A novel scaffold for in situ electrochemical detection of cell biomarkers was developed using electrospun nanofibers and commercial adhesive polymeric membranes. The electrochemical sensing of cell biomarkers requires the cultivation of the cells on/near the (bio)sensor surface in a manner to preserve an appropriate electroactive available surface and to avoid the surface passivation and sensor damage. This can be achieved by employing biocompatible nanofiber meshes that allow the cells to have a normal behavior and do not alter the electrochemical detection. For a better mechanical stability and ease of handling, nylon 6/6 nanofibers were collected on commercial polymeric membranes, at an optimal fiber density, obtaining a double-layered platform. To demonstrate the functionality of the fabricated scaffold, the screening of cellular stress has been achieved integrating melanoma B16-F10 cells and the (bio)sensor components on the transducer whereas the melanin exocytosis was successfully quantified using a commercial electrode. Either directly on the surface of the (bio)sensor or spatially detached from it, the integration of cell cultures in biosensing platforms based on electrospun nanofibers represents a powerful bioanalytical tool able to provide real-time information about the biomarker release, enzyme activity or inhibition, and monitoring of various cellular events.
Electrospun nanofibres based on poly(styrene sulfonate) doped poly(3,4-ethylenedioxythiophene) (PEDOT:PSS) were fabricated using a straightforward procedure which combines electrospinning, sputtering deposition and electrochemical synthesis. In general, electrospun fibre meshes based on conducting polymers are prepared by mixing the conducting polymer with a carrier polymer or chemically coverage of the fibres. In contrast, freestanding nylon 6/6 nanofibre webs were prepared through electrospinning and were coated by sputtering with gold in order to make them conductive. Further, a PEDOT:PSS layer was electrochemically deposited onto the metalized nanofibre meshes and the synthesis parameters were chosen in such a way to preserve the high active area of the fibres. The prepared material was morphologically characterized and the formation of PEDOT:PSS was also demonstrated. The PEDOT:PSS coated nanofibres revealed remarkable electrical properties (sheet resistance of about 3.5 Omega cm(-2)), similar to those of metalized nanofibres (sheet resistance around 3 Omega cm2). The in vitro studies using L929 fibroblast mouse cells showed that the bioactive material has no cytotoxic effect and allows proliferation. Moreover, after 72 h of incubation, the fibroblasts shrunk their nuclei and spread suggesting that a differentiation in myofibroblast occurs without application of any kind of external stimuli. These results will be helpful for developing efficient materials for wound healing applications that work without energy consumption.
The bioavailability of the administered drugs that reach the systemic circulation is the first point in resolving the pathology of patients. Albumin-based nanoparticles represent an increasingly used strategy to deliver cancer drugs into cells that otherwise cannot overcome biological barriers. In this work, rutin (Ru), a flavonoid with anticancer and antioxidant potential, was incorporated into bovine serum albumin nanoparticles (BSA-Ru NPs), developed using the desolvation method, and the entire system was characterized and evaluated by scanning electron microscopy (SEM), atomic force microscopy (AFM), and UV–Vis absorption spectroscopy. The results showed that BSA and BSA–Ru NPs are uniformly distributed, have relatively large sizes, and have a time stability of more than 60%. Furthermore, the effect of these nanohybrids on the thermal stability of liposomal membranes was evaluated by surface plasmon resonance (SPR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The viability evaluation was assessed by the tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) protocol in the fibroblast L929 line and a high level of biocompatibility, confirmed by SEM results, was found.
