times, and dried in vacuo to obtain the product.

实施例9可自修复纤维素水凝胶的制备Example 9 Preparation of self-healing cellulose hydrogel

将240mg氨基葡萄糖修饰的纤维素溶解在2mL pH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将400mg的苯硼酸修饰的聚乙二醇加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小40μm，孔隙率约为82％，杨氏模量1.5kPa。240 mg of glucosamine-modified cellulose was dissolved in 2 mL of pH 7.4 PBS buffer (0.01 mol/L) to prepare a solution with a mass concentration of 12%, and then 400 mg of phenylboronic acid-modified polyethylene glycol was added to the solution. In the above solution, ultrasonically dispersed uniformly and stood still for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 40 μm, a porosity of about 82%, and a Young's modulus of 1.5 kPa.

实施例10可自修复纤维素水凝胶的制备Example 10 Preparation of self-healing cellulose hydrogel

将240mg氨基葡萄糖修饰的纤维素溶解在2mL pH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的聚谷氨酸加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小25μm，孔隙率约为80％，杨氏模量2.0kPa。纤维素水凝胶的扫描电镜照片及实物照片如图3所示。由扫描电镜照片可以看出该水凝胶孔径较为均一，平均孔径大小约25μm。240 mg of glucosamine-modified cellulose was dissolved in 2 mL of pH 7.4 PBS buffer (0.01 mol/L) to prepare a solution with a mass concentration of 12%, and then 120 mg of phenylboronic acid-modified polyglutamic acid was added to the solution. In the above solution, ultrasonically dispersed uniformly and stood still for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 25 μm, a porosity of about 80%, and a Young's modulus of 2.0 kPa. The SEM photos and real photos of the cellulose hydrogel are shown in Figure 3. It can be seen from the scanning electron microscope pictures that the pore size of the hydrogel is relatively uniform, and the average pore size is about 25 μm.

实施例11可自修复纤维素水凝胶的制备Example 11 Preparation of self-healing cellulose hydrogel

将240mg氨基葡萄糖修饰的纤维素溶解在2mL pH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的聚赖氨酸加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小20μm，孔隙率约为78％，杨氏模量2.8kPa。240 mg of glucosamine-modified cellulose was dissolved in 2 mL of pH 7.4 PBS buffer (0.01 mol/L) to prepare a solution with a mass concentration of 12%, and then 120 mg of phenylboronic acid-modified polylysine was added to the solution. In the above solution, ultrasonically dispersed uniformly and stood still for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 20 μm, a porosity of about 78%, and a Young's modulus of 2.8 kPa.

实施例12可自修复纤维素水凝胶的制备Example 12 Preparation of self-healing cellulose hydrogel

将240mg多巴胺修饰的纤维素溶解在2mL pH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将150mg的苯硼酸修饰的β-环糊精加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小25μm，孔隙率约为78％，杨氏模量2.3kPa。240 mg of dopamine-modified cellulose was dissolved in 2 mL of pH 7.4 PBS buffer (0.01 mol/L) to prepare a solution with a mass concentration of 12%, and then 150 mg of phenylboronic acid-modified β-cyclodextrin was added to the solution. In the above solution, ultrasonically dispersed uniformly and stood still for 10 minutes to form a hydrogel. The average pore size of the hydrogel is 25 μm, the porosity is about 78%, and the Young's modulus is 2.3 kPa.

实施例13可自修复纤维素水凝胶的制备Example 13 Preparation of self-healing cellulose hydrogel

将240mg多巴胺修饰的纤维素溶解在2mLpH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的BSA加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小20μm，孔隙率约为76％，杨氏模量2.5kPa。240 mg of dopamine-modified cellulose was dissolved in 2 mL of PBS buffer (0.01 mol/L) with a pH of 7.4 to prepare a solution with a mass concentration of 12%, and then 120 mg of phenylboronic acid-modified BSA was added to the above solution, and ultrasonicated. Disperse uniformly and stand for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 20 μm, a porosity of about 76%, and a Young's modulus of 2.5 kPa.

实施例14可自修复纤维素水凝胶的制备Example 14 Preparation of self-healing cellulose hydrogel

将240mg多巴胺修饰的纤维素溶解在2mLpH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的壳聚糖加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小20μm，孔隙率约为78％，杨氏模量2.3kPa。240 mg of dopamine-modified cellulose was dissolved in 2 mL of PBS buffer (0.01 mol/L) with a pH of 7.4 to prepare a solution with a mass concentration of 12%, and then 120 mg of phenylboronic acid-modified chitosan was added to the above solution. , ultrasonically dispersed uniformly, and static for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 20 μm, a porosity of about 78%, and a Young's modulus of 2.3 kPa.

