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Targeted drug delivery is one of the attractive ways in which cancer treatment can significantly reduce side effects. In the last two decades, the use of antibodies as a tool for accurate detection of cancer has been noted. On the other hand, the binding of drugs and carriers containing drugs to the specific antibodies of cancer cells can specifically target only these cells. However, the use of whole antibodies brings challenges, including their large size, the complexity of conjugation, the high cost of production, and the creation of immunogenic reactions in the body. The use of nanobodies, or VHHs, which are a small part of camel heavy chain antibodies, is very popular due to their small size, high craftsmanship, and low production cost. In this article, in addition to a brief overview of the structure and characteristics of nanobodies, the use of this molecule in the targeted drug delivery of breast cancer has been reviewed.
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DNA-based analytical techniques have provided an advantageous sensing assay in the realm of biotechnology. Bio-inspired fluorescent nanodots are a novel type of biological staining agents with excellent optical properties widely used for cellular imaging and diagnostics. In the present research, we successfully synthesized bio-dots with excellent optical properties and high-quantum yield from DNA sodium salt through the hydrothermal method. We conjugated the bio-dots with 3' Eclipse Dark Quencher (Eclipse)-labeled single-strand oligodeoxyribonucleotide according to carbodiimide chemistry, to design a fluorescence resonance energy transfer (FRET) probe. The results confirmed the prosperous synthesis and surface functionalization of the bio-dot. Analysis of size, zeta potential, and FTIR spectroscopy verified successful bioconjugation of the bio-dots with probes. UV-visibility analysis and fluorescence intensity profile of the bio-dot and bio-dot@probes represented a concentration-dependent quenching of fluorescent signal of bio-dot by Eclipse after probe conjugation. The results demonstrated that TaqMan PCR was not feasible using the designed bio-dot@probes. Our results indicated that bio-dot can be used as an efficient fluorescent tag in the design of fluorescently labeled oligonucleotides with high biocompatibility and optical features.
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Transferencia Resonante de Energía de Fluorescencia , Puntos Cuánticos , Transferencia Resonante de Energía de Fluorescencia/métodos , Estudios de Factibilidad , ADN/genética , Colorantes , Colorantes Fluorescentes/químicaRESUMEN
BackgroundAcute leukemia involving lymphocytic and myeloid cells is cancer with a high mortality rate. Swift and timely diagnosis might be a potential approach to improving patient prognosis and survival. The microRNA (miRNA) signatures are emerging nowadays for their promising diagnostic potential. MiRNA levels from bone marrow can be used as prognostic biomarkers.MethodsThe current study was designed to evaluate if the microRNAs and tumor suppressor genes (TSGs) profiling of hematopoietic bone marrow could help in acute leukemia early detection. Also, we assessed the DNA methyltransferase 3A (DNMT3A) expression and its possible epigenetic effects on miRNAs plus TSGs expression levels. The expression levels of ten miRNAs and four TSGs involved in acute lymphocytic leukemia (ALL) as well as acute myeloid leukemia (AML) were quantified in 43 and 40 bone marrow samples of ALL and AML patients in comparison with cancer-free subjects via real-time quantitative PCR (RT-qPCR). The receiver-operating-characteristic (ROC) analysis of miRNAs was performed in the study groups. Further, the correlation between the DNMT3A and TSGs was calculated.ResultsSignificant differences were detected in the bone marrow expression of miRNAs and TSGs (P < 0.05) between acute leukemia patients and healthy group. ROC analysis confirmed the ability of miR-30a, miR-101, miR-132, miR-129, miR-124, and miR-143 to discriminate both ALL and AML patients with an area under the ROC curve of ≥ 0.80 (P < 0.001) and high accuracy. The correlation between DNMT3A and P15/P16 TSGs revealed that DNMT3A plays a vital role in epigenetic control of TSGs expression. Our findings indicated that the downregulation of bone marrow miRNAs and TSGs was accompanied by acute leukemia development.ConclusionsThe authors conclude that this study could contribute to introducing useful biomarkers for acute leukemia diagnosis.
