MicroRNAs (miRNAs) are small non-coding RNAs (18-22 nucleotides) that regulate gene expression and are associated with various diseases, including Laryngeal Cancer (LCa), which has a high mortality rate due to late diagnosis. Traditional methods for miRNA detection present several drawbacks (time-consuming steps, high cost and high false positive rate). Early-stage diagnosis and selective detection of miRNAs remain challenging. This study proposes a 3D flexible biosensor that combines nanofibers (NFs), gold nanoparticles (AuNPs), and an inverse molecular sentinel (iMS) for enzyme-free, SERS-based detection of miRNA-223-3p, evaluated as a potential LCa biomarker. The electrospun flexible nanofibers decorated with AuNPs enhance Raman signal. Selective detection of miRNA-223-3p is achieved by immobilizing an iMS-DNA probe labeled with a Raman reporter (Cyanine 3) on the AuNPs. The iMS distinctive stem-and-loop structure undergoes a conformational change upon interaction with the miRNA-223-3p, producing an "on to off" SERS signal. The proposed sensor demonstrated a linear detection range from 10 to 250 fM, with a limit of detection (LOD) of 19.50 ± 0.05 fM. The sensor selectivity was confirmed by analyzing the SERS signal behaviour in the presence of both Non-complementary miRNA and miRNA with three mismatched base pairs. This easily fabricable sensor requires no amplification and offers key advantages, including sensitivity, flexibility, and cost-effectiveness.
- Klíčová slova
- Flexible sensors, Laryngeal Cancer, Nanofiber, SERS, miRNA-223-3p,
- MeSH
- biosenzitivní techniky * metody MeSH
- časná detekce nádoru * metody MeSH
- kovové nanočástice chemie MeSH
- lidé MeSH
- limita detekce MeSH
- mikro RNA * analýza genetika MeSH
- nádory hrtanu * diagnóza genetika MeSH
- nanovlákna * chemie MeSH
- Ramanova spektroskopie * metody MeSH
- zlato chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- mikro RNA * MeSH
- MIRN223 microRNA, human MeSH Prohlížeč
- zlato MeSH
In electrospun scaffolds, coaxial electrospinning is gaining increased attention due to its potential for biocomponent encapsulation and controlled delivery. However, the encapsulation of biocomponents, such as liposomes, remains challenging because of their low stability in commonly used electrospinning solvents. This study, therefore, aims to develop a novel coaxial electrospinning formulation for crafting a liposome-encapsulated, rapid-release coaxial fiber. Liposomes demonstrated desirable stability in fish gelatin/phosphate-buffered saline (PBS) solutions, which remain liquid at room temperature and exhibit exceptional spinnability at concentrations exceeding 80 w/v% due to the reduction in surface tension. Fluorescent labelling examinations confirmed the successful encapsulation of liposomes within coaxial fibers electrospun from a 160 w/v% gelatin/PBS core and a 20 w/v% PCL/chloroform/N,N-dimethylformamide (DMF) shell. The gelatin/PBS core solution formed solid ends at the tips of the core-shell fiber post-spinning, while maintaining a liquid state within the shell, thereby enabling the encapsulation of liposomes within the PCL coaxial fiber. Upon exposure to medium, the solid ends dissolve, enabling the rapid release of liposomes. The successful development of this liposome-loaded electrospun coaxial fiber, using fish gelatin, highlights its potential for creating advanced liposome delivery systems.
- Klíčová slova
- PCL, coaxial electrospinning, fish gelatin, liposomes,
- Publikační typ
- časopisecké články MeSH
Development of electrospun nanofibers with suitable properties to promote wound healing is an advantage in developing non-invasive skin treatments. We showed the potential application of Polyvinyl acetate (PVA) and Polyvinylpyrrolidone (PVP) combined with Helichrysum italicum oil (HO) in wound healing. During this process, Tight junctions (TJs) play a crucial role in maintaining skin integrity. TJs are intercellular junctions composed of a variety of transmembrane proteins, including Occludin (OCLN), observed also in migrating epithelial cells. Changes in OCLN expression affect epidermal permeability, indicating an active role in the healing process. Within this context, we studied the OCLN expression during healing after scratch assay on Keratinocytes (HaCaT), by a confocal microscopic analysis. In addition, we evaluated the effect of treatment after scratch on cell elasticity by Atomic Force Microscopy (AFM) analysis. All results show a positive trend in cell proliferation and viability on HaCaT treated with functionalized nanofibers. These results were confirmed by the expression of genes involved in the early stages of the regenerative process. Understanding the cell mechanisms involved in skin changes during repair process would allow future application of nanomaterials combined with HO in vivo.
