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Polysaccharides, complex carbohydrates composed of long chains of residues of sugar molecules, have garnered significant attention in recent years due to their diverse applications across various industries [...].
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Polisacáridos , Polisacáridos/química , Humanos , Materiales Biocompatibles/químicaRESUMEN
Hypoxia is a significant feature of solid tumors and frequently poses a challenge to the effectiveness of tumor-targeted chemotherapeutics, thereby limiting their anticancer activity. Hypoxia-activated prodrugs represent a class of bio-reductive agents that can be selectively activated in hypoxic compartments to unleash the toxic warhead and thus, eliminate malignant tumor cells. However, their applicability can be further elevated by installing fluorescent modalities to yield hypoxia-activated theragnostic prodrugs (HATPs), which can be utilized for the simultaneous visualization and treatment of hypoxic tumor cells. The scope of this review is to summarize noteworthy advances in recent HATPs, highlight the challenges and opportunities for their further development, and discuss their potency to serve as personalized medicines in the future.
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Lipid droplets (LDs) act as an energy reservoir in cancer cells; on the other hand, mitochondria are hyperactive to fulfill the energy demand to accelerate cell proliferation. We are interested in unfolding the relationship between the cellular energy reservoir and energy producer through fluorescence labeling. Thus, a dual organelle-targeted fluorescent probe MLD-1 has been rationally developed. It visualized the crosstalk between mitochondrial dysfunction and the fluctuation of LDs in live cells. Its two-photon ability allowed us to acquire deep tissue images. For the first time, we have shown that the probe has the ability to track the accumulation of LDs in different mouse organs during pancreatic inflammation. MLD-1, being a selectively polarity-driven, chemo- and photostable LD probe, may offer great possibilities for studying LD-associated biology in due course.
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Colorantes Fluorescentes , Pancreatitis , Animales , Ratones , Colorantes Fluorescentes/metabolismo , Gotas Lipídicas/metabolismo , Enfermedad Aguda , Pancreatitis/metabolismo , MitocondriasRESUMEN
The redox regulator glutathione (GSH) migrates to the nucleus to give a safeguard to DNA replication in the S-phase. The fluctuation of GSH dynamics in the cell cycle process may help to understand cancerogenesis or other abnormalities in DNA replication. For the first time, we attempted to track the time-dependent S-phase change using the newly developed ratiometric fluorescent probe Nu-GSH. This probe is highly chemoselective towards glutathione and shows an emission intensity shift from 515 nm to 455 nm. It has shown fluorescence reversibility from blue to green channels while scavenging reactive oxygen species H2O2. Both ratiometric fluorescence images and FACS analysis have provided quantitative information on the GSH levels in the nucleoli during DNA replication in the S-phase. Furthermore, GSH fluctuation reciprocated the decay of the S-phase on a time scale. Additionally, its two-photon ability guaranteed its capability to study GSH dynamics in live cells/tissues noninvasively. We envision that the probe Nu-GSH can be used to get high-throughput quantitative information on glutathione dynamics and give an opportunity to monitor its perturbation during the course of cell division.
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Colorantes Fluorescentes , Peróxido de Hidrógeno , Humanos , Células HeLa , Replicación del ADN , Glutatión/metabolismoRESUMEN
A ß-galactosidase activatable fluorescent turn-on theranostic Gal-CGem exhibits gemcitabine release specifically in ß-galactosidase overexpressing hepatic carcinoma cells. The cytotoxicity of Gal-CGem in cancer cells is achieved through the apoptotic cell death pathway. Overall, Gal-CGem is a new frontline prodrug in cancer therapy that has provided antineoplastic information through fluorescence imaging.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Carcinoma Hepatocelular/diagnóstico por imagen , Carcinoma Hepatocelular/tratamiento farmacológico , Colorantes Fluorescentes/metabolismo , Colorantes Fluorescentes/farmacología , Humanos , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/tratamiento farmacológico , Imagen Óptica/métodos , Medicina de Precisión , Nanomedicina Teranóstica/métodos , beta-Galactosidasa/metabolismoRESUMEN
The flavin adenine dinucleotide (FAD) is an indispensable coenzyme in live cells. It acts as a catalyst in many redox responsive metabolic reactions, including oxidative phosphorylation in mitochondria. The real-time monitoring of flavin is important to understand the disorder in the metabolic process, redox system, etc. Thus, we have developed a fluorescent probe CPy-1 that noncovalently binds with flavin to exhibit the FRET process. 1H- NMR and docking study indicated that there is a strong hydrophobic interaction between flavins and CPy-1. Also, a π-π stacking between isoalloxazine ring in flavin and quinoline and coumarin moieties of CPy-1 favors self-assembly. The nontoxic probe CPy-1 could distinguish cancer cells from normal cells based on expressions of endogenous FAD.
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Flavina-Adenina Dinucleótido , Colorantes Fluorescentes , Dinitrocresoles , Mononucleótido de Flavina , Flavina-Adenina Dinucleótido/química , Flavinas/química , Flavinas/metabolismo , Transferencia Resonante de Energía de FluorescenciaRESUMEN
The endogenous H2S-driven theranostic H2S-Gem has been invented. The theranostic prodrug H2S-Gem is selectively activated in cancer cells, releasing active gemcitabine with a simultaneous fluorescence turn-on. H2S-Gem selectively inhibited cancer cell growth compared to the mother chemotherapeutic gemcitabine. Overall, it is a unique protocol for tracking and transporting chemotherapeutic agents to tumor areas without the guidance of tumor-directive ligands.
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Antimetabolitos Antineoplásicos/farmacología , Desoxicitidina/análogos & derivados , Sulfuro de Hidrógeno/farmacología , Profármacos/farmacología , Antimetabolitos Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Desoxicitidina/química , Desoxicitidina/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Fluorescencia , Células HeLa , Humanos , Sulfuro de Hidrógeno/química , Ligandos , Profármacos/química , Nanomedicina Teranóstica , GemcitabinaRESUMEN
Biodegradable polymeric microparticles are widely used in drug delivery systems with prolonged-release profiles and/or cell microcarriers. Their fabrication via the oil/water emulsion solvent evaporation technique has normally required emulsifiers in the aqueous phase. The present work aims to evaluate the effectiveness of various polysaccharides, such as chitosan, hyaluronic acid, cellulose, arabinogalactan, guar and their derivatives, as an alternative to synthetic surfactants for polylactide microparticle stabilization during their fabrication. Targeted modification of the biopolymer's chemical structure was also tested as a tool to enhance polysaccharides' emulsifying ability. The transformation of biomacromolecules into a form of nanoparticle via bottom-up or top-down methods and their subsequent application for microparticle fabrication via the Pickering emulsion solvent evaporation technique was useful as a one-step approach towards the preparation of core/shell microparticles. The effect of polysaccharides' chemical structure and the form of their application on the polylactide microparticles' total yield, size distribution and morphology was evaluated. The application of polysaccharides has great potential in terms of the development of green chemistry and the biocompatibility of the formed microparticles, which is especially important in biomedicine application.
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We have synthesized new magnetic resonance imaging (MRI) T1 contrast agents (CA1 and CA2) that permit the activatable recognition of the cellular vicinal thiol motifs of the protein thioredoxin. The contrast agents showed MR relaxivities typical of gadolinium complexes with a single water molecule coordinated to a Gd3+ center (i.e., ~4.54 mM-1s-1) for both CA1 and CA2 at 60 MHz. The contrast agent CA1 showed a ~140% relaxivity enhancement in the presence of thioredoxin, a finding attributed to a reduction in the flexibility of the molecule after binding to thioredoxin. Support for this rationale, as opposed to one based on preferential binding, came from 1H-15N-HSQC NMR spectral studies; these revealed that the binding affinities toward thioredoxin were almost the same for both CA1 and CA2. In the case of CA1, T1-weighted phantom images of cancer cells (MCF-7, A549) could be generated based on the expression of thioredoxin. We further confirmed thioredoxin expression-dependent changes in the T1-weighted contrast via knockdown of the expression of the thioredoxin using siRNA-transfected MCF-7 cells. The nontoxic nature of CA1, coupled with its relaxivity features, leads us to suggest that it constitutes a first-in-class MRI T1 contrast agent that allows for the facile and noninvasive monitoring of vicinal thiol protein motif expression in live cells.
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Rastreo Celular/métodos , Medios de Contraste , Espectroscopía de Resonancia Magnética , Compuestos de Sulfhidrilo , Tiorredoxinas , Línea Celular Tumoral , Medios de Contraste/síntesis química , Medios de Contraste/química , Humanos , Fantasmas de Imagen , Compuestos de Sulfhidrilo/metabolismo , Tiorredoxinas/metabolismoRESUMEN
Urgency in finding a suitable therapy in tumor hypoxia strives to develop hypoxia-targeted activatable theranostic. A strategic theranostic prodrug (Azo-M) has been synthesized. Its azo-linker scission under the hypoxia condition has released an near-infrared (NIR)-reporter to determine the extent of chemotherapeutic (melphalan analogue) activation. Under an artificial hypoxia condition, a large shift from 520 to 590 nm in UV absorption was observed in Azo-M. Alongside, the emission maxima had appeared at 625 nm under the said condition. The Azo-M post-incubated HeLa cells have shown upregulation of various apoptotic factors under oxygen deprivation (3%) condition. Azo-M has shown antiproliferative activity under hypoxia conditions in various cancer cells. An ex-vivo biodistribution study indicated that theranostic Azo-M only activated in tumor tissue and to some extent in the liver. The therapeutic activity study in vivo indicated that Azo-M effectively reduced the tumor size and volume (about 2-fold) without the change of bodyweight of mice. The theranostic Azo-M can be a cornerstone to suppress tumor hypoxia and tracking its extent of suppression.
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Hipoxia , Nanomedicina Teranóstica , Animales , Línea Celular Tumoral , Células HeLa , Humanos , Ratones , Distribución TisularRESUMEN
We have developed an amphiphilic pH probe (P1CS) to detect pH levels in the plasma membrane in cancer cells. An elevated fluorescence signal at 550 nm at the cell surface of cancer cells (MDA-MB-231, HeLa cells) prompted the application of P1CS as a pH marker for the cancer cell surface, discriminating it from normal cells (WI-38). Moreover, the probe enables labeling of the surface of multilayered tumor spheroids, which promotes its use as a marker for the surface of tumor tissue.
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Membrana Celular/química , Fluorescencia , Colorantes Fluorescentes/análisis , Colorantes Fluorescentes/química , Neoplasias/química , Tensoactivos/análisis , Tensoactivos/química , Línea Celular , Humanos , Concentración de Iones de Hidrógeno , Estructura Molecular , Neoplasias/patologíaRESUMEN
Alkaline phosphatase (ALP) is an enzyme that actively plays a significant role in the various metabolic processes by transferring a phosphate group to the protein, nucleic acid, etc. The elevated level of ALP in blood plasma is the hallmark of inflammation/cancer. The hyperactive mitochondria in cancer cells produce an excess of ATP to fulfill the high energy demand. Thus, we have developed a fluorescent probe Mito-Phos for ALP, which can detect phosphatase expression in mitochondria in live cells. The probe Mito-Phos has shown ~15-fold fluorescence intensity increments at 450 nm in the presence of 500 ng/mL of ALP. It takes about 60 min to consume the whole amount of ALP (500 ng/mL) in physiological buffer saline. It can selectively react with ALP even in the presence of other probable cellular reactive components. It is highly biocompatible and nontoxic to the live cells. It has shown ALP expression in a dose-dependent manner by providing concomitant fluorescence images in the blue-channel region. It has localized exclusively in the mitochondria in live cells. The probe Mito-Phos is highly biocompatible with the ability to assess ALP expression in mitochondria in live cells.
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Fosfatasa Alcalina/metabolismo , Materiales Biocompatibles/química , Colorantes Fluorescentes/química , Mitocondrias/enzimología , Fosfatasa Alcalina/química , Supervivencia Celular , Regulación Enzimológica de la Expresión Génica , Células HeLa , Humanos , CinéticaRESUMEN
The 1,4-dihydronicotinamide adenine dinucleotide (NADH) is one of the key coenzymes that participates in various metabolic processes including maintaining the redox balance. Early information on the imbalance of NADH is crucial in the context of diagnosing the pathogenic conditions. Thus, a dual-channel fluorescent probe (MQN) is developed for tracking of NADH/NAD(P)H in live cells. In the presence of NADH, only it showed emission signals at 460 and 550 nm upon excitation at 390 and 450 nm, respectively. The probe could provide accurate information on NADH levels in cancer cells (HeLa) and normal cells (WI-38). We observed that the NADH level in cancer cells (HeLa) is relatively higher than that in normal WI-38 cells. We received similar information on NADH upon calibrating with a commercial NADH kit. Moreover, we evaluated substrate-specific NADH expression in the glycolysis pathway and oxidative phosphorylation process. Also, the dual-channel probe MQN has visualized NADH manipulation in the course of depletion of GSH to maintain cellular redox balance. This dual-channel molecular probe MQN comes out as a new detection tool for NADH levels in live cells and tumor mimic spheroids.
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Color , Colorantes Fluorescentes/química , NAD/metabolismo , Esferoides Celulares/metabolismo , Línea Celular , Células HeLa , Humanos , NAD/química , Esferoides Celulares/químicaRESUMEN
In modern society, the use of cosmetics has increased extensively; unfortunately, so-called several toxic metal salts are present as the colorant or filler in cosmetics. The ferrous ion (Fe2+) is one of the metal ions used in cosmetics as a colorant. Ferrous ion (Fe2+) is a vital component in live cells. Considering the adverse effect of high doses of ferrous ions in cosmetics and live cells, we developed a turn-on fluorescent probe PFe(II) for quantitative estimation of ferrous ion (Fe2+) in cosmetics and monitoring of labile ferrous (Fe2+) ion in live cells. The fluorescent probe PFe(II) showed a visual color change from colorless to orange in the presence of ferrous ion (Fe2+) in the cosmetics. We observed that UV-absorption increased at 390 nm upon incubation with ferrous ion (Fe2+). The probe PFe(II) has provided quantitative information on ferrous ion (Fe2+) in various cosmetics, kajol, lip balm, face foundation, mascara, eyeliner, lipliner, face makeup, sindoor, lipstick, nail polish in ppm level through the fluorescence signaling at 460 nm.The probe PFe(II) provided information on labile Fe2+ ion pool via a fluorescence imaging. It is a new addition to the diagnostic inventory for detecting ferrous ion in live cells and cosmetics.
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Cosméticos/análisis , Compuestos Ferrosos/análisis , Colorantes Fluorescentes/química , Células HeLa , Humanos , Límite de Detección , Análisis Espectral/métodosRESUMEN
The redox-regulator glutathione (GSH) maintains a specific redox potential to sustain routine cellular activity from oxidative damage. In the early stage of the cell cycle process, the glutathione levels increase in the nuclei for protecting the DNA replication process from reactive oxygen species (ROS). In the first attempt, we developed a new ratiometric fluorescent probe that has provided information about glutathione levels in the nuclei. The UV-vis. absorption of probe GScp has shown a hypsochromic shift from 410 to 350 nm in the presence of GSH. In fluorescence titration, we observed that fluorescence emission of the GScp switched from 510 to 460 nm in the presence of GSH. The self-calibrated probe GScp has shown nearly optimal reversibility in GSH redox dynamics with the dissociation constant 2.47 mM. The probe is ideal for GSH tracking in live cells, as its toxicity has within the safe zone. The probe GScp has validated GSH levels in nucleoli by providing fluorescence images in blue-channel. This finding inspires us to use for validation of GSH dynamics in the nucleoli in the cell cycle process.
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Nucléolo Celular/metabolismo , Colorantes Fluorescentes/química , Glutatión/análisis , Glutatión/metabolismo , Animales , Línea Celular Tumoral , Cumarinas/síntesis química , Cumarinas/química , Colorantes Fluorescentes/síntesis química , Humanos , Límite de Detección , Ratones , Microscopía Confocal , Microscopía FluorescenteRESUMEN
Graft copolymers of chitosan with cellulose ether have been obtained by the solid-state reactive mixing of chitin, sodium hydroxide and hydroxyethyl cellulose under shear deformation in a pilot twin-screw extruder. The structure and composition of the products were determined by elemental analysis and IR spectroscopy. The physicochemical properties of aqueous solutions of copolymers were studied as a function of the composition, and were correlated to the mechanical characteristics of the resulting films to assess the performance of new copolymers as coating materials, non-woven fibrous materials or emulsifiers for interface stabilization during the microparticle fabrication process.
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Abnormal anaerobic metabolism leads to a lowering of the pH of many tumours, both within specific intracellular organelles and in the surrounding extracellular regions. Information relating to pH-fluctuations in cells and tissues could aid in the identification of neoplastic lesions and in understanding the determinants of carcinogenesis. Here we report an amphiphilic fluorescent pH probe (CS-1) that, as a result of its temporal motion, provides pH-related information in cancer cell membranes and selected intracellular organelles without the need for specific tumour targeting. Time-dependent cell imaging studies reveal that CS-1 localizes within the cancer cell-membrane about 20 min post-incubation. This is followed by migration to the lysosomes at 30 min before being taken up in the mitochondria after about 60 min. Probe CS-1 can selectively label cancer cells and 3D cancer spheroids and be readily observed using the green fluorescence channel (λ em = 532 nm). In contrast, CS-1 only labels normal cells marginally, with relatively low Pearson's correlation coefficients being found when co-incubated with standard intracellular organelle probes. Both in vivo and ex vivo experiments provide support for the suggestion that CS-1 acts as a fluorescent label for the periphery of tumours, an effect ascribed to proton-induced aggregation. A much lower response is seen for muscle and liver. Based on the present results, we propose that sensors such as CS-1 may have a role to play in the clinical and pathological detection of tumour tissues or serve as guiding aids for surgery.
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Concurrently, manipulation of mitochondrial activity and its monitoring have enormous significance in cancer therapy and diagnosis. In this context, a fluorescent probe MitoDP has been developed for validating H2S mediated protonophore (2,4-dinitrophenol, DNP) induced mitochondrial membrane potential change, ROS formation and ATP depletion in cancer cells. The extent of protonophore activation for mitochondrial dysfunction is monitored through fluorescence signalling at 450 nm. The current study provides a proof for the concept of endogenous H2S-mediated controlled and spatial release of bioactive agents, or toxins specifically in mitochondria of cancer cells.
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2,4-Dinitrofenol/farmacología , Colorantes Fluorescentes/farmacología , Sulfuro de Hidrógeno/farmacología , Mitocondrias/efectos de los fármacos , 2,4-Dinitrofenol/química , Células 3T3 , Animales , Proliferación Celular/efectos de los fármacos , Colorantes Fluorescentes/química , Células HCT116 , Células HeLa , Humanos , Sulfuro de Hidrógeno/química , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones , Mitocondrias/metabolismo , Estructura Molecular , Imagen Óptica , Especies Reactivas de Oxígeno/metabolismo , Espectrometría de FluorescenciaRESUMEN
We have developed a FRET-based fluorescent probe (PHS1) as a combination of two different fluorophores (coumarin and naphthalimide); which can detect both exogenous and endogenous H2S and H2O2 in live cells through multicolor images. The precise overlap between UV-absorption of naphthalimide and the emission band of coumarin in probe PHS1 allows the acquisition of the self-calibrated information of dual analytes through FRET-based imaging. The UV-Vis absorption (λabs 390â¯nm) and fluorescence emission (λem 460â¯nm) of probe PHS1 in the presence of H2O2 are increased â½35- fold and â½15-fold respectively. It also allows the estimation of the levels of H2S through enhancement of emission intensity at 550â¯nm. The probe PHS1 exhibits high stability against various analytes, including various pH (4-9.5). The cell viability assay data indicate that the probe is not harmful to the cancer cells. The nontoxic nature of the probe PHS1 encourages application for cancer cell labeling. The probe PHS1 can detect the level of endogenous H2O2, H2S, and H2O2/H2S in cancer cells through blue, green and FRET-based green channel imaging. PHS1 is a unique probe, has potential application for diagnosing cancer by providing information on the level of dual analytes (H2S, H2O2) in cancer cells.
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Transferencia Resonante de Energía de Fluorescencia/métodos , Colorantes Fluorescentes/química , Peróxido de Hidrógeno/análisis , Sulfuro de Hidrógeno/análisis , Imagen Molecular/métodos , Línea Celular Tumoral , Supervivencia Celular , Color , Cumarinas , Humanos , Naftalimidas , Neoplasias/diagnóstico por imagenRESUMEN
Herein, we report a rhodamine-based redox probe (MQR) to visualize cytosolic NADH in the cellular milieu. Its high sensitivity and selectivity allowed it to track the alteration of the NADH level under metabolic perturbation, suggesting its potential as a useful tool to study the association between the NADH level and metabolic abnormalities with clinical significance.