Engineering multi-layered tissue constructs using acoustic levitation.

Engineering tissue structures that mimic those found in vivo remains a challenge for modern biology. We demonstrate a new technique for engineering composite structures of cells comprising layers of heterogeneous cell types. An acoustofluidic bioreactor is used to assemble epithelial cells into a sheet-like structure. On transferring these cell sheets to a confluent layer of fibroblasts, the epithelial cells cover the fibroblast surface by collective migration maintaining distinct epithelial and fibroblast cell layers. The collective behaviour of the epithelium is dependent on the formation of cell-cell junctions during levitation and contrasts with the behaviour of mono-dispersed epithelial cells where cell-matrix interactions dominate and hinder formation of discrete cell layers. The multilayered tissue model is shown to form a polarised epithelial barrier and respond to apical challenge. The method is useful for engineering a wide range of layered tissue types and mechanistic studies on collective cell migration.

Biocompatibility of poly(2-alkyl-2-oxazoline) brush surfaces for adherent lung cell lines.

Development of synthetic surfaces that are highly reproducible and biocompatible for in vitro cell culture offers potential for development of improved models for studies of cellular physiology and pathology. They may also be useful in tissue engineering by removal of the need for biologically-derived components such as extracellular matrix proteins. We synthesised four types of 2-alkyl-2-oxazoline polymers ranging from the hydrophilic poly(2-methyl-2-oxazoline) to the hydrophobic poly(2-n-butyl-2-oxazoline). The polymers were terminated using amine-functionalised glass coverslips, enabling the synthetic procedure to be reproducible and scaleable. The polymer-coated glass slides were tested for biocompatibility using human epithelial (16HBE14o-) and fibroblastic (MRC5) cell lines. Differences in adhesion and motility of the two cell types was observed, with the poly(2-isopropyl-2-oxazoline) polymer equally supporting the growth of both cell types, whereas poly(2-n-butyl-2-oxazoline) showed selectivity for fibroblast growth. In summary, 2-alkyl-2-oxazoline polymers may be a useful tool for building in vitro model cell culture models with preferential adhesion of specific cell types.

A thin-reflector microfluidic resonator for continuous-flow concentration of microorganisms: a new approach to water quality analysis using acoustofluidics.

An acoustofluidic device has been developed for concentrating vegetative bacteria in a continuous-flow format. We show that it is possible to overcome the disruptive effects of acoustic streaming which typically dominate for small target particles, and demonstrate flow rates compatible with the testing of drinking water. The device consists of a thin-reflector multi-layered resonator, in which bacteria in suspension are levitated towards a glass surface under the action of acoustic radiation forces. In order to achieve robust device performance over long-term operation, functional tests have been carried out to (i) maintain device integrity over time and stabilise its resonance frequency, (ii) optimise the operational acoustic parameters, and (iii) minimise bacterial adhesion on the inner surfaces. Using the developed device, a significant increase in bacterial concentration has been achieved, up to a maximum of ~60-fold. The concentration performance of thin-reflector resonators was found to be superior to comparable half-wave resonators.