Chemotherapy agents reduce protein synthesis and ribosomal capacity in myotubes independent of oxidative stress.

Chemotherapeutic agents (CAs) are first-line antineoplastic treatments against a wide variety of cancers. Despite their effectiveness in halting tumor progression, side effects associated with CAs promote muscle loss by incompletely understood mechanisms. To address this problem, we first identified how oxidative stress impairs protein synthesis in C2C12 myotubes. Transient elevations in reactive oxygen species (ROS) resulted in protein synthesis deficits and reduced ribosomal (r)RNA levels. Oxidative stress did not reduce rRNA gene (rDNA) transcription, but it caused an increase in rRNA and protein oxidation. To determine whether CAs affect protein synthesis independent of oxidative stress, we exposed myotubes to Paclitaxel (PTX), Doxorubicin (DXR), or Marizomib (Mzb) at doses that did result in elevated ROS levels (sub-ROS). Exposure to CAs reduced protein synthesis and rRNA levels, but unlike oxidative stress, sub-ROS exposures impaired rDNA transcription. These results indicate that although oxidative stress disrupts protein synthesis by compromising ribosomal quantity and quality, CAs at sub-ROS doses compromise protein synthesis and ribosomal capacity, at least in part, by reducing rDNA transcription. Therefore, CAs negatively impact protein synthesis by causing oxidative stress in addition to directly reducing the ribosomal capacity of myotubes in a ROS-independent manner.

Molecular and physical technologies for monitoring fluid and electrolyte imbalance: A focus on cancer population.

Several clinical examinations have shown the essential impact of monitoring (de)hydration (fluid and electrolyte imbalance) in cancer patients. There are multiple risk factors associated with (de)hydration, including aging, excessive or lack of fluid consumption in sports, alcohol consumption, hot weather, diabetes insipidus, vomiting, diarrhea, cancer, radiation, chemotherapy, and use of diuretics. Fluid and electrolyte imbalance mainly involves alterations in the levels of sodium, potassium, calcium, and magnesium in extracellular fluids. Hyponatremia is a common condition among individuals with cancer (62% of cases), along with hypokalemia (40%), hypophosphatemia (32%), hypomagnesemia (17%), hypocalcemia (12%), and hypernatremia (1-5%). Lack of hydration and monitoring of hydration status can lead to severe complications, such as nausea/vomiting, diarrhea, fatigue, seizures, cell swelling or shrinking, kidney failure, shock, coma, and even death. This article aims to review the current (de)hydration (fluid and electrolyte imbalance) monitoring technologies focusing on cancer. First, we discuss the physiological and pathophysiological implications of fluid and electrolyte imbalance in cancer patients. Second, we explore the different molecular and physical monitoring methods used to measure fluid and electrolyte imbalance and the measurement challenges in diverse populations. Hydration status is assessed in various indices; plasma, sweat, tear, saliva, urine, body mass, interstitial fluid, and skin-integration techniques have been extensively investigated. No unified (de)hydration (fluid and electrolyte imbalance) monitoring technology exists for different populations (including sports, elderly, children, and cancer). Establishing novel methods and technologies to facilitate and unify measurements of hydration status represents an excellent opportunity to develop impactful new approaches for patient care.

Evaluation of supercritical CO2 sterilization efficacy for sanitizing personal protective equipment from the coronavirus SARS-CoV-2.

The purpose of this research is to evaluate the supercritical carbon dioxide (scCO2) sterilization-based NovaClean process for decontamination and reprocessing of personal protective equipment (PPE) such as surgical masks, cloth masks, and N95 respirators. Preliminarily, Bacillus atrophaeus were inoculated into different environments (dry, hydrated, and saliva) to imitate coughing and sneezing and serve as a "worst-case" regarding challenged PPE. The inactivation of the microbes by scCO2 sterilization with NovaKill or H2O2 sterilant was investigated as a function of exposure times ranging from 5 to 90 min with a goal of elucidating possible mechanisms. Also, human coronavirus SARS-CoV-2 and HCoV-NL63 were inoculated on the respirator material, and viral activity was determined post-treatment. Moreover, we investigated the reprocessing ability of scCO2-based decontamination using wettability testing and surface mapping. Different inactivation mechanisms have been identified in scCO2 sanitization, such as membrane damage, germination defect, and dipicolinic acid leaks. Moreover, the viral sanitization results showed a complete inactivation of both coronavirus HCoV-NL63 and SARS-CoV-2. We did not observe changes in PPE morphology, topographical structure, or material integrity, and in accordance with the WHO recommendation, maintained wettability post-processing. These experiments establish a foundational understanding of critical elements for the decontamination and reuse of PPE in any setting and provide a direction for future research in the field.

