This mini-review aims at gaining knowledge on basic aspects of plant nanotechnology. While in recent years the enormous progress of nanotechnology in biomedical sciences has revolutionized therapeutic and diagnostic approaches, the comprehension of nanoparticle-plant interactions, including uptake, mobilization and accumulation, is still in its infancy. Deeper studies are needed to establish the impact of nanomaterials (NMs) on plant growth and agro-ecosystems and to develop smart nanotechnology applications in crop improvement. Herein we provide a short overview of NMs employed in plant science and concisely describe key NM-plant interactions in terms of uptake, mobilization mechanisms, and biological effects. The major current applications in plants are reviewed also discussing the potential use of polymeric soft NMs which may open new and safer opportunities for smart delivery of biomolecules and for new strategies in plant genetic engineering, with the final aim to enhance plant defense and/or stimulate plant growth and development and, ultimately, crop production. Finally, we envisage that multidisciplinary collaborative approaches will be central to fill the knowledge gap in plant nanotechnology and push toward the use of NMs in agriculture and, more in general, in plant science research.

Poly(N-isopropylacrylamide) (PNIPAm) is widely used to fabricate cell sheet surfaces for cell culturing, however copolymer and interpenetrated polymer networks based on PNIPAm have been rarely explored in the context of tissue engineering. Many complex and expensive techniques have been employed to produce PNIPAm-based films for cell culturing. Among them, spin coating has demonstrated to be a rapid fabrication process of thin layers with high reproducibility and uniformity. In this study, we introduce an innovative approach to produce anchored smart thin films both thermo- and electro-responsive, with the aim to integrate them in electronic devices and better control or mimic different environments for cells in vitro. Thin films were obtained by spin coating of colloidal solutions made by PNIPAm and PAAc nanogels. Anchoring the films to the substrates was obtained through heat treatment in the presence of dithiol molecules. From analyses carried out with AFM and XPS, the final samples exhibited a flat morphology and high stability to water washing. Viability tests with cells were finally carried out to demonstrate that this approach may represent a promising route to integrate those hydrogels films in electronic platforms for cell culture applications.

Parylene C is a well-known polymer and it has been mainly employed as a protective layer for implantable electronics. In this paper, we propose a new approach to use Parylene C as a versatile template for patterning soft materials potentially applicable as scaffolds in cardiac tissue engineering (TE). Parylene C substrates were anisotropically patterned through standard lithographic process with hydrophilic channels separating raised hydrophobic strips. Ridges and grooves of the template are 10 µm width and depth ranging from 1 to 17 µm. Polydimethylsiloxane (PDMS) and Polyacrylamide (PAm) hydrogel have been chosen as soft polymers to be moulded. Thanks to their chemical and physical properties PDMS and PAm hydrogel mimic the extracellular matrix (ECM). PDMS was spin coated on micropatterned Parylene C obtaining composite substrates with 460 nm and 1.15 µm high grooves. The Young’s modulus of the composite Parylene C/PDMS was evaluated and it was found to be almost half when compared to PDMS. PAm hydrogel was also printed using collagen coated micro-grooved Parylene C. Optical micrographs and fluorescence analysis show the successful topographic and protein pattern transfer on the hydrogel.

In this work, soft microgels of Poly(N-Isopropylacrylamide) (PNIPAm) at two different sizes and of interpenetrated polymer network (IPN) composed of PNIPAm and Poly(Acrylic Acid) (PAAc) were synthesized. Then, solutions of these different types of microgels have been spin-coated on glass substrates with different degrees of hydrophobicity. PNIPAm particles with a larger diameter form either patches or a continuous layer, where individual particles are still distinct, depending on the dispersion concentration and spin speed. On the other, PNIPAm particles with a smaller diameter and IPN particles form a continuous and smooth film, with a thickness depending on the dispersion concentration and spin-speed. The difference in morphology observed can be explained if one considers that the microgels may behave as colloidal particles or macromolecules, depending on their size and composition. Additionally, the microgel size and composition can also affect the stability of the depositions when rinsed in water. In particular, we find that the smooth and continuous films show a stimuli-dependent stability on parameters such as temperature and pH, while large particle layers are stable under any condition except on hydrophilic glass by washing at 50 °C.

We recently demonstrated that patterned Parylene C films could be effectively used as a mask for directly copolymerizing proteins on polyacrylamide hydrogel (PAm). In this work, we have proved the applicability of this technique for studying the effect such platforms render on neonatal rat ventricular myocytes (NRVMs). Firstly, we have characterised topographically and mechanically the scaffolds in liquid at the nano-scale level. We thus establish that such platforms have physical properties that closely mimics the in vivo extracellular environment of cells. We have then studied the cell morphology and physiology by comparing cultures on flat uniformly-covered and collagen-patterned scaffolds. We show that micro-patterns promote the elongation of cells along the principal axis of the ridges coated with collagen. In several cases, cells also tend to create bridges across the grooves. We have finally studied cell contraction, monitoring Ca2+ cycling at a certain stimulation. Cells seeded on patterned scaffolds present significant responses in comparison to the isotropic ones.

In cardiac tissue engineering (TE), in vitro models are essential for the study of healthy and pathological heart tissues in order to understand the underpinning mechanisms. In this scenario, scaffolds are platforms that can realistically mimic the natural architecture of the heart, and they add biorealism to in vitro models. This paper reports a novel and robust technique to fabricate cardiovascular-mimetic scaffolds based on Parylene C and Polydimethylsiloxane (PDMS). Parylene C is employed as a mask material for inducing hybrid and nonhybrid micropatterns to the PDMS layer. Hybrid architectures present striped hydrophobic/hydrophilic surfaces, whereas non-hybrid scaffolds only corrugated topographies. Herein, we demonstrate that wavy features on PDMS can be obtained at the micro-and nanoscale and that PDMS can be integrated into the microfabrication process without changing its intrinsic physical properties. A study of the effects of these scaffolds on the growth of Neonatal Rat Ventricular Myocytes (NRVMs) cultures reveals that cell alignment occurs only for the case of hybrid architectures made of hydrophilic PDMS and hydrophobic Parylene C.

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