Spatiotemporal Regulated Click Hydrogels for 3D Cell Culture

用于 3D 细胞培养的时空调节点击水凝胶

• 批准号: 1006711
• 负责人: Kristi Anseth
• 金额: $ 42万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-15 至 2013-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1006711&HistoricalAwards=false
• 关键词: Spatiotemporal; Regulated; Click; Hydrogels; 3D

The aim of this proposal is to fully develop click chemistry reactions to create a dynamic in vitro 3D cell culture platform that will enable researchers to explore how cell-material interactions influence important cellular functions. These biomaterials will give the user control of specific physical, chemical, and biological cues comprising the cell microenvironment in both time and space. To accomplish this significant challenge, the proposed work will exploit the classic alkyne/azide click chemistry, modified to enable a cytocompatible copper-free polymerization, to synthesize basic hydrogels as a simplified mimic of the extracellular matrix in the presence of cells. The biochemical functionality of the gels will then be tuned using a new variant of click chemistry, a thiol-ene photoaddition reaction, that is fully compatible with peptide chemistry and allows spatiotemporally regulated introduction of biological epitopes. Finally, a photodegradable linker will be integrated into the crosslinks of the base gel formulation to allow light directed degradation of the material. Using these three independent and cytocompatible reactions for gelation, photodegradation, and photocoupling, dynamic biomaterials systems will be created that will provide newfound opportunities to probe the dynamic exchange of information between cells and their microenvironment. The specific aims of this work are to: (1) synthesize click-based hydrogels using orthogonal chemistries that allow manipulation of the material properties through photodegradation and introduction of biological epitopes through a photocoupling reaction, (2) manipulate the properties of the above hydrogels through photodegradation, photocoupling, and combined photodegradation and photocoupling, (3) culture human mesenchymal cells (hMSCs) on 2D gel surfaces and characterize morphology, cytoskeletal organization, and focal adhesions as a function of elasticity and patterned adhesive ligand presentation, (4) encapsulate hMSCs in 3D and examine their response to material-directed morphological changes and spatially varying adhesive ligand distribution, (5) train a diverse group of undergraduate and graduate students at the interface of polymer chemistry and molecular biology, and (6) disseminate these results to the industrial and academic public to assure maximum impact on fundamental and applied biomaterials chemistry.BROADER IMPACTS: This new class of multifunctional hydrogels will help overcome numerous problems with existing 3D cell culture platforms by providing a facile means for manipulating material functionality and mechanics with spatial and temporal control. These new materials will enable researchers to probe fundamental biological questions via a variety of previously unattainable 3D cell studies. The efficient nature of the gel chemistry will allow its adoption by a wide array of non-experts. These material systems should find applications for basic 3D cell culture, design of tissue engineering matrices, platforms for drug delivery and screening, stand-alone biomaterials implants, as well as other non-biological applications. The multidisciplinary team environment in the Anseth laboratory, coupled with long-standing collaborations with world-class clinical and biological laboratories, will provide an exceptional educational environment for multiple graduate and undergraduate students. The PI and her research group have a history of extensive outreach to high school students and the general public, and the team will work diligently through multiple mechanisms to highlight the impact of biomaterial science in benefiting society.

该提案的目的是充分开发点击化学反应，以创建动态的体外3D细胞培养平台，使研究人员能够探索细胞-材料相互作用如何影响重要的细胞功能。 这些生物材料将使用户控制特定的物理，化学和生物线索，包括细胞微环境在时间和空间。 为了完成这一重大挑战，拟议的工作将利用经典的炔/叠氮化物点击化学，修改，使细胞相容的无铜聚合，合成基本的水凝胶作为一个简化的模拟细胞外基质的存在下的细胞。 然后将使用点击化学的新变体（硫醇-烯光加成反应）来调整凝胶的生物化学功能性，所述硫醇-烯光加成反应与肽化学完全相容，并且允许时空调节的生物表位的引入。 最后，可光降解的连接体将被整合到基础凝胶制剂的交联中，以允许材料的光定向降解。 使用这三个独立的和细胞相容的反应凝胶化，光降解和光耦合，动态生物材料系统将被创建，这将提供新的发现的机会，以探测细胞和它们的微环境之间的动态信息交换。 这项工作的具体目标是：（1）使用正交化学合成基于点击的水凝胶，所述正交化学允许通过光降解和通过光偶联反应引入生物表位来操纵材料性质，（2）通过光降解、光偶联和组合的光降解和光偶联来操纵上述水凝胶的性质，（3）在2D凝胶表面上培养人间充质细胞（hMSC），并表征形态、细胞骨架组织和作为弹性和图案化粘附配体呈递的函数的粘着斑，（4）在3D中包封hMSC并检查它们对材料定向的形态变化和空间变化的粘附配体分布的响应，（5）在高分子化学和分子生物学的界面上培养一批多样化的本科生和研究生，（6）将这些结果传播给工业界和学术界，以确保对基础和应用生物材料化学产生最大的影响。这类新的多功能水凝胶将通过提供一种简便的方法来帮助克服现有3D细胞培养平台的许多问题，通过空间和时间控制来操纵材料功能和力学。 这些新材料将使研究人员能够通过各种以前无法实现的3D细胞研究来探索基本的生物学问题。 凝胶化学的高效性质将允许其被广泛的非专家采用。这些材料系统应该可以应用于基本的3D细胞培养，组织工程基质的设计，药物输送和筛选平台，独立的生物材料植入物以及其他非生物应用。在安赛斯实验室的多学科团队环境，再加上与世界一流的临床和生物实验室的长期合作，将为多个研究生和本科生提供一个特殊的教育环境。PI和她的研究小组有着广泛接触高中生和公众的历史，该团队将通过多种机制努力工作，以突出生物材料科学在造福社会方面的影响。