Concentrations Of Doxorubicin Research Articles

Polysaccharide based supramolecular injectable hydrogels for in situ treatment of bladder cancer

Implantable system maximizes drug concentration and continuously releases drugs near the tumor, which is an effective tool to solve the difficult retention of chemotherapy drugs in bladder cancer. In this work, a novel polysaccharide supramolecular injectable hydrogel (CCA hydrogels for short) is rapidly constructed by simply mixing cationic chitosan, anionic sulfobutyl ether β-cyclodextrin (SBE-β-CD) and a trace amount of silver ions. The injected hydrogel reconstituted and regained its shape in less than 1 h, and it can still maintain the elasticity suitable for the human body. By packaging the drug directly, the gel achieves a high concentration of doxorubicin, an anticancer drug. Using MB49-luc cells as the model of bladder tumor for anti-tumor in vivo, the CCA-DOX gel has obvious inhibitory effect on bladder tumor, and its inhibitory effect is much greater than that of free DOX. Therefore, this self-healing injectable hydrogel has great potential for in situ treatment of bladder cancer.

Occupational Exposure during Intraperitoneal Pressurized Aerosol Chemotherapy Using Doxorubicin in a Pig Model

BackgroundThis study evaluated occupational exposure levels of doxorubicin in healthcare workers performing rotational intraperitoneal pressurized aerosol chemotherapy (PIPAC) procedures. MethodsAll samples were collected during PIPAC procedures applying doxorubicin to an experimental animal model (pigs). All procedures were applied to seven pigs, each for approximately 44 min. Surface samples (n = 51) were obtained from substances contaminating the PIPAC devices, surrounding objects, and protective equipment. Airborne samples were also collected around the operating table (n = 39). All samples were analyzed using ultra-high performance liquid chromatography-mass spectrometry. ResultsAmong the surface samples, doxorubicin was detected in only five samples (9.8%) that were directly exposed to antineoplastic drug aerosols in the abdominal cavity originating from PIPAC devices. The telescopes showed concentrations of 0.48–5.44 ng/cm2 and the trocar showed 0.98 ng/cm2 in the region where the spraying nozzles were inserted. The syringe line connector showed a maximum concentration of 181.07 ng/cm2, following a leakage. Contamination was not detected on the surgeons' gloves or shoes. Objects surrounding the operating table, including tables, operating lights, entrance doors, and trocar holders, were found to be uncontaminated. All air samples collected at locations where healthcare workers performed procedures were found to be uncontaminated. ConclusionsMost air and surface samples were uncontaminated or showed very low doxorubicin concentrations during PIPAC procedures. However, there remains a potential for leakage, in which case dermal exposure may occur. Safety protocols related to leakage accidents, selection of appropriate protective equipment, and the use of disposable devices are necessary to prevent occupational exposure.

Abstract 2527: Investigation of doxorubicin interference in flow cytometry measurements of apoptosis and necrosis

Abstract Background: Doxorubicin (DOX) is frequently used for investigations of breast cancer using flow cytometry with fluorescent stains for apoptosis and necrosis. DOX interference, however, can occur with these fluorescent measurements which is not reported in many published reports. The objectives of this study were to identify such interference and to assess its extent. Methods: MDA-MB-231 and MCF-7 cells were incubated for 24 h with 0.1, 1, and 10 µM DOX. Cells were then stained for apoptosis and necrosis using two commercial assay kits. Fluorescence was measured with FL1 (apoptosis), FL2 (primary for DOX), and FL3 (necrosis) detectors in cells with DOX alone, with DOX plus stains, and in control cells without DOX. Measurements were repeated two to five times using replicates and trials. Statistics were evaluated by t-tests. Results: Overall, DOX fluorescence interference increased with concentration in both cell lines. After DOX incubation, cells stained with caspase 3/7 and SYTOX showed fluorescence in the FL1 detector indicating caspase 3/7 activation in apoptosis. But the DOX component of fluorescence with this detector ranged from 11 to 95%. Dox fluorescence in the FL3 detector for SYTOX measurements indicating necrosis ranged from 26 to 96% of total fluorescence. Cells stained with Annexin V (with the FITC label) and propidium iodide (PI) showed fluorescence in the FL1 detector for Annexin V indicating apoptosis. However, the DOX component of fluorescence ranged from 11 to 25%. DOX fluorescence in the FL3 detector for PI indicating necrosis ranged from 87 to 106%. Conclusions: Without correction for DOX interference, and even at low concentrations, substantial errors can occur for apoptosis and necrosis measurements using flow cytometry with these stains in breast cancer cells. At high DOX concentrations, the interference can erroneously indicate an extensive presence of these cell death processes. Corrections for DOX interference include background subtraction and flow cytometer compensation. Acknowledgments: The authors gratefully acknowledge financial support from the Agnes Varis Trust for Women in Science (REN), the Milton Lev Memorial Faculty Research Fund (CMO) as well as Saint Joseph’s University support for student research. The authors also thank Bin Chen, PhD, and Richard Howley for assistance with flow cytometry. Citation Format: Rachel E. Nicoletto, Richard Wilkes, Clyde M. Ofner. Investigation of doxorubicin interference in flow cytometry measurements of apoptosis and necrosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2527.

