Application of sequential cyclic compression on cancer cells in a flexible microdevice.

Mechanical forces shape physiological structure and function within cell and tissue microenvironments, during which cells strive to restore their shape or develop an adaptive mechanism to maintain cell integrity depending on strength and type of the mechanical loading. While some cells are shown to experience permanent plastic deformation after a repetitive mechanical tensile loading and unloading, the impact of such repetitive compression on deformation of cells is yet to be understood. As such, the ability to apply cyclic compression is crucial for any experimental setup aimed at the study of mechanical compression taking place in cell and tissue microenvironments. Here, we demonstrate such cyclic compression using a microfluidic compression platform on live cell actin in SKOV-3 ovarian cancer cells. Live imaging of the actin cytoskeleton dynamics of the compressed cells was performed for varying pressures applied sequentially in ascending order during cell compression. Additionally, recovery of the compressed cells was investigated by capturing actin cytoskeleton and nuclei profiles of the cells at zero time and 24 h-recovery after compression in end point assays. This was performed for a range of mild pressures within the physiological range. Results showed that the phenotypical response of compressed cells during recovery after compression with 20.8 kPa differed observably from that for 15.6 kPa. This demonstrated the ability of the platform to aid in the capture of differences in cell behaviour as a result of being compressed at various pressures in physiologically relevant manner. Differences observed between compressed cells fixed at zero time or after 24 h-recovery suggest that SKOV-3 cells exhibit deformations at the time of the compression, a proposed mechanism cells use to prevent mechanical damage. Thus, biomechanical responses of SKOV-3 ovarian cancer cells to sequential cyclic compression and during recovery after compression could be revealed in a flexible microdevice. As demonstrated in this work, the observation of morphological, cytoskeletal and nuclear differences in compressed and non-compressed cells, with controlled micro-scale mechanical cell compression and recovery and using live-cell imaging, fluorescent tagging and end point assays, can give insights into the mechanics of cancer cells.

Microdevice-based mechanical compression on living cells.

Compressive stress enables the investigation of a range of cellular processes in which forces play an important role, such as cell growth, differentiation, migration, and invasion. Such solid stress can be introduced externally to study cell response and to mechanically induce changes in cell morphology and behavior by static or dynamic compression. Microfluidics is a useful tool for this, allowing one to mimic in vivo microenvironments in on-chip culture systems where force application can be controlled spatially and temporally. Here, we review the mechanical compression applications on cells with a broad focus on studies using microtechnologies and microdevices to apply cell compression, in comparison to off-chip bulk systems. Due to their unique features, microfluidic systems developed to apply compressive forces on single cells, in 2D and 3D culture models, and compression in cancer microenvironments are emphasized. Research efforts in this field can help the development of mechanoceuticals in the future.

Wavelength Selective Solar Cells Using Triple Cation Perovskite.

Perovskite materials offer high-efficiency low-cost solar cells and applications versatility. We report on cesium-based hybrid perovskite solar cells with wavelength-selective properties ranging from 500 nm (UV-VIS) to 800 nm (IR). The band gap tuning was achieved through composition changes of mainly lead(II) iodide PbI2 and lead(II) bromide PbBr2. The optical spectra of the developed materials were studied, including the photoluminescence (PL), optical transparency, X-ray diffraction and external quantum efficiency for samples prepared under different compositions. It was found that a high content of iodine displayed a photoluminescence (PL) peak at 790 nm, whereas a high content of bromine showed a PL peak at 548 nm. The combined composition mixture of PbI2 and PbBr2 can be fine-tuned to prepare materials that absorbed light in the visible range (640-660 nm) or other selective wavelengths in the range from 500 to 800 nm. The illuminated current-voltage characteristics of the solar cells were carried out under the AM 1.5 condition using an ABET solar simulator with a reference solar cell for comparison and control. The average efficiency of the fabricated solar cells ranged from 3.5% to 15.5%, depending on perovskite composition. Wavelength-selective solar cells have potential applications in smart windows, building of integrated PVs and solar-operated greenhouses.

Finite-size and disorder effects on 1D unipartite and bipartite surface lattice resonances.

Optical resonances in bipartite metal nanostructure lattices are more resilient to finite size-effects than equivalent unipartite lattices, but the complexities of their behaviour in non-ideal settings remain relatively unexplored. Here we investigate the quality factor and extinction efficiency of 1D Ag and Au unipartite and bipartite lattices. By modelling finite size lattices over a range of periods we show that the quality factor of Ag bipartite lattices is significantly better than unipartite lattices. This improvement is less pronounced for Au bipartite lattices. We also show that bipartite lattices are dramatically affected by structure size variations at scales that are typically seen in electron beam lithography fabrication in contrast to unipartite lattices, which are not as sensitive.

