基于甲基乙烯基醚—马来酸交替共聚物智能水凝胶的制备及性能研究

发布时间：2018-10-08 18:19

【摘要】：为了满足生物医学研究需求及实现应用目的,需要开发具有各种功能的智能水凝胶。智能水凝胶因具有广阔的应用前景而成为生物医学领域的研究热点之一。本论文以具有良好生物相容性的合成高分子聚合物甲基乙烯基醚-马来酸交替共聚物P(MVE-alt-MA)为主要原料,采用不同的方法制备了四种基于P(MVE-alt-MA)的智能水凝胶,并系统地研究了这些智能水凝胶的物理和化学性质。主要内容包括：1. P(MVE-alt-MA)-g-β-CD/P(MVE-alt-MA)-g-Azo超分子水凝胶的制备及刺激响应性。首先通过酰胺化反应将环糊精衍生物(EDA-6-β-CD)接枝到P(MVE-alt-MA),得到主体聚合物P(MVE-alt-MA)-g-β-CD;接着通过酰胺化反应将对氨基偶氮苯(Azo)接枝到P(MVE-alt-MA),得到客体聚合物P(MVE-alt-MA)-g-Azo。最后,在室温下,将主体聚合物P(MVE-aIt-MA)-g-β-CD水溶液与客体聚合物P(MVE-alt-MA)-g-Azo水溶液混合,通过主链上p-CD与Azo主-客体间包结络合作用形成P(MVE-alt-MA)-g-β-CD/P(MVE-alt-MA)-g-Azo超分子水凝胶。该超分子水凝胶呈现相互连通的多孔结构和易于调节的流变学特性,它具有光响应性和pH值响应性。利用CCK-8检测水凝胶对人卵巢癌细胞SKOV3细胞的毒性,结果显示该超分子水凝胶没有明显的细胞毒性。采用激光扫描共聚焦显微镜观察了经DiO染色的SKOV3细胞分布在该超分子水凝胶内部不同深度。经培养在该水凝胶中一段时间的SKOV3细胞通过紫外光照射致水凝胶向溶胶转变而被成功脱附,且从水凝胶中分离出的SKOV3细胞仍能存活。此细胞分离研究工作为P(MVE-alt-MA)智能水凝胶应用于细胞后续研究奠定实验和理论基础。2. P(MVE-alt-MA)-g-β-CD/P(MVE-alt-MA)-g-Ad超分子水凝胶的制备及刺激响应性。首先,以LiH作反应助剂,通过酯化反应将β-CD接枝到P(MVE-alt-MA),得到主体聚合物P(MVE-alt-MA)-g-β-CD;接着以DCC作催化剂,通过酰胺化反应将盐酸金刚烷乙胺(Ad)接枝到P(MVE-alt-MA),得到客体聚合物P(MVE-alt-MA)-g-Ad。最后,在室温下,将主体聚合物P(MVE-alt-MA)-g-β-CD水溶液与客体聚合物P(MVE-alt-MA)-g-Ad水溶液混合,通过主链上β-CD与Ad间的包结络合作用组装形成P(MVE-alt-MA)-g-β-CD/P(MVE-alt-MA)-g-Ad超分子水凝胶。该超分子水凝胶具有pH值响应性和自愈合性。该水凝胶被证明具良好生物相容性,可被用作细胞三维培养支架。此研究工作为功能化细胞三维培养支架的制备提供新途径和新材料。3. P(MVE-alt-MA)/CS复合纳米凝胶的制备及性能。首先通过P(MVE-alt-MA)中的羧基与壳聚糖(CS)中的氨基发生静电相互作用形成聚电解质络合物,接着通过交联剂戊二醛(GA)中的醛基与CS中的氨基形成席夫碱获得纳米凝胶。该纳米凝胶具有pH值敏感性。以盐酸阿霉素为模型药物,研究了该纳米凝胶的体外药物释放行为。体外药物释放实验结果显示,该纳米凝胶有望被用作药物载体。4. P(MVE-alt-MA)-crosslinked-AChS水凝胶的制备及性能。以EDC/NHS作催化剂,ADH为交联剂通过化学交联制备了P(MVE-alt-MA)-crosslinked-AChS水凝胶。该水凝胶具有pH值敏感性,在适宜浓度条件下对细胞无明显细胞毒性,且细胞在水凝胶中能够良好的增殖,有望作为细胞培养支架。此研究工作为天然聚合物与人工合成聚合物复合型水凝胶的制备及应用于细胞三维培养提供研究基础。总的来说,本工作制备了一系列基于P(MVE-alt-MA)的智能水凝胶。这类智能水凝胶可为生物医学研究的应用提供强有力的平台,尤其是在三维细胞培养等方面。
[Abstract]:In order to meet the needs of biomedical research and achieve application, intelligent hydrogels with various functions need to be developed. Intelligent hydrogel has become one of the hot spots in the field of biomedicine because of its wide application prospect. Four kinds of intelligent hydrogels based on P (MVE-MAA-MA) were prepared by different methods, and the physical and chemical properties of these intelligent hydrogels were studied systematically. The main content includes: 1. Preparation and stimulation response of P (MVE-MAA-MA)-g-MAA-CD/ P (MVE-MAA-MA)-g-Azo supermolecular hydrogels. The cyclodextrin derivatives (EDA-6-MAA-CD) were first grafted onto P (MVE-MAA-MA) by a chlorosulfonation reaction to obtain a bulk polymer P (MVE-MAA-MA)-g-AZO-CD; followed by grafting of p (MVE-MAA-MA) to P (MVE-MAA-MA) by a chlorosulfonation reaction to give the guest polymer P (MVE-MAA-MA)-g-Azo. Finally, at room temperature, the main polymer P (MVE-aIt-MA)-g-MAA-CD aqueous solution was mixed with the guest polymer P (MVE-MAA-MA)-g-Azo aqueous solution, and P (MVE-MAA-MA)-g-MAA-CD/ P (MVE-MAA-MA)-g-Azo supermolecular hydrogels were formed by the complexation of p-CD on the main chain and the Azo main-guest. The ultra-molecular hydrogel exhibits a porous structure in communication with each other and a rheological property which is easy to adjust, and has light responsiveness and pH responsiveness. Using CCK-8 to detect the toxicity of hydrogel on SKOV3 cells in human ovarian cancer