Bioengineering Single Crystal Growth

ID：MPS/DMR/BMAT(7623) 0805313 PI：Joester, Derk 组织：西北大学 标题：生物工程单晶生长 智力优点：尽管生物矿化组织具有极具吸引力的特性，并且近期在仿生材料合成方面取得了巨大进展，但生物晶体生长的许多特征尚未在体外重现：多晶型控制、弯曲和/或分支单晶以及纳米尺度控制有机-无机复合材料。通过开发一种生物技术替代散装材料合成可以获得很多好处。在这项提案中，PI 将探索海胆胚胎原代间充质细胞 (PMC) 的微图案细胞培养，作为指导负责单晶合成的细胞机制的一种手段。通过这种方式，该项目将利用功能齐全的合成引擎并尝试对其进行重新编程。这将为研究和最终在体外对其进行逆向工程提供基础。将采取以下具体任务来探索和发展通过引导生物沉积工程化单晶形状、连接性和更大复合结构的范围和限制：（1）开发微图案以控制 PMC 在表面上的放置。通过这种方式，引导方解石（CaCO3）单晶的生物沉积。特别是，建立血管内皮生长因子 (VEGF) 诱导的图案形貌、粘附性、趋化性以及限制细胞数量和密度。 (2) 探索连接模式以再现生物三辐射分支或两个单独的单晶元件的融合。将简单的形状和连接结合起来，形成日益复杂的晶体形状和 2D 和 3D 连接性。 (3) 通过活细胞成像、红外和拉曼显微镜以及电子显微镜成像等互补技术，进一步表征生物晶体生长机制的矿物/蛋白质运输途径、无定形前体相以及无定形到晶体的转变。 更广泛的影响：拟议的研究有望在利用生物合成矿化途径创建有用的陶瓷结构方面取得重大进展。 该项目还包括研究和教育一体化的全面计划。 PI 将开发一个完整季度的生物矿化本科课程，旨在培训学生的沟通技巧，让他们接受专业同行评审，并评估他们的分析能力。 学生将通过在线公共访问 WIKI 传播所获得的知识，该 WIKI 将由学生撰写、审阅和编辑。 将举办设计竞赛，开发用于引导晶体生长的新型细胞培养装置，从而吸引学生参与 PI 实验室积极研究的问题，并在跨学科项目中运用他们的工程技能。 PI实验室的本科生研究机会将在海胆饲养、细胞培养和活细胞成像等生物相关领域提供高度跨学科的交叉培训，这些活动将补充洁净室光刻和先进成像技术材料表征的培训。面向高中生的外展活动将针对芝加哥、罗杰斯公园和埃文斯顿的多元化社区。由于实验室目前所有研究生都是女性，因此将实现对积极女性榜样的特别集中的投射。将与高中科学教师、工程和生物材料课程模块、西北校区参观以及暑期实习计划结合起来。

ID: MPS/DMR/BMAT(7623) 0805313 PI: Joester, Derk ORG: Northwestern UniversityTitle: Bioengineering Single Crystal GrowthINTELLECTUAL MERIT: Despite the highly attractive properties of biological mineralized tissues and great recent progress in bio-inspired materials synthesis, many of the hallmarks of biological crystal growth have yet to be reproduced in vitro: polymorph control, curving and/or branching single crystals, and nm scale control of organic-inorganic composites. Much could be gained by developing a biotechnological alternative to bulk materials synthesis. In this proposal the PI will explore micro-patterned cell culture of sea urchin embryonic primary mesenchyme cells (PMCs) as a means to guide the cellular machinery responsible for single crystal synthesis. In this way the project will take advantage of a fully capable synthetic engine and attempt to reprogram it. This will then provide the basis for investigating and ultimately reverse engineering it in vitro. The following specific tasks will be pursued to explore and develop the range and limits of engineering single crystal shape, connectivity, and larger composite structures by guided biological deposition: (1) Develop micro-patterning to control the placement of PMCs on a surface. In this way, guide the biological deposition of single calcite (CaCO3) crystals. In particular, establish pattern topography, adhesiveness, chemotaxis induced by vascular endothelial growth factor (VEGF), and limiting cell numbers and densities. (2) Explore junction patterns to reproduce the biological tri-radiate branching or fusion of two individual single crystalline elements. Combine simple shapes and junctions for increasingly complex crystal shape and connectivity in 2D and 3D. (3) Further characterize the biological crystal growth machinery with respect to routes of mineral/protein trafficking, amorphous precursor phases, and amorphous-to crystalline transitions by a complementary suite of techniques including live cell imaging, IR and Raman microscopy, and electron microscopic imaging. BROADER IMPACTS: The proposed research promises a major advance in harnessing biosynthetic mineralization pathways to create useful ceramic structures. The project also includes a thorough plan for integration of research and education. The PI will develop a full quarter undergraduate course in biomineralization designed to train students in communications skills, expose them to professional peer review, and assess their analytical skills. Students will disseminate the knowledge gained through an online, public access WIKI that will be written, reviewed, and edited by students. A design competition will be held to develop new cell culture devices for guided crystal growth and thus draw students into questions actively researched in the PI?s laboratory and engage their engineering skills in a trans-disciplinary project. Undergraduate research opportunities in the PI?s lab will provide highly interdisciplinary cross training in bio-related areas such as sea urchin husbandry, cell culture, and live cell imaging, and these activities will complement training in clean room photolithography and materials characterization by advanced imaging techniques. Outreach to high school students will be targeted to ethnically diverse neighborhoods of Chicago, Rogers Park, and Evanston. As all current graduate students in the lab are female, an especially focused projection of positive female role models will be achieved. In conjunction with high school science teachers, engineering and biomaterials class modules, visits on the Northwestern Campus, and a summer internship program will be organized.