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Microfluidics is a highly useful platform for culturing, monitoring, and testing biological cells. The integration of electrodes into microfluidic channels extends the functionality, sensing, and testing capabilities of microfluidic systems. By employing an electrochemical impedance spectroscopy (EIS) technique, the non-invasive, label-free detection of the activities of cells in real-time can be achieved. To address the movement toward spatially resolving cells in cell culture, we developed a sensory system capable of electro-addressing cell location within a microfluidic channel. This simple system allows for real-time cell location, integrity monitoring (of barrier producing cells), and confluency sensing without the need for frequent optical evaluation—saving time. EIS results demonstrate that cells within microfluidic channels can be located between various pairs of electrodes at different positions along the length of the device. Impedance spectra clearly differentiates between empty, sparse, and confluent microfluidic channels. The system also senses the level of cell confluence between electrode pairs—allowing for the relative quantification of cells in different areas of the microfluidic channel. The system’s electrode layout can easily be incorporated into other devices. Namely, organ-on-a-chip devices, that require the monitoring of precise cell location and confluency levels for understanding tissue function, modeling diseases, and for testing therapeutics.

The blood brain barrier (BBB) is formed by neurovascular units (NVUs) that shield the central nervous system (CNS) from a range of factors found in the blood that can disrupt delicate brain function. As such, the BBB is a major obstacle to the delivery of therapeutics to the CNS. Accumulating evidence suggests that the BBB plays a key role in the onset and progression of neurological diseases. Thus, there is a tremendous need for a BBB model that can predict penetration of CNS-targeted drugs as well as elucidate the BBB's role in health and disease. We have recently combined organ-on-chip and induced pluripotent stem cell (iPSC) technologies to generate a BBB chip fully personalized to humans. This novel platform displays cellular, molecular, and physiological properties that are suitable for the prediction of drug and molecule transport across the human BBB. Furthermore, using patient-specific BBB chips, we have generated models of neurological disease and demonstrated the potential for personalized predictive medicine applications. Provided here is a detailed protocol demonstrating how to generate iPSC-derived BBB chips, beginning with differentiation of iPSC-derived brain microvascular endothelial cells (iBMECs) and resulting in mixed neural cultures containing neural progenitors, differentiated neurons, and astrocytes. Also described is a procedure for seeding cells into the organ chip and culturing of the BBB chips under controlled laminar flow. Lastly, detailed descriptions of BBB chip analyses are provided, including paracellular permeability assays for assessing drug and molecule permeability as well as immunocytochemical methods for determining the composition of cell types within the chip.

In this paper, a multilayer barrier thin film, based on polyvinylidene difluoride (PVDF)–silicon dioxide (SiO2), has been fabricated on a PET substrate through a novel method of joint fabrication techniques. The inorganic SiO2 thin film was deposited using a roll-to-roll atmospheric atomic layer deposition system (R2R-AALD), while the organic PVDF layer was deposited on the surface of SiO2 through the electrohydrodynamic atomization (EHDA) technique. The multilayer barrier thin films exhibited very good surface morphology, chemical composition, and optical properties. The obtained values for arithmetic surface roughness and water contact angle of the as-developed multilayer barrier thin film were 3.88 nm and 125°, respectively. The total thickness of the multilayer barrier thin film was 520 nm with a high optical transmittance value (85–90%). The water vapor transmission rate (WVTR) of the barrier thin film was ~ 0.9 × 10−2 g m−2 day−1. This combination of dual fabrication techniques (R2R-AALD and EHDA) for the development of multilayer barrier thin films is promising for gas barrier applications.

Titanium dioxide (TiO2) thin film was fabricated using titanium isopropoxide as a precursor through an atmospheric low-temperature roll-to-roll chemical vapor deposition method. TiO2 was deposited on the PET substrate in the temperature range of room temperature to 100°C, and the working pressure was 740 Torr. The surface morphology of TiO2 thin film was analyzed by field emission scanning electron microscopy and a 2D surface profiler. The results revealed that the growth rate of TiO2 film was 31 nm/min at 100°C, and it also showed that the surface is uniform and smooth. Moreover, the lowest root mean square roughness (R q) value of 1.87 nm was obtained for TiO2 film prepared at 100°C. The composition of TiO2 film was confirmed by X-ray photoelectron spectroscopy (XPS) analysis. The film showed very good chemical and optical properties while increasing the substrate deposition temperature. The UV–Vis spectroscopy analysis revealed that TiO2 films exhibited excellent optical transmittance, more than 91% observed in the visible region.

A highly sensitive body motion sensor has been fabricated based on a composite active layer of zinc stannate (ZnSnO3) nano-cubes and poly(vinylidene fluoride) (PVDF) polymer. The thin film-based active layer was deposited on polyethylene terephthalate flexible substrate through D-bar coating technique. Electrical and morphological characterizations of the films and sensors were carried out to discover the physical characteristics and the output response of the devices. The synergistic effect between piezoelectric ZnSnO3 nanocubes and β phase PVDF provides the composite with a desirable electrical conductivity, remarkable bend sensitivity, and excellent stability, ideal for the fabrication of a motion sensor. The recorded resistance of the sensor towards the bending angles of −150° to 0° to 150° changed from 20 MΩ to 55 MΩ to 100 MΩ, respectively, showing the composite to be a very good candidate for motion sensing applications.

