Multidrug resistance (MDR) in colorectal cancer (CRC) arises from interacting tumour-cell, pharmacological and microenvironmental programmes that undermine both drug activity and delivery. Chemotherapy-associated MDR is the principal focus of this review, while resistance to targeted therapy and immunotherapy is considered where it directly affects delivery design or patient selection. Bioactive compounds can modulate several resistance pathways, but their translational value is constrained by poor solubility, instability, rapid metabolism and inadequate exposure at resistant lesions. This review critically examines when nanomedicine can make such mechanistic activity pharmacologically and translationally meaningful. We link resistance biology to delivery functions, evaluate bioactive chemosensitiser candidates against human exposure, compare major nanomedicine design strategies, and assess nano-bio interactions, repeat-dose safety, manufacturing and clinical positioning. Across the evidence, three limitations recur. First, concentrations associated with chemosensitisation for curcumin, resveratrol, epigallocatechin gallate and quercetin generally exceed measured human parent-analyte exposure. Second, many resistance claims rely on parental or poorly characterised models, whereas studies combining defined resistance provenance with functional mechanism and exposure confirmation remain uncommon. Third, human studies demonstrate feasibility, tissue exposure or treatment response, but not mechanism-specific reversal of CRC MDR by a bioactive compound or bioactive nanomedicine. Progress therefore depends less on adding new particle classes than on matching a necessary formulation function to a defined resistance or spatial barrier, quantifying active exposure in the relevant compartment, validating mechanism in appropriate models, and integrating repeat-dose safety, scalable manufacturing and biomarker-guided clinical development.

Obesity exacerbates rheumatoid arthritis (RA). However, the underlying mechanisms remain incompletely defined. Elucidating these mechanisms can help the identification of novel therapeutic targets. Herein, we used high-fat diet (HFD)-induced obese collagen-induced arthritis (CIA) mice to investigate these mechanisms. Immunohistochemistry revealed that obesity exacerbated joint inflammation and cartilage degradation. Next, integrated label-free quantitative proteomics and cytometry by time-of-flight (CyTOF) were used to characterize lymphocyte subsets. Proteomic profiling identified 26 differentially expressed proteins in obese versus lean CIA mice, including the transcription factors EOMES and KLF2, the TGFβ receptor (TGFβR) signaling component TGFBR2, and the tissue-resident memory (TRM) T cell marker CD103. CyTOF analysis revealed a robust 3.0-fold increase (P = 0.0043) in the proportion of CD103⁺ TRM cells among CD3⁺ T cells in obese CIA mice, characterized by a large effect size. Immunofluorescence results confirmed this increase in synovial tissues. Treatment with asiaticoside (a TGF-β/Smad-suppressing triterpenoid) significantly reduced TRM cell proportions (P < 0.05) and ameliorated symptoms in obese CIA mice. Collectively, these findings establish a novel mechanistic axis in which obesity-induced TGFβR-hyperactivation promotes TRM cell accumulation, which exacerbates arthritis severity in this RA model. Our findings provide a preclinical rationale for targeting TGFβR/TRM in human RA with obesity as a comorbidity.

Molecular glues(MGs)are small molecules that modulate protein-protein interactions by inducing or stabilizing ternary complexes,thereby reprogramming cellular networks rather than inhibiting single targets.Natural products,shaped by evolutionary selection and characterized by exceptional structural diversity,constitute a rich and underexplored source of glue-like compounds.Here,we propose a conceptual framework for rationalizing the multi-target efficacy of traditional medicines through molecular glue mechanisms.Diverse classes of natural products,including polyketides,terpenoids,steroids,lignans,organic acids,and alkaloids,have been shown to promote proximity-driven regulation of signaling pathways and targeted protein degradation.This interaction-based mode of action provides a molecular explanation for the systems-level therapeutic effects of traditional formulations that act through multiple pathways simultaneously.Integration of molecular glue theory with network pharmacology and modern chemical biology approaches offers a strategy to decode the complex pharmacology of herbal medicines and to expand the druggable proteome.Advances in proximity-based screening technologies and machine learning are expected to further accelerate the discovery of natural product-derived molecular glues for next-generation therapeutics.