Hyperthermia Improves Doxorubicin-Based Chemotherapy by Activating Mitochondrial Apoptosis in Bladder Cancer

Purpose: Although intravesical chemotherapy has several antitumoral benefits, it can also have severe side effects. The development of novel therapeutic approaches for bladder cancer (BC) is thus warranted. Hyperthermia (HT) is a widely applicable adjuvant therapy in various cancers. Therefore, this study investigated the effect of HT on improving the chemosensitivity of BC. Materials and Methods: The BC cell lines 5637 and T24 were cultured and treated with HT (43°C) for 24 h. Then, cell viability and survival were assessed using resazurin reagent and colony formation assay, respectively. Western blot assay was used to analyze the levels of Bax, Bcl-2, cleaved caspase-3, and cleaved poly (ADP-ribose) polymerase (PARP) protein expression. Mitochondria degradation was observed by MitoTracker Green staining. Results: In BC cells, HT co-administered with various concentrations of doxorubicin significantly inhibited cell viability and survival. Moreover, HT combined with doxorubicin promoted mitochondrial apoptosis, which caused Bax upregulation and Bcl-2 downregulation. Levels of cleaved caspase-3 and PARP protein expression were also elevated after co-treatment. Conclusion: Taken together, HT improved the chemosensitivity of BC cells to doxorubicin. HT combined with chemotherapy further activated mitochondrial apoptosis in BC cells. The findings suggested that HT may serve as a potential adjunctive treatment for BC that is ready to be applied clinically.

Reduced Graphene Oxide and Gold Nanoparticles-Modified Electrochemical Aptasensor for Highly Sensitive Detection of Doxorubicin

Doxorubicin (DOX) is the most clinically important antibiotic in cancer treatment, but its severe cardiotoxicity and other side effects limit its clinical use. Therefore, monitoring DOX concentrations during therapy is essential to improve efficacy and reduce adverse effects. Here, we fabricated a sensitive electrochemical aptasensor for DOX detection. The sensor used gold wire as the working electrode and was modified with reduced graphene oxide (rGO)/gold nanoparticles (AuNPs) to improve the sensitivity. An aptamer was used as the recognition element for the DOX. The 5' end of the aptamer was modified with a thiol group, and thus immobilized to the AuNPs, and the 3' end was modified with methylene blue, which acts as the electron mediator. The combination between the aptamer and DOX would produce a binding-induced conformation, which changes the electron transfer rate, yielding a current change that correlates with the concentration of DOX. The aptasensor exhibited good linearity in the DOX concentration range of 0.3 μM to 6 μM, with a detection limit of 0.1 μM. In addition, the aptasensor was used for DOX detection in real samples and results, and showed good recovery. The proposed electrochemical aptasensor will provide a sensitive, fast, simple, and reliable new platform for detecting DOX.

Polyherbal formulation conjugated to gold nanoparticles induced ferroptosis in drug-resistant breast cancer stem cells through ferritin degradation.