Conductive Bioimprint Using Soft Lithography Technique Based on PEDOT:PSS for Biosensing.

Culture platform surface topography plays an important role in the regulation of biological cell behaviour. Understanding the mechanisms behind the roles of surface topography in cell response are central to many developments in a Lab on a Chip, medical implants and biosensors. In this work, we report on a novel development of a biocompatible conductive hydrogel (CH) made of poly (3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) and gelatin with bioimprinted surface features. The bioimprinted CH offers high conductivity, biocompatibility and high replication fidelity suitable for cell culture applications. The bioimprinted conductive hydrogel is developed to investigate biological cells' response to their morphological footprint and study their growth, adhesion, cell-cell interactions and proliferation as a function of conductivity. Moreover, optimization of the conductive hydrogel mixture plays an important role in achieving high imprinting resolution and conductivity. The reason behind choosing a conducive hydrogel with high resolution surface bioimprints is to improve cell monitoring while mimicking cells' natural physical environment. Bioimprints which are a 3D replication of cellular morphology have previously been shown to promote cell attachment, proliferation, differentiation and even cell response to drugs. The conductive substrate, on the other hand, enables cell impedance to be measured and monitored, which is indicative of cell viability and spread. Two dimensional profiles of the cross section of a single cell taken via Atomic Force Microscopy (AFM) from the fixed cell on glass, and its replicas on polydimethylsiloxane (PDMS) and conductive hydrogel (CH) show unprecedented replication of cellular features with an average replication fidelity of more than 90%. Furthermore, crosslinking CH films demonstrated a significant increase in electrical conductivity from 10-6 S/cm to 1 S/cm. Conductive bioimprints can provide a suitable platform for biosensing applications and potentially for monitoring implant-tissue reactions in medical devices.

Tumour Initiation: a Discussion on Evidence for a "Load-Trigger" Mechanism.

Appropriate mechanical forces on cells are vital for normal cell behaviour and this review discusses the possibility that tumour initiation depends partly on the disruption of the normal physical architecture of the extracellular matrix (ECM) around a cell. The alterations that occur thence promote oncogene expression. Some questions, that are not answered with certainty by current consensus mechanisms of tumourigenesis, are elegantly explained by the triggering of tumours being a property of the physical characteristics of the ECM, which is operative following loading of the tumour initiation process with a relevant gene variant. Clinical observations are consistent with this alternative hypothesis which is derived from studies that have, together, accumulated an extensive variety of data incorporating biochemical, genetic and clinical findings. Thus, this review provides support for the view that the ECM may have an executive function in induction of a tumour. Overall, reported observations suggest that either restoring an ECM associated with homeostasis or targeting the related signal transduction mechanisms may possibly be utilised to modify or control the early progression of cancers. The review provides a coherent template for discussing the notion, in the context of contemporary knowledge, that tumourigenesis is an alliance of biochemistry, genetics and biophysics, in which the physical architecture of the ECM may be a fundamental component. For more definitive clarification of the concept there needs to be a phalanx of experiments conceived around direct questions that are raised by this paper.

Nanostructures for Light Trapping in Thin Film Solar Cells.

Thin film solar cells are one of the important candidates utilized to reduce the cost of photovoltaic production by minimizing the usage of active materials. However, low light absorption due to low absorption coefficient and/or insufficient active layer thickness can limit the performance of thin film solar cells. Increasing the absorption of light that can be converted into electrical current in thin film solar cells is crucial for enhancing the overall efficiency and in reducing the cost. Therefore, light trapping strategies play a significant role in achieving this goal. The main objectives of light trapping techniques are to decrease incident light reflection, increase the light absorption, and modify the optical response of the device for use in different applications. Nanostructures utilize key sets of approaches to achieve these objectives, including gradual refractive index matching, and coupling incident light into guided modes and localized plasmon resonances, as well as surface plasmon polariton modes. In this review, we discuss some of the recent developments in the design and implementation of nanostructures for light trapping in solar cells. These include the development of solar cells containing photonic and plasmonic nanostructures. The distinct benefits and challenges of these schemes are also explained and discussed.

The extracellular topographical environment influences ovarian cancer cell behaviour.

