Bioengineering Single Crystal Growth

ID：MPS/DMR/BMAT（7623）0805313 PI：Joester，Derk ORG：西北大学名称：生物工程单晶生长智力优势：尽管生物矿化组织具有非常吸引人的特性，并且最近在生物启发材料合成方面取得了很大进展，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，以及有机-无机复合材料的纳米尺度控制。通过开发一种替代散装材料合成的生物技术，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）的微模式细胞培养，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，该项目将充分利用一个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进行逆向工程提供基础。以下具体任务将通过引导生物沉积来探索和开发工程单晶形状、连接性和较大复合结构的范围和限制：（1）开发微图案化以控制PMC在表面上的放置。通过这种方式，引导生物沉积单方解石（CaCO 3）晶体。特别是，建立模式拓扑结构，血管内皮生长因子（VEGF）诱导的趋化性，并限制细胞数量和密度。(2)探索连接模式来重现生物三辐射分支或两个独立单晶元素的融合。结合联合收割机简单的形状和结，以实现日益复杂的晶体形状和2D和3D连接。(3)通过一套互补的技术，包括活细胞成像、IR和拉曼显微镜和电子显微镜成像，进一步表征生物晶体生长机制中的矿物质/蛋白质运输路线、无定形前体相和无定形到结晶转变。 更广泛的影响：这项研究有望在利用生物合成矿化途径创造有用的陶瓷结构方面取得重大进展。 该项目还包括一个全面的研究和教育一体化计划。 PI将开发一个完整的四分之一的生物矿化本科课程，旨在培养学生的沟通技巧，让他们接触专业的同行评审，并评估他们的分析能力。 学生将通过一个在线的、公共访问的维基百科来传播所获得的知识，该维基百科将由学生撰写、审查和编辑。 一个设计比赛将举行，以开发新的细胞培养装置，指导晶体生长，从而吸引学生到积极研究的问题，在PI？的实验室和从事他们的工程技术在一个跨学科项目。 PI的本科研究机会？的实验室将提供高度跨学科的交叉培训，在生物相关领域，如海胆养殖，细胞培养，活细胞成像，这些活动将补充培训，在洁净室光刻和材料表征先进的成像技术。对高中生的宣传将针对芝加哥、罗杰斯公园和埃文斯顿的种族多样化社区。由于实验室目前所有的研究生都是女性，因此将实现积极的女性榜样的特别集中的投射。与高中科学教师，工程和生物材料类模块，在西北校区参观，并将组织暑期实习计划。

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.