A microengineered human corneal epithelium-on-a-chip for eye drops mass transport evaluation.

Animals are commonly used for pharmacokinetic studies which are the most frequent events tested during ocular drug development and preclinical evaluation. Inaccuracy, cost, and ethical criticism in these tests have created a need to construct an in vitro model for studying corneal constraints. In this work, a porous membrane embedded microfluidic platform is fabricated that separates a chip into an apical and basal side. After functionalizing the membrane surface with fibronectin, the membrane's mechanical and surface properties are measured to ensure correct modeling of in vivo characteristics. Immortalized human corneal epithelial cells are cultured on the membrane to create a microengineered corneal epithelium-on-a-chip (cornea chip) that is validated with experiments designed to test the barrier properties of the human corneal epithelium construct using model drugs. A pulsatile flow model is used that closely mimics the ocular precorneal constraints and is reasonable for permeability analysis that models in vivo conditions. This model can be used for preclinical evaluations of potential therapeutic drugs and to mimic the environment of the human cornea.

Electrospun Polythiophene Phenylenes for Tissue Engineering.

This research focuses on the design of biocompatible materials/scaffold suitable for use for tissue engineering. Porous fiber mats were produced through electrospinning of polythiophene phenylene (PThP) conducting polymers blended with poly(lactide- co-glycolic acid) (PLGA). A peptide containing an arginylglycylaspartic acid (RGD) fragment was synthesized using solid phase peptide synthesis and subsequently grafted onto a PThP polymer using azide-alkyne "click" chemistry. The obtained RGD functionalized PThP was also electrospun into a fiber mat. The electrospun mats' morphology, roughness and stiffness were studied by means of scanning electron microscopy (SEM) and atomic force microscopy (AFM) and their electroactivity by cyclic voltammetry. The fibers show excellent cytocompatibility in culture assays with human dermal fibroblasts-adult (HDFa) and human epidermal melanocytes-adult (HEMa) cells. The electrospun fibers' roughness and stiffness changed after exposing the fiber mats to the cell culture medium (measured in dry state), but these changes did not affect the cell proliferation. The cytocompatibility of our porous scaffolds was established for their applicability as cell culture scaffolds by means of investigating mitochondrial activity of HDFa and HEMa cells on the scaffolds. The results revealed that the RGD moieties containing PThP scaffolds hold a promise in biomedical applications, including skin tissue engineering.

Lipid-polymer hybrid nanoparticle-mediated therapeutics delivery: advances and challenges.

With rapid advances in nanomedicine, lipid-polymer hybrid nanoparticles (LPHNPs) have emerged as promising nanocarriers for several biomedical applications, including therapeutics delivery and biomedical imaging. Significant research has been dedicated to biomimetic or targeting functionalization, as well as controlled and image-guided drug-release capabilities. Despite this research, the clinical translation of LPHNP-mediated therapeutics delivery has progressed incrementally. In this review, we discuss the recent advances in and challenges to the development and application of LPHNPs, present examples to demonstrate the advantages of LPHNPs in therapeutics delivery and imaging applications, and discuss the translational obstacles to LPHNP technology.

An ultra-sensitive biophysical risk assessment of light effect on skin cells.

