Bioengineering Single Crystal Growth

ID: MPS/DMR/BMAT(7623) 1106208 PI: Joester， Derk ORG: Northwestern university标题：生物工程单晶生长知识价值：生物矿化组织（例如骨骼，牙齿，外壳）是复杂的有机-无机复合材料，具有纳米级的分层结构。它们的功能作用包括机械强化、光学元件、重力或磁场传感。经过数亿年的进化优化，生物体已经取得了非凡的工程壮举，如低重量高骨骼韧性，自磨牙齿，自我修复能力，低能量足迹，可持续合成。生物晶体生长的许多特征尚未在体外重现：弯曲和/或分支单晶，对多晶的控制，非晶前体的使用，以及有机-无机复合材料的纳米尺度控制。该项目研究了控制细胞培养中单晶协同生长的方法，目的是真正地培养出符合其预期用途的材料。海胆生物矿化系统为这些研究提供了平台。以下具体任务将通过引导生物沉积来探索和发展生物工程单晶形状，连通性和更大的复合结构的范围和局限性：(1)通过多维活细胞显微镜探索微模式上的细胞动力学，以更好地理解粘块之间的桥接现象，细胞在凝集素模式上的运动，以及分支和/或融合位点的形成。(2)描述了针状物、有机基质和细胞机制在针状物发育关键阶段的相互作用，即初始沉积、线状针状物、分支位点和节理。(3)在原代间充质细胞（PMC）培养中，过表达和纯化内源性信号因子SpVEGF-3，作为一种普遍的细刺发生诱导剂，并探索固定化VEGF，研究其作为化学引诱剂的作用。(4)研究PMCs中针状基质蛋白分选信号，确定将蛋白靶向到针状细胞室所需的最小信号肽。(5)利用融合蛋白研究针状基质，并尝试通过表达外源多态性开关（如Starmaker）影响多晶选择。更广泛的影响：在环境温度下从海水中生长新型功能材料的能力可以创造出全新的可持续材料合成方法。此外，该项目还将向大一和大二学生介绍海胆生物矿化和软光刻技术。发现实验室吗?作为入门级材料科学课程一部分的模块。他将通过这个班和MRSEC等中心的少数族裔招募渠道，为学年和夏季研究项目招募本科生研究人员。这些学生在生物相关领域（如分子生物学、生物化学和细胞培养）接受高度跨学科的培训，这将补充洁净室培训、照相和软光刻以及尖端技术的材料表征。该项目将继续为移动实验室开发模块，该实验室由之前的奖项启动，将该研究的各个方面带到当地的高中和中学。此外，正在与芝加哥植物园开发的谋杀神秘活动将适用于谢德水族馆。最后，PI将利用西北大学的资源，通过参加包括实验室参观和公开讲座在内的一系列活动，与整个社会接触。

ID: MPS/DMR/BMAT(7623) 1106208 PI: Joester, Derk ORG: Northwestern UniversityTitle: Bioengineering Single Crystal GrowthINTELLECTUAL MERIT: Biological mineralized tissues (e.g. bones, teeth, shells) are sophisticated organic-inorganic composites with hierarchical architecture down to the nm-scale. Their functional roles include mechanical strengthening, optical elements, and gravity or magnetic field sensing. Optimized in hundreds of millions of years of evolution, organisms have achieved remarkable engineering feats such as high bone toughness at low weight, self-sharpening teeth, self-repair capability, and low energy footprint, sustainable syntheses. Many of the hallmarks of biological crystal growth have yet to be reproduced in vitro: curving and/or branching single crystals, control over polymorph, use of amorphous precursors, and nm scale control of organic-inorganic composites. This project investigates approaches to control the cooperative growth of single crystals in cell culture with the goal to quite literally grow materials with properties optimized for their intended use. The biomineralization system in sea urchin serves as the platform for these studies. The following specific tasks will be pursued to explore and develop the range and limits of bioengineering single crystal shape, connectivity, and larger composite structures by guided biological deposition: (1) Explore, by multi-dimensional live cell microscopy, cellular dynamics on micro-patterns to better understand phenomena of bridging between sticky patches, motility of cells on lectin patterns, and formation of branch and/or fusion sites. (2) Characterize interplay of spicule mineral, organic matrix, and cellular machinery at key points in the spicule development, namely the initial deposit, linear spicules, branch sites, and joints. (3) Overexpress and purify the endogenous signaling factor SpVEGF-3 as a universal inducer of spiculogenesis in primary mesenchyme cell (PMC) cultures, and explore immobilization of the VEGF to study its role as a chemo-attractant. (4) Investigate spicule matrix protein sorting signals in PMCs and determine minimum signal peptide required to target proteins to the spicule compartment. (5) Use fusion proteins to study spicule matrix, and attempt to influence polymorph selection by expression of exogenous polymorph switches such as Starmaker. BROADER IMPACTS: The ability to grow novel functional materials at ambient temperatures from seawater could create entirely new approaches to sustainable materials synthesis. In addition, the PI will expose freshmen and sophomores to sea urchin biomineralization and soft lithography in the context of new ?Discovery Lab? modules that are part of an entry-level materials science class. From this class and through minority recruitment channels at centers such as the MRSEC and others, he will recruit undergraduate researchers for academic year and summer research projects. Such students receive highly interdisciplinary training in bio-related areas (e.g. molecular biology, biochemistry, and cell culture) that will complement clean room training, photo and soft lithography, and materials characterization by cutting-edge techniques. The project will continue to develop modules for the mobile lab jump-started with the previous award to carry aspects of this research to local high and middle schools. In addition, a murder mystery activity that is being developed with the Chicago Botanic Garden will be adapted for use at the Shedd Aquarium. Finally, the PI will leverage resources at Northwestern University to reach out to society at large through participation in a range of events that include lab tours and public lectures.