The importance of the biophysical cellular environment in cancer development has been increasingly recognised but so far has been only superficially studied. Here we investigated the effect of cell-like substrate topography on ovarian cancer cell behaviour and potential underlying signalling pathways. We observed changes in cell morphology in response to substrate topography, which implies modification of structure-function associations. Differences in focal adhesion signalling and Rho/ROCK activity suggested their involvement in the biomechanically-driven cellular responses. Cell-like topography was also shown to modulate the MAPK pathway and hence potentially regulate cell proliferation. The selective regulation of the cells by the mechanotransduction pathways that we noted has wide ranging implications for understanding cancer development. We established that the physical architecture of cell culture substrate is sufficient to influence cancer cell behaviour, independent of genetic composition or biochemical milieu.

Direct Writing and Characterization of Three-Dimensional Conducting Polymer PEDOT Arrays.

Direct writing is an effective and versatile technique for three-dimensional (3D) fabrication of conducting polymer (CP) structures. It is precisely localized and highly controllable, thus providing great opportunities for incorporating CPs into microelectronic array devices. Herein we demonstrate 3D writing and characterization of poly(3,4-ethylenedioxythiophene)-polystyrenesulfonate (PEDOT:PSS) pillars in an array format, by using an in-house-constructed variant of scanning ion conductance microscopy (SICM). CP pillars with different aspect ratios were successfully fabricated by optimizing the writing parameters: pulling speed, pulling time, concentration of the polymer solution, and the micropipette tip diameter. Especially, super high aspect ratio pillars of around 7 µm in diameter and 5000 µm in height were fabricated, indicating a good capability of this direct writing technique. Additions of an organic solvent and a cross-linking agent contribute to a significantly enhanced water stability of the pillars, critical if the arrays were to be used in biologically relevant applications. Surface morphologies and structural analysis of CP pillars were characterized by scanning electron microscopy and Raman spectroscopy, respectively. Electrochemical properties of the individual pillars of different heights were examined by cyclic voltammetry using a double-barrel micropipette as an electrochemical cell. Exceptional mechanical properties of the pillars, such as high flexibility and robustness, were observed when bent by applying a force. The 3D pillar arrays are expected to provide versatile substrates for functionalized and integrated biological sensing and electrically addressable array devices.

Cell-like features imprinted in the physical nano- and micro-topography of the environment modify the responses to anti-cancer drugs of endometrial cancer cells.

Topographical features of cells at nanometre resolution were fabricated in polystyrene. The study investigated the effect of physical topography on the response of cancer cells to the common anticancer drugs, paclitaxel and doxorubicin. Human endometrial cancer cells (Ishikawa) were incubated on substrates containing cell-like features that had been fabricated using our bioimprint methodology to create moulds of cells with positive (convex) and negative (concave) topography. Control cultures were performed on flat substrates. Effects of the drugs on caspase-3 expression, proliferating nuclear antigen (PCNA) expression, cell number and vascular endothelial growth factor (VEGF) secretion were determined. Results revealed that the topography influenced the cell responses in a drug-dependent manner i.e. paclitaxel effects were sensitive to topography differently to those of doxorubicin. In addition, function signalling pathways were sensitive to the detailed topography i.e. positive imprint and negative imprint induced distinct response patterns. The results in this study show for the first time that a culture surface with cell-like topography, that has both nano- and micro-resolution, influences endometrial cancer cell responses to chemotherapy drugs. The effects are dependent on the topography and also on the chemotherapy drug. In particular, the platforms described have potential to provide substrates with high physical relevancy on which to undertake preclinical testing of new drugs. The method also allows for use of different cell types to provide cell-specific topography. The results imply that physical architecture of the cancer cell environment may be a suitable prospective target to enhance clinical activity of traditional drugs. Additionally or alternatively we provide compelling support for the notion that understanding the physical component of the nano- and micro-environment may encourage a redirection of drug development. Further, our observation that the cells distinguish between the different cell-like topographies (positive and negative bioimprints) indicates that a realistic topography is advantageous as growth platforms in experiment design.

The characteristics of Ishikawa endometrial cancer cells are modified by substrate topography with cell-like features and the polymer surface.