Doxorubicin (Dox) is a potent chemotherapeutic agent used in the treatment of cancer. In the present study, pH responsive chitosan polymer coated Dox nanoparticle (Composite) was developed to investigate targeted drug delivery against breast cancer. The anticancer drug DOX-ZnO QDs was loaded to the chitosan nanoparticles. The synthesized free and drug loaded nanoparticle were analyzed using Fourier transmission electron microscopy (FTIR) and UV-Visible spectroscopy(UV-Vis). The particle size was measured using Transmission Electron Microscopy (TEM). Further, the composite was evaluated for its anticancer effects. Drug release analysis showed significantly larger amount of drug released in acidic pH of 5.0 compared to pH 7.4. The composite was significantly more cytotoxic to the breast cancer cells MCF-7 and SKBR-3. The composite was however, less toxic to HEK-293 human embryonic kidney cells confirming minimum side effects on normal cells andcytotoxic to tumor cells. DAPI staining showed nuclear degradation in composite treated breast cancer cells. The cellular uptake of the composite was analyzed by confocal microscopy. The composite induced a G0/G1 phase arrest in breast cancer cells and the number of colonies formed by the composite treated breast cancer cells formed less number of colonies compared to free NP. Our results showed that our composite could serve as a promising therapeutic approach to improve clinical outcomes against various malignancies.

Most cancer patients die as a consequence of distant metastases, which are frequently unresponsive to cancer therapy. This study focuses on the anti-tumorigenic and anti-metastatic properties of tangeretin-zinc oxide quantum dots (Tan-ZnO QDs) against the NCI-H358 cell line. Tan-ZnO QDs are pH-sensitive and capitalize on the acidic pH maintained in the tumor microenvironment; therefore, targeted drug delivery is directed specifically to cancer cells, leaving the normal cells less affected. Tan was loaded into synthesized ZnO QDs, and drug loading was analyzed using Fourier transform infrared (FTIR) spectroscopy and ultraviolet-visible (UV–Vis) spectrometry. Crystalline phase and particle size were measured using transmission electron microscopy (TEM) and X-ray diffraction (XRD). Drug release was evaluated in buffered solutions with differing pH for up to 15 h. The results confirmed stable drug release (80%) in an acidic pH. Tan-ZnO QDs induced significant cytotoxicity in NCI-H358 metastatic cells, while not markedly affecting HK-2 human normal cells. Morphology of treated H358 cells analyzed via atomic force microscopy (AFM) showed an increased surface roughness and pores. Further, the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells increased after treatment with Tan-ZnO QDs. DNA fragmentation was also induced after treatment with increasing concentrations of Tan-ZnO QDs in H358 cells. We also confirmed regulation of apoptosis via expression levels of Bax and Bcl-2 proteins; G2/M phase cell cycle arrest was observed. Additionally, cell proliferation and migration drastically decreased, and cell invasion and migration, hallmarks of metastasis, were significantly inhibited in H358 cells. Matrix metalloproteinase (MMP)2 and MMP9, markers of metastasis, as well as vascular endothelial growth factor (VEGF), a marker of angiogenesis, were significantly downregulated upon treatment with Tan-ZnO QDs. In conclusion, our novel formulation destabilized H358 cells by using its acidic tumor microenvironment, thereby regulating cell apoptosis, proliferation, and metastatic properties.

Current trends in therapeutic research are the application of nanomaterial carriers for cancer therapy. One such molecule, ZnO, originally used in diagnosis and as a drug carrier, is gaining importance for its biological properties. Here, we report for the first time, the scope of ZnO QDs for enhanced cytotoxicity against MCF-7 and metastatic MDA-MB-231 human breast cancer cells. Unlike other ZnO nanostructures, ZnO QDs are dispersed and small sized (8–10 nm) which is believed to greatly increase the cellular uptake. Furthermore, the acidic tumor microenvironment attracts ZnO QDs enhancing targeted therapy while leaving normal cells less affected. Results from MTT assay demonstrated that ZnO QDs induced cytotoxicity to MCF-7 and metastatic MDA-MB-231 breast cancer cells at very low concentrations (10 and 15 μg/ml) as compared to other reported ZnO nanostructures. HEK-293 cells showed less toxicity at these concentrations. Confocal microscope images from DAPI staining and TUNEL assay demonstrated that ZnO QDs induced nuclear fragmentation and apoptosis in MCF-7 and MDA-MB-231. FACS results suggested ZnO QDs treatment induced cell cycle arrest at the G0/G1 phase in these cells. ZnO QDs drastically decreased the proliferation and migration of MCF-7 and MDA-MB-231 as seen from the results of the clonogenic and wound healing assays respectively. Furthermore, our data suggested that ZnO QDs regulated apoptosis via Bax and Bcl-2 proteins as validated by immunofluorescence and western blot. Taken together, our findings demonstrate that these ultra-small sized ZnO QDs destabilize cancer cells by using its acidic tumor microenvironment thereby inducing apoptosis and controlling the cell proliferation and migration at low dosages.