Aim: Due to their minimal side effects, the anti-cancer properties of the polyherbal formulation are being investigated. However, due to their low absorption potential, the administration of polyherbal formulations is restricted. Loading the polyherbal formulation into gold nanoparticles enhances the bioavailability of the polyherbal formulation (PHF) accompanied by reducing the concentration of doxorubicin (dox). Ferroptosis is one of the novel pathways that specifically target cancer stem cells due to high ferritin levels. Hence, in the present study, we conjugated polyherbal formulation with gold nanoparticles and studied its effect on inducing ferroptosis in drug-resistant breast cancer cell lines. Materials and methods: PHF and dox conjugated to gold nanoparticles were characterized using FTIR, UV-Vis spectrophotometer, DLS, particle size analyzer, and XRD. The drug entrapment and efficiency studies were performed to assess the biodegradable potential of the synthesized gold nanoparticles. Paclitaxel-resistant breast cancer stem cells were generated, and an MTT assay was performed to evaluate the cytotoxicity potential of AuNP-PHF and AuNP-dox. Scratch assay and clonogenic assay were performed to assess the migration and proliferation of the cells after treatment with chosen drug combinations. The ability of PHF and dox conjugated to gold nanoparticles to induce ferritinophagy was evaluated by RT-PCR. Finally, image analysis was performed to check apoptosis and cellular ROS using inverted fluorescent microscope. The ability to induce cell cycle arrest was assessed by cell cycle analysis using flow cytometer. Results and conclusion: PHF and dox conjugated to gold nanoparticles showed high stability and showed to induce ferritin degradation in drug resistant breast cancer stem cells through ferritin degradation. AuNP-PHF in combination with low dose of AuNP-Dox nanoconjugate could be used as an effective cancer therapeutic agent, by targeting the autophagy necroptosis axis.

Anticancer effect and safety of doxorubicin and nutraceutical sulforaphane liposomal formulation in triple-negative breast cancer (TNBC) animal model

Female breast cancer is the most deadly cancer in women worldwide. The triple-negative breast cancer subtype therapies, due to the lack of specific drug targets, are still based on systemic chemotherapy with doxorubicin, which is burdened with severe adverse effects. To enhance therapeutic success and protect against systemic toxicity, drug carriers or combination therapy are being developed. Thus, an innovative liposomal formulation containing doxorubicin and the main nutraceutical, sulforaphane, has been developed. The anticancer efficacy and safety of the proposed liposomal formulation was evaluated in vivo, in a 4T1 mouse model of triple-negative breast cancer, and the mechanism of action was determined in vitro, using triple-negative breast cancer MDA-MB-231 and non-tumorigenic breast MCF-10A cell line. The elaborated drug carriers were shown to efficiently deliver both compounds into the cancer cell and direct doxorubicin to the cell nucleus. Incorporation of sulforaphane resulted in a twofold inhibition of tumor growth and the potential of up to a fourfold reduction in doxorubicin concentration due to the synergistic interaction between the two compounds. Sulforaphane was shown to increase the accumulation of doxorubicin in the nuclei of cancer cells, accompanied by inhibition of mitosis, without affecting the reactive oxygen species status of the cell. In normal cells, an antagonistic effect resulting in less cytotoxicity was observed. In vivo results showed that sulforaphane incorporation yielded not only cardioprotective, but also nephro- and hepatoprotective effects. The results of the research revealed the prospects of applying sulforaphane as a component of liposomal doxorubicin in triple-negative breast cancer chemotherapy.

Stable and Reusable Lace-like Black Silicon Nanostructures Coated with Nanometer-Thick Gold Films for SERS-Based Sensing.

We propose a simple, fast, and low-cost method for producing Au-coated black Si-based SERS-active substrates with a proven enhancement factor of 106. Room temperature reactive ion etching of silicon wafer followed by nanometer-thin gold sputtering allows the formation of a highly developed lace-type Si surface covered with homogeneously distributed gold islands. The mosaic structure of deposited gold allows the use of Au-uncovered Si domains for Raman peak intensity normalization. The fabricated SERS substrates have prominent uniformity (with less than 6% SERS signal variations over large areas, 100 × 100 μm2). It has been found that the storage of SERS-active substrates in an ambient environment reduces the SERS signal by less than 3% in 1 month and not more than 40% in 20 months. We showed that Au-coated black Si-based SERS-active substrates can be reused after oxygen plasma cleaning and developed relevant protocols for removing covalently bonded and electrostatically attached molecules. Experiments revealed that the Raman signal of 4-MBA molecules covalently bonded to the Au coating measured after the 10th cycle was just 4 times lower than that observed for the virgin substrate. A case study of the reusability of the black Si-based substrate was conducted for the subsequent detection of 10-5 M doxorubicin, a widely used anticancer drug, after the reuse cycle. The obtained SERS spectra of doxorubicin were highly reproducible. We demonstrated that the fabricated substrate permits not only qualitative but also quantitative monitoring of analytes and is suitable for the determination of concentrations of doxorubicin in the range of 10-9-10-4 M. Reusable, stable, reliable, durable, low-cost Au-coated black Si-based SERS-active substrates are promising tools for routine laboratory research in different areas of science and healthcare.

