Nanoemulsion in cancer therapy: Enhancing the therapeutic efficacy of capecitabine and 5-fluorouracil in colorectal cancer with thymoquinone nanoemulsion.

<h2>Abstract</h2> <p><strong>Background: </strong>Myocardial infarction (MI) is a cardiovascular disease that is the leading cause of death at all ages. Inflammation and oxidation processes constitute the basic pathophysiology of MI development. C-reactive protein (CRP) and transforming growth factor β (TGF-β) are markers that are often used to evaluate the level of inflammation, especially in MI.</p> <p><strong>Objective: </strong>This study aimed to evaluate the anti-inflammatory potential of thymoquinone (TQ), the major bioactive compound of <em>Nigella sativa</em>, by assessing its binding affinity through molecular docking, in which TQ exhibited more favorable binding energies compared to the native ligand.</p> <p><strong>Methods: </strong>Using the VegaZZ, PyMOL, and BIOVIA Discovery Studio tools, AutoDock Vina software was used for in silico research to test the active molecule TQ and produce visual profiles of native CRP and TGF-β ligands. Using the pkCSM method, pharmacokinetic predictions were carried out.</p> <p><strong>Results:</strong> Thymoquinone (2-methyl-5-propan-2-ylcyclohexa-2,5-diene-1,4-dione) showed favorable binding affinity to both CRP and TGF-β, with docking scores of –3.60 and –4.15 kcal/mol, respectively, which are more favorable than those of the native ligands (–2.39 and –2.73 kcal/mol) and comparable to enalapril (–4.84 and –6.13 kcal/mol). The RMSD value for CRP was 1.421 Å, while the value for TGF-β was 0.253 Å, indicating excellent structural alignment and validating the docking approach.</p> <p><strong>Conclusions: </strong>These in silico findings suggest that TQ warrants further investigation in vitro and in vivo as a potential modulator of inflammatory pathways in MI.</p>

The present study reports the presence of thymoquinone (TQ), a promising benzoquinone compound, in the lichen Dirinaria frostii (strain DF_MSCBU, NCBI-accession no. PP955960), collected from Similipal National Park, India. The extract of D. frostii was subjected to GCMS analysis and detected the presence of thymoquinone. Further, purification of the TQ fraction was carried out using HPLC against a standard thymoquinone reference. FTIR analysis identified the key functional groups. In addition, Nuclear Magnetic Resonance (¹H NMR and ¹³C NMR) spectroscopy was performed to determine the structure and composition of the molecule. The purified TQ was then used for antibacterial assays against pathogenic bacteria Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Vibrio cholerae following the MIC method showed pathogen inhibition at different incubation time, of both Gram +ve and Gram -ve bacteria. Thymoquinone showed significant bactericidal activity compared to the standard antibiotics azithromycin, streptomycin, and erythromycin in a dose-dependent manner. This study demonstrates occurrence of thymoquinone from D. frostii as a novel finding and opens new possibilities for its therapeutic applications in the future.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, with resistance to 5-fluorouracil (5-FU) representing a major therapeutic challenge. Thymoquinone (TQ), a bioactive constituent of Nigella sativa, exhibits anticancer activity; however, the mechanisms underlying TQ-induced cell death appear to be highly context dependent. This study aimed to characterize cell line-specific death pathways triggered by TQ in colorectal cancer models with distinct molecular backgrounds and differential responsiveness to 5-FU. Human CRC cell lines RKO (5-FU-sensitive) and SW1116 (poorly responsive), along with normal colon epithelial cells (CCD-841CoN), were treated with TQ, 5-FU, or their combination for 24 h. Cell viability, DNA fragmentation, caspase-3/7, -8, and -9 activity, cell death phenotypes, and expression of apoptosis- and necroptosis-related genes were evaluated using MTT assays, ELISA, luminescent assays, flow cytometry, and RT-qPCR. TQ significantly reduced viability in both CRC cell lines while exerting minimal cytotoxicity toward normal cells. In RKO cells, characterized by microsatellite instability (MSI), TQ induced DNA fragmentation, caspase activation, and transcriptional upregulation of pro-apoptotic genes, consistent with apoptosis-associated signaling. In contrast, SW1116 cells, which exhibit chromosomal instability (CIN) and reduced responsiveness to 5-FU, displayed decreased viability accompanied by suppressed caspase activity and predominant features of caspase-independent necrotic cell death. This differential response may be attributed to the CIN phenotype, which has been associated with impaired apoptotic signaling and enhanced tolerance to cytotoxic stress. Combined TQ and 5-FU treatment did not produce synergistic cytotoxicity, as confirmed by Bliss independence analysis, but revealed distinct, cell line-dependent death programs. These findings demonstrate that TQ modulates cell death execution in a molecular context-dependent manner rather than enhancing 5-FU efficacy through pharmacological synergy.