This study characterized β-lactoglobulin/low-methoxyl-pectin formulations containing oxalipalladium, compared preparations with and without folic acid, and evaluated pH-buffer release and HCT116 cell responses. Formulations were assessed by dynamic light scattering, zeta-potential measurement, spectrophotometric encapsulation-efficiency estimation, scanning electron microscopy, and atomic-force microscopy. Dialysis-based release was assessed in buffers at pH 1.2, 4.5, 7.0, and 7.5. HCT116 responses were examined by MTT, Annexin V/propidium iodide flow cytometry, and RT-qPCR. At pH 4.5, particle sizes were 40 nm with folic acid and 164 nm without folic acid; corresponding zeta potentials were − 10.53 and − 8.88 mV, PDI values were 0.10 and 0.14, and operational encapsulation-efficiency estimates were 75.2

Molecular glues (MGs) are a new class of small molecules that can modulate protein-protein interactions, leading to selective protein degradation, stabilization or activation. This breakthrough opens new possibilities for treating diseases such as cancer and neurodegenerative disorders. By targeting proteins previously considered "undruggable" due to the lack of conventional binding pockets, MGs are transforming drug discovery and delivery. By modifying protein interactions, MGs have potential not only in oncology but also in other biomedical applications. This review delves into how MGs induce targeted protein interactions, describes some of their therapeutic uses and recent advancements in this rapidly evolving field.

Colorectal cancer (CRC) remains a significant public health concern, emphasizing the need for innovative therapeutic strategies to improve patient outcomes. This study aimed to develop a highly efficient nanocarrier for targeted drug delivery, enhancing drug efficacy while minimizing concentrations and limiting adverse effects. We synthesized protein-based beta-lactoglobulin (beta lg) nanoparticles (NPs), loaded with 5-fluorouracil (5-FU) and sodium butyrate (NaB), and further functionalized with folic acid (FA) for specific targeting of folate receptor- positive CRC cells. The beta lg-5-FU-NaB-FA nanoplatforms exhibited a well-defined size of 208 nm with a narrow size distribution (PDI approximate to 0.5). Zeta potential measurements showed a value of -11.4 mV, indicating stability and suitability for drug delivery. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) confirmed the nanocarrier's spherical morphology and efficient distribution. Drug release profiles demonstrated that the NPs released more drugs at neutral to alkaline pH levels, attributed to pectin's ionization properties. The efficacy of the prepared beta lg-5-FU-NaB-FA nanoplatforms was investigated on HCT116 and Caco2 CRC cells, along with the normal cell line CRL-1831. The beta lg-5-FU-NaB-FA nanoplatforms exhibited remarkable cytotoxicity against both HCT116 and Caco2 CRC cells compared to free drugs, highlighting the efficacy of targeted delivery in folate receptor-positive cells. These NPs induce cell apoptosis and cell cycle arrest more effectively than free drugs, demonstrating their potential for targeted cancer therapy. Furthermore, a decrease in the expression of crucial genes involved in the Wnt signaling pathway was observed, which offers a valuable understanding of their underlying mechanism. Collectively, our results suggest that the FA-targeted beta lg nano- carriers represent a promising platform for the efficient and targeted delivery of 5-FU and NaB in folate receptor- positive CRC. This novel nanocarrier holds the potential to enhance therapeutic outcomes while minimizing side effects, providing a new avenue for the treatment of CRC.

