Design of Cell-Recognizable Nano-device and Application to Biosensing and Tissue-engineering

在我们的研究中，我们设计了细胞可识别的纳米探针来分析细胞在体外的行为，并像在体内一样调节它们。本课题的最终目标是利用所设计的纳米探针，在体外创造细胞的纳米调控环境，具有再生医学和细胞靶向诊断和治疗的潜力。生物纳米技术的研究目标是基于此的。本课组建立了细胞-基质工程技术，利用新设计的载糖聚合物和嵌合蛋白，如E-cadherin Fc、EGF-Fc、liff -Fc等，调控es细胞和肝细胞的功能。纳米/微图像化、探针/载体技术和生物材料被用于细胞的动态测量和调控。建立了碳酸盐磷灰石纳米载体技术，用于将dna、rna、蛋白质和药物高效地递送到窖内结构域。我们成功实现了三大目标。他们是，1。新型细胞识别探针的设计及其在细胞处理中的应用[j]。生物相容性纳米载体系统在靶向细胞治疗中的应用；采用双光子聚合法设计纳米/微三维图案培养基质。首先，我们设计了具有细胞可识别结构域和IgG-Fc结构域的嵌合蛋白。我们发现Fc结构域可以成功地将纳米探针锚定在任何类型的疏水性衬底上，从聚苯乙烯盘子到纳米纤维。对于细胞识别域，我们测试的范围从细胞粘附蛋白（如E-cadherin和N-cadherin）到细胞因子（如EGF和LIF）。我们设计的任何一种配体，细胞都成功地识别了它们，我们发现它们可以按照我们的设计来调节细胞。下一步，我们发现每一种基因工程的嵌合蛋白都可以用作纳米探针。我们已经将细胞可识别蛋白应用于靶向细胞治疗的药物/基因传递系统，以及对培养细胞的局部刺激。我们发现我们的纳米探针可以实现每一个目标。最后，我们利用双光子引发聚合的方法设计了纳米调控的培养底物。在我们的项目中，可以制作分辨率为180nm的培养基质，也可以在基质上培养细胞。所制备的衬底可以用于细胞动力学行为的分析。少

In our research, we have designed cell-recognizable nano-probe to analyze cellular behavior in vitro and regulate thme as in vivo. Final goal of our project is to create nano-regulated circumstances of the cells in vitro by using designed nano-probe which have a potential for regenerative medicine and cell-targetting diagnosis and therapy. Bionanothechnology are based on the research objectives. Cell-Matrices engineering are established by our group to regulate the functions of ES-cells as well as hepatocytes using newly-engineered sugar-carrying polymers and chimeric proteins such as E-cadherin Fc, EGF-Fc, LIF-Fc and so on. Nano/micro patterning, probes/carriers technology and biomaterials were developed for dynamic measurements and regulations of cells. The nano carriers technology of carbonate apatite was also established for highly efficient delivery of DNAs, RNAs, proteins and drugs into intra-cellar domain.We have successfully achieved three major goal. They are, 1. Designing nov … More el cell-recognizable probe as genetically-engineered matrices and application to the cell processing, 2. Designing biocompatible nano-carrier system for targeted cell therapy, 3. Designing nano/ micro 3D patterned culture substrate using two photon polymerization method. For first step, we have designed chimeric proteins which owns cell-recognizable domain and IgG-Fc domain. We found Fc domain successfully allows to anchore nano-probe to any kide of hydrophobic substrate ranging from polystyrene dish to the nano-fiobers. For cell-recognizable domain we tested ranging from cell-cell adhesion proteins such E-cadherin and N-cadherin to cytokines such as EGF and LIF. Any type of ligand we design, the cells successfully recognized them and we found they could regulate cells as we designed. For the next step, we found each of genetically engineered such chimeric proteins were possible to use as nano-probes. We have applied the cell-recognizable proteins for drug/ gene delivery system for targeted cell therapy, and for local stimulation to thte cultured cells. We found our nano-probes could achieve each of the goal. Finally, we desinged nano-regulated culture substrate by using two-photon initiate polymerizatin method. In our project, culture substrate could be fabricated with resolution of 180nm, and also cell could be cultured on the substrate. The substrate we fabricated could achieved to analyze cellular dynamic behavior. Less