Biogenic thermoresponsive polyelectrolyte multilayers as potential substrata for the generation of cell sheets for tissue engineering

生物热响应聚电解质多层作为组织工程细胞片生成的潜在基质

• 批准号: 369505795
• 负责人: Professor Dr. Thomas Groth
• 金额: --
• 依托单位: Abteilung Holztechnologie und Holzwerkstoffe
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/369505795?language=en
• 关键词: Biogenic; thermoresponsive; polyelectrolyte; multilayers; potential

A novel approach for the formation of thermoresponsive surface coatings based on derivatives from cellulose and chitosan as biogenic, biocompatible and environmentally-friendly biopolymers from renewable resources will be explored for making culture substrata for mammalian cells to permit non-enzymatic release of cells and cell sheets by temperature change. Three different thermoresponsive elements together with sulfate as anionic groups will be introduced into the backbone of these biopolymers. The resulting polyelectrolytes will be systematically characterized regarding their chemical structures, molecular weights, zeta potentials and lower critical solution temperatures (LCST), in order to achieve thermoresponsive compounds with a sufficient charge density as well as a transition temperature around 37°C. After the selection of candidate derivatives with suitable LCST and charge density, these polyelectrolytes will be used for the formation of multilayers via the layer-by-layer (LbL) technique, which will be studied by surface plasmon resonance, quartz micro balance, ellipsometry and contact angle measurements. The thermoresponsive effects shall be analyzed by changes of water content and wetting properties of the multilayer coatings in the temperature range from 5°C-37°C. Protein adsorption is a prerequisite for the adhesion of cells and has been shown to be a determinant of cell adhesion on thermoresponsive surface coatings, which will be studied in this project regarding the binding of fibronectin and serum proteins. Multilayers with thermoresponsive properties that permit reversible adsorption of proteins will be further used to study the adhesion and growth of cells from three different germ layers. The thermoresponsive effect will be studied by changes in cell shape and detachment of single cells during decrease of temperature from 37°C to 5°C as well as by the release of complete cell sheet in response to temperature changes. Altogether, the development of the new system provides essential advantages over existing fully synthetic thermoresponsive polymers like poly(N-isopropylacrylamide) because of the inherent bioactivity of sulfated cellulose/chitosan towards mitogenic and morphogenic growth factors, the excellent biocompatibility as well as the potential long-term degradability. These advantages allow these systems not only to be feasible for in vitro applications to generate cell sheets for engineering various tissues for transplantation including skin, cornea, myocardium, etc., but also to be potentially useful for diverse in vivo applications, where the thermoresponsive release of proteins and/or cells is desirable.

一种新的方法，用于形成的温度响应性表面涂层的基础上衍生物的纤维素和壳聚糖作为生物，生物相容性和环境友好的生物聚合物从可再生资源将探索用于制造培养基质的哺乳动物细胞，允许非酶释放的细胞和细胞片的温度变化。将三种不同的温敏元素与硫酸根作为阴离子基团一起引入这些生物聚合物的主链中。将对所得聚电解质的化学结构、分子量、zeta电位和低临界溶解温度（LCST）进行系统表征，以获得具有足够电荷密度以及约37°C转变温度的热敏化合物。在选择具有合适的LCST和电荷密度的候选衍生物之后，这些聚电解质将用于通过逐层（LbL）技术形成多层膜，该技术将通过表面等离子体共振、石英微天平、椭圆偏振仪和接触角测量来研究。应通过多层涂层在5°C-37°C温度范围内含水量和润湿性能的变化来分析温敏效应。蛋白质吸附是细胞粘附的先决条件，并已被证明是一个决定因素的细胞粘附在温敏表面涂层，这将是在这个项目中研究的纤连蛋白和血清蛋白的结合。具有温度响应特性的多层膜允许蛋白质的可逆吸附，将进一步用于研究来自三个不同胚层的细胞的粘附和生长。通过在温度从37°C降低至5°C期间细胞形状的变化和单细胞的脱离以及通过响应于温度变化的完整细胞片的释放来研究温度响应效应。总而言之，新系统的开发提供了比现有的完全合成的温敏聚合物如聚（N-异丙基丙烯酰胺）的基本优势，因为硫酸化纤维素/壳聚糖对促有丝分裂和形态发生生长因子的固有生物活性，优异的生物相容性以及潜在的长期降解性。这些优点使得这些系统不仅可用于体外应用以产生用于工程化各种移植组织（包括皮肤、角膜、心肌等）的细胞片，而且还可潜在地用于需要蛋白质和/或细胞的温度响应性释放的各种体内应用。