The aim of this study was to analyze photo-dynamic and photo-pathology changes of different color light radiations on human adult skin cells. We used a real-time biophysical and biomechanics monitoring system for light-induced cellular changes in an in vitro model to find mechanisms of the initial and continuous degenerative process. Cells were exposed to intermittent, mild and intense (1-180 min) light with On/Off cycles, using blue, green, red and white light. Cellular ultra-structural changes, damages, and ECM impair function were evaluated by up/down-regulation of biophysical, biomechanical and biochemical properties. All cells exposed to different color light radiation showed significant changes in a time-dependent manner. Particularly, cell growth, stiffness, roughness, cytoskeletal integrity and ECM proteins of the human dermal fibroblasts-adult (HDF-a) cells showed highest alteration, followed by human epidermal keratinocytes-adult (HEK-a) cells and human epidermal melanocytes-adult (HEM-a) cells. Such changes might impede the normal cellular functions. Overall, the obtained results identify a new insight that may contribute to premature aging, and causes it to look aged in younger people. Moreover, these results advance our understanding of the different color light-induced degenerative process and help the development of new therapeutic strategies.

Microengineered biomimetic ocular models for ophthalmological drug development.

Current ophthalmological drug discovery and testing methods have limitations and concerns regarding reliability, ethicality, and applicability. These drawbacks can be mitigated by developing biomimetic eye models through mathematical and experimental methods which are often referred to as "eye-on-a-chip" or "eye chip". These eye chip technologies emulate ocular physiology, anatomy, and microenvironmental conditions. Such models enable understanding of the fundamental biology, pharmacology, and toxicology mechanisms by investigating the pharmacokinetics and pharmacodynamics of various candidate drugs under ocular anatomical and physiological conditions without animal models. This review provides a comprehensive overview of the latest advances in theoretical and in vitro experimental models of the anterior segment of the eye and its microenvironment, including eye motions and tear film dynamics. The current state of ocular modeling and simulation from predictive models to experimental models is discussed in detail with their advantages and limitations. The potential for future eye chip models to expedite new ophthalmic drug discoveries is also discussed.

Evaluation of UV radiation-induced toxicity and biophysical changes in various skin cells with photo-shielding molecules.

Ultraviolet radiation (UVR) triggers many complex events in different types of skin cells, including benign, malignant and normal cells. Chromophores present in these cells play a crucial role in various cellular processes. Unprecedented methods are required for the real-time monitoring of changes in an in vitro model exposed to intermittent mild and intense UVR to determine the mechanisms underlying cell degeneration and the effects of unexpected toxic, agonist and antagonist agents. This study reports the analytical application of a whole cell-based sensor platform for examining the biophysical effects of UVR. We used human keratinocyte, melanocyte and fibroblast cell lines to determine the normal, pathological and protective roles of UVR. In addition, we examined the real-time morphological, biophysical and biomechanical changes associated with cell degeneration induced by UVR at 254 and 365 nm. Information on UVR-induced changes in the cytoskeleton ultrastructure, cellular integrity, cell spreading area, actin microfilament distribution inflammation, microtubule damage, membrane damage, rupture and death was characterized by examining the loss or increase in biophysical and biomechanical properties of these cells. All cells exposed to UVR at 254 and 365 nm showed a significant increase in surface roughness and stiffness in a time-dependent manner. UVR-induced toxicity in differently pigmented skin cells was compared with that in cells pretreated with melanin, keratin and basic fibroblast growth factor to analyze the shielding efficiency of these agents. Melanin exerted a significant shielding effect compared to the other two agents. The biophysical and biomechanical information obtained in this study could advance our understanding of the UVR-induced degeneration process, and help in developing new interventions strategies.

