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However, the roles of osteocytes in cancer cell invasion and metastasis remain unknown. Materials and Methods: In this study, we investigated the effects of early osteocytes on the behaviour of breast and prostate cancer cells. The proliferation of cultured cells was assessed using the AlamarBlue assay. The electric cell-substrate impedance sensing ECIS system was used to measure spreading, attachment and migratory behaviour of cancer cells in response to conditioned medium CM from mouse osteocytes.

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Conclusion: Osteocytes play diverse roles in the proliferative and migratory potential of breast and prostate cancer cells that may be associated with cancer-specific bone metastasis and requires further investigation. User Name Password Sign In. Correspondence to: Dr. Select reports are outlined, and not all the work that has been performed in the field of drug screening and development is covered. Keywords: screening of bioactive agents, impedance-based cell study, electric cell-substrate impedance sensing ECIS , high-throughput screening, real-time drug evaluation.

Intensive research on the development of new drugs for safer therapeutic interventions for various diseases, based on scientific advancements in high-throughput screening HTS , has so far resulted in the development of synthetic, semisynthetic, and natural compounds.

Metastasis: How Cancer Spreads

The bioimpedance method has proved to be an ideal technique for whole cellular behavior detection without radioactive or fluorescence-labeled substrates to quantify cell morphology changes. Following this development, Giaever and Keese developed electric cell-substrate impedance sensing ECIS as a continuous monitoring system for studying cell behaviors using a noninvasive, real-time, and label-free method. Bioelectrical impedance cell culture platforms are capable of analyzing all cellular events, including cell attachment, spreading, migration, growth, mitosis, cell-cell contacts behavior, cell-matrix contacts behavior, and death of adhering cells, in a quantitative manner.

ECIS has several benefits over other cell-based biosensing methods, in areas including analysis of cell behavior under flow, metastatic potential, and endothelial barrier functions, and toxicological and pharmacological drug screening. The ECIS system can monitor the impedance signal changes that occur due to electric current between the cells and electrodes and has the capability to detect, analyze, and aid investigation of the effects of bioactive and inactive compounds on cells.

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The output signal changes can provide statistical values for behaviors of the cell on each electrode, which result from cellular changes caused by external stimuli such as chemical compounds, drugs and or any analytes. Compared to other statistical data measurement systems, ECIS has the greatest potential in drug discovery and screening of new drugs for new diseases and disorders. Great advances have been made in recent decades in the areas of drug discovery and screening, with numerous upgrades to bioimpedance platforms and much innovation in techniques. As a result, treatment efficacy and quality of life have improved.

A hybrid bioimpedance system that uses microfluidic channels has recently emerged. This system is capable of measuring living biological adherent or nonadherent cells as an in vitro model for studying the cellular behavior changes caused by external and internal stimuli.

This system offers possible new directions for biomedical and therapeutic applications such as new drug evaluation, testing, and screening as well as toxin identification for various diseases, including various forms of oxidative stress-induced damage. This system has many advantages: rapidity, enhance sensitivity, bio-recognition on-the-spot and early-stage detection, and targeting are important features of this system.

In brief, it can detect any forms of substances, including chemicals, microorganisms, and environmental toxins, because the cell periphery has different types of receptors which can produce different responses for different substances. Because of these important features, ECIS is recognized as a unique and reliable way to determine the physiological characters of living cells.

This review focuses on identification of various bioactive therapeutic molecules and screening for various diseases during various cellular events using ECIS. Furthermore, this review will provide enriched information about ECIS-based tools for bioactive molecule screening and drug development.

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  • ECIS is an advanced system that is based on the principles of bioimpedance and that uses alternating current AC to evaluate cellular behavior in physiological conditions. Some of the typical ECIS electrode array types, which are used in different assays for cell study, are shown in Figure 1C. Notes: A Current flow before cell attachment.

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    B After cell attachment, the current flows with cells from the surface of Au-sensing electrodes. Cells are grown to confluence on electrodes. The current flow between working electrodes and counter electrodes through cell culture medium, which acted as electrolyte. The structural design of the ECIS system comprises two electrodes: one is a small working electrode and the other a large counter electrode on the bottom of the culture plate.


    The lock-in amplifier is connected to a personal computer for data acquisition, storage, and interpretation. After cell seeding, cells drift downward and attach to the stratum of the electrode, which then passes the impeded current directly into the bulk electrolyte as the result of anchored plasma membrane intrusion above the electrode surface. This represents the passage of current through the available cell substrate space as well as between available intercellular junctions. This can result in changes in impedance, and the rate of changes of the impedance increases until the cells reach confluent on electrode.

    Initially, resistance increases due to attachment and spreading, followed by confluence, of the cells, and the changes in impedance are measured. The changes in electrical impedance of this real-time system are a function of AC frequency, and the resulting voltage and resistance are measured with a lock-in amplifier. This resulting impedance is monitored every second at various suitable preset frequencies 1— kHz , and data collection and processing are carried out with suitable software. The impedance measurement is noninvasive.

