Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Physicochemical characterization of the Se–Ag/chitosan hybrid nanocomposite.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques:

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Survival analysis demonstrating the protective effect of Se/Ag chitosan nanocomposite treatment in diethylnitrosamine-induced hepatocellular carcinoma. (A) Kaplan–Meier survival curves showing survival probability over the 14 weeks experimental period. Control and Se/Ag alone groups maintained 100% survival. Early HCC and Late HCC groups exhibited progressive mortality with final survival rates of 55% and 20%, respectively. Treatment with Se/Ag nanocomposite significantly improved survival outcomes in both earlystage (85% survival) and late-stage (70% survival) HCC groups compared to their respective untreated counterparts. (B) Bar graph depicting final survival rates at the experimental endpoint (week 14). The graph demonstrates complete survival in control and Se/Ag alone groups, severe mortality in untreated HCC groups (particularly late-stage HCC at 20%), and substantial protective effects of Se/Ag treatment in both HCC progression stages. (C) Cumulative mortality curves illustrating the temporal progression of death events. Late HCC group showed the earliest and steepest mortality increase, while Se/Ag treatment significantly delayed and reduced cumulative mortality in both HCC groups throughout the observation period. (D) Statistical comparison table presenting log-rank test results for key group comparisons. All comparisons between HCC groups and controls, as well as between treated and untreated HCC groups, showed highly significant differences ( p < 0.01 to p < 0.0001).

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Control, Comparison

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Biochemical, inflammatory, and multivariate assessment of therapeutic response to Se–Ag–CS nanocomposites in hepatocellular carcinoma. (A) Serum liver enzyme activities (ALT, AST, and ALP) across healthy, HCC, early-treatment (Early Tx), and late-treatment (Late Tx) groups, showing significant enzyme elevation in HCC and partial to substantial normalization following treatment. (B) Oxidative stress markers, malondialdehyde (MDA) and reduced glutathione (GSH), indicating pronounced oxidative imbalance in HCC and restoration toward redox homeostasis after therapy. (C) Pro-inflammatory cytokines TNF-α and IL-6, markedly elevated in HCC and significantly reduced upon treatment, with greater suppression in early intervention. (D) Serum alpha-fetoprotein (AFP) levels relative to the clinical threshold, demonstrating strong AFP reduction following treatment, particularly in the early-treatment group. (E) Superoxide dismutase (SOD) activity, reflecting impaired antioxidant defense in HCC and recovery after nanocomposite administration. (F) Oxidative stress index (OSI; MDA/GSH × 100), summarizing redox imbalance and highlighting effective oxidative stress mitigation with treatment. (G) Therapeutic efficacy quantified by AFP reduction, showing superior response in early-stage HCC compared with late-stage disease. (H) Multi-biomarker efficacy analysis illustrating coordinated reductions in liver enzymes and inflammatory cytokines, with enhanced benefit in early HCC. (I) Principal component analysis (PCA) integrating all biomarkers, demonstrating clear separation between healthy, HCC, and treated groups, and confirming systemic biochemical normalization following Se–Ag–CS nanocomposite therapy.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Clinical Proteomics, Activity Assay, Drug discovery, Biomarker Discovery

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Multiscale pharmacokinetic–pharmacodynamic and therapeutic response modeling of green-synthesized Se–Ag–CS nanocomposites in hepatocellular carcinoma. Simulated plasma and peripheral tissue concentration–time profiles following systemic administration illustrate rapid absorption, systemic distribution, and sustained exposure above the effective concentration threshold. Hepatic accumulation kinetics demonstrate preferential liver targeting and prolonged retention relative to plasma, with stage-specific effective concentration (EC 50 ) thresholds distinguishing early and late hepatocellular carcinoma (HCC). Concentration–response relationships link nanocomposite exposure to tumor cell kill, highlighting enhanced therapeutic sensitivity in early-stage HCC compared with advanced disease. Time-dependent pharmacodynamic modeling further confirms sustained antitumor activity, with reduced efficacy observed in late-stage HCC. Alpha-fetoprotein (AFP) dynamics serve as a biomarker of therapeutic response, showing marked suppression under early treatment conditions relative to late treatment and untreated controls. Liver enzyme normalization profiles ALT and AST indicate transient treatment-associated perturbations followed by recovery toward physiological ranges, supporting a favorable hepatic safety profile. Tumor growth inhibition modeling over a 30 days period demonstrates significant regression with early intervention and growth stabilization under late treatment compared with unchecked progression in untreated tumors. Key pharmacokinetic parameters—including maximum concentration ( C _max), time to peak concentration ( T _max), area under the curve (AUC), clearance, mean residence time, and liver targeting index—quantitatively support enhanced hepatic delivery efficiency and therapeutic performance of the Se–Ag–CS nanocomposites.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Clinical Proteomics, Synthesized, Concentration Assay, Activity Assay, Biomarker Discovery, Inhibition