Three new Cu(II) Schiff-base complexes {[Cu(SBn)]ClO4, (n = 1,2,3)} were synthesized and characterized by spectroscopic techniques and single-crystal x-ray crystallography (SCXRC). The N3O-type unsymmetrical Schiffbase ligands (SB1-3) were formed from the stepwise reaction of one equivalent of 2,2-dimethyl-1,3-propanediamine with one equivalent of two distinct aldehydes, i.e., salicylaldehyde, 5-bromosalicylaldehyde, or 3-methoxysalicyaldehyde (n = 1,2,3, respectively) while keeping the second aldehyde as pyridine-2-carbaldehyde in the three complexes. Three human carcinoma cell lines were used to test the complexes' in-vitro antiproliferative properties; i.e., human colorectal (HCT116), lung (A549), and breast (MCF7) malignant cells. All of the complexes showed good cytotoxic effects that varied with the dose of the synthesized complexes. Molecular docking studies were performed to study the interactions between the synthesized complexes against MCF-7 human breast cancer cells (PDB id: 4zvm), A549 lung carcinoma epithelia (PDB id: 5xnv), and human colorectal cancer cells HCT116 (PDB id: 6gue). The docking score of [Cu(SB3)]ClO4 with 4xvm (-8.46 kcal.mol- 1) was superior, while [Cu(SB2)]ClO4 had a better docking score with 5xnv (-7.25 kcal.mol- 1) and 6gue (-8.15 kcal.mol- 1).

Lenalidomide (LNA), a potent immunomodulatory drug, acts as a molecular glue that enhances drug-protein interaction. This study investigates the interaction between LNA and human serum albumin (HSA), focusing on structural modifications and protein aggregation. Employing a combination of spectroscopic techniques, including intrinsic fluorescence, temperature scanning, and circular dichroism (CD), alongside molecular docking and molecular dynamics (MD) simulations, we explored the drug–protein interaction dynamics and thermodynamic parameters. Our findings reveal that LNA induces both dynamic and static quenching mechanisms, primarily driven by hydrophobic interactions. Temperature scanning fluorescence showed no significant change in HSA’s melting temperature (Tm) with LNA. CD analysis indicated an increase in beta-sheet content, suggesting enhanced protein aggregation. Additionally, our analyses confirmed the increased size and stability of HSA aggregates in the presence of LNA. Docking simulations identified subdomain IIA as the primary binding site for the HSA-LNA complex, with a binding energy of −7.8 kcal·mol−1. Key interactions, including hydrogen bonding and van der Waals forces, were driven by residues such as His 146, Asp 108, and Ala 194, ensuring stable and specific ligand binding. Molecular dynamics (MD) simulations revealed the temperature-dependent dynamics of HSA and its ligand-bound complex. Analyses of root mean square deviation (RMSD) and root mean square fluctuation (RMSF) indicated minor structural changes and variations in flexibility at both 298 and 310 K, with ligand binding providing slight stabilization to the protein at lower temperatures. Additional studies are warranted to fully elucidate the conformational effects induced by drug binding. Radius of gyration analysis suggested a subtle expansion of the protein upon ligand binding. Hydrogen bond analysis demonstrated stable interactions, averaging 3.0 bonds at 298 K and 3.4 bonds at 310 K, underscoring their significance in complex stabilization. Solvent-accessible surface area (SASA) analysis indicated that HSA-ligand complexes possess less dense microstructures compared to free HSA, with increased solvent accessibility at both temperatures. Interaction energy calculations and MM-PBSA studies revealed stronger binding affinities at 310 K, primarily driven by van der Waals forces, which is consistent with experimental findings. These insights into the molecular interactions between LNA and HSA provide valuable implications for drug development and therapeutic applications, highlighting the potential of LNA as a molecular glue in modulating protein interactions.

Volatile organic compounds (VOCs), recognized as critical contributors to atmospheric pollution, demand energy-intensive degradation processes typically exceeding 200 degrees C for complete mineralization. The development of cost-effective catalytic systems capable of efficient VOC removal under low-temperature conditions remains a pressing global challenge. Herein, an ozone- assisted catalytic oxidation (OACO) strategy is proposed utilizing a copper-doped alpha-MnO2 catalyst (Cu/MnO2) to achieve high-performance VOC degradation at mild temperatures. The Cu-doped alpha-MnO2 catalyst, characterized by a distinct rod-like nanostructure, exhibited a significant enhancement in its catalytic activity. This meticulously designed approach capitalizes on the beneficial crystal phase of alpha-MnO2 and incorporates strategic Cu modification, thereby achieving enhanced catalytic performance. Mechanistically, ozone activation generates reactive oxygen species, which synergistically mediate VOC oxidation pathways. Systematic evaluation demonstrated that the OACO system achieves > 80% conversion efficiency for diverse VOCs (toluene, trimethylamine, ethyl acetate, and ethanol) at 80-100 degrees C, a temperature significantly lower than conventional thermal catalysis. This work establishes a scalable and energy-efficient paradigm for VOC abatement, bridging the gap between catalytic activity and operational feasibility in environmental remediation.