Although the effect of thymoquinone (TQ), the biologically active component of black cumin seeds, has been investigated in many types of cancer, its effects on molecular pathways of cancer cells further need to be elucidated. This study aimed to investigate the effects of TQ on genotoxicity, oxidative stress, apoptosis, and inflammatory pathways in SW620 cells treated with irinotecan (IR), celecoxib (Clx), individually or both. DNA damage, oxidative stress/inflammatory response, cell cycle, apoptosis, and expression of carcinogenesis-related genes were determined using the Comet assay, ELISA assay, flow cytometry, and RT-PCR techniques, respectively. TQ (8 µM) combination with IR (8 µM) + Clx (4 µM) significantly increased cytotoxicity and genotoxicity of IR with/without Clx, while decreasing oxidative stress and inflammatory responses. TQ upregulated p21 and Bax, while downregulating Bcl-2 and Bcl-XL, increasing Bax/Bcl-2 ratio, which indicates an induction of apoptotic pathways. TQ also increased the percentage of cells in G2/M phase and decreased those in G0/G1 phase, thereby affecting cell cycle checkpoints in IR-treated cells with or without Clx. In conclusion, this study shows that the combination of TQ with IR, Clx, or both exerts significant effects on SW620 colon cancer cells, such that by enhancing DNA damage, TQ may induce G2/M cell cycle arrest and apoptosis, while reducing inflammatory responses, oxidative stress, and G0/G1 cell cycle arrest. These in vitro findings indicate that TQ may enhance the chemotherapeutic effects of IR and act as a potential adjuvant therapy; however, further in vivo studies are required to verify its suggested effects.

Spinal cord injury (SCI) results in rapid bone loss and osteoporosis development. This study investigated the synergistic effects of a combination of thymoquinone (TQ) and coenzyme Q10 (CoQ10) on SCI-induced osteoporosis. Thirty male Wistar rats underwent complete spinal cord transection at T10 and were divided into five groups: sham, untreated SCI, TQ-treated (100mg/kg/day), CoQ10-treated (10mg/kg/day), and combination therapy (TQ + CoQ10). After 21 days of treatment, combination therapy significantly improved BBB locomotor scores (12.1±0.7vs. 2.1±0.4 in untreated SCI; p < 0.001) and increased femoral BMD and BMC by 32.8% and 37.9%, respectively. Micro-CT analysis revealed 45.2% higher bone volume/total volume ratio and enhanced trabecular architecture. The combination therapy significantly downregulated SOST expression (76.2% reduction) while upregulating periostin (184.7% increase). Osteoclast markers decreased (CTR: 57.3%, cathepsin K: 63.1%) while osteoblast markers increased (ALP: 143.8%, osterix: 127.4%). Oxidative stress markers improved substantially, with 47.3% lower MDA levels and 86.5% higher SOD activity. TQ + CoQ10 combination therapy produces both synergistic and additive effects that ameliorates SCI-induced osteoporosis in association with altered expression of Wnt/β-catenin signaling components, enhancing bone formation, and reducing oxidative stress, demonstrating superior therapeutic efficacy compared to monotherapies.

Tofacitinib citrate nanoemulgel induced repigmentation in vitiligo: Assessment of oxidative stress, protein, immunohistochemistry, and mRNA expressions.

Multifunctional bilayered nanofibrous scaffold of chitosan/poly(caprolactone) enriched with simvastatin and thymoquinone for accelerated diabetic wound healing.

A sprayable TQ/Ce6@SAB/F-gel for accelerating wound healing via hypoxia-tolerant photodynamic therapy and immune-metabolic pathway.

Thymoquinone (TQ), the principal active constituent of Nigella stativa, has demonstrated numerous biological properties and therapeutic effects on various diseases. However, its therapeutic potential against cardiac hypertrophy remains uncertain. This study aims to investigate the protective effects of TQ on stress-induced cardiac hypertrophy and elucidate the underlying mechanisms. Our findings reveal that TQ mitigates stress-induced cardiac hypertrophy in mice and AngII-induced hypertrophy in H9c2 cells. Moreover, TQ inhibits cardiomyocyte ferroptosis and apoptosis by downregulating PTGS2, Bax, and upregulating GPX4, Bcl-2, thereby alleviating cardiac hypertrophy and dysfunction. Mechanistically, the protective effects of TQ against ferroptosis and apoptosis in cardiac hypertrophy were reversed by the PPAR-γ inhibitor (GW9662). In addition, TQ treatment led to increased protein expression levels of P-PI3K and P-AKt. Taken together, our findings suggest that TQ could attenuate cardiac hypertrophy through activation of the PPAR-γ/PI3K/Akt signalling pathway.

