EAGER: Interfacial Design of Modular, Multifunction Peptide Amphiphile Hydrogels for Tissue Engineering

1253913/ KokkoliIntellectual Merit：在许多情况下，单一的生物活性配体或蛋白质被用来功能化生物材料，因此，它不能准确地模拟必要的复杂性，以支持或促进所需的生物现象，如长期细胞粘附、存活和表型维持。因此，设计多功能界面的能力变得越来越重要，因为许多生物材料需要不同的配体和化学线索。到目前为止，还没有研究表明支架功能化与多类配体的好处是显著的还是附加的。在这个探索性的EAGER提案中，PI建议设计肽类两亲体，模拟各种细胞结合结构域（纤维连接蛋白和I型和III型胶原）、生长因子（如碱性成纤维细胞生长因子（bFGF））和生长因子结合肽，以模块化方式组合并产生明确的多组分水凝胶界面，优化以支持不同细胞的培养。这种新型的细胞外基质(ECM)-模拟肽两亲性水凝胶支架，能够通过多种信号生物分子的模拟肽的模块化组合来复制足够的生化复杂性，将产生紧急特性。不同的肽两亲体将被评估为水凝胶单独，成对和不同浓度的混合物。所有凝胶将被表征流变测量，以评估其模量和扫描电子显微镜（SEM）图像。原代人肝细胞将被用作模型细胞系统，并将被播种在水凝胶的界面上以及水凝胶内部（2D和3D评估）。细胞增殖、细胞活力以及肝细胞功能将在不同的时间点使用不同的检测方法进行评估。更广泛的影响：设计能够呈现更复杂信号的接口将最终允许探索方法，反过来，这可能对生物传感器、组织工程和再生医学中潜在应用的功能生物材料领域的进步做出重大贡献。教学计划与研究计划紧密结合。教育活动将侧重于通过实验室实习、参加小组会议和国家会议以及在科学期刊上发表文章来培训研究生和本科生。PI的目标是吸引和留住女性和其他代表性不足的群体进入科学和工程领域。PI在这一领域有着良好的记录。

1253913/ KokkoliIntellectual Merit: In many cases, a single bioactive ligand or protein is used to functionalize a biomaterial, and as a result, it fails to accurately mimic the complexity necessary to support or promote desired biological phenomena such as long term celladhesion, survival and phenotype maintenance. The ability to design multifunctional interfaces is therefore becoming increasingly important as many biomaterials require the presence of different ligands and chemical cues. Thus far, there is no study that has investigated whether the benefits of scaffold functionalization with multiple classes of ligands are significant or additive. In this exploratory EAGER proposal, the PI proposes to design peptide amphiphiles that will mimic various cell binding domains (fibronectin and type I and type III collagen), growth factors, such as the basic fibroblast growth factor (bFGF), and a growth factor binding peptide, to be combined in a modular fashion and produce defined, multicomponent hydrogel interfaces, optimized to support the culture of different cells. This novel extracellular matrix (ECM)-mimetic peptide amphiphile hydrogel scaffold, capable of reproducing sufficient biochemical complexity via modular combinations of peptide mimetics from multiple classes of signaling biomolecules will give rise to emergent properties. The different peptide amphiphiles will be evaluated as hydrogels individually, in pairs and in mixtures at different concentrations. All gels will be characterized with rheology measurements to evaluate their modulus and scanning electron microscope (SEM) images. Primary human hepatocytes will be used as the model cell system and will be seeded on the interface of the hydrogels as well as within the hydrogels (2D and 3D evaluation). Cell proliferation, cell viability as well as hepatocyte function will be evaluated using different assays at different time points.Broader Impact: Design of interfaces capable of presenting more complex signals will ultimately allow for exploration of approaches which could, in turn, significantly contribute to advances in the field of functional biomaterials with potential applications in biosensors, tissue engineering and regenerative medicine. The educational plan is strongly integrated with the research plan. Educational activities will focus on training of graduate and undergraduate students though lab exposure, participation in group meetings and national meetings as well as publishing in scientific journals. The goal of the PI is to attract and retain women and other underrepresented groups to science and engineering. The PI has a strong record in this area.