Aptamer-based Upconversion Fluorescence Sensor for Doxorubicin Detection.

Doxorubicin is a common chemotherapeutic drug used to treat a variety of cancers. Monitoring the concentration of doxorubicin in human biological fluids is vital for treatment. In this work, we report an aptamer-functionalized, 808nm-excited core-shell upconversion fluorescence sensor for specific detection of doxorubicin (DOX). Upconversion nanoparticles and DOX are used as energy donors and energy acceptors respectively. Aptamers immobilized on the surface of upconversion nanoparticles act as the molecular recognition element for DOX. The binding of DOX to the immobilized aptamers results in the fluorescence quenching of the upconversion nanoparticles via a fluorescence resonance energy transfer process. The relative fluorescence intensity exhibits a good linear response to DOX concentration in the range of 0.5μM to 55μM with a detection limit of 0.5μM. The aptasensor displays high specificity and anti-interference against other antibiotics, common ions, and biomolecules owing to strong and specific interactions of aptamers towards DOX. The sensor is further applied for the detection of DOX in urine with spike recoveries of nearly 100%.

Bone marrow stromal cell-derived exosomes improve oxidative stress and pyroptosis in doxorubicin-induced myocardial injury in vitro by regulating the transcription of GSDMD through the PI3K-AKT-Foxo1 pathway.

Doxorubicin (DOX) can contribute to severe myocardial injury, and bone marrow stromal cells (BMSC)-exosomes (Exos) improves acute myocardial infarction. Hence, this research investigated whether BMSC-Exos alleviated DOX-induced myocardial injury. BMSC-derived Exos were isolated and identified, and the optimal concentration of DOX was confirmed. H9C2 cells were treated with DOX and BMSC-Exos or in combination with the protein kinase B (AKT) inhibitor. Reactive oxygen species (ROS) and JC-1 were detected to assess oxidative stress (OS) and mitochondrial membrane damage, respectively. In addition, the expression of pyroptosis-related molecules was measured. The expression of phosphatidylinositol 3 kinase (PI3K)-AKT pathway-related proteins and the phosphorylation and acetylation of forkhead box O1 (Foxo1) in the cell nucleus and cytoplasm were tested. Last, interactions between Foxo1 and gasdermin D (GSDMD) were assessed. BMSC-Exo treatment increased viability and mitochondrial membrane potential and reduced lactic dehydrogenase release and ROS levels in DOX-treated H9C2 cells. Furthermore, the addition of BMSC-Exos suppressed DOX-induced activation and upregulation of NLRP3 and apoptosis-associated speck-like protein containing A CARD (ASC) and in vitro cleavage of caspase-1, GSDMD, interleukin (IL)-1β, and IL-18 proteins. Additionally, BMSC-Exo treatment enhanced the expression of phosphorylated (p)-PI3K, p-AKT, and p-mTOR in DOX-treated H9C2 cells and the levels of phosphorylated Foxo1 in the cytoplasm of DOX-treated H9C2 cells. Foxo1 was enriched in the promoter region of GSDMD. Moreover, the AKT inhibitor API-2 annulled the effects of BMSC-Exos on OS, pyroptosis, and Foxo1 phosphorylation in DOX-treated H9C2 cells. BMSC-Exos phosphorylated Foxo1 and inactivated Foxo1 transcription via the PI3K-AKT pathway to diminish GSDMD expression, thus restraining DOX-induced pyroptosis and OS of myocardial cells.