Lead is a widespread environmental hazard that can adversely affect multiple biological functions. Blood cells are the initial targets that face lead exposure. However, a systematic assessment of lead dynamics in blood cells at single-cell resolution is still absent. Herein, C57BL/6 mice were fed with lead-contaminated food. Peripheral blood was harvested at different days. Extracted red blood cells and leukocytes were stained with 19 metal-conjugated antibodies and analyzed by mass cytometry. We quantified the time-lapse lead levels in 12 major blood cell subpopulations and established the distribution of lead heterogeneity. Our results show that the lead levels in all major blood cell subtypes follow lognormal distributions but with distinctively individual skewness. The lognormal distribution suggests a multiplicative accumulation of lead with stochastic turnover of cells, which allows us to estimate the lead lifespan of different blood cell populations by calculating the distribution skewness. These findings suggest that lead accumulation by single blood cells follows a stochastic multiplicative process.

In this study, we employed Ajwain seed extract to synthesize oxalipalladium (OX) NPs and systematically investigated their physicochemical properties and biological activities. Characterization studies revealed that the OX NPs exhibited an average size of approximately 31.2 nm with a stable zeta potential of -26 mV, indicating colloidal stability conducive to drug delivery applications. We confirmed the homogeneous and spherical nature of the NPs, with FTIR spectra highlighting the presence of functional groups consistent with OX and Ajwain extract. Notably, OX NPs demonstrated potent anticancer activity against HCT116 colon cancer cells, inducing dose-dependent apoptosis. Compared to free oxaliplatin, OX NPs exhibited enhanced cytotoxicity. Flow cytometry analysis further elucidated the apoptotic pathway induced by OX NPs, confirming their efficacy as anticancer agents. Additionally, investigation into the molecular interactions between human serum albumin (HSA) and NPs revealed structural alterations in the protein upon interaction with the ligand. Analysis using three-dimensional fluorescence spectroscopy revealed alterations in the surroundings of Tyr and Trp residues, suggesting the influence of nanoparticles (NPs) on the protein's structural integrity. In a nutshell, our study contributes to the advancement of natural product-based synthesis procedures and therapeutic strategies for colorectal cancer treatment. Ajwain-assisted OX NPs exhibit enhanced cytotoxicity against colorectal cancer cells, leveraging HSA interactions for improved drug delivery and overcoming drug resistance.

Expansion microscopy (ExM) is an innovative super-resolution imaging technique that utilizes physical expansion to magnify biological samples, facilitating the visualization of cellular structures that are challenging to observe using traditional optical microscopes. The fundamental principle of ExM revolves around employing a specialized hydrogel to uniformly expand biological samples, thereby achieving super-resolution imaging under conventional optical imaging conditions. This technology finds application not only in various biological samples such as cells and tissue sections, but also enables super-resolution imaging of large biological molecules including proteins, nucleic acids, and metabolite molecules. In recent years, numerous researchers have delved into ExM, resulting in the continuous development of a range of derivative technologies that optimize experimental protocols and broaden practical application fields. This article presents a comprehensive review of these derivative technologies, highlighting the utilization of ExM for anchoring nucleic acids, proteins, and other biological molecules, as well as its applications in biomedicine. Furthermore, this review offers insights into the future development prospects of ExM technology.

