Bioengineering Single Crystal Growth

ID：MPS/DMR/BMAT（7623）1106208 主要研究者：Joester，Derk ORG：西北大学名称：生物工程单晶生长智力优势：生物矿化组织（如骨骼，牙齿，贝壳）是复杂的有机-无机复合材料，具有纳米级的层次结构。 它们的功能作用包括机械强化、光学元件和重力或磁场传感。 经过数亿年的进化优化，生物体已经取得了卓越的工程壮举，例如重量轻的高骨骼韧性，自磨牙齿，自我修复能力，以及低能量足迹，可持续合成。 生物晶体生长的许多标志尚未在体外重现：弯曲和/或分支单晶，多晶型物的控制，无定形前体的使用，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体协同生长的方法，目标是真正生长出具有针对其预期用途优化的特性的材料。 海胆中的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探索和开发生物工程单晶形状、连接性和更大复合结构的范围和限制：（1）通过多维活细胞显微镜探索微观模式上的细胞动力学，以更好地理解粘性斑块之间的桥接现象、凝集素模式上的细胞运动性以及分支和/或融合位点的形成。 (2)描述骨针矿物、有机基质和细胞机械在骨针发育关键点的相互作用，即初始存款、线性骨针、分支位点和关节。 (3)过表达和纯化内源性信号传导因子SpVEGF-3作为原代间充质细胞（PMC）培养物中毛刺形成的通用诱导剂，并探索VEGF的固定化以研究其作为化学引诱剂的作用。 (4)研究骨针基质蛋白分选信号在PMC和确定所需的最小信号肽靶蛋白的骨针隔室。 (5)利用融合蛋白研究骨针基质，并尝试通过表达外源性多晶型开关（如Starmaker）来影响多晶型选择。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为可持续材料合成创造全新的方法。 此外，PI将暴露新生和新生海胆生物矿化和软光刻的背景下，新的？探索实验室？这些模块是入门级材料科学课程的一部分。 从这个班，并通过少数民族招聘渠道在中心，如MRSEC和其他人，他将招募本科研究人员学年和夏季研究项目。 这些学生在生物相关领域（例如分子生物学，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，照片和软光刻以及尖端技术的材料表征。 该项目将继续为移动的实验室开发模块，该实验室是由上一个奖项启动的，将这项研究的各个方面带到当地的高中和中学。 此外，一个正在与芝加哥植物园开发的谋杀之谜活动将被改编为在谢德水族馆使用。 最后，PI将利用西北大学的资源，通过参加一系列活动，包括实验室图尔斯和公开讲座，向社会广泛宣传。

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.