实施例15可自修复纤维素水凝胶的制备Example 15 Preparation of self-healing cellulose hydrogel

将240mg多巴胺修饰的纤维素溶解在2mLpH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的聚赖氨酸加入到上述溶液中，超声分散均匀，静止10分钟，形成水凝胶。该水凝胶平均孔径大小16μm，孔隙率约为76％，杨氏模量3.2kPa。240 mg of dopamine-modified cellulose was dissolved in 2 mL of PBS buffer (0.01 mol/L) with a pH of 7.4 to prepare a solution with a mass concentration of 12%, and then 120 mg of phenylboronic acid-modified polylysine was added to the above solution. In the middle, ultrasonically dispersed uniformly and stood still for 10 minutes to form a hydrogel. The hydrogel has an average pore size of 16 μm, a porosity of about 76%, and a Young's modulus of 3.2 kPa.

实施例16共价交联纤维素水凝胶的制备Example 16 Preparation of covalently cross-linked cellulose hydrogel

将240mg多巴胺修饰的纤维素溶解在2mL pH为7.4的PBS缓冲液(0.01mol/L)中，配制成质量浓度为12％的溶液，然后将120mg的苯硼酸修饰的聚赖氨酸，77mg EDC·HCL(0.4mmol)和46mg NHS(0.4mmol)加入到上述溶液中，超声分散均匀，振摇2小时，形成共价交联的水凝胶。加入3mL去离子水，震荡后倒出，如此洗涤三次，以除去未反应的EDC·HCL和NHS，得到纯水凝胶。该水凝胶平均孔径大小15μm，孔隙率约为75％，杨氏模量3.8MPa。240 mg of dopamine-modified cellulose was dissolved in 2 mL of pH 7.4 PBS buffer (0.01 mol/L) to prepare a solution with a mass concentration of 12%, then 120 mg of phenylboronic acid-modified polylysine, 77 mg of EDC ·HCL (0.4 mmol) and 46 mg NHS (0.4 mmol) were added to the above solution, dispersed uniformly by ultrasonic, and shaken for 2 hours to form a covalently cross-linked hydrogel. 3 mL of deionized water was added, poured out after shaking, and washed three times to remove unreacted EDC·HCl and NHS to obtain a pure hydrogel. The average pore size of the hydrogel is 15 μm, the porosity is about 75%, and the Young's modulus is 3.8 MPa.

Claims

Translated fromChinese

1.一种含苯硼酸的纤维素水凝胶的制备方法，其特征在于包括如下步骤：1. a preparation method of the cellulose hydrogel containing phenylboronic acid, is characterized in that comprising the steps:

(1)将具有柔性二元或多元醇或顺式二醇以及氨基的化合物在缩合剂存在下通过酰胺化反应与具有羧基的纤维素盐在水介质中共价偶联制得具有柔性二元或多元醇或顺式二醇的纤维素，(1) Covalently coupling a compound with a flexible dihydric or polyhydric alcohol or cis-diol and an amino group with a cellulose salt having a carboxyl group in an aqueous medium through an amidation reaction in the presence of a condensing agent to obtain a flexible dihydric or Cellulose of polyol or cis-diol,

所述具有柔性二元或多元醇以及氨基的化合物结构通式如下：The general structural formula of the compound with flexible dihydric or polyhydric alcohol and amino group is as follows:

A代表饱和或不饱和的环，两个羟基为邻位或间位顺式取代；

A represents a saturated or unsaturated ring, and the two hydroxyl groups are cis-substituted at ortho or meta positions;

2.如权利要求1所述的制备方法，其特征在于所述步骤(1)具有羧基的纤维素盐选自羧甲基纤维素钠、羧乙基纤维素钠或羧甲基羟丙基纤维素钠中的一种或几种。2. preparation method as claimed in claim 1 is characterized in that the cellulose salt with carboxyl group of described step (1) is selected from sodium carboxymethyl cellulose, sodium carboxyethyl cellulose or carboxymethyl hydroxypropyl cellulose One or more kinds of sodium.

