Real-time monitoring of skin wound healing on nano-grooves topography using electric cell-substrate impedance sensing (ECIS)

Yao, Chen; An, Yu; Jin, Tongyu; Zhang, Fan; He, Pingang

doi:10.1016/j.snb.2017.04.183

Cited by 34 publications

(22 citation statements)

References 36 publications

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“…The curve shown in Figure 5b was obtained at time 17 h of the experiment, but the same tendency can be extracted from the different points of curve in Figure 5a. Thus, it can be considered that the cell index value mirrors the total number of adhered cells initially seeded, in agreement with what has been previously reported [43,44].…”

Section: Demonstration Of the Use Of Inkjet-printed Sensors For Impedsupporting

confidence: 90%

Real-Time Impedance Monitoring of Epithelial Cultures with Inkjet-Printed Interdigitated-Electrode Sensors

Mojena-Medina

Hubl²,

Bäuscher

et al. 2020

Sensors

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From electronic devices to large-area electronics, from individual cells to skin substitutes, printing techniques are providing compelling applications in wide-ranging fields. Research has thus fueled the vision of a hybrid, printing platform to fabricate sensors/electronics and living engineered tissues simultaneously. Following this interest, we have fabricated interdigitated-electrode sensors (IDEs) by inkjet printing to monitor epithelial cell cultures. We have fabricated IDEs using flexible substrates with silver nanoparticles as a conductive element and SU-8 as the passivation layer. Our sensors are cytocompatible, have a topography that simulates microgrooves of 300 µm width and ~4 µm depth, and can be reused for cellular studies without detrimental in the electrical performance. To test the inkjet-printed sensors and demonstrate their potential use for monitoring laboratory-growth skin tissues, we have developed a real-time system and monitored label-free proliferation, migration, and detachment of keratinocytes by impedance spectroscopy. We have found that variations in the impedance correlate linearly to cell densities initially seeded and that the main component influencing the total impedance is the isolated effect of the cell membranes. Results obtained show that impedance can track cellular migration over the surface of the sensors, exhibiting a linear relationship with the standard method of image processing. Our results provide a useful approach for non-destructive in-situ monitoring of processes related to both in vitro epidermal models and wound healing with low-cost ink-jetted sensors. This type of flexible sensor as well as the impedance method are promising for the envisioned hybrid technology of 3D-bioprinted smart skin substitutes with built-in electronics.

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Section: Demonstration Of the Use Of Inkjet-printed Sensors For Impedsupporting

confidence: 90%

Real-Time Impedance Monitoring of Epithelial Cultures with Inkjet-Printed Interdigitated-Electrode Sensors

Mojena-Medina

Hubl²,

Bäuscher

et al. 2020

Sensors

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“…ECIS is now capable of becoming an effective and noninvasive analytical method that facilitates a more direct and continuous early assessment of the cytotoxicity of drugs and environmental toxins in vitro more conveniently (Xie et al, 2012). An ECIS device that was developed using nano-grooves to simulate the internal extracellular matrix, with grooves 75 nm in depth and 200 nm in width, generated less intense resistance signals in human foreskin fibroblast (HFF) cell migration and proliferation, and HaCaT cell migration, but more intense ones in HaCaT cell proliferation (Cui et al, 2017). FKBP51-mediated inhibition of I SOC leads to decreased calcium entry-induced inter-endothelial gap formation and, thus, preserved the endothelial barrier by decreasing actin stress fiber and inter-endothelial cell gap formation in addition to attenuating the decrease in resistance observed with control cells using ECIS (Hamilton et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Mechanisms of silver nanoparticles-induced cytotoxicity and apoptosis in rat tracheal epithelial cells

Tang

Chen

et al. 2019

J. Toxicol. Sci.

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Silver nanoparticles (AgNPs) are increasingly utilized in a number of applications. This study was designed to investigate AgNPs induced cytotoxicity, oxidative stress and apoptosis in rat tracheal epithelial cells (RTE). The RTE cells were treated with 0, 100 μg/L and 10,000 μg/L of the AgNPs with diameters of 10 nm and 100 nm for 12 hr. The cell inhibition level, apoptosis ratio, reactive oxygen species (ROS), malondialdehyde (MDA) and metallothionein (MT) content were determined. The mRNA expression of cytoc, caspase 3, and caspase 9 was measured by quantitative real-time polymerase chain reaction (qRT-PCR). In addition, we also analyzed the cytoc, caspase 3, pro-caspase 3, caspase 9, and pro-caspase 9 protein expression by western blotting. Electric cell-substrate impedance sensing (ECIS) analysis showed that the growth and proliferation of RTE cells were significantly inhibited in a dose-dependent manner under AgNPs exposure. The cell dynamic changes induced by 10 nm AgNPs were more severe than that of the 100 nm AgNPs exposure group. The intracellular MT, ROS, and MDA content increased when the exposure concentration increased and size reduced, whereas Ca 2+-ATPase activity and Na + /K +-ATPase activity changed inversely. The relative expression of protein of cytoc, caspase 3, and caspase 9 were upregulated significantly, which indicated that AgNPs induced apoptosis of RTE cells through the caspase-dependent mitochondrial pathway. Our results demonstrate that AgNPs caused obvious cytotoxicity, oxidative stress, and apoptosis in RTE cells, which promoted the releasing of cytochrome C and pro-apoptotic proteins into the cytoplasm to activate the caspase cascade and finally led to apoptosis.

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“…This technique is suitable to study the paracrine signals (conditioned media) produced by stem [ 10 ] or other types of cells under either mechanical [ 7 ] or electrical stimuli [ 11 ]. Furthermore, it is possible to study the response of free cells and those seeded on scaffolds made of both synthetic polymers and natural extracellular matrices [ 11 , 12 – 14 ]. In this context, the in vitro scratch wound healing assay is useful to evaluate the proliferation and migration process when exposing cells to metabolites present in the conditioned media.…”

Section: Introductionmentioning

confidence: 99%

An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays

et al. 2020

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In vitro scratch wound healing assay, a simple and low-cost technique that works along with other image analysis tools, is one of the most widely used 2D methods to determine the cellular migration and proliferation in processes such as regeneration and disease. There are open-source programs such as imageJ to analyze images of in vitro scratch wound healing assays, but these tools require manual tuning of various parameters, which is time-consuming and limits image throughput. For that reason, we developed an optimized plugin for ima-geJ to automatically recognize the wound healing size, correct the average wound width by considering its inclination, and quantify other important parameters such as: area, wound area fraction, average wound width, and width deviation of the wound images obtained from a scratch/ wound healing assay. Our plugin is easy to install and can be used with different operating systems. It can be adapted to analyze both individual images and stacks. Additionally, it allows the analysis of images obtained from bright field, phase contrast, and fluorescence microscopes. In conclusion, this new imageJ plugin is a robust tool to automatically standardize and facilitate quantification of different in vitro wound parameters with high accuracy compared with other tools and manual identification.

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Real-time monitoring of skin wound healing on nano-grooves topography using electric cell-substrate impedance sensing (ECIS)

Cited by 34 publications

References 36 publications

Real-Time Impedance Monitoring of Epithelial Cultures with Inkjet-Printed Interdigitated-Electrode Sensors

Real-Time Impedance Monitoring of Epithelial Cultures with Inkjet-Printed Interdigitated-Electrode Sensors

Mechanisms of silver nanoparticles-induced cytotoxicity and apoptosis in rat tracheal epithelial cells

An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays

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