Diagnosis of diseases with low facilities, speed, accuracy and sensitivity is an important matter in treatment. Bioprobes based on iron oxide nanoparticles are a good candidate for early detection of deadly and infectious diseases such as tetanus due to their high reactivity, biocompatibility, low production cost and sample separation under a magnetic field. In this study, silane groups were coated on surface of iron oxide nanoparticles using tetraethoxysilane (TEOS) hydrolysis. Also, NH2groups were generated on the surface of silanized nanoparticles using 3-aminopropyl triethoxy silane (APTES). Antibody was immobilized on the surface of silanized nanoparticles using TCT trichlorothriazine as activator. Silanization and stabilized antibody were investigated by using of FT-IR, EDX, VSM, SRB technique. UV/vis spectroscopy, fluorescence, agglutination test and ELISA were used for biosensor performance and specificity. The results of FT-IR spectroscopy showed that Si-O-Si and Si-O-Fe bonds and TCT chlorine and amine groups of tetanus anti-toxoid antibodies were formed on the surface of iron oxide nanoparticles. The presence of Si, N and C elements in EDX analysis confirms the silanization of iron oxide nanoparticles. VSM results showed that the amount of magnetic nanoparticles after conjugation is sufficient for biological applications. Antibody stabilization on nanoparticles increased the adsorption intensity in the uv/vis spectrometer. The fluorescence intensity of nano bioprobe increased in the presence of 10 ng ml-1. Nanobio probes were observed as agglomerates in the presence of tetanus toxoid antigen. The presence of tetanus antigen caused the formation of antigen-nanobioprobe antigen complex. Identification of this complex by HRP-bound antibody confirmed the specificity of nanobioprobe. Tetanus magnetic nanobioprobe with a diagnostic limit of 10 ng ml-1of tetanus antigen in a short time can be a good tool in LOC devices and microfluidic chips.

Purpose This study aimed to determine if Ki-67, a commonly used marker to measure tumor proliferation, is a reliable prognostic factor in various types of gastrointestinal (GI) cancers based on current high-quality multivariable evidence. Methods A comprehensive search was conducted in PubMed, Embase, Scopus, and ISI Web of Science databases to investigate the association between Ki-67 positivity and overall survival (OS) and disease/recurrence-free survival (DFS/RFS) in GI cancers. Heterogeneity was assessed using Chi-square-based Q and I 2 analyses and publication bias using funnel plots and Egger’s analysis. In addition, Ki-67 levels in different GI cancers were examined by different platforms. The prognostic capability of Ki-67, gene ontology (GO), and pathway enrichment analysis were obtained from GEPIA2 and STRING. Results Totally, 61 studies, involving 13,034 patients, were deemed eligible for our evaluation. The combined hazard ratios (HRs) demonstrated the prediction ability of overexpressed Ki-67 for a worse OS (HR: 1.67, P < 0.001; HR: 1.37, P = 0.021) and DFS/RFS (HR: 2.06, P < 0.001) in hepatocellular and pancreatic malignancies, respectively, as confirmed by multi-omics databases. However, similar correlation was not found in esophageal, gastric, and colorectal cancers. Furthermore, most of the associations were identified to be robust based on different subcategories and publication bias assessment. Finally, enriched Ki-67-related genes were found to be involved in various important signaling pathways, such as cell cycle, P53 signaling network, and DNA damage responses. Conclusion This study supports that Ki-67 can serve as an independent prognostic biomarker for pancreatic and hepatocellular malignancies in clinical settings.

This article delves into the interaction between HSA protein and synthesized platinum complexes, with formula: [Pt(Propyl-NH2)2(Propylglycine)]NO3 and [Pt(Tertpentyl-NH2)2(Tertpentylglycine)]NO3, through a range of methods, including spectroscopic (UV-visible, fluorescence, synchronous fluorescence and CD) analysis and computational modeling (molecular docking and MD simulation). The binding constants, the number of binding sites, and thermodynamic parameters were obtained at 25 to 37 degrees C. The study found that both complexes could bind with HSA (moderate affinity for Tertpentyl and strong affinity for Propyl derivatives) and occupied one binding site in HSA (validated with, Stern-Volmer, Job-plots, and molecular docking investigations) located in subdomain IIA. The binding mechanisms of both mentioned Pt(II) agents were different, with the Propyl derivative predominantly using van der Waals forces and hydrogen bond interactions with a static quenching mechanism and the Tertpentyl derivative mainly utilizing hydrophobic force with a dynamic quenching mechanism. However, the two ligands affected protein differently; the Tertpentyl complex did not significantly alter the protein structure upon binding, as evidenced by synchronous fluorescence spectroscopy (SFS), CD spectroscopy, and MD analysis. The outcome helps in understanding the binding mechanisms and structural modifications induced by the ligands, which could aid in the innovation of more effective and stable Pt(II)-based drugs.