Burn injuries represent a worldwide public health concern. Nigella sativa (NS), a herbal medicinal plant, widely used in Ayurveda, Unani, and Siddha holds potential to address this issue. This review evaluated the potential of NS extract and its key component, Thymoquinone (TQ), in accelerating burn wound healing, focusing on their mechanisms, safety, and efficacy. Studies were retrieved from PubMed, Google Scholar, and ScienceDirect using keywords "Nigella sativa," "Thymoquinone," "burns," and "wound healing." Reports were screened for suitability, and the scientific name was verified on www.worldfloraonline.org. NS extract and TQ accelerate burn wound healing in vitro and in vivo through various mechanisms involving anti-inflammatory, anti-oxidative, anti-microbial, and tissue regenerative effects. TQ achieved 91.35% wound closure in NIH/3T3 cells (mouse fibroblasts) and wound closure rate of 2.35 ± 0.05% in 3T3-CCL92 cells (fibroblasts from mouse embryos). It also reduced oxidative stress and inflammation in murine burn models. Nanoformulations of TQ showed higher safety (LD₅₀: 300-2000 mg/kg) than pure TQ (LD₅₀: 50-300 mg/kg). A Phase I trial confirmed the safety of black cumin oil (5% TQ) at 200 mg/day for 90 days in healthy adults. Therefore, formulations based on NS extract and TQ can manage burn injuries and be extrapolated into clinical settings.

Cancer therapy is often hindered by poor solubility, low bioavailability, drug resistance, and tumor microenvironmental barriers associated with conventional chemotherapeutics. Polymeric nano–drug delivery systems offer a promising strategy to overcome these limitations, particularly for synergistic multi-drug delivery. In this study, a biodegradable and biocompatible PLGA copolymer (70:30, M̅w ≈ 14,500) was synthesized by ring-opening polymerization using zinc proline complex as an initiator through a green route. The copolymer’s potential for delivering topotecan (TPT), a water-soluble chemotherapeutic, thymoquinone (TQ), a poorly water-soluble chemotherapeutic, and their combination (TPT+TQ) for cancer treatment was investigated. These nanoparticles demonstrateda consistent particle size < 200 nm high encapsulation efficiency along with desirable controlled-release attributes. Moreover, they exhibited specific release characteristics and cytotoxic effects against HeLa cells, achieving an IC50 value of 20.88 µM for the combination therapy (TPT+TQ). Additionally, cytocompatibility testing on L929 fibroblasts confirmed over 98% cell viability for blank PLGA nanoparticles. Additionally, confocal imaging studies confirmed efficient cellular uptake and nuclear localization of the nanoparticles. Overall, the PLGA based dual drug loaded nanoparticles presents a promising approach for targeted, synergistic co-delivery, potentially improving efficacy and reducing toxicity in cancer therapy.

High-dose subacute and acute thymoquinone treatments alleviate LPS-induced depressive-like behavior in mice by reducing inflammation via the IDO/KYN pathway.

ABSTRACT Cyclophosphamide (CYP) is a widely‐used chemotherapeutic drug known to induce ovarian damage and infertility in women. The present study evaluated the potential protective roles of zofenopril, a sulfhydryl‐containing angiotensin‐converting enzyme inhibitor, and thymoquinone (ThQ), alone and in combination, against CYP‐induced ovarian damage in rats. Thirty adult female rats were divided into five groups: a control group that received normal saline; a CYP‐Induced group given a single intraperitoneal (IP) dose of 200 mg/kg CYP on day 17 of the experiment; and three groups pre‐treated with zofenopril, ThQ, or Zofenopril+ThQ for 17 days before CYP‐administration, and continuing for 2 days after CYP exposure. Animals were euthanized, and blood and ovarian tissues were collected 48 h after CYP administration. Serum follicle‐stimulating hormone (FSH), luteinizing hormone (LH), anti‐Müllerian hormone (AMH), and estradiol (E2) levels were measured, as well as the apoptotic biomarker caspase‐3, total antioxidant capacity (TAOC), and tumor necrosis factor (TNF‑α). Ovarian histomorphometry was also assessed. CYP injection induced ovarian injury, characterized by reduced serum AMH and E2 levels, increased FSH and LH levels, oxidative stress, and increased levels of inflammatory and apoptotic markers. In addition, severe ovarian atrophy, regression and degradation of ovarian follicles, and atrophic follicles were observed. Zofenopril and ThQ administration significantly mitigated these CYP‐mediated effects, restoring hormone levels, reducing oxidative stress and inflammation, and improving ovarian histology compared with the CYP‐Induced group. Based on the present study, zofenopril and ThQ have protective effects against CYP‐induced ovarian injury; while co‐administration of zofenopril+ThQ was more effective at inhibiting CYP‐induced effects on TAOC, TNF‑α, and caspase‐3 levels.