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Humanos , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Médula Ósea/patología , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Pronóstico , MicroARNs/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismoRESUMEN
BACKGROUND: Acute leukemia involving lymphocytic and myeloid cells is cancer with a high mortality rate. Swift and timely diagnosis might be a potential approach to improving patient prognosis and survival. The microRNA (miRNA) signatures are emerging nowadays for their promising diagnostic potential. MiRNA levels from bone marrow can be used as prognostic biomarkers. METHODS: The current study was designed to evaluate if the microRNAs and tumor suppressor genes (TSGs) profiling of hematopoietic bone marrow could help in acute leukemia early detection. Also, we assessed the DNA methyltransferase 3A (DNMT3A) expression and its possible epigenetic effects on miRNAs plus TSGs expression levels. The expression levels of ten miRNAs and four TSGs involved in acute lymphocytic leukemia (ALL) as well as acute myeloid leukemia (AML) were quantified in 43 and 40 bone marrow samples of ALL and AML patients in comparison with cancer-free subjects via real-time quantitative PCR (RT-qPCR). The receiver-operating-characteristic (ROC) analysis of miRNAs was performed in the study groups. Further, the correlation between the DNMT3A and TSGs was calculated. RESULTS: Significant differences were detected in the bone marrow expression of miRNAs and TSGs (P < 0.05) between acute leukemia patients and healthy group. ROC analysis confirmed the ability of miR-30a, miR-101, miR-132, miR-129, miR-124, and miR-143 to discriminate both ALL and AML patients with an area under the ROC curve of ≥ 0.80 (P < 0.001) and high accuracy. The correlation between DNMT3A and P15/P16 TSGs revealed that DNMT3A plays a vital role in epigenetic control of TSGs expression. Our findings indicated that the downregulation of bone marrow miRNAs and TSGs was accompanied by acute leukemia development. CONCLUSIONS: The authors conclude that this study could contribute to introducing useful biomarkers for acute leukemia diagnosis.
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Leucemia Mieloide Aguda , MicroARNs , Leucemia-Linfoma Linfoblástico de Células Precursoras , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Médula Ósea/patología , Detección Precoz del Cáncer , Genes Supresores de Tumor , Humanos , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , MicroARNs/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , PronósticoRESUMEN
Nanotechnology has taken novel approaches to advance cancer therapeutic efficacy employing multifunctional nanocarriers with drug delivery and imaging function. In this work, we designed a biocompatible and affordable fluorescence nanocarrier called chitosan (CS)-carbon dot (CD) hybrid nanogel for cell imaging. The green CDs were synthesized using tomato juice through a simple single-step hydrothermal method. Chitosan polymer was used as a carrier for co-delivery CDs and the anti-cancer drug with low solubility, silibinin (Sil), to design the Sil-chitosan carbon dots hybrid nanogels (CCHNs) system. After optimizing the physicochemical properties of nanostructure by DLS, FTIR, FESEM, TEM, and UV-visible techniques, the successful uptake of the fluorescent nanoparticle conjugates into MCF-7 breast cancer cells occurred. Then we embedded CDs in chitosan nanogel. The resultant CCHNs demonstrated optical properties similar to free-CDs, a desirable size distribution (55.22 nm) with a positive surface charge, a suitable loading capacity for Sil (35%), and drug release vulnerable to pH changes. The fluorescent nanocarrier could transfer Sil to MCF-7 cancer cells without remarkable toxicity. The results of the fluorescent microscope indicated that after 4 h, the solid fluorescent signal was received from cells containing CCHNs compared to free CDs and confirmed the ability of hybrid nanogels to high cellular uptake. This study demonstrates a multifunctional nanocarrier containing therapeutic compounds and fluorescent agents that provide cellular imaging to enhance therapeutic efficacy.
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Nanopartículas , Neoplasias , Preparaciones Farmacéuticas , Fotoquimioterapia , Carbono , Liberación de Fármacos , Colorantes Fluorescentes , Humanos , Nanopartículas/química , Fotoquimioterapia/métodosRESUMEN
Aptamer-Carbon Dot (CD) bioconjugation is an attractive target-tracking strategy in detecting cell surface antigens. This study describes an effective imaging paradigm for CA-125 antigen imaging. Our experience encompasses green CD synthesis and characterization, CD-capture probe conjugation through covalent bonding, the hybridization linkage of CD-probe to aptamer and their coupling confirmation, and fluorescent targeted imaging of ovarian cancer cells. As a result, the synthesized CDs from lemon extract by hydrothermal reaction show average size of 2 nm with maximum fluorescence intensity at excitation/emission 360/450 nm. CD-probe construction was provided by functional group interactions of CD and probe via EDC/NHS chemistry. The linkage of CD-probe to aptamer was conducted by Watson-Crick nucleotide pairing. The assessment of CD-probe and CD-probe-aptamer fabrication was validated by the increase in surface roughness through AFM analysis, the diminish of fluorescence intensity of CD after bioconjugation, and particle size growth of the construct. Conjugates with negligible cytotoxicity, appropriate zeta potential, and good aptamer release were applied in cellular imaging. This targeted diagnosis method was employed the four reported DNA aptamers toward fluorescence intensity. The DOV-3 aptamer showed more qualified detection over other aptamer conjugates during fluorescent microscopy analysis. In conclusion, the CD-probe-aptamer conjugate applications as toxic-free method can open new horizons in fluorescent nano-imaging in the field of targeted cancer cell diagnosis.