    Figure 2 A typical electric cell-substrate impedance sensing ECIS measurement graph of normal HDFn cell growth response for 20 hours, showing various cellular morphological changes. Abbreviation: HDFn, human dermal fibroblasts, neonatal cells. To elucidate impedance date, several researchers have derived calculations for quantifying cell parameters, including impedance changes and cell population via different mathematical models. Using least squares optimization, Giaever and Keese derived a mathematical approximation for the total impedance of cells covering the electrode, as illustrated by Equation The measurement of capacitance shows that higher frequencies are most suited for cell spreading because they increase cell coverage of the electrode.

    At high frequency ranges, measurement is more sensitive to changes in cell attachment and spreading at the electrode. However, 10—40 kHz is the main frequency range used for measuring impedance.

    WO2016139566A1 - Electrical cell-substrate impedance sensor (ecis) - Google Patents

    In most of the ECIS based studies, the large-sized integrated microelectrode array platforms has been utilized, this system can provide data for the combined physical responses of cells. The ECIS system has been developed based on various approaches and methods used to evaluate the cellular properties, including microelectrode array chip pattern, positioning, sensing device, and flow. Some cell-based biosensing chip fabrication designs are depicted in Figure 3. Figure 3 Schematic diagram showing various devices and microelectrode chip fabrication types for different studies using bioimpedance platform.

    Notes: A Real-time optical imaging and impedance measurements. The camera is located above the cell culture chip, which enables provision of real-time imaging.

    Drug and bioactive molecule screening based on a bioelectrical impedance cell culture platform

    B Microfluidic based cell culture sensing system: interdigitated array of electrodes on glass for impedance sensing, a polydimethylsiloxane PDMS layer for gradient generation and cell culture, which can provide the concentration dependent cellular behavior. C Three-dimensional depiction of a hydrogel chamber of a diffusion cell culture chip integrated with electric cell-substrate impedance sensing ECIS.

    D Qualitative and quantitative data acquisition using a computer interface with a data monitoring and storage system. Recently, Xu et al and Tran et al developed an integrated device with microfluidic channels to prevent unwanted flow based on a hydrogel-based diffusion cell chip to produce a gradient concentration with prolonged stability for high-throughput assays.

    Electric cell-substrate impedance sensing and cancer metastasis | UTS Library

    For further continuous development, pneumatic microvalves, air-bubble valves, 17 and thermopneumatic gas-bubble microvalves for temperature control 18 were integrated into the microfluidic chips to control the flow. Study of complex and simultaneous cellular responses to various substances using HTS platforms has been improved by the integration of a hydrogel-based diffusion microfluidic channel chip.

    To achieve further continuous development of cell-based biosensing and drug screening techniques, researchers developed two- and three-dimensional microfluidic-based gradient generators for HTS, which can provide a synergistic effect for combined administration of multiple-drug doses. This system can provide fine detectability on individual electrodes, compared to many cells on a large-sized electrode, and has potential in environmental toxin identification and drug estimation.

    The real-time optical imaging and impedance measurements are perhaps better for qualitative and quantitative studies. The gold electrode has been shown to be a safe, nontoxic, and nonreactive material for use as a bioimpedance sensor chip; however, for certain analyses, a custom-made specific array could be a better choice. HTS is an important method in drug discovery, as it allows rapid pharmacological testing of various chemical compounds in order that safe, nontoxic, and maximally tolerated drugs with no adverse side effects to normal cell activities can be developed.

    Quantitative impedance measurement has been successfully applied to a variety of cell types using different kinds of chemicals, therapeutic compounds, and nanoparticles, including polymer, metal, and metal oxide nanoparticles. The drug screening test is used to find out the various effects of a drug prior in the development of effective treatment for various health problems. This system can be used in various cellular assays, including cytotoxicity, wound-healing, anticancer, antiviral, protectivity, migration, and barrier-disruptive assays, as well as assays for many other cellular activities.

    Examples of its application in cellular assays are shown in Figure 4. For the evaluation of health-related problems, we are pointing out some examples of health risk effects at the molecular level of analysis; in particular, chemokine attraction occurs during chronic inflammation upon binding of adhesion molecules from leukocytes to the activated endothelium. Figure 4 Applications of the ECIS system for analysis of cellular behaviors and activity for drug screening studies.

    Notes: A Barrier function analysis for paracellular pathway and permeability study. B Ion channel activity analysis for studying ion transport mechanisms and whether the compound is able to block the channel. D Cell metabolism analysis for studying the differences in growth and metabolic status of cells. E Cytotoxicity screening for studying analyte toxicity responses to the cells. F Cancer metastasis analysis for studying cancer cell behavior including the potentiality of drugs effects on cells.

    G Photoprotectivity analysis for studying photodamaging and photoprotective effects. H Drug resistance analysis for studying drug resistance capacity in various cells, including cancer cells, including cancer cells. In therapeutics, it is important to identify suitable compounds with the ability to control the interaction between leukocytes and endothelial compartments.