Modulation of coagulation activity by thymoquinone (TQ), a 1,4-benzoquinone derivative of Nigella sativa, prompted the identification and validation of a molecular mechanism for hemostasis. Consequently, this research utilizes network pharmacology, molecular docking, and dynamics approaches to investigate the role of TQ for improved hemostasis. Initially, protein targets related to hemostasis and TQ were extracted, where the common targets were subjected to KEGG pathway enrichment and gene ontology analysis to determine the top pathways responsible for inducing hemostasis. The key protein targets from the identified pathways were selected for molecular docking and MM-GBSA calculations, where the targets with the highest absolute Glide docking and ΔGbind scores were further considered for molecular dynamics simulation. The results indicated that TQ exerts potential effects on hemostasis by modulating cMet, PDGFR, and PI3KA proteins involved in the EGFR-TKIR pathway. The nontoxic concentration of TQ was determined using the MTT assay, which revealed a safe concentration (<2.5µM) of TQ for biological applications. In practice, TQ was found to induce the formation of stable fibrin clots in platelet-poor plasma, confirming its hemostatic potential. Finally, in vivo studies in a rat tail hemorrhage model confirmed its clotting activity, where modulation of cMet and PDGFR indicates the potential of TQ in inducing rapid hemostasis. Thus, this study integrated in silico, in vitro, and in vivo approaches to investigate the role of TQ in hemostasis through the EGFR-TKIR pathway, a mechanism that remains largely unexplored in blood clotting.

INTRODUCTION: Chronic Myeloid Leukaemia (CML) is a hematopoietic malignancy caused by the BCR-ABL1 fusion oncoprotein, originating from Philadelphia chromosome translocation that enhances leukemic cell survival and therapeutic resistance. Aberrant MAPK pathway activation promotes proliferation and inhibits apoptosis, contributing to CML progression and resistance to tyrosine kinase inhibitors like imatinib with prolonged use. Thymoquinone (TQ), a bioactive molecule, has attracted considerable interest Z for its anticancer characteristics that are worth investigating. MATERIALS AND METHODS: K562 CML cells were divided into an untreated group and a TQ-treated group and observed for 24 and 48 hours. Different TQ concentrations were administered to the TQ-treated group. Dose and time dependent effects on cell growth were assessed to evaluate cytotoxicity and determine the IC50 value in both groups. RNA was extracted from K562 CML cells based on the IC50 value and proceeded with RT-qPCR analysis on 7 genes involved, assigned as Raf1, B-Raf, ERK1, ERK2, K-Ras, H-Ras, and N-Ras genes, while the beta-actin gene was used as a housekeeping gene. Protein was extracted for the determination of protein and phosphorylation levels of Raf, MEK1/2, and ERK1/2, and assessed using the Jess Simple Western protocol. The Wilcoxon Signed-Rank test was performed using IBM SPSS, with p&lt;0.05 considered statistically significant. Results: TQ treatment significantly reduced the expression of all genes analysed in K562 cells. It also decreased protein and phosphorylation levels of Raf, MEK1/2, and ERK1/2. Conclusion: These findings suggest that TQ effectively inhibits MAPK signalling in K562 CML cells, highlighting its potential as a future treatment for CML.

Design and development of a low-cost potentiostat for thymoquinone (TQ) detection in black cumin using a graphite-based TQ-imprinted polymer sensor