CYTOMORPHOLOGICAL CHANGES IN MCF7-DOX CELLS AND THEIR CORRELATION WITH THE ACTIVITY OF THE CELL CYCLE REGULATOR P21 AFTER COMBINED EXPOSURE TO INFRARED LASER AND DOXORUBICIN

Introduction: In recent years, studies of infrared laser irradiation effect on tumor cells in combination with various chemotherapeutic agents have been developing dynamically. Objective: To study the effect of infrared laser in combination with low doses of doxorubicin on cytomorphological characteristics of MCF-7DOX culture cells and p21 expression level. Materials and Methods: MCF-7DOX tumor cells were cultured in DMEM medium with 10% fetal calf serum (FCT) and 40 μg/ml gentamicin. Doxorubicin was added to the cells to a final concentration of 2 μg/ml and 0.5 μg/ml. Cells were irradiated with infrared laser (Fotonika-Plus, Ukraine) with a wavelength of 810 nm (irradiation time — 5 min, power density — 50 mV/cm², irradiation dose 15 J/cm²). Photomicrographs of cells were taken using a Carl Zeiss microscope, Germany. An immunocytochemical study was also conducted using monoclonal antibodies to the factor p21 (Thermo Scientific, USA). Results: The evaluation of morphological characteristics in micropreparations testifies to the antitumor effectiveness of the combined effect of infrared laser irradiation and doxorubicin. It is noteworthy that the cytotoxic effects of chemotherapy in combination with photobiomodulation were observed when using different doses of doxorubicin: 2 μg/ml and 0.5 μg/ml. When studying the biological activity of cells using immunocytochemistry, it was established that the combination of doxorubicin and infrared laser irradiation led to an increase in the expression of p21, and it is important to note that a similar effect was observed at both concentrations of doxorubicin: as at 2.0 μg/ ml and 0.5 μg/ml. The simultaneous destructive effect on tumor cells and the increase in p21 expression allows us to make a preliminary cautious assumption that the proposed technique of combination of infrared laser irradiation with doxorubicin determines the effect of the p21 factor in the role of a tumor suppressor. Conclusions: The results of the study allow us to consider the use of doxorubicin in combination with infrared laser as a promising method of cytotoxic effect on tumor cells.

Doxorubicin-Loaded Polyelectrolyte Multilayer Capsules Modified with Antitumor DR5-Specific TRAIL Variant for Targeted Drug Delivery to Tumor Cells

Recently, biodegradable polyelectrolyte multilayer capsules (PMC) have been proposed for anticancer drug delivery. In many cases, microencapsulation allows to concentrate the substance locally and prolong its flow to the cells. To reduce systemic toxicity when delivering highly toxic drugs, such as doxorubicin (DOX), the development of a combined delivery system is of paramount importance. Many efforts have been made to exploit the DR5-dependent apoptosis induction for cancer treatment. However, despite having a high antitumor efficacy of the targeted tumor-specific DR5-B ligand, a DR5-specific TRAIL variant, its fast elimination from a body limits its potential use in a clinic. A combination of an antitumor effect of the DR5-B protein with DOX loaded in the capsules could allow to design a novel targeted drug delivery system. The aim of the study was to fabricate PMC loaded with a subtoxic concentration of DOX and functionalized with the DR5-B ligand and to evaluate a combined antitumor effect of this targeted drug delivery system in vitro. In this study, the effects of PMC surface modification with the DR5-B ligand on cell uptake both in 2D (monolayer culture) and 3D (tumor spheroids) were studied by confocal microscopy, flow cytometry and fluorimetry. Cytotoxicity of the capsules was evaluated using an MTT test. The capsules loaded with DOX and modified with DR5-B demonstrated synergistically enhanced cytotoxicity in both in vitro models. Thus, the use of the DR5-B-modified capsules loaded with DOX at a subtoxic concentration could provide both targeted drug delivery and a synergistic antitumor effect.

Increased Intracellular Free Zinc Has Pleiotropic Effects on Doxorubicin-Induced Cytotoxicity in hiPCS-CMs Cells.