- Klíčová slova
- Bioactive molecules, Cellular mechanism, Keratinocytes, Nanofibers, Regenerative medicine, Tissue regeneration, Wound healing,
- MeSH
- buněčné linie MeSH
- hojení ran účinky léků MeSH
- keratinocyty * účinky léků metabolismus cytologie MeSH
- kůže účinky léků metabolismus MeSH
- lidé MeSH
- mikroskopie atomárních sil MeSH
- nanovlákna * chemie MeSH
- okludin metabolismus genetika MeSH
- oleje rostlin farmakologie chemie MeSH
- polyvinylalkohol chemie MeSH
- povidon * chemie farmakologie MeSH
- proliferace buněk * účinky léků MeSH
- pružnost účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- okludin MeSH
- oleje rostlin MeSH
- polyvinylalkohol MeSH
- povidon * MeSH
PAN fibers are characterized by having a large surface-to-volume ratio and small pores, which are beneficial for applications in filtration and specific molecular detection systems. Naturally, larger items are filtered, and a lower ratio between specific and nonspecific binding is expected since small pores do not allow larger elements to penetrate through membranes; thus, nonspecific binding is enhanced. We prepared and tested fiber membranes (diameter cca 700 nm) functionalized with a specific antibody to prove that even microscopic systems such as bacteria could be specifically identified. In addition, we established a methodology that enabled the effective binding of bacteria in not only an aqueous environment but also air. Our data clearly prove that even large systems such as bacteria could be specifically identified by fiber membranes surface-functionalized with a specific antibody. This research opens the door to the construction of biosensors for the fast, inexpensive, and sensitive identification of airborne bacterial contaminants and other airborne pollutants.
- Klíčová slova
- Air detection, Airborne pathogen detection, Antibody-based nanobiosensor, Functionalized PAN nanofibers, Nanofiber biosensors,
- MeSH
- Bacteria izolace a purifikace MeSH
- biologické markery analýza MeSH
- biosenzitivní techniky metody MeSH
- membrány umělé * MeSH
- mikrobiologie vzduchu * MeSH
- monitorování životního prostředí metody přístrojové vybavení MeSH
- povrchové vlastnosti MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- membrány umělé * MeSH
MicroRNAs (miRNAs) are involved in post-transcriptional gene expression regulation and in mechanisms of cancer growth and metastases. In this light, miRNAs could be promising therapeutic targets and biomarkers in clinical practice. Therefore, we investigated if specific miRNAs and their target genes contribute to laryngeal squamous cell carcinoma (LSCC) development. We found a significant decrease of miR-449a in LSCC patients with nodal metastases (63.3%) compared with patients without nodal involvement (44%). The AmpliSeq Transcriptome of HNO-210 miR-449a-transfected cell lines allowed the identification of IL6-R as a potential target. Moreover, the downregulation of IL6-R and the phosphorylation reduction of the downstream signaling effectors, suggested the inhibition of the IL-6 trans-signaling pathway. These biochemical effects were paralleled by a significant inhibition of invasion and migration in vitro and in vivo, supporting an involvement of epithelial-mesenchymal transition. These findings indicate that miR-449a contributes to suppress the metastasization of LSCC by the IL-6 trans-signaling block and affects sensitivity to external stimuli that mimic pro-inflammatory conditions.
- Klíčová slova
- IL-6 trans-signaling, LSCC, MT: non-coding RNAs, gene expression, metastases miR-449a, microRNAs,
- Publikační typ
- časopisecké články MeSH
The skin is the primary tissue affected by wounds and aging, significantly impacting its protective function. Natural products are widely used in cosmetics, representing a new approach to preventing age-related damage. Nanomedicine combines nanotechnology and traditional treatments to create innovative drugs. The main targets of nanotechnological approaches are wound healing, regeneration, and rejuvenation of skin tissue. The skin barrier is not easily permeable, and the creation of modern nanodevices is a way to improve the passive penetration of substances. In this study, Helichrysum italicum oil (HO) was combined with different types of electrospun nanofibers to study their protective activity on the skin and to evaluate their future application for topical treatments. In the present research, we used biodegradable polymers, including polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), which were characterized by a scanning electron microscope (SEM). All results show a positive trend in cell proliferation and viability of human skin stem cells (SSCs) and BJ fibroblasts pre-treated with combined nanofibers and then exposed to UV stress. Gene expression analysis revealed the activation of a molecular rejuvenation program in SSCs treated with functionalized nanofibers before UV exposure. Understanding the mechanisms involved in skin changes during aging allows for the future application of nanomaterials combined with HO directly to the patients.
- Klíčová slova
- Helichrysum italicum, bioactive molecules, drug delivery, molecular mechanisms, nanofibers, nanosystem, skin aging, stem cells,
- MeSH
- biologické přípravky * farmakologie MeSH
- hojení ran MeSH
- kůže MeSH
- lidé MeSH
- nanovlákna * MeSH
- polyvinylalkohol MeSH
- stárnutí kůže * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické přípravky * MeSH
- polyvinylalkohol MeSH
The paper aimed to prepare quaternary chitosan-based nanofibers as bioabsorbable wound dressings. To this aim, fully biodegradable chitosan/N,N,N-trimethyl chitosan (TMC) nanofibers were designed and prepared via electrospinning, using poly(ethylene glycol) as sacrificial additive. The new biomaterials were structurally and morphologically characterized by FTIR and NMR spectroscopy, thermogravimetric analysis, X-ray diffraction and scanning electron microscopy, and their properties required for wound dressings application were investigated and discussed in detail. Thus, the nanofiber behavior was investigated by swelling, dynamic vapor sorption, and in vitro biodegradation in media mimicking the wound exudate. The mechanical properties were analysed from the stress-strain curves, the bioadhesivity from the texture analysis and the mucoadhesivity from the Zeta potential and transmittance measurements. The antimicrobial activity was assessed against S. aureus and E. coli strains, and the biocompatibility was tested in vitro on normal human dermal fibroblasts, and in vivo on rats. The application of the fiber mats with the best balance of properties as dressings on deep burn wound models in rats showed wound closure and active healing, with fully restoration of epithelia. It was concluded that the combination of chitosan with TMC into nanofibers provides new potential bioabsorbable wound dressing, opening new perspectives in regenerative medicine.
