Layered Double Hydroxide Nanotransporter for Molecule Delivery to Intact Plant Cells.

Wenlong Bao,Qiang Wang,Junya Wang,Yinglang Wan,Dermot O'Hare

doi:10.1038/srep26738

Abstract

Here we report a powerful method that facilitates the transport of biologically active materials across the cell wall barrier in plant cells. Positively charged delaminated layered double hydroxide lactate nanosheets (LDH-lactate-NS) with a 0.5‒2 nm thickness and 30‒60 nm diameter exhibit a high adsorptive capacity for negatively charged biomolecules, including fluorescent dyes such as tetramethyl rhodamine isothiocyanate (TRITC), fluorescein isothiocyanate isomer I(FITC) and DNA molecules, forming neutral LDH-nanosheet conjugates. These neutral conjugates can shuttle the bound fluorescent dye into the cytosol of intact plant cell very efficiently. Furthermore, typical inhibitors of endocytosis and low temperature incubation did not prevent LDH-lactate-NS internalization, suggesting that LDH-lactate-NS penetrated the plasma membrane via non-endocytic pathways, which will widen the applicability to a variety of plant cells. Moreover, the absence of unwanted side effects in our cytological studies, and the nuclear localization of ssDNA-FITC suggest that nano-LDHs have potential application as a novel gene carrier to plants.

Highlights

Nanoparticle-based delivery technologies have unique advantages to transport exogenous molecules across the hydrophobic plasma membrane
The X-ray diffraction (XRD) patterns of a dried bulk Mg-Al-lactate sample is given in Fig. 1A, the XRD pattern exhibits the typical Bragg reflections which may be indexed as the 00l reflections in addition to several broader and asymmetrically shaped hk[0] Bragg reflections at higher 2θvalues
A clear Tyndall light scattering effect was observed for the colloidal suspension of Layered double hydroxide (LDH)-lactate-NS, which indicates the presence of well-dispersed exfoliated nanosheets of the layered LDH-lactate (Fig. 1D)

Summary

Introduction

Nanoparticle-based delivery technologies have unique advantages to transport exogenous molecules across the hydrophobic plasma membrane. A suitable nanotransporter for living plant cells must have the following specific properties: (i) nanometer size to penetrate the cell wall, (ii) cause low damage to the PM, and (iii) exhibit a high absorbance capacity for the target molecule(s). Several research groups have developed LDHs as efficient molecule transporter for the delivery of functional nucleotides and bio-active drugs[29,30,31,32] These studies demonstrate that selected LDHs can be excellent transporters to living cells and show good biocompatibility and low cytotoxicity. We found that [Mg0.75Al0.25(OH)2](C3H6O3)0.25 (Mg-Al-lactate LDH) loaded DNA molecules with high efficiency and facilitated delivery of DNA into human cells (293T) with low cytotoxicity[35] These studies encouraged us to investigate the application of delaminated lactate-containing LDH nanosheets to deliver biologically active molecules to intact plant cells

Methods

Results

Conclusion