Johnson Retnaraj Samuel Selvan Christyraj

Piperine (PIP), a plant alkaloid with anti-inflammatory and antioxidant effects, has poor solubility and bioavailability, limiting its therapeutic potential in macrophage-mediated inflammatory and oxidative stress conditions. Despite various nanocarrier systems being explored for bioactive compounds, the specific combination of mannose-functionalized chitosan with dual stabilizers (TPGS and Tween 80) for enhanced macrophage targeting and piperine delivery has not been investigated. We hypothesized that this novel formulation would significantly enhance piperine solubility, macrophage uptake, and anti-inflammatory/antioxidant effects compared to conventional systems, while modulating apoptosis-related pathways. This study evaluated targeted and non-targeted nanoparticles synthesized by ionic gelation and emulsification using RAW 264.7 and THP-1 macrophages. FTIR, UV–Vis, XRD, and CHNS confirmed mannose conjugation, while SEM, TEM, and AFM revealed morphology. Physicochemical properties were assessed by DLS, encapsulation efficiency (EE%), drug loading (DL%), and stability. Biological evaluations included drug release, cytotoxicity (MTT), apoptosis analysis (Annexin V–FITC/PI staining), cellular uptake (fluorescence microscopy with coumarin-6), anti-inflammatory assays (extracellular and intracellular NO inhibition, cytokine suppression), antioxidant activity (DPPH, ABTS, FRAP, TAC), intracellular ROS/RNS, and apoptosis-related markers. Targeted nanoparticles showed larger mean size (162 nm) versus non-targeted ones (78 nm). EE% was 82% (targeted) and 92% (non-targeted). Both demonstrated sustained 72 h release. Cellular uptake was significantly greater for targeted nanoparticles. Both formulations reduced NO and pro-inflammatory cytokines, regulated apoptosis-associated markers, and induced controlled apoptosis at higher concentrations, with stronger effects observed for targeted particles. Antioxidant activity increased dose-dependently, with targeted nanoparticles showing superior intracellular ROS/RNS suppression. This novel multi-functional platform efficiently encapsulates PIP, enhances macrophage targeting, modulates apoptosis pathways, and demonstrates superior therapeutic promise for inflammation-related disorders.

Background: Piper nigrum L. (PNL) is a rich source of piperine, a bioactive alkaloid with pharmaceutical, cosmetic, nutritional supplement, and agricultural applications, yet efficient and sustainable extraction methods remain underexplored. Methods: This study compared ultrasonic bath extraction (UBE) and ultrasonic probe extraction (UPE) using natural deep eutectic solvents (NADES) for isolating piperine from PNL fruits. Six NADES formulations were screened, with NADES-5 (choline chloride:glycerin:urea, 1:1:1) showing superior performance. Response surface methodology with a Box–Behnken design optimized extraction parameters, including liquid-to-solid ratio, extraction time, temperature, and water content, for both UBE and UPE. Results: Optimized UPE consistently outperformed UBE, yielding 49.97 mg/g of piperine versus 25.67 mg/g under identical NADES conditions. Comprehensive characterization using TLC, HPTLC, UV, FTIR, Raman, HPLC, NMR, XRD, SEM, and EDX confirmed the successful isolation and structural integrity of piperine, with samples obtained via UPE exhibiting higher purity (98.7% vs. 95.2%) and enhanced crystallinity. In vitro cytotoxicity assays demonstrated that piperine extracted by UPE showed stronger activity against C2C12 myoblasts (IC₅₀: 24.3 μg/mL vs. 40.6 μg/mL) and greater anticancer effects in MCF-7 and HT-29 cells compared to piperine extracted by UBE. Antioxidant evaluation via DPPH, ABTS, FRAP, and TAC assays, along with intracellular reactive oxygen and nitrogen species suppression in THP-1 and RAW 264.7 macrophages, further confirmed the superior biological potential of the UPE-derived piperine sample. Conclusions: These findings indicate that UPE using NADES is a sustainable approach for high-yield piperine extraction with enhanced purity and bioactivity, supporting its potential for pharmaceutical applications.

Acquisition and consolidation of recognition memory involves dopaminergic activity, which is determined to be dysregulated in Alzheimer disease (AD). Aromatic turmerone (Ar-tur), one of the major bioactive components of Curcuma longa, has been identified to protect dopaminergic neurons from various pathological insults. We have computationally investigated the drug-like nature and the therapeutic potential of Ar-tur in treating AD and have compared its targets with FDA-approved AD drugs. Cheminformatics, pharmacokinetics, toxicity, and system pharmacology studies were performed in predicting the drug-like properties, protein targets, toxicity profiles, target's functional association, and enrichment analysis of Ar-tur. Notably, Ar-tur had targeted 94 proteins, and 29 proteins of these proteins were among the top 15 targets of the 20 FDA-approved drugs. These targets were identified to be dysregulated in the hippocampus and entorhinal cortex in the AD brain, signifying that targets of Ar-tur play roles in disease modification and regulation. Based on gene ontology analysis, KEGG, Reactome, and WikiPathways, we identified that Ar-tur regulates dopamine neurotransmitter receptor activity and dopamine binding activity. Dopamine receptors (DRs), DR2 and DR4, were among its top 15 targets of Ar-tur. Molecular docking of DRs with dopamine, cariprazine, brexpiprazole, RO-10-5824, CP226269, and donepezil connotes that Ar-tur behaves like an agonist of DR2 and DR4 by binding to the active site residues like the dopamine and DR agonists. This is the first report stating the impressive drug-like properties of Ar-tur and holds the potential to rescue the vulnerable dopaminergic system by regulating DRs.

Exosomes are the smallest of the extracellular vesicles, essential in intercellular communication by transporting lipids, proteins, and RNA, thereby influencing the recipient cells. The exosomes from cerebrospinal fluid, central nervous system, and other body fluids have substantial modifications in their molecular composition under various disease conditions. Recent research suggests their potential use as biomarkers for early and prompt detection of neurodegenerative disorders, particularly Parkinson's disease (PD), which is characterized by the misfolded aggregates of the α-synuclein protein. Notably, exosome-based dysregulated microRNAs (miRNAs) are emerging as potential biomarkers for accurate Parkinson's disease diagnosis. Exosomal miRNAs' potential and limitations as early diagnostic biomarkers for Parkinson's disease are discussed comprehensively in this review.

Selenium, an essential trace element, plays a critical role in antioxidant defence and cellular homeostasis. Selenium injection (500 nM to 300 µM) significantly enhanced anterior blastema regeneration in the earthworm Perionyx excavatus, with maximal blastema length observed at 300 µM (4.30 ± 0.12 mm vs. 3.09 ± 0.08 mm in controls). Histological analysis revealed concentration-dependent tissue restoration, including longitudinal muscle layer thickness (40.9 ± 0.43 µm at 300 µM vs. 29.5 ± 0.38 µm in controls) and expansion of epithