DEVELOPMENT OF FEASIBILITIES FOR IMMUNE CELL-MEDIATED NANOPARTICLE TRANSPORT THROUGH THE ENDOTHELIAL BARRIER UNDER FLOW CONDITIONS
Open Access
- Author:
- Anumba, Nmachi Chinenye
- Millennium Scholars Program:
- Biomedical Engineering (BME)
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisor:
- Cheng Dong, Thesis Supervisor
- Keywords:
- Drug delivery
Nanoparticles
Cell migration
Blood-Brain Barrier - Abstract:
- The blood-brain barrier (BBB) has consistently proved itself to be a roadblock in the development and administration of drugs for central nervous system (CNS) disorders and brain tumors. The cellular barrier is notorious for its highly selective permeability which limits the number of effective CNS drugs that have direct access to the brain. For this reason, study was done on the feasibility of using T lymphocyte Jurkat cells as a carrier to uptake Biodegradable Photoluminescent Polylactic Acid (BPLP-PLA) nanoparticles as they migrate through the BBB, carrying these nanoparticles to the other side of the tight endothelial barrier. This study builds on previous work in which this drug delivery mechanism was studied under static conditions by adding a flow component to the experiments. A new bottom flow chamber plate was designed and made to accommodate for the use of suspensive Jurkat cells and facilitate a more accurate data collection process through use of flow cytometry. A reliable protocol was also developed for making a tight, confluent in vitro BBB model comprised of Bovine Brain Microvascular Endothelial Cells (BBMVEC). The migration of Jurkat cells was studied statically in the newly designed chamber plate at a concentration of 2 x 105 cells/mL and a CXCL12 chemoattractant concentration of 500 ng/mL. Flow migration studies of the Jurkat cells were then performed under a shear rate of 50 sec-1. Tumor Necrosis Factor – α (TNF- α) was used to treat the endothelial cells and upregulate the expression of ICAM-1 in an effort to increase the migration of the Jurkat immune cells under flow conditions. However, the increase in migration shown by the use of TNF- α was proven to be statistically insignificant by way of the t-test. The BPLP-PLA nanoparticles were successfully conjugated to the Jurkat cells and an experiment comparing nanoparticles being carried by Jurkat cells to a control experiment in which Jurkat cells were not used was performed. The results of this experiment showed an increase in migrated molecules with the assistance of Jurkat cells, however there was little difference in the sample fluorescence reading of the two experiments. This indicates that a significant amount of nanoparticles crossed the barrier in both cases and that further research needs to be done in the optimization of this process for more accurate results.