Key Laboratory of Functional Polymer Materials, Ministry of Education, Institute of Polymer Chemistry, College of Chemistry, Nankai University, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300071, PR China.
Key Laboratory of Functional Polymer Materials, Ministry of Education, Institute of Polymer Chemistry, College of Chemistry, Nankai University, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300071, PR China.
Acta Biomater. 2017 May;54:259-270. doi: 10.1016/j.actbio.2017.03.009. Epub 2017 Mar 8.
A thermoresponsive polymer-protein biodynamer was prepared via the bioconjugation of an aliphatic aldehyde-functionalized copolymer to hydrazine-modified bovine serum albumin (BSA) through reversible pyridylhydrazone linkages. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography (SEC) results indicated that the pyridylhydrazone linkages cleaved in an intracellular-mimicking acidic milieu, thus leading to the release of BSA. The dynamic character of the protein biodynamer was demonstrated by exchange reactions with aldehyde-containing molecules. The biodynamer self-assembled into spherical micelles at a temperature above its lower critical solution temperature (LCST). Subsequently, BSA molecules within the hydrophilic coronae of the micelles were readily cross-linked via reaction with cystamine at 45°C, and multi-stimuli-responsive nanoparticles were generated. The biohybrid nanoparticles reversibly swelled and shrank as the cores of the nanoparticles were solvated below the LCST and desolvated above the LCST. The accessible reversibility of the pyridylhydrazone bonds imparts pH-responsive and dynamic characteristics to the nanoparticles. The nanoparticles displayed glutathione (GSH) responsiveness, and the synergistic effects of pH and GSH resulted in complete disintegration of the nanoparticles under the intracellular-mimicking acidic and reductive conditions. The nanoparticles were also enzyme-responsive and disintegrated rapidly in the presence of protease. In vitro cytotoxicity and cell uptake assays demonstrated that the nanoparticles were highly biocompatible and effectively internalized by HepG2 cells, which make them interesting candidates as vehicles for drug delivery application and biomimetic platforms to investigate the biological process in nature.
In this research, we report the synthesis of a temperature and pH dual-responsive polymer-protein biodynamer through reversible pyridylhydrazone formation. The prepared biodynamer can offer a potential platform for intracellular protein delivery. The multi-stimuli-responsive biohybrid nanoparticles containing disulfide functionalities are constructed by cross-linking albumin coronae of the biodynamer micelles. With the combination of a thermoresponsive polymer, protein and reversible covalent bonds, the biohybrid nanoparticles are endowed with highly biocompatible, environmentally responsive and adaptive features. These nanoparticles present the ability to undergo changes in their constitution, hydrodynamic size and nanostructure in response to physical, chemical and biological stimuli, which make them interesting candidates as vehicles for drug delivery application and a biomimetic platform to investigate the biological process in nature.
通过将脂肪族醛官能化共聚物与通过可逆吡啶腙键合到肼改性牛血清白蛋白（BSA）上，制备了温敏聚合物-蛋白质生物动力学剂。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和尺寸排阻色谱（SEC）结果表明，在细胞内模拟的酸性环境中，吡啶腙键断裂，导致 BSA 释放。通过与含醛分子的交换反应证明了蛋白质生物动力学剂的动态特性。生物动力学剂在其下临界溶液温度（LCST）以上自组装成球形胶束。随后，在 45°C 下，胶束亲水性冠层内的 BSA 分子通过与胱胺反应很容易交联，并生成多刺激响应性纳米颗粒。当纳米颗粒的核在 LCST 以下被溶剂化且在 LCST 以上被去溶剂化时，生物杂交纳米颗粒会可逆地溶胀和收缩。吡啶腙键的可及性可逆性赋予纳米颗粒 pH 响应和动态特性。纳米颗粒显示出谷胱甘肽（GSH）响应性，并且 pH 和 GSH 的协同作用导致在细胞内模拟的酸性和还原条件下纳米颗粒完全解体。纳米颗粒还对蛋白酶具有响应性，并在存在蛋白酶的情况下迅速分解。体外细胞毒性和细胞摄取实验表明，纳米颗粒具有高度的生物相容性，并被 HepG2 细胞有效内化，这使它们成为药物输送应用的有前途的载体和仿生平台，以研究自然界中的生物过程。
在这项研究中，我们通过可逆吡啶腙形成报告了一种温度和 pH 双重响应的聚合物-蛋白质生物动力学剂的合成。所制备的生物动力学剂可为细胞内蛋白质递药提供潜在平台。通过交联生物动力学剂胶束的白蛋白冠，构建了含有二硫键功能的多刺激响应性生物杂交纳米颗粒。通过将温敏聚合物、蛋白质和可逆共价键结合，生物杂交纳米颗粒具有高度的生物相容性、环境响应性和适应性。这些纳米颗粒具有根据物理、化学和生物刺激改变其组成、流体动力尺寸和纳米结构的能力，这使它们成为药物输送应用的有前途的载体和仿生平台，以研究自然界中的生物过程。