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Aptámeros de Nucleótidos , Neoplasias Ováricas , Aptámeros de Nucleótidos/química , Antígeno Ca-125 , Carbono/química , Colorantes Fluorescentes/química , Humanos , Imagen Óptica/métodos , Neoplasias Ováricas/diagnóstico por imagenRESUMEN
Fluorescence resonance energy transfer (FRET)-based biosensors are effective analytical tools extensively used in fields of biomedicine, pharmacology, toxicology, and food sciences. Ratiometric imaging of substantial cellular processes, molecular components, and biological interactions is widely performed by these biosensors. A variety of FRET-based biosensors have provided comprehensive insights into underlying mechanisms of pathological conditions in live cells, tissues, and organisms. Moreover, integration of FRET-based biosensors with the current bioanalytical techniques allows for accurate, rapid, and sensitive diagnosis and proposes the advanced strategies for treatment. Precise analysis of ligand-receptor interactions by FRET-based biosensors has presented a basis for determination of novel therapeutic agents. Therefore, this study was designed to review the recent developments in FRET-based biosensors and their biomedical applications. In addition, characteristics, challenges, and outlooks of these biosensors were discussed.
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Investigación Biomédica , Técnicas Biosensibles , Transferencia Resonante de Energía de Fluorescencia , Colorantes Fluorescentes/química , HumanosRESUMEN
The increasing prevalence of cancer has led to expanding traditional medicine objectives for developing novel drug delivery systems. A wide range of plant-derived polyphenol bioactive substances have been investigated in order to explore the anti-cancer effects of these natural compounds and to promote the effective treatment of cancer through apoptosis induction. In this regard, plant-derived polyphenol compounds, including curcumin, silibinin, quercetin, and resveratrol, have been the subject of intense interest for anti-cancer applications due to their ability to regulate apoptotic genes. However, some limitations of pure polyphenol compounds, such as poor bioavailability, short-term stability, low-cellular uptake, and insufficient solubility, have restricted their efficiency. Nanoscale formulations of bioactive agents have provided a novel platform to address these limitations. This paper reviews recent advances in nanoformulation approaches of polyphenolic drugs and their effects on improving the delivery of chemotherapy agents to cancer cells.
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Curcumina , Nanopartículas , Neoplasias , Preparaciones Farmacéuticas , Curcumina/farmacología , Sistemas de Liberación de Medicamentos , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Polifenoles , TranscriptomaRESUMEN
DNA-protein bioconjugation is an appealing target-tracking strategy. The new capability of DNA molecule as a biological nanomaterial endows unique fluorescence and physicochemical properties to be applied in bioimaging. Progression in targeted imaging is contingent on the conjugation of diagnostic nanoparticles to biomolecular signatures, particularly antibody-based ligands. Here, we have reported our recent experience, DNA-dot synthesis and characterization, the covalent conjugation of DNA-dot to goat F(ab')2 IgG and Epidermal Growth Factor Receptor (EGFR) antibodies, DNA-dot@antibody coupling confirmation, and fluorescent targeted imaging of lung cancer cell line. As a result, the average size of DNA-dot was 4.5-5 nm which was conjugated to amine-rich antibodies with returned PO4-1 groups on the DNA-dot surface via PN bond. The synthetic DNA-dots were conjugated to the goat F(ab')2 IgG and tested for fluorescent detection usability by indirect Dot-blot assay. Also, DNA-dot@EGFR conjugates identified lung cancer cells with 84-92% specificity and 100% sensitivity in five concentrations, associated with 0.0025 to 0.04 g 100 µL-1 DNA-dot. The results demonstrated that bioconjugated DNA-dot can do the diagnosis profiling of molecular biomarkers. Generally, DNA-dot bioconjugation with antibody is implemented within two days and biomarker detection takes one day. Consequently, DNA-dot@antibody is potentially a toxic-free, swift, and efficient method of antibody labeling that opens up new horizons in fluorescent nanoimaging in the field of cancer cell detection.