Technological progress has led to the development of analytical tools that promise a huge socio-economic impact on our daily lives and an improved quality of life for all. The use of plant extract synthesized nanoparticles in the development and fabrication of optical or electrochemical (bio)sensors presents major advantages. Besides their low-cost fabrication and scalability, these nanoparticles may have a dual role, serving as a transducer component and as a recognition element, the latter requiring their functionalization with specific components. Different approaches, such as surface modification techniques to facilitate precise biomolecule attachment, thereby augmenting recognition capabilities, or fine tuning functional groups on nanoparticle surfaces are preferred for ensuring stable biomolecule conjugation while preserving bioactivity. Size optimization, maximizing surface area, and tailored nanoparticle shapes increase the potential for robust interactions and enhance the transduction. This article specifically aims to illustrate the adaptability and effectiveness of these biosensing platforms in identifying precise biological targets along with their far-reaching implications across various domains, spanning healthcare diagnostics, environmental monitoring, and diverse bioanalytical fields. By exploring these applications, the article highlights the significance of prioritizing the use of natural resources for nanoparticle synthesis. This emphasis aligns with the worldwide goal of envisioning sustainable and customized biosensing solutions, emphasizing heightened sensitivity and selectivity.
A novel electrochemical biosensor was developed to monitor fibroblast cells stress levels for the first time in situ under external stimuli based on the recognition of superoxide anion released upon cell damage. The biosensor comprised metallized polycaprolactone electrospun fibers covered with zinc oxide for improved cell adhesion and signal transduction, whilst stable bioconjugates of mercaptobenzoic acid-functionalized gold nanoparticles/superoxide dismutase were employed as recognition bioelements. Biosensors were first tested and optimized for in situ generated superoxide detection by fixed potential amperometry at +0.3 V, with minimal interferences from electroactive species in cell culture media. L929 fibroblast cells were then implanted on the optimized biosensor surface and the biosensor morphologically characterized by scanning electron microscopy (SEM) and fluorescence microscopy, which illustrated the network-type pattern of fibroblasts adjacent to the fiber scaffold. Fibroblast stress was induced by zymosan and monitored at the cells integrated biosensor using fixed potential amperometry (CA) with a sensitivity of 26 nA cm-2 μg mL-1 zymosan and electrochemical impedance spectroscopy (EIS), with similar sensitivity of the biosensor considering the Rs and Z' parameters of around 0.13 Ω cm2 μg-1 mL and high correlation factors R2 of 0.9994. The obtained results underline the applicability of the here developed biosensor for the electrochemical screening of the fibroblast cells stress. The concept in using low-cost biocompatible polymeric fibers as versatile scaffolds for both enzyme immobilization and cell adhesion, opens a new path in developing biosensors for the in-situ investigation of a variety of cellular events.
Hematologic malignancies represent cancer diseases that affect the bone marrow and blood cells and include various subtypes depending mostly on the morphology of the cells. It is well known that an early diagnosis could be very useful for increasing survival rates in cancer, especially in the aggressive forms which can quickly progress to untreatable forms. The development of an easy, fast, and sensitive analytical tool with indicative applications in diagnosis and follow-up care, that could bring benefits in the discovery of new malignant diseases or relapses is reported. Oncofetal antigen/immature laminin receptor protein (OFA/iLRP) is an immunogenic protein found in fetal cells as well as overexpressed on the surface of some malignant tumors, including some hematologic malignancies. An aptamer, AB3, was selected and reported in the literature, having as a target the immature laminin receptor protein. Using the AB3 aptamer and its affinity to immature laminin receptor protein-positive cells an aptasensor was developed and tested on Jurkat cells. For the immobilization of the aptamer, graphene oxide modified screen printed electrodes were used and subjected to an activation procedure. The aptasensor development and cell caption were evaluated using electrochemical methods as well as microscopic techniques. The limit of detection of the developed aptasensor was 3.3 x 10(3) cells mL(-1), meaning 16 cells in the 5 mu L of suspension tested.
Albumin-based nanoparticles used as drug delivery systems are an important research topic in nanomedicine due to the special properties of albumin, such as biocompatibility, biodegradability, and well-tolerance without any side effects. Herein, human serum albumin (HSA) nanoparticles (NPs) were synthesized via the nanoprecipitation method, analyzed through spectroscopy and microscopy, and evaluated in vitro. UV-Vis absorption spectroscopy was used in order to investigate their stability over time, scanning electron microscopy (SEM), fluorescence microscopy and atomic force microscopy (AFM) were used for morphological characterization and the MTS protocol was assessed for the biocompatibility evaluation. The AFM and SEM results for HSA-NPs on a solid surface indicated a uniform distribution and sizes around 50 - 70 nm. Also, FM images obtained for HSA-NPs labeled with the fluorescent indicator Thioflavin T showed that HSA-NPs have the propensity to form large aggregates that tend to structure into amyloid fibrils. The in vitro studies on the fibroblast L929 and colon HT-29 cell lines demonstrated that HSA NPs did not reduce cell viability, and the microscopy images revealed the tendency of HSA NPs to concentrate on the cell surface. These results could be exploited in the development of new folic acid-containing drug delivery systems that target cancer cells that have membrane receptors for folic acid.