cells, the results showed that the supermolecular hydrogel had no obvious cytotoxicity. Laser scanning confocal microscopy was used to observe the distribution of SKOV3 cells stained by DiO in different depths inside the hydrogel. SKOV3 cells cultured in the hydrogel for a period of time were successfully desorbed by UV irradiation of the hydrogel to the sol, and SKOV3 cells separated from the hydrogel remained viable. This cell separation study lay an experimental and theoretical foundation for cell-based follow-up study of P (MVE-MAA-MA) intelligent hydrogel. Preparation and stimulation response of P (MVE-MAA-MA)-g-MAA-CD/ P (MVE-MAA-MA)-g-Ad supermolecular hydrogels. At first, LiH is used as the reaction additive to graft the MO-CD to P (MVE-MAA-MA) through esterification reaction to obtain the main polymer P (MVE-MAA-MA)-g-MAA-CD; followed by DCC as the catalyst, and the adamantane ethylamine (Ad) is grafted to P (MVE-MAA-MA) through the esterification reaction to obtain the guest polymer P (MVE-MAA-MA)-g-Ad. Finally, at room temperature, the main polymer P (MVE-MAA-MA)-g-MAA-CD aqueous solution was mixed with the guest polymer P (MVE-MAA-MA)-g-Ad aqueous solution, and P (MVE-MAA-MA)-g-MAA-CD/ P (MVE-MAA-MA)-g-Ad supermolecular hydrogels were assembled by the ligation and complexation of CdSe-CD on the main chain and Ad. The super-molecular hydrogel has a pH value response and a self-healing property. The hydrogel is demonstrated to have good biocompatibility and can be used as a cell three-dimensional culture scaffold. This study provides a new approach and new material for the preparation of functional cell three-dimensional culture scaffold. Preparation and Properties of P (MVE-MAA-MA)/ CS Composite Nanogels The polyelectrolyte complex is first formed by electrostatic interaction with the amino groups in the chitosan (CS) in P (MVE-MAA-MA), and then the nanogel is obtained by the formation of the aldehyde groups in the aldehyde groups in the cross-linking agent glutaraldehyde (GA) and the amino groups in the CS. the nanogel has a ph sensitivity. In vitro drug release behavior of the nano-gel was studied by using adriamycin as a model drug. In vitro drug release experiments showed that the nanogel was expected to be used as a drug carrier. Preparation and properties of P (MVE-MAA-MA)-Crosslinked-Sepharose hydrogel. A P (MVE-MAA-MA)-Crosslinked-Sepharose hydrogel was prepared by chemical crosslinking with EDC/ MAA as catalyst. The hydrogel has the sensitivity of pH value, has no obvious cytotoxicity to the cells under suitable concentration conditions, and the cell can be well proliferated in the hydrogel, and is expected to be used as a cell culture support. The research work provides a foundation for the preparation of natural polymer and synthetic polymer composite hydrogel and its application in three-dimensional culture of cells. In general, this work prepares a series of intelligent hydrogels based on P (MVE-MAA-MA). This intelligent hydrogel can provide a powerful platform for biomedical research applications, especially in three-dimensional cell culture.
【学位授予单位】：东南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R943

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