3.如权利要求1所述的制备方法，其特征在于所述步骤(1)所述具有柔性二元或多元醇或顺式二醇以及氨基的化合物选自氨基葡萄糖或多巴胺。3 . The preparation method according to claim 1 , wherein the compound with flexible dihydric or polyhydric alcohol or cis diol and amino group in the step (1) is selected from glucosamine or dopamine. 4 .

4.如权利要求1所述的制备方法，其特征在于步骤(1)所述的缩合剂选自1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和/或N-羟基琥珀酰亚胺。4. preparation method as claimed in claim 1 is characterized in that the described condensing agent of step (1) is selected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and /or N-hydroxysuccinimide.

5.如权利要求1所述的制备方法，其特征在于步骤(1)所述的水介质为酸性或中性，步骤(2)所述水介质为中性或碱性。5 . The preparation method according to claim 1 , wherein the water medium described in step (1) is acidic or neutral, and the water medium described in step (2) is neutral or basic. 6 .

6.如权利要求1所述的制备方法，其特征在于步骤(1)所述

A代表饱和五元环、饱和六元环或不饱和环状基团。6. preparation method as claimed in claim 1 is characterized in that described in step (1)

A represents a saturated five-membered ring, a saturated six-membered ring or an unsaturated cyclic group.

7.如权利要求6所述的制备方法，其特征在于步骤(1)所述

A代表环戊烯、环己烯、苯环、萘、蒽或喹啉。7. preparation method as claimed in claim 6 is characterized in that described in step (1)

A represents cyclopentene, cyclohexene, benzene ring, naphthalene, anthracene or quinoline.

8.如权利要求1所述的制备方法，其特征在于步骤(1)反应温度为10-50℃，反应时间为1-24小时，步骤(2)反应温度为10-50℃，反应时间为10min到1小时，反应条件为静置。8. preparation method as claimed in claim 1 is characterized in that step (1) reaction temperature is 10-50 ℃, and reaction times is 1-24 hours, and step (2) reaction temperature is 10-50 ℃, and reaction times is From 10 min to 1 hour, the reaction condition is to stand.

9.如权利要求1所述的制备方法，其特征在于步骤(2)所述具有柔性二元或多元醇或顺式二醇的纤维素的质量百分浓度为5％-20％，所述的多苯硼酸化合物的质量百分浓度为1％-20％。9 . The preparation method according to claim 1 , wherein the mass percentage concentration of the cellulose with flexible dihydric or polyhydric alcohol or cis diol described in step (2) is 5%-20%, and the The mass percentage concentration of the polyphenylboronic acid compound is 1%-20%.

10.如权利要求1所述的制备方法，其特征在于步骤(2)所述多苯硼酸化合物通过具有两个或两个以上官能团的化合物与带有氨基或羧基的苯硼酸或苯硼酸衍生物偶联制得，所述的具有两个或两个以上官能团的化合物为具有羧基、氨基、羟基、卤素、炔基和叠氮中的一种或几种的聚乙二醇、环糊精、聚氨基酸、蛋白质或多聚糖。10. The preparation method according to claim 1, wherein the polyphenylboronic acid compound described in step (2) passes through a compound having two or more functional groups and a phenylboronic acid or a phenylboronic acid derivative with an amino group or a carboxyl group. Coupling and preparation, the compound with two or more functional groups is polyethylene glycol, cyclodextrin, Polyamino acids, proteins or polysaccharides.

11.如权利要求10所述的制备方法，其特征在于步骤(2)所述具有两个或两个以上官能团的化合物选自二氨基聚乙二醇、二羟基聚乙二醇、α-、β-、γ-环糊精、聚氨基酸、蛋白质、多聚糖中的一种或几种。11. preparation method as claimed in claim 10 is characterized in that the compound with two or more functional groups described in step (2) is selected from diamino polyethylene glycol, dihydroxy polyethylene glycol, α-, One or more of β-, γ-cyclodextrin, polyamino acid, protein, and polysaccharide.

12.一种含苯硼酸的纤维素水凝胶，其特征在于采用如权利要求1-11任一项所述方法制备而成。12. A cellulose hydrogel containing phenylboronic acid, characterized in that it is prepared by the method according to any one of claims 1-11.

13.如权利要求12所述的含苯硼酸的纤维素水凝胶在制备组织工程材料中的应用。13. The application of the phenylboronic acid-containing cellulose hydrogel according to claim 12 in the preparation of tissue engineering materials.