Azathioprine (AZA) is a pharmacologic immunosuppressive agent administrated in various conditions such as autoimmune disease or to prevent the rejection of organ transplantation. The mechanism of action is based on its biologically active metabolite 6-mercaptopurine (6-MP), which is converted, among others, into thioguanine nucleotides capable of incorporating into replicating DNA, which may act as a strong UV chromophore and trigger DNA oxidation. The interaction between azathioprine and DNA, before and after exposure to solar simulator radiation, was investigated using UV–vis spectrometry and differential pulse voltammetry at a glassy carbon electrode. The results indicated that the interaction of AZA with UV radiation was pH-dependent and occurred with the formation of several metabolites, which induced oxidative damage in DNA, and the formation of DNA-metabolite adducts. Moreover, the viability assays obtained for the L929 cell culture showed that both azathioprine and degraded azathioprine induced a decrease in cell proliferation.
PC-12 cells have been widely used as a neuronal line study model in many biosensing devices, mainly due to the neurogenic characteristics acquired after differentiation, such as high level of secreted neurotransmitter, neuron morphology characterized by neurite outgrowth, and expression of ion and neurotransmitter receptors. For understanding the pathophysiology processes involved in brain disorders, PC-12 cell line is extensively assessed in neuroscience research, including studies on neurotoxicity, neuroprotection, or neurosecretion. Various analytical technologies have been developed to investigate physicochemical processes and the biosensors based on optical and electrochemical techniques, among others, have been at the forefront of this development. This article summarizes the application of different biosensors in PC-12 cell cultures and presents the modern approaches employed in neuronal networks biosensing.
Romania is going through a period of economic transition, subject to the pressures of globalization that affect the evolution of the family, at the micro social level, structurally, from the behaviour and relational point of view. The continuous process of changes in the labour market, the inefficiency of the association between vocational training and job satisfaction, the financial difficulties felt by most families but also the challenge of modernity have emphasized the phenomenon of migration in the last decade. The departure of parents who have to work abroad has become a worrying phenomenon with a higher incidence in the eastern half of the country. It has got complex effects on the evolution of the family, especially on the children left at home with one of their parents or their tutors. Nowadays, the studies show more and more situations of neglect in which children become victims and suffer emotionally and physically. They also suffer various abuses, they are exploited through work or sex. In schools, there is a new profile of special educational requirements (not deficiencies), the profile of children left at home without parental support. It is worrying the migration phenomenon seen as a value model by the young generation and its negative effects at school level: decrease of motivation for learning or school abandonment. The present study discusses a review of the current scientific literature objectively, which examines the impact of breaking attachment relationships between children and parents on socio-emotional development and school outcomes. The Romanian society knows an important socio-economic phenomenon, which has grown since 1990: migration. In 2017, a study carried out at the request of the Romanian Government recorded more than 85,000 children left home alone with one of the parents or without parental supervision. We aim to analyse what effects at school and socio-emotional level have the loss of attachment ties having as moderators the gender of the migrant parent, the duration of the separation, the age at which the separation occurs. When these relationships are interrupted, the child’s emotional development is affected, his emotional balance having repercussions in his social life. The purpose of this study is to identify, monitor the dimensions of the phenomenon in intensely affected areas (Braila and Galati counties), the psycho-pedagogical aspects of children with migrant parents exposed to situations of vulnerability, marginalization and to propose a program of educational strategies in order to optimize school motivation. The main objective of the research is to identify, evaluate and involve them into adaptive actions that have as their objective the rebalancing of the socio-affective relations