Colorectal cancer (CRC) ranks third in malignant tumor mortality worldwide. Oxaliplatin, a third-generation chemotherapeutic drug, treats several malignancies, including colorectal cancer, after cisplatin and carboplatin. However, reducing side effects and optimizing efficacy is challenging. The utilization of plant-assisted green synthesis for the production of nanoparticles is a cost-effective and environmentally sustainable approach. This method could be at the forefront of chemotherapy due to its simplicity and efficiency. The ginger extract has been well-introduced with anti-cancer and antioxidant properties. The present study involved the green synthesis and characterization of oxaliplatin nanoparticles (OxPt NPs) through dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), inductively coupled plasma spectroscopy (ICP), field emission scanning electron microscopy (FESEM), and atomic force microscopy (AFM). This study investigated the effectiveness of OxPt NPs, ginger extract, and conventional oxaliplatin (first-line drug of CRC) on the HCT116 colorectal cancer cell line through MTT assay comparatively. The findings exhibited an outstanding efficiency of 72

The binding of drugs to plasma proteins, such as human serum albumin (HSA), encompasses two main functions: depot and transport, which are performed by these proteins. This binding has significant pharmacodynamic and pharmacokinetic consequences. 2-(furan-2-yl)-1H-imidazo[4,5-f][1,10]phenanthroline 1,2-diamino cyclohexane platinum (Pt(II)) or palladium (Pd(II)) complexes, have a similar structure. These new synthetic complexes have desirable anticancer activity, and the binding of these complexes with HSA was investigated. Spectroscopic and thermodynamic analyses indicate that the interactions between HSA and the complexes occur spontaneously with the static-dynamic quenching mechanism. However, the interactions are primarily driven by noncovalent forces such as hydrophobic interactions and π–π stacking, as evidenced by the positive enthalpy and entropy values associated with these complexes. In the circular dichroism (CD) results, HSA had fewer structural changes with the addition of the Pt(II) than the Pd(II) complex. Docking studies were performed at three Sudlow sites, I, II, and III, to identify the complex’s binding site on HSA. The reactive sites of HSA to Pt(II) and Pd(II) complexes mainly reside within its hydrophobic cavity in domain II (site II). Furthermore, molecular dynamics simulations showed low RMSD, Rg, and binding energy values during the 400 ns for the Pt(II) complex. The results suggest that the connection between the Pt(II) complex and HSA is reversible. The Pt(II) complex was a more desirable candidate than the Pd(II) complex. The results of the experimental methods are consistent with those of dynamic simulation and molecular docking methods, indicating that HSA is more stable in the presence of the Pt(II) complex.

Imaging mass cytometry (IMC) permits high-dimensional single-cell spatial proteomics by harnessing mass tags to replace conventional fluorescence tags. However, the current IMC technique commonly adopts metal-chelated polymer (MCP) tags, which are limited in sensitivity, multiplicity and data acquisition speed. Here, we demonstrate nanometal-organic framework (NMOF) tags, which could concurrently augment IMC's sensitivity, multiplicity, and acquisition speed. We designed and synthesized uniform-sized Zr-NMOFs (similar to 31 nm, PDI < 0.1) and then functionalized them with heterobifunctionalized aptamers containing phosphate groups and fluorescent moieties to generate Zr-NMOF_Aptamer probes. Such functionalization enabled direct ligand exchange with zirconium ions on Zr-NMOFs, thus allowing for concurrent fluorescence and mass signal acquisitions. The fluorescence signal enabled large-scale rapid imaging to quickly locate the region-of-interest, therefore significantly reducing IMC's blind scanning time and compensating for IMC's lower resolution. Meanwhile, the Zr-NMOF_Aptamer probe exhibited specific molecular recognition and a fourfold enhancement in signal amplification over the commercial MCP probe. Additionally, we showed that Zr-NMOF_Aptamer probes were compatible with commercial MCP probes for high-multiplex co-staining in IMC analysis. The Zr-NMOF_Aptamer probe represents a promising development of next-generation molecular probes for spatial proteomics with IMC.