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

This review will present a brief discussion on the recent advancements of bioelectrical impedance cell-based biosensors, especially the electric cell-substrate impedance sensing (ECIS) system for screening of various bioactive molecules. The different technical integrations of various chip types, working principles, measurement systems, and applications for drug targeting of molecules in cells are highlighted in this paper. Screening of bioactive molecules based on electric cell-substrate impedance sensing is a trial-and-error process toward the development of therapeutically active agents for drug discovery and therapeutics. In general, bioactive molecule screening can be used to identify active molecular targets for various diseases and toxicity at the cellular level with nanoscale resolution. In the innovation and screening of new drugs or bioactive molecules, the activeness, the efficacy of the compound, and safety in biological systems are the main concerns on which determination of drug candidates is based. Further, drug discovery and screening of compounds are often performed in cell-based test systems in order to reduce costs and save time. Moreover, this system can provide more relevant results in in vivo studies, as well as high-throughput drug screening for various diseases during the early stages of drug discovery. Recently, MEMS technologies and integration with image detection techniques have been employed successfully. These new technologies and their possible ongoing transformations are addressed. Select reports are outlined, and not all the work that has been performed in the field of drug screening and development is covered.

Effects of agmatine and resveratrol on RGC-5 cell behavior under light stimulation.

A light radiation causes dysfunction and death of retinal cells and leads to degeneration. Present study, investigated the light-induced cell dysfunction, and their activity. Further, the effects of agmatine and resveratrol on light-induced damage and these underlying photo-oxidative and protective mechanisms were monitored by real-time bio-impedance system. After light exposure retinal ganglion cells underwent death in a time dependent manner. During light exposure the cells elevate free radicals and Ca(2+), followed by nitric oxide (NO) and tumor necrosis factor-α (TNF-α), which can be facilitated to cell demise. The results revealed that these drugs can control the elevation of free radical, calcium gating, NO level, and increased TNF-α, which could diminish cell photo-damage. In summary, resveratrol helps more to rescue damaged cells compared to agmatine. The proposed system suggested mechanism could meet to identify the photo-toxic effects in retinal cells, and provides high throughput screening for early stages photo-damage.

Photoprotective effects of apple peel nanoparticles.

Plants contain enriched bioactive molecules that can protect against skin diseases. Bioactive molecules become unstable and ineffective due to unfavorable conditions. In the present study, to improve the therapeutic efficacy of phytodrugs and enhance photoprotective capability, we used poly(D,L-lactide-co-glycolide) as a carrier of apple peel ethanolic extract (APETE) on permeation-enhanced nanoparticles (nano-APETE). The in vitro toxicity of nano-APETE-treated dermal fibroblast cells were studied in a bioimpedance system, and the results coincided with the viability assay. In addition, the continuous real-time evaluations of photodamage and photoprotective effect of nano-APETE on cells were studied. Among three different preparations of nano-APETE, the lowest concentration provided small, spherical, monodispersed, uniform particles which show high encapsulation, enhanced uptake, effective scavenging, and sustained intracellular delivery. Also, the nano-APETE is more flexible, allowing it to permeate through skin lipid membrane and release the drug in a sustained manner, thus confirming its ability as a sustained transdermal delivery. In summary, 50 µM nano-APETE shows strong synergistic photoprotective effects, thus demonstrating its higher activity on target sites for the treatment of skin damage, and would be of broad interest in the field of skin therapeutics.

A transdermal delivery system to enhance quercetin nanoparticle permeability.

Targeting oxidative stress with inhibiting or boosting the endogenous levels of antioxidants potentially has an outstanding effect in the treatment of oxidative-stress-related diseases. The present work demonstrates the synthesis of quercetin nanoemulsion as one of the potential antioxidants for the treatment of oxidative- stress-related diseases. A quercetin nanoemulsion was prepared using poly(lactic-co-glycolic acid) (PLGA), hyaluronic acid (HA) and emulsifier (Tween-20) through a solvent evaporation technique. The efficiency of the nanoemulsion was evaluated with and without chemical permeation enhancer (CPE). The FT-IR result shows no interaction between quercetin and polymer that proves excellent compatibility. The transdermal delivery ability was evaluated using in vitro release and ex vivo permeation analysis. The transdermal drug-release mechanism was studied by the mathematical model and was found to obey a zero-order, diffusion-controlled mechanism. In vitro toxicity and cell behavior, including cell adhesion, proliferation and cell death of quercetin-nanoemulsion-treated L929 cells, were elucidated by the electrical cell-substrate impedance sensing (ECIS) technique. The produced nanoemulsion showed a high encapsulation efficiency, less toxicity, controlled delivery with enhanced transdermal drug permeation and effective scavenging of free radicals.

