Biotechnological engineering of turnip mosaic virus-derived nanoparticles by functionalization with natural compounds of different biological activities

Abstract

ABSTRACT In this thesis, plant-nanoparticles derived from Turnip mosaic virus (TuMV) where evaluated as technological platforms. These Viral Nanoparticles (VNPs) can be modified by chemically attaching molecules to its surface in order to cater different purposes which make them valuable biomaterials. The VNPs were produced by inoculation of Indian mustard (Brassica juncea). TuMV was evaluated for biomedical applitcations by testing cytotoxicity to cell cultures. Furthermore, TuMV was coupled with fluorescent molecules Alexa555 and Cy5.5 to assess interaction of TuMV with cells. Interaction of TuMV with mammal cell lines was found at both extracellular and intracellular levels. TuMV-Cy5.5 was injected to immunodeficient mice (Foxn1nu), followed in vivo for four days and organs were harvested to determine organ distribution of VNP distribution. It was found that liver and tumor tissue showed the highest accumulation of TuMV-Cy5.5. In this thesis, the chemical conjugation of epigallocatechin gallate (EGCG) was performed via Mannich reaction. TuMV-EGCG showed enhanced antimicrobial, antibiofilm and antitumor activity in comparison to free EGCG. In vivo assays of Pseudomonas syringae against tomato plants showed that EGCG-TuMV was capable of diminishing bacterial virulence. Also, antiviral activity against zika virus (ZIKV) was tested finding low antiviral activity. TuMV was tested as an additive for gelatyn methacryloil (GelMa). Hydrogels resemble some of the properties of the extracellular matrix in mammals and have become a key biomaterial in the tissue engineering arena. Assays with TuMV-GelMA formulations showed that this can be use as a bioink for 3-D printing and as a scaffold for cell proliferation enhancing enlongation. Furthermore the epithelial growth factor (EGF) was chemically attached to TuMV surface and used as an functionalized additive for GelMa bioinks. Tests showed that EGF retained its activity and its addition to a gel matrix allowed cellular prolifetation. In this thesis, the possibility of chemically attach different molecules while retaining its activity is proven. Also, it tests the versatility of TuMV as a biomaterial that can be used in an array of different scenarios.