Plant secondary metabolites are valuable therapeutics not readily synthesized by traditional chemistry techniques. Although their enrichment in plant cell cultures is possible following advances in biotechnology, conventional methods of recovery are destructive to the tissues. Nanoharvesting, in which nanoparticles are designed to bind and carry biomolecules out of living cells, offers continuous production of metabolites from plant cultures. Here, nanoharvesting of polyphenolic flavonoids, model plant-derived therapeutics, enriched in Solidago nemoralis hairy root cultures, is performed using engineered mesoporous silica nanoparticles (MSNPs, 165 nm diameter and 950 m2/g surface area) functionalized with both titanium dioxide (TiO2, 425 mg/g particles) for coordination binding sites, and amines (NH2, 145 mg/g particles) to promote cellular internalization. Intracellular uptake and localization of the nanoparticles (in Murashige and Skoog media) in hairy roots were confirmed by tagging the particles with rhodamine B isothiocyanate, incubating the particles with hairy roots, and quenching bulk fluorescence using trypan blue. Nanoharvesting of biologically active flavonoids was demonstrated by observing increased antiradical activity (using 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay) by nanoparticles after exposure to hairy roots (indicating general antioxidant activity), and by the displacement of the radio-ligand [3H]-methyllycaconitine from rat hippocampal nicotinic receptors by solutes recovered from nanoharvested particles (indicating pharmacological activity specific to S. nemoralis flavonoids). Post-nanoharvesting growth suggests that the roots are viable after nanoharvesting, and capable of continued flavonoid synthesis. These observations demonstrate the potential for using engineered nanostructured particles to facilitate continuous isolation of a broad range of biomolecules from living and functioning plant cultures.
|Journal||Materials Science and Engineering C|
|State||Published - Jan 2020|
Bibliographical noteFunding Information:
This research was supported by United States National Institutes of Health (NIH Grant nos. R41AT008312 and 2R44AT008312-02 ) and Kentucky Science and Engineering Foundation ( KSEF-2929-RDE-016 ). We thank Dr. Jacob Lilly and Dr. Calvin Cahall for their help and training in fluorescence microscopy.
This research was supported by United States National Institutes of Health (NIH Grant nos. R41AT008312 and 2R44AT008312-02) and Kentucky Science and Engineering Foundation (KSEF-2929-RDE-016). We thank Dr. Jacob Lilly and Dr. Calvin Cahall for their help and training in fluorescence microscopy.
© 2019 Elsevier B.V.
- Cellular internalization
- Engineered mesoporous silica
ASJC Scopus subject areas
- Materials Science (all)
- Condensed Matter Physics
- Mechanics of Materials
- Mechanical Engineering