NanoEngineering and MicroPhysiological Systems

National Institute Of Biomedical Imaging And Bioengineering, Bethesda

Investigators

Abstract

Development of Microfluidic Thyroid Organoid-on-a-chip We are interested in developing a microfluidic model of the thyroid for use in studying thyroid cancer and thyroid autoimmunity. We have established a protocol for generation of thyroid organoids from a human thyroid follicular cell line. We have also established a protocol for generation of endothelial cell microvascular networks using primary human thyroid endothelial cells. We have conducted extensive experiments to optimize an antibody staining protocol for the thyroid cells and organoids. Our end goal is to incorporate the thyroid organoids and microvascular networks within a microfluidic device to enable detailed investigation of cellular interactions in thyroid tissue. Evaluation of Immunomodulatory Properties of Bacterial Extracellular vesicles Extracellular vesicles are small biological nanoparticles that are used as a form of communication for bacterial and mammalian cells. They incorporate proteins and other biological molecules and are exchanged between different cells. We have set up a BSL-1 bacterial culture space within the Section on Immunoengineering lab space, where we have established a workflow and protocols for the isolation of bacterial extracellular vesicles, as well as their characterization through nanoparticle tracking analysis (NTA), tunable resistive pulse sensing (TRPS), and gel electrophoresis. We have also evaluated the effects of different storage conditions and times on bacterial extracellular vesicle integrity and stability. In addition, we have initiated proteomic analysis (through the NIDDK Advanced Mass Spectrometry Core) to identify bacterial proteins and bacterial extracellular vesicle proteins with high sequence homology to thyroid protein epitopes. Through these efforts, we aim to evaluate the potential role of bacterial extracellular vesicles in thyroid immune responses. Development of a Thyroid Extracellular Matrix Hydrogel for Enhanced Thyroid Cell Culture In collaboration with Dr. Kaitlyn Sadtler (NIBIB Section on Immunoengineering), we are developing a hydrogel derived from thyroid extracellular matrix for use in phenotypic cell culture of thyroid cells. Preliminary decellularization protocols have been established, and experiments for hydrogel synthesis are being initiated. Evaluation of Thyroid Autoimmunity in Mice with chronic IFN-gamma expression In collaboration with Dr. Howard Young (NCI), we are evaluating whether chronic IFN-gamma expression in mice, which leads to other forms of autoimmunity, also results in a thyroid autoimmune phenotype. We have established protocols for thyroid tissue isolation and histological staining and have conducted preliminary ELISA assays as part of this work. Establishment of Nanoparticle Synthesis and Characterization Capabilities within NIBIB We have established a nanoparticle characterization space with capability for evaluation of nanoparticle size, zeta potential, and concentration through multiple instruments. This can enable direct evaluation of nanoparticle properties immediately after synthesis or biogenesis. We have optimized instrument protocols for characterization of extracellular vesicles.