(1) the mechanisms and outcomes of doxorubicin (DOX)-dependent toxicity upon changed intracellular zinc (Zn) concentrations in the cardiomyocytes obtained from human-induced pluripotent stem cells (hiPCS-CMs) were investigated; (2) cells exposed to the DOX were pretreated or cotreated with zinc pyrythione (ZnPyr) and various cellular endpoints and mechanisms were analyzed via cytometric methods; (3) both DOX concentrations (0.3 and 1 µM) induced a concentration-dependent loss of viability, an activation of autophagy, cell death, and the appearance of senescence. These phenotypes were preceded by an oxidative burst, DNA damage, and a loss of mitochondrial and lysosomal integrity. Furthermore, in DOX-treated cells, proinflammatory and stress kinase signaling (in particular, JNK and ERK) were upregulated upon the loss of free intracellular Zn pools. Increased free Zn concentrations proved to have both inhibitory and stimulatory effects on the investigated DOX-related molecular mechanisms, as well as on signaling pathways on the resulting cell fates; and (4) free intracellular Zn pools, their status, and their elevation might have, in a specific context, a pleiotropic impact upon DOX-dependent cardiotoxicity.

Curcuma xanthorrhiza Rhizome Extract Induces Apoptosis in HONE-1 Nasopharyngeal Cancer Cells Through Bid

BACKGROUND: Curcuma xanthorrhiza rhizomes have been demonstrated to have anticancer properties toward various types of cancer cells. The effect of C. xanthorrhiza rhizome extract (CXRE) on nasopharyngeal cancer (NPC) cells, including HONE-1 cell line has not been elucidated yet. Therefore, the effect of CXRE on the apoptosis of HONE-1 cells and its possible underlying mechanism are necessary to be explored.METHODS: C. xanthorrhiza rhizomes were minced, dried, extracted with distilled ethanol, filtered, and evaporated to produce CXRE. HONE-1 cells were seeded, starved, and treated with dimethyl sulfoxide (DMSO), Doxorubicin, or various concentrations of CXRE. Treated HONE-1 cells were stained with 4',6'-diamidino-2-phenylindole (DAPI) and the number of viable cells was counted. HONE-1 cells were also collected, lysed, and further processed for immunoblotting analysis to measure Bid activity.RESULTS: The number of viable HONE-1 cells decreased in concentration- and time-dependent manner. The number of viable cells in 50 and 250 μg/mL CXRE-treated groups were significantly lower compared with that in the DMSO-treated group after 24 h. At 48 h incubation period, the number of viable cells in 10, 50 and 250 μg/mL CXRE-treated groups were significantly lower compared with that in the DMSO-treated group. The number of viable cells in 250 μg/mL CXRE-treatment group was not significantly different compared with that in the Doxorubicin-treated group after 48 h. Bid expression levels in CXRE-treated groups were lower compared with that in the DMSO-treated group.CONCLUSION: CXRE could induce apoptosis via Bid activation, hence reducing the viability of HONE-1 cells.KEYWORDS: Curcuma xanthorrhiza, nasopharyngeal cancer, HONE-1 cells, apoptosis, Bid

Formulation development of lipid polymer hybrid nanoparticles of doxorubicin and its in-vitro, in-vivo and computational evaluation.

The purpose of this study was to assess the parameters of doxorubicin (DOX) loaded lipid polymer hybrid nanoparticles (LPHNs) formulation development, and then the bioavailability of DOX were determined in the rabbit model, in order to evaluate the intrinsic outcome of dosage form improvement after the oral administration. LPHNs were prepared by combine approach, using both magnetic stirring and probe sonication followed by its characterization in terms of size-distribution (Zeta Size), entrapment efficiency (EE), loading capacity, and the kinetics of DOX. LPHNPs were further characterized by using scanning electron microscopy (SEM), powder X-Ray diffractometry (P-XRD), Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), in vitro and in vivo studies. The molecular modeling was determined through the density functional theory (DFT) simulations and interactions. DOX loaded and unloaded LPHNs were administered orally to the rabbits for bioavailability and pharmacokinetic parameters determinations. The plasma concentration of DOX was determined through high performance liquid chromatography (HPLC). The average size of DOX-loaded LPHNs was 121.90 ± 3.0nm. The drug loading of DOX was 0.391% ± 0.01 of aqueous dispersion, where its encapsulation efficiency was 95.5% ± 1.39. After oral administration of the DOX-LPHNs, the area under the plasma drug concentration-time curve (AUC) improved about 2-folds comparatively (p < 0.05). DFT simulations were used to understand the interactions of polymers with different sites of DOX molecule. The larger negative binding energies (-9.33 to -18.53kcal/mol) of the different complexes evince that the polymers have stronger affinity to bind with the DOX molecule while the negative values shows that the process is spontaneous, and the synthesis of DOX-LPHNs is energetically favorable. It was concluded that DOX-LPHNs provides a promising new formulation that can enhance the oral bioavailability, which have optimized compatibilities and improve the pharmacokinetic of DOX after oral administration.