This paper focuses on the synthesis of nano-oxali-palladium coated with turmeric extract (PdNPs) using a green chemistry technique based on the reduction in the Pd (II) complex by phytochemicals inherent in turmeric extract. PdNPs were examined and characterized using Field Emission Scanning Electron Microscopy (FESEM), Dynamic Light Scattering (DLS), Fourier Transform Infrared (FTIR), and Atomic Force Microscopy (AFM). Using different spectroscopic and molecular dynamics simulations, a protein-binding analysis of the produced nanoparticle was conducted by observing its interaction with human serum albumin (HSA). Lastly, the cytotoxic effects and apoptotic processes of PdNPs were studied against the HCT116 human colorectal cell line using the MTT assay and flow cytometry tests. According to the findings, PdNPs with spherical and homogenous morphology and a size smaller than 100 nm were generated. In addition, they can induce apoptosis in colorectal cancer cells in a dose-dependent manner with a lower Cc 50 (78 µL) than cisplatin and free oxali-palladium against HCT116 cells. The thermodynamic characteristics of protein binding of nanoparticles with HSA demonstrated that PdNPs had a great capacity for quenching and interacting with HSA through hydrophobic forces. In addition, molecular dynamics simulations revealed that free oxali-palladium and PdNP attach to the same area of HSA via non-covalent interactions. It is conceivable to indicate that the synthesized PdNPs are a potential candidate for the construction of novel, nature-based anticancer treatments with fewer side effects and a high level of eco-friendliness. Graphical Abstract

Background: The use of nanoparticle drug delivery systems to enhance the therapeutic efficacy and reduce the side effects of anticancer drugs is taken into consideration. Astaxanthin (ATX) is a natural xanthophyll carotenoid with antioxidant, anti-inflammatory, and antiapoptotic properties used to prevent and treat some cancers. Objectives: In the present study, the antioxidant effect of beta-lactoglobulin (β-LG) nanocapsules containing ATX and 5-fluorouracil (5-FU; the first-line therapy for colorectal cancer) on the antioxidant enzymes activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in HCT116 colorectal cancer cell line was examined. Methods: In this experimental study, HCT116 cells were treated with different treatments of encapsulation of ATX in β-LG, encapsulation 5-FU in β-LG, co-encapsulation of ATX and 5-FU in β-LG, free ATX, free 5-FU, free ATX and free 5-FU, or β-LG nanocapsules without drugs for 24, 48 and 72 hours. There is a control group in which HCT116 cells were not treated with any drug. Then, 50% inhibitory concentration (IC50) and cell viability were determined using an MTT assay. The antioxidant enzyme activity of SOD, CAT, and GPX was measured by a colorimetric method in HCT116 cells. Results: Different treatments reduced the cell viability and increased apoptotic cells in a timedependent manner, which was significant for beta-lactoglobulin nanocapsules treatment (P<0.05). It means receiving more 5-FU or ATX in the encapsulated form by HCT116 cells. The antioxidant enzyme activity of SOD, CAT, and GPX in HCT116 cells treated with beta-lactoglobulin nanocapsule treatment significantly increased compared to the control group (P<0.001). Moreover, the antioxidant activity of these enzymes in different treatments containing ATX (free or encapsulation) was significantly higher than in other treatments (P<0.05). The most increase in the activity of antioxidant enzymes is recorded in the treatment of nanocapsules containing ATX and 5-FU simultaneously. Conclusion: Increased activity of antioxidant enzymes in addition to the induction of apoptosis in colorectal cancer cells by various treatments of beta-lactoglobulin nanocapsules indicates more effective drug administration in encapsulated form as well as synergistic thera[peutic effects of ATX and 5-FU. Moreover, the main increase in antioxidant enzyme activity may be related to ATX.