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Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.

ABSTRACT: Microscale biopatterning enables regulation of cell-material interactions and cell shape, and enables multiplexed high-throughput studies in a cell- and reagent-efficient manner. The majority of available techniques rely on physical contact of a stamp, pin, or mask with mainly a dry surface. Inkjet and piezoelectric printing is carried out in a non-contact manner but still requires a substantially dry substrate to ensure fidelity of printed patterns. These existing methods, therefore, are limited for patterning onto delicate surfaces of living cells because physical contact or substantially dry conditions are damaging to them. Microfluidic patterning with laminar streams does enable non-contact patterning in fully aqueous environments but with limited throughput and reagent diffusion across interfacial flows. Here, we describe a polymeric aqueous two-phase system that enables patterning nanolitres of a reagent-containing aqueous phase, in arbitrary shapes, within a second aqueous phase covering a cell monolayer. With the appropriate medium formulation, reagents of interest remain confined to the patterned phase without significant diffusion. The fully aqueous environment ensures high reagent activity and cell viability. The utility of this strategy is demonstrated with patterned delivery of genetic materials to mammalian cells for phenotypic screening of gene expression and gene silencing.

SUBMITTER: Tavana H

PROVIDER: S-EPMC2782768 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

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Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.

Tavana H H Jovic A A Mosadegh B B Lee Q Y QY Liu X X Luker K E KE Luker G D GD Weiss S J SJ Takayama S S

Nature materials 20090816 9

Microscale biopatterning enables regulation of cell-material interactions and cell shape, and enables multiplexed high-throughput studies in a cell- and reagent-efficient manner. The majority of available techniques rely on physical contact of a stamp, pin, or mask with mainly a dry surface. Inkjet and piezoelectric printing is carried out in a non-contact manner but still requires a substantially dry substrate to ensure fidelity of printed patterns. These existing methods, therefore, are limite ...[more]

PMID: 19684584

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Project description:In the current era of human life, we have been facing an increased consumption of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs). Nevertheless, NSAIDs are not completely metabolized by humans and are further excreted into domestical effluents. Several studies have been showing that a wide variety of pharmaceuticals are present in water effluents and are thus a matter of serious concern in the public health. Although treatment plants use sophisticated technologies for pollutants/contaminants removal, none of these processes was particularly designed for NSAIDs. In this perspective, this work addresses the use of a liquid-liquid extraction approach, employing ionic liquids (ILs), for the removal of NSAIDs from aqueous media. In particular, aqueous biphasic systems (ABS) composed of ILs and aluminium-based salts, which are already used in water treatment plants, were tested for the removal of diclofenac, ibuprofen, naproxen and ketoprofen. With these systems, extraction efficiencies of NSAIDs up to 100% were obtained in a single-step. The recovery of NSAIDs from the IL medium and the recyclability of the IL-rich phase were then ascertained to guarantee the development of a more sustainable and cost-effective strategy. Based on the remarkable increase in the solubility of NSAIDs in the IL-rich phase (from a 300- to a 4100-fold when compared with pure water), water was then studied as an effective anti-solvent, and where single-step recovery percentages of NSAIDs from the IL-rich phase up to 91% were obtained. After the "cleaning" of the IL-rich phase by the induced precipitation of NSAIDs, the phase-forming components were recovered and reused in four consecutive cycles, with no detected losses on both the extraction efficiency and recovery of NSAIDs by induced precipitation. Finally, an integrated process is here proposed, which comprises the (i) removal of NSAIDs from aqueous media, (ii) the cleaning of the IL-rich phase by the recovery of NSAIDs by induced precipitation, and (iii) the phase-forming components recycling and reuse, aiming at unlocking new doors for alternative treatment strategies of aqueous environments.

Project description:We have investigated the phase behavior of self-assembled lyotropic liquid crystals (LC) formed by ternary mixtures of oleoylethanolamide (OEA), water and arginine. OEA, a natural analog of the endogenous cannabinoid anandamide involved in the peripheral regulation of feeding, was selected as a main component due to its capacity to induce efficient decreases in food intake and gains in body mass. Arginine was selected as representative hydrophilic amino acid and added to the OEA-water mixture at different concentrations. The phase diagrams were determined by combining cross-polarized optical microscopy and small angle x-ray scattering. First, the phase diagram for the OEA-water system was determined. It was shown that these two compounds give rise to reverse Ia3d double gyroid and reverse Pn3m double diamond cubic phases existing in bulk over a large window of temperature and composition, and that for water content beyond 25% Pn3m coexisted with excess water. Successively, the influence of arginine as guest molecule in the water channels of the reverse LC was investigated. For the sake of comparison, results for the OEA-water-arginine system were compared with analog series of OEA-water-glucose. The results showed that, at a fixed water content and temperature, the phase behavior of the liquid crystalline phases is strongly dependent on arginine concentration. In more detail, arginine could be encapsulated in the bulk OEA-water LC up to 2.0% wt, whereas transitions from Ia3d to Pn3m cubic phase were observed with increasing arginine concentration. Interestingly, upon an increase of water concentration beyond 20-25%, Pn3m phase started to coexist with excess water releasing the arginine in external water solution. Quantitative measurements of arginine content inside the LC water channels and in the excess external water solution revealed a complete release of the amino acid, demonstrating that the investigated lyotropic liquid crystalline systems can be used as ideal vehicles for the delivery of functional hydrophilic active molecules in aqueous environment.

Dataset Information

Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.

Publications

Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.

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