- Klíčová slova
- Bioabsorbable, Burn wound healing, N,N,N-trimethyl chitosan nanofibers,
- MeSH
- antibakteriální látky farmakologie chemie MeSH
- chitosan * chemie MeSH
- Escherichia coli MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- nanovlákna * chemie MeSH
- obvazy MeSH
- Staphylococcus aureus MeSH
- vstřebatelné implantáty MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- chitosan * MeSH
Nanodiamonds are innovative nanocrystalline carbon particles able to deliver chemically conjugated miRNAs. In oncology, the use of miRNA-based therapies may represent an advantage, based on their ability to simultaneously target multiple intracellular oncogenic targets. Here, nanodiamonds were tested and optimized to deliver miR-34a, a miRNA playing a key role in inhibiting tumor development and progression in many cancers. The physical-chemical properties of nanodiamonds were investigated suggesting electrical stability and uniformity of structure and size. Moreover, we evaluated nanodiamond cytotoxicity on two breast cancer cell models and confirmed their excellent biocompatibility. Subsequently, nanodiamonds were conjugated with miR-34a, using the chemical crosslinker polyethyleneimine; real-time PCR analysis revealed a higher level of miR-34a in cancer cells treated with the different formulations of nanodiamonds than with commercial transfectant. A significant and early nanodiamond-miR-34a uptake was recorded by FACS and fluorescence microscopy analysis in MCF7 and MDA-MB-231 cells. Moreover, nanodiamond-miR-34a significantly inhibited both cell proliferation and migration. Finally, a remarkable anti-tumor effect of miR-34a-conjugated nanodiamonds was observed in both heterotopic and orthotopic murine xenograft models. In conclusion, this study provides a rationale for the development of new therapeutic strategies based on use of miR-34a delivered by nanodiamonds to improve the clinical treatment of neoplasms.
- Klíčová slova
- MT: Delivery Strategies, MiR-34a, MicroRNA, breast cancer, gene delivery, gene therapy, miRNA replacement therapy, nanodiamonds, nanomedicine, nanotechnology,
- Publikační typ
- časopisecké články MeSH
Breast cancer is the most prevalent cancer type in women worldwide. It proliferates rapidly and can metastasize into farther tissues at any stage due to the gradual invasiveness and motility of the tumor cells. These crucial properties are the outcome of the weakened intercellular adhesion, regulated by small guanosine triphosphatases (GTPases), which hydrolyze to the guanosine diphosphate (GDP)-bound conformation. We investigated the inactivating effect of ARHGAP1 on Rho GTPases involved signaling pathways after treatment with a high dose of doxorubicin. Label-free quantitative proteomic analysis of the proteome isolated from the MCF-7 breast cancer cell line, treated with 1 μM of doxorubicin, identified RAC1, CDC42, and RHOA GTPases that were inactivated by the ARHGAP1 protein. Upregulation of the GTPases involved in the transforming growth factor-beta (TGF-beta) signaling pathway initiated epithelial-mesenchymal transitions. These findings demonstrate a key role of the ARHGAP1 protein in the disruption of the cell adhesion and simultaneously allow for a better understanding of the molecular mechanism of the reduced cell adhesion leading to the subsequent metastasis. The conclusions of this study corroborate the hypothesis that chemotherapy with doxorubicin may increase the risk of metastases in drug-resistant breast cancer cells.
- Klíčová slova
- breast cancer, cell adhesion, doxorubicin, mass spectrometry, metastases, proteomics,
- MeSH
- cdc42 protein vázající GTP metabolismus MeSH
- doxorubicin farmakologie MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- nádory prsu * farmakoterapie MeSH
- proteiny aktivující GTPasu * metabolismus MeSH
- proteomika MeSH
- rac1 protein vázající GTP metabolismus MeSH
- Rho proteiny vázající GTP * metabolismus MeSH
- rhoA protein vázající GTP metabolismus MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- ARHGAP1 protein, human MeSH Prohlížeč
- cdc42 protein vázající GTP MeSH
- doxorubicin MeSH
- proteiny aktivující GTPasu * MeSH
- rac1 protein vázající GTP MeSH
- Rho proteiny vázající GTP * MeSH
- rhoA protein vázající GTP MeSH