Synergistic effects of concurrent photodynamic therapy with indocyanine green and chemotherapy in hepatocellular carcinoma cell lines and mouse models.

Photodynamic therapy (PDT) using an 808nm laser irradiation with indocyanine green (ICG) has shown tumoricidal effects in a hepatocellular (HCC) orthotopic xenograft model. Recently, combining PDT with concurrent chemotherapy has shown synergistic outcomes and a better therapeutic effect for cancer treatment. In the present study, we utilized a combination of chemotherapy drugs and PDT using ICG in vitro and in vivo in a patient-derived orthotopic xenograft (PDoX) model. We independently performed PDT and chemotherapy with sorafenib or doxorubicin in the Huh-7 and Hep3b cell lines by increasing the sorafenib or doxorubicin concentration and increasing the total energy of 808nm light. Subsequently, we combined the two treatments to confirm the effects on cell viability. The combination index (CI) was evaluated in vitro, and thereafter, in the HCC PDoX mouse model, 808nm laser irradiation with intravenously injected ICG and chemotherapy using doxorubicin were performed for twelve days. The viability of the Huh-7 and Hep3B cell lines decreased rapidly as the concentration of sorafenib or doxorubicin increased and as the total energy of 808nm light increased. The cell viability of the Huh-7 and Hep3b cell lines with combined PDT and chemotherapy was less than that with PDT or chemotherapy alone. The CI was <1 in the sorafenib- or doxorubicin-treated Huh-7 and Hep3b cell lines. In the HCC PDoX mouse model, tumor size was markedly decreased, and complete remission achieved compared to that of the single chemotherapy or PDT and control groups. The synergistic effect of concurrent PDT and chemotherapy in the HCC cell line and PDoX model was confirmed with no definite adverse effect. Concurrent PDT and chemotherapy could be applied in further preclinical studies.

One‐Pot Synthesis of Doxorubicin‐Bioactive Glass‐Ceramic Hybrid Nanoparticles through a Bio‐Inspired Route for Anti‐Cancer Therapy

AbstractIn this study, we report doxorubicin preparation with bioactive glass‐ceramic as a novel hybrid nanoparticle formulation. Synthesis was carried out through a bio‐inspired route where different precursors of bioactive glass are added to a Tris(hydroxymethyl)aminomethane buffer adjusted to slightly alkaline pH containing doxorubicin. With increasing concentration of doxorubicin, loading increased in quantity with comparatively higher drug release in acidic pH than neutral pH. Without synthetic surfactant or high temperature calcination, bioactive glass‐ceramic nanoparticles demonstrated significantly superior cytotoxic behavior towards osteosarcoma cell line when compared to equivalent free drug or its action towards non‐cancerous cell line. The particles exhibited hydroxyapatite deposition when immersed in simulated body fluid for 7 days. The bio‐inspired route for synthesis of doxorubicin‐bioactive glass‐ceramic hybrid nanoparticles is an efficient cost ‐effective synthesis mechanismwhich is environmentally friendly without hampering the stability or activity of the drug.

Effect of Cellular and Microenvironmental Multidrug Resistance on Tumor-Targeted Drug Delivery in Triple-Negative Breast cancer.