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Statistical, correlative, and predictive analysis of biomarker modulation following Se–Ag–CS nanocomposite therapy in hepatocellular carcinoma. (A) Biomarker correlation matrix illustrating strong positive associations among liver injury markers (ALT, AST, ALP, bilirubin), inflammatory cytokines (TNF-α, IL-6), oxidative stress indicators (MDA), and AFP, with pronounced negative correlations between antioxidant defenses (GSH, SOD) and pathological biomarkers, indicating coordinated disease-driven dysregulation. (B) One-way ANOVA statistical analysis confirming significant intergroup differences across all measured biomarkers, with highly significant F-statistics and p -values supporting robust treatment effects. (C) Distribution of serum AFP levels across healthy, HCC, early-treatment, and late-treatment groups, demonstrating marked elevation in untreated HCC and substantial reduction following therapy, particularly with early intervention. (D) Projected AFP time-course profiles over 14 days, showing progressive AFP increase in untreated tumors, accelerated decline with early treatment, and partial response under late treatment relative to the normal reference range. (E) Comparative treatment efficacy based on percentage reduction of key biomarkers (ALT, AST, AFP, and TNF-α), highlighting superior therapeutic benefit in early-stage HCC compared with late-stage disease.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Biomarker Discovery

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of HCC and therapeutic response to Se–Ag/chitosan nanocomposite treatment. (A) Preservation of hepatic tissue architecture (% normal structure). Nanocomposite treatment substantially preserved liver architecture compared with untreated early and late HCC. (B) Inflammatory infiltration scored on a 0–10 scale, indicating reduced immune cell infiltration in treated groups. (C) Hepatic tissue necrosis (% area), showing marked attenuation following treatment. (D) Overall HCC severity score combining architectural disruption, necrosis, and inflammation. (E) p53 immunoexpression levels (% positive cells and intensity), demonstrating enhanced apoptotic signaling in treated tumors. (F) Ki-67 proliferation index (% positive cells), illustrating suppressed tumor proliferation posttreatment. (G) Correlation between proliferation and apoptosis indices, highlighting treatment-induced shift toward apoptosis in both early and late HCC. (H) Histological treatment response score (% improvement relative to untreated HCC), with higher efficacy in early intervention. (I) Radar plot summarizing multiple histological features (architecture, hepatocyte integrity, sinusoidal structure), showing normalization trends in treated livers. (J) Heatmap of normalized histopathological features across groups, illustrating treatmentmediated modulation of proliferation, apoptosis, inflammation, and necrosis. All data represent mean ± SD. Statistical significance: p < 0.05, p < 0.01, p < 0.001.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Immunohistochemical staining, Clinical Proteomics, Preserving, Disruption

Journal: RSC Advances

Article Title: Chitosan-confined selenium–silver nanohybrids enable redox-modulated mitochondrial apoptosis and survival improvement in experimental hepatocellular carcinoma

doi: 10.1039/d6ra01968d

Figure Lengend Snippet: Multiscale computational framework elucidating the physicochemical and biological mechanisms of Se–Ag chitosan nanocomposites in hepatocellular carcinoma therapy. (A) Representative molecular dynamics trajectory (5 ns, 80 atoms) showing structural stability of Se–Ag nanocomposites. (B) Energy conservation profile illustrating stable kinetic, potential, and total energies over the simulation time. (C) Temperature equilibration under Berendsen thermostat control at 310 K. (D) Radial distribution function (RDF) highlighting preferential interatomic distances and structural ordering. (E) Potential of mean force (PMF) depicting the thermodynamic favorability of nanoparticle–protein interactions. (F) Summary of binding parameters, including optimal distance and dissociation constant, confirming strong electrostatic and van der Waals contributions. (G) Dose-dependent kinetics of reactive oxygen species (ROS) generation. (H) Time-dependent glutathione (GSH) depletion indicating tumor-selective oxidative stress. (I) Quantum confinement effect on band gap energy as a function of nanoparticle radius. (J) Size-dependent work function variation. (K) Optical absorption properties across UV-visible spectra. (L) Density of states (DOS) illustrating size-dependent electronic structure modulation. (M) Langmuir adsorption isotherm describing nanoparticle–surface interactions. (N) Surface reaction kinetics based on the Langmuir–Hinshelwood model. (O) Arrhenius plot showing temperature dependence of reaction rate constants. (P) Pourbaix diagram indicating pH-dependent selenium speciation and thermodynamic stability under physiological conditions.

Article Snippet: Youssef M. Hassan: Performed all experimental work, nanocomposite synthesis, computational modeling (quantum chemistry, molecular dynamics, PBPK, systems biology, machine learning), data analysis, and manuscript writing.

Techniques: Control, Binding Assay, Adsorption