Impedance-based cell culture platform to assess light-induced stress changes with antagonist drugs using retinal cells.

This Article describes an unprecedented, simple, and real-time in vitro analytical tool to measure the luminous effect on the time responses function of retinal ganglion cells (RGC-5) by electric cell substrate impedance sensing (ECIS) system. The ECIS system was used for the continuous measurement of different color light-induced effects on the response of cells that exposed to protective drugs. The measurement suggests that the association of photo-oxidative stress was mediated by reactive oxygen species (ROS), which plays a critical role that leads to cell stress, damages, and retinopathy, resulting in eye degenerative diseases. Continuous light radiation caused time-dependent decline of RGC-5 response and resulted in photodamage within 10 h due to adenosine 5'-triphosphate depletion and increased ROS level, which is similar to in vivo photodamage. The ECIS results were correlated with standard cell viability assay. ECIS is very helpful to determine the protective effects of analyzed drugs such as ß-carotene, quercetin, agmatine, and glutathione in RGC-5 cells, and the maximum drug activity of nontoxic safer drug concentrations was found to be 0.25, 0.25, 0.25, and 1.0 mM, respectively. All drugs show protection against light radiation toxicity in a dose-dependent manner; the most effective drug was found to be glutathione. The proposed system identifies the phototoxic effects in RGC-5 and provides high throughput drug screening for photo-oxidative stress during early stages of drug discovery. This study is convenient and potential enough for the direct measurements of the photoprotective effect in vitro and would be of broad interest in the field of therapeutics.

Light-induced anatomical alterations in retinal cells.

Exploring the cellular behavior and real-time evaluation of light-induced stress in retinal cells requires advanced sensitive methods. Here, an analytical application of a bioimpedance system was developed to quantify cell morphology changes in light-induced anatomical response. Hence, an unprecedented light setup was established using a bioimpedance system to generate artificial light that stresses retinal ganglion cells (RGCs), and the corresponding impedance was recorded. The dynamics of initial light-induced stress and the morphological changes were examined. This examination showed impedance drop, and the normalized variance ratios were used to quantify the changes. A rapid impedance drop was measured after exposure to the red and blue lights, whereas it was found that a minimum impedance drop was analyzed under the green light exposure. These data have revealed a strong, unconventional reliable model to detect the onset changes for photo-oxidative stress-related studies by the morphological changes of neurite retraction and cell body shrinkage. The data revealed that functions of the RGC-5 cell line under light exposure were diminished in cell strength and activity was hindered. These reduced functions might be related to the declined levels of some protein markers and stress factors. This is an effective model to study stress, morphological changes, and photodamage.

Dual drug-loaded nanoparticles on self-integrated scaffold for controlled delivery.

Antioxidant (quercetin) and hypoglycemic (voglibose) drug-loaded poly-D,L-lactideco-glycolide nanoparticles were successfully synthesized using the solvent evaporation method. The dual drug-loaded nanoparticles were incorporated into a scaffold film using a solvent casting method, creating a controlled transdermal drug-delivery system. Key features of the film formulation were achieved utilizing several ratios of excipients, including polyvinyl alcohol, polyethylene glycol, hyaluronic acid, xylitol, and alginate. The scaffold film showed superior encapsulation capability and swelling properties, with various potential applications, eg, the treatment of diabetes-associated complications. Structural and light scattering characterization confirmed a spherical shape and a mean particle size distribution of 41.3 nm for nanoparticles in the scaffold film. Spectroscopy revealed a stable polymer structure before and after encapsulation. The thermoresponsive swelling properties of the film were evaluated according to temperature and pH. Scaffold films incorporating dual drug-loaded nanoparticles showed remarkably high thermoresponsivity, cell compatibility, and ex vivo drug-release behavior. In addition, the hybrid film formulation showed enhanced cell adhesion and proliferation. These dual drug-loaded nanoparticles incorporated into a scaffold film may be promising for development into a transdermal drug-delivery system.