Multidrug resistance (MDR) reduces the efficacy of chemotherapy. Besides inducing the expression of drug efflux pumps, chemotherapy treatment alters the composition of the tumor microenvironment (TME), thereby potentially limiting tumor-directed drug delivery. To study the impact of MDR signaling in cancer cells on TME remodeling and nanomedicine delivery, we generated multidrug-resistant 4T1 triple-negative breast cancer (TNBC) cells by exposing sensitive 4T1 cells to gradually increasing doxorubicin concentrations. In 2D and 3D cell cultures, resistant 4T1 cells are presented with a more mesenchymal phenotype and produced increased amounts of collagen. While sensitive and resistant 4T1 cells showed similar tumor growth kinetics in vivo, the TME of resistant tumors was enriched in collagen and fibronectin. Vascular perfusion was also significantly increased. Fluorophore-labeled polymeric (∼10nm) and liposomal (∼100nm) drug carriers were administered to mice with resistant and sensitive tumors. Their tumor accumulation and penetration were studied using multimodal and multiscale optical imaging. At the whole tumor level, polymers accumulate more efficiently in resistant than in sensitive tumors. For liposomes, the trend was similar, but the differences in tumor accumulation were insignificant. At the individual blood vessel level, both polymers and liposomes were less able to extravasate out of the vasculature and penetrate the interstitium in resistant tumors. In a final in vivo efficacy study, we observed a stronger inhibitory effect of cellular and microenvironmental MDR on liposomal doxorubicin performance than free doxorubicin. These results exemplify that besides classical cellular MDR, microenvironmental drug resistance features should be considered when aiming to target and treat multidrug-resistant tumors more efficiently.

Demonstration of doxorubicin's cardiotoxicity and screening using a 3D bioprinted spheroidal droplet-based system.

Doxorubicin (DOX) is a highly effective anthracycline chemotherapy agent effective in treating a broad range of life-threatening malignancies but it causes cardiotoxicity in many subjects. While the mechanism of its cardiotoxic effects remains elusive, DOX-related cardiotoxicity can lead to heart failure in patients. In this study, we investigated the effects of DOX-induced cardiotoxicity on human cardiomyocytes (CMs) using a three-dimensional (3D) bioprinted cardiac spheroidal droplet based-system in comparison with the traditional two-dimensional cell (2D) culture model. The effects of DOX were alleviated with the addition of N-acetylcysteine (NAC) and Tiron. Caspase-3 activity was quantified, and reactive oxygen species (ROS) production was measured using dihydroethidium (DHE) staining. Application of varying concentrations of DOX (0.4 μM-1 μM) to CMs revealed a dose-specific response, with 1 μM concentration imposing maximum cytotoxicity and 0.22 ± 0.11% of viable cells in 3D samples versus 1.02 ± 0.28% viable cells in 2D cultures, after 5 days of culture. Moreover, a flow cytometric analysis study was conducted to study CMs proliferation in the presence of DOX and antioxidants. Our data support the use of a 3D bioprinted cardiac spheroidal droplet as a robust and high-throughput screening model for drug toxicity. In the future, this 3D spheroidal droplet model can be adopted as a human-derived tissue-engineered equivalent to address challenges in other various aspects of biomedical pre-clinical research.

Comparison of Characterization in Two-Dimensional and Three-Dimensional Canine Mammary Gland Tumor Cell Models.

Canine mammary gland tumors can be used as predictive models for human breast cancer. There are several types of microRNAs common in human breast cancer and canine mammary gland tumors. The functions of microRNAs in canine mammary gland tumors are not well understood. We compared the characterization of microRNA expression in two-dimensional and three-dimensional canine mammary gland tumor cell models. We evaluated the differences between two- and three-dimensional cultured canine mammary gland tumor SNP cells by assessing microRNA expression levels, morphology, drug sensitivity, and hypoxia. The expression of microRNA-210 in the three-dimensional-SNP cells was 10.19 times higher than that in the two-dimensional-SNP cells. The intracellular concentrations of doxorubicin in the two- and three-dimensional-SNP cells were 0.330 ± 0.013 and 0.290 ± 0.048 nM/mg protein, respectively. The IC50 values of doxorubicin for the two- and three-dimensional-SNP cells were 5.2 and 1.6 μM, respectively. Fluorescence of the hypoxia probe, LOX-1, was observed inside the sphere of three-dimensional-SNP cells without echinomycin but not in two-dimensional-SNP cells. The three-dimensional-SNP cells treated with echinomycin showed weak LOX-1 fluorescence. The present study showed a clear difference in microRNA expression levels in cells cultured in a two-dimensional adherent versus a three-dimensional spheroid model.