Conventional in vitro culture studies on flat surfaces do not reproduce tissue environments, which have inherent topographical mechanical signals. To understand the impact of these mechanical signals better, we use a cell imprinting technique to replicate cell features onto hard polymer culture surfaces as an alternative platform for investigating biomechanical effects on cells; the high-resolution replication of cells offers the micro- and nanotopography experienced in typical cell-cell interactions. We call this platform a Bioimprint. Cells of an endometrial adenocarcinoma cell line, Ishikawa, were cultured on a bioimprinted substrate, in which Ishikawa cells were replicated on polymethacrylate (pMA) and polystyrene (pST), and compared to cells cultured on flat surfaces. Characteristics of cells, incorporating morphology and cell responses, including expression of adhesion-associated molecules and cell proliferation, were studied. In this project, we fabricated two different topographies for the cells to grow on: a negative imprint that creates cell-shaped hollows and a positive imprint that recreates the raised surface topography of a cell layer. We used two different substrate materials, pMA and pST. We observed that cells on imprinted substrates of both polymers, compared to cells on flat surfaces, exhibited higher expression of ß1-integrin, focal adhesion kinase, and cytokeratin-18. Compared to cells on flat surfaces, cells were larger on imprinted pMA and more in number, whereas on pST-imprinted surfaces, cells were smaller and fewer than those on a flat pST surface. This method, which provided substrates in vitro with cell-like features, enabled the study of effects of topographies that are similar to those experienced by cells in vivo. The observations establish that such a physical environment has an effect on cancer cell behavior independent of the characteristics of the substrate. The results support the concept that the physical topography of a cell's environment may modulate crucial oncological signaling pathways; this suggests the possibility of cancer therapies that target pathways associated with the response to mechanical stimuli.

Bioimprinted polymer platforms for cell culture using soft lithography.

BACKGROUND: It is becoming recognised that traditional methods of culture in vitro on flat substrates do not replicate physiological conditions well, and a number of studies have indicated that the physical environment is crucial to the directed functioning of cells in vivo. In this paper we report the development of a platform with cell-like features that is suitable for in vitro investigation of cell activity. Biological cells were imprinted in hard methacrylate copolymer using soft lithography. The cell structures were replicated at high nanometre scale resolution, as confirmed by atomic force microscopy. Optimisation of the methacrylate-based co-polymer mixture for transparency and biocompatibility was performed, and cytotoxicity and chemical stability of the cured polymer in cell culture conditions were evaluated. Cells of an endometrial adenocarcinoma cell line (Ishikawa) were cultured on bioimprinted substrates. RESULTS: The cells exhibited differential attachment on the bioimprint substrate surface compared to those on areas of flat surface and preferentially followed the pattern of the original cell footprint. CONCLUSIONS: The results revealed for the first time that the cancer cells distinguished between behavioural cues from surfaces that had features reminiscent of themselves and that of flat areas. Therefore the imprinted platform will lend itself to detailed studies of relevant physical substrate environments on cell behaviour. The material is not degraded and its permanency allows reuse of the same substrate in multiple experimental runs. It is simple and does not require expensive or specialised equipment. In this work cancer cells were studied, and the growth behaviour of the tumour-derived cells was modified by alterations of the cells' physical environment. Implications are also clear for studies in other crucial areas of health, such as wound healing and artificial tissues.

On-chip analysis of C. elegans muscular forces and locomotion patterns in microstructured environments.

The understanding of force interplays between an organism and its environment is imperative in biological processes. Noticeably scarce from the study of C. elegans locomotion is the measurement of the nematode locomotion forces together with other important locomotive metrics. To bridge the current gap, we present the investigation of C. elegans muscular forces and locomotion metrics (speed, amplitude and wavelength) in one single assay. This assay uses polydimethylsiloxane (PDMS) micropillars as force sensing elements and, by variation of the pillar arrangement, introduces microstructure. To show the usefulness of the assay, twelve wild-type C. elegans sample worms were tested to obtain a total of 4665 data points. The experimental results lead to several key findings. These include: (1) maximum force is exerted when the pillar is in contact with the middle part of the worm body, (2) C. elegans locomotion forces are highly dependent on the structure of the surrounding environment, (3) the worms' undulation frequency and locomotion speed increases steadily from the narrow spacing of 'honeycomb' design to the wider spacing of 'lattice' pillar arrangement, and (4) C. elegans maintained their natural sinusoidal movement in the microstructured device, despite the existence of PDMS micropillars. The assay presented here focuses on wild type C. elegans, but the method can be easily applied to its mutants and other organisms. In addition, we also show that, by inverting the measurement device, worm locomotion behaviour can be studied in various substrate environments normally unconducive to flexible pillar fabrication. The quantitative measurements demonstrated in this work further improve the understanding of C. elegans mechanosensation and locomotion.