Breast cancer is the most commonly diagnosed cancer globally, predominantly affecting older women. Thymoquinone (TQ) is a natural therapeutic agent that exerts anticancer efficacy. However, its pharmacological effects are restricted by inadequate aqueous solubility and bioavailability. The present study aimed to develop TQ-encapsulated polyethyleneimine/poly(lactic acid) nanoparticles (TQ-PEI/PLA-NPs) to enhance the delivery of TQ into breast cancer cells. The solvent evaporation-emulsification method was employed to synthesize TQ-PEI/PLA-NPs, and their physicochemical properties were subsequently examined. TQ-PEI/PLA-NPs had a crystalline structure, a zeta potential of +1 mV, and a spherical shape with a diameter of 80-90 nm. The encapsulation efficiency was 85.77 ± 2.21% (w/w), while the drug loading capacity was 9.32 ± 1.14% (w/w). The release rate of TQ from TQ-PEI/PLA-NPs was marginally elevated at pH: 5.8 (81.21 ± 0.87%) compared to pH 3.5 and 7.2. MTT assay using TQ-PEI/PLA-NPs confirmed concentration-dependent cytotoxicity against MCF-7 cells after 24 h of treatment, and IC50 was found to be 21.99 μg/mL. The intracellular delivery of TQ in MCF-7 cells caused cell death, as indicated by AO/EBr staining, mitochondrial transmembrane potential assay, and Caspase-3 and -9 investigations. The observed MCF-7 cell death attributed to TQ was induced by reactive oxygen species (ROS) and impaired mitochondrial membrane potential. The ROS potentially damaged the mitochondrial membrane, and further studies supported the induction of apoptosis. Our results indicated that TQ-PEI/PLA-NPs, which cause cytotoxicity to breast cancer cells, as evidenced by the decreased MCF-7 cell counts, may exhibit significant therapeutic potential for breast cancer therapies.

Abstract Objectives Structural, molecular and immunohistochemical changes in the liver of rats after acute cadmium (Cd) exposure and the effects of carvacrol (Car) and thymoquinone (TQ) were studied in this research. Methods Control, DMSO, Cd-treated (3.5 mg/kg by subcutane), Car (50 mg/kg gavage)-Cd treated, TQ (5 mg/kg by gavage)- Cd treated, carvacrol, and thymoquinone groups were formed.H&amp;E and Masson’s Trichrome stainings were performed. Histological scoring was performed in liver. On the liver sections TNF-a, IL-6, and NF-kB immunostainings were carried out. Also sections of liver tissue were evaluated by electron microscopy. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and triglyceride (TG), oxidative stress parameters, MDA and GPx, were evaluated. Tnf-a, Nfkb1, IL6, mTor and Casp3 mRNA gene expression levels were examined using RT-PCR. The obtained data was statistically analyzed using GraphPad Prism software. Results Treatment groups showed significant reduction in ALT and TG when compared to cadmium group.In the cadmium group, MDA levels increased significantly and GPX1 levels decreased compared to the control group. As a results of the microscopic studies, cellular and structural defects by cadmium were decreased in the presence of carvacrol and thymoquinone. NF-kB, TNF-a and IL-6 immunostained surface area increased in cadmium compared to the control group. Cd-Car and Cd+Thymoquinone alleviated these inflammatory expressions. In treatment groups, Nfkb1, Tnf-a, IL6, mTor, Casp3 expressions reduced compared to cadmium alone. Conclusions The study results indicate that carvacrol and thymoquinone alleviate the toxic effects of liver tissue induced by cadmium.

Bisphenol A (BPA) is an industrial and environmental toxin. Chronic BPA exposure-mediated apoptosis and Ca2+ influx have been linked to the development of certain reproductive problems and granulosa cell damage in humans by raising cytosolic (cROS) and mitochondrial (mROS) free reactive oxygen species. However, these effects were modulated by upregulating glutathione (GSH) and glutathione peroxidase (GSHPx), while downregulating the TRPM2 channel activity. In human KGN granulosa cells, we aimed to investigate the protective function of the antioxidant thymoquinone (TQ) against BPA-induced TRPM2 channel activation, oxidative, and apoptotic changes. Five experimental groups were established: control, TQ, BPA, BPA + TQ, and BPA + TRPM2 antagonist [2-aminoethoxydiphenyl borate (2APB) and (N-p-Amylcinnamoyl)anthranilic acid (ACA)]. TRPM2 agonists (H2O2 and ADP-ribose) and BPA-induced TRPM2 stimulations (Ca2+ influx and TRPM2 current density) were downregulated after incubation with TQ and ACA/2APB. BPA-mediated increases in Zn2+, Fe2+, oxidants (mROS and cROS), apoptotic markers (caspase-3, caspase-8, and caspase-9), and cell death were decreased by TQ and ACA therapy. On the other hand, BPA-induced decreases in GSH and GSHPx levels were reversed after the TQ and ACA incubations. In conclusion, TQ reduced the BPA-induced increase in Ca2+ influx, Fe2+, Zn2+, apoptosis, and oxidative granulosa cell injury via the suppression of TRPM2.