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Anticuerpos/química , ADN/química , Neoplasias/diagnóstico , Imagen Óptica/métodos , Puntos Cuánticos/química , Biomarcadores/análisis , Estudios de Factibilidad , Humanos , Técnicas In Vitro , Neoplasias/químicaRESUMEN
The hydrothermal treatment of green carbon dots (CDs) is an appropriate fluorescent probe synthesis method. CDs are exploited as biological staining agents, especially for cellular detection and imaging. The nitrogen-doped green carbon dots (N-CDs) formation can improve the fluorescence intensity property in a one-step process. Here, we report two N-CDs from lemon and tomato extraction in the presence of hydroxylamine. Lemon and tomato N-CDs showed the blue fluorescence under ultraviolet radiation of about 360 nm. The characterization of CDs and N-CDs showed the presence of N-H and C-N bonds which enhanced the fluorescence efficiency. The mean size of lemon and tomato N-CDs were about 2 and 3 nm with an increased quantum yield (QY) of 5% and 3.38%, respectively. The CDs and N-CDs cytotoxicity assay exhibited high cell viability approximately 85% and 73%, respectively. N-CDs show superior fluorescent intensity in different solvents and significant stability under long-time UV irradiation, different PH and high ionic strength. Our results indicated that the use of N-CDs in cell imaging can lead to fluorescence intensity enhancement as well as proper biocompatibility. Therefore, the safe and high fluorescence intensity of green N-CDs can be utilized for fluorescent probes in biolabeling and bioimaging applications.
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Carbono/química , Fluorescencia , Nanopartículas/química , Colorantes Fluorescentes/química , Células HeLa , Humanos , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Microscopía Fluorescente , Nitrógeno/química , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Fluorescence imaging via carbon dots (CDs) has found multifarious applications in the biomedical sciences including biosensing, cancer cell bioimaging, drug delivery and tracking therapeutic response. Presently, the latest generation of fluorescence CDs known as green-CDs has attracted ever-increasing attention due to the use of natural sources, low-cost synthesis, nanoscale size, promising biocompatibility, superior photoluminescence, and ease of functionalization for versatile applications, which in turn could have higher priority over the traditional toxic fluorescent agents. In this review, we aim to have a new insight into the engineering green-CDs and their physicochemical properties. Moreover, we discuss the possible applications of green-CDs in self and active targeting, therapeutics delivery, and finally their promising future in cancer theranostics.
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Carbono/análisis , Colorantes Fluorescentes/análisis , Neoplasias/diagnóstico , Neoplasias/terapia , Puntos Cuánticos/análisis , Nanomedicina Teranóstica/métodos , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/uso terapéutico , Portadores de Fármacos/análisis , Sistemas de Liberación de Medicamentos/métodos , Técnicas de Transferencia de Gen , Humanos , Imagen Óptica/métodosRESUMEN
DNA molecule engineering has become an attractive discipline in various research scopes. The profound influence of selective and sensitive sensing of DNA molecules in disease diagnosis and molecular imaging is established. In this perspective, we try to shed light on the state-of-the-art technology of DNA bioconjugation assays in DNA biosensor, DNA barcode, DNA nanostructures, and DNA ultra-resolution fluorescence imaging. Non-invasive, simple, and swift biotechniques benefit molecular diagnosis, evaluation of disease stages, and also play a central role in fundamental researches. We discuss the limitations of traditional procedures and the eminence impacts of the advanced methods with clinical applications in timely detection and management of diseases like cancer, genetic disorders, and recognition of microbial pathogens. The predictable and programmable DNA strands have paved the way for cellular and molecular imaging with the ability of single-molecule switching nanoscopy. Consequently, the DNA conjugation tool as an identification paradigm of biological agents in interaction with bio-spesific components is at the heart of biological processes.
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Técnicas Biosensibles , ADN/química , Imagen Molecular/métodos , Nanotecnología , Investigación Biomédica/tendencias , Humanos , Nanoestructuras/químicaRESUMEN
Streptokinase is widely used as an anticoagulant drug for the treatment of heart attacks. Because of antibody production against injected drug in individuals consuming streptokinase and causing allergic reactions, streptokinase treatment effects become neutral. Recombinant mutant type of streptokinase was prepared by removing of 42 amino acids from the C terminal region. ELISA plates were coated by natural and mutant streptokinase as antigen. Ninety-six normal serum samples as well as 27 streptokinase consumer serum samples (patients with acute myocardial infraction) were analyzed. The results showed that serum antibodies against natural streptokinase were three times more than those against the mutated streptokinase. In case of preserving thrombolytic activity, mutated streptokinase can be used as an alternative of the natural form.