Microengineered Platforms for High-throughput Characterization of Cellular Microenvironments

用于细胞微环境高通量表征的微工程平台

• 批准号: RGPIN-2014-04010
• 负责人: Kim, Keekyoung
• 金额: $ 1.68万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=679329
• 关键词: Microengineered; Platforms; throughput; Characterization; Cellular

Osteoarthritis is the most common form of arthritis, affecting 1 out of every 10 people in Canada. The total cost of arthritis is estimated at $33 billion dollars per year. Similarly, millions of patients in Canada undergo procedures to correct bone deformities each year. Tissue engineering seeks to provide an alternate therapeutic approach using stem cells. Mesenchymal stem cells derived from bone marrow retain a multi-lineage potential to differentiate to fat cells, heart muscle cells, bone cells, and cartilage cells. The physical properties of local microenvironments will play an important role in the differentiation of mesenchymal stem cell. A number of studies have been aimed at combining stem cells with biomaterials for cartilage and bone regeneration, but to date, no study has systematically examined the combinatorial factors to control the physical properties for stem cell differentiation in a 3-D microenvironment. In the 3-D cellular microenvironment, cells interact with their surroundings by processing various chemical and physical signals. To control cellular microenvironments for stem cells, hydrogels have attracted great interests due to high water content, biocompatibility, and mechanical properties resembling natural tissues. However, various parameters of hydrogels are associated with controlling stem cell fate; the optimization of the parameters is essential. Therefore, a high-throughput screening technology to test many parameters in one experiment will facilitate the systematic examination and the optimization of cellular microenvironments generated by hydrogels.**High-throughput screening technology is a method of using robotic liquid handling devices to quickly fabricate microarrays of chemical, genetic, and pharmacological materials and conduct tests in a high-throughput manner. In this research program, we adopt this technology to develop a hydrogel microarray with hundreds of micrometer-sized functional hydrogel spots (diameter: ~500µm; thickness: ~100µm) printed on a standard microscope slide. We will use alginate hydrogels that can be polymerized by ultra violet light. We will test 27 combinations of alginate hydrogels with three different parameters and three different conditions of each parameter that control physical properties. We will characterize the physical properties, such as microstructure, stiffness, and adhesion, using advanced characterization techniques such as atomic force microscopy and scanning electron microscopy. The developed alginate microarray will be used to optimize the effects of physical properties on mesenchymal stem cell differentiation.**Although the molecular mechanisms associated with the biological responses have yet to be clarified, such technology may be widely applicable in cell-microenvironment research. Also, the physical properties identified in this program could be used as design parameters for engineering new biomaterials and microenvironments for tissue regeneration. In this program, we propose to engineer and apply high-throughput screening technology for the identification of cell-microenvironment interactions that increase stem cell differentiation into the bone and cartilage. The results of this research will have high potential to positively affect the large number of patients that undergo procedures to repair bone and cartilage each year. Thus, the orthopedic community in Canada would benefit from the new treatment techniques composed of osteogenic and chondrogenic biomaterials that not only support, but also induce tissue formation. The developed platforms will also be applicable to numerous tissue regeneration applications, such as cardiovascular and nerve tissues and beneficial for biomaterials and tissue engineering research in Canada.

骨关节炎是关节炎的最常见形式，在加拿大每10人中，每年的总成本为33美元。使用干细胞向脂肪细胞提供治疗方法。在3-D微环境中，细胞通过处理各种AL和物理信号来控制干细胞的细胞微环境，由于较高的水含量而引起了极大的兴趣，但类似于自然组织水凝胶的参数与控制参数的圆锥形相关。 - 在此研究计划中，我们采用了数百个微米大小的功能水凝胶点（直径：〜500µm厚度：〜100µm）的水凝胶微阵列。通过超紫色的光聚合，我们将使用高级特征（例如原子力显微镜和扫描），测试每个参数的三个不同参数和三种不同的条件。显微镜。开发的藻酸盐将优化与生物学反应的分化的物理性质的裂痕，尚未被固定。组织的微环境。这不是组织组织，开发的平台也将用于加拿大生物材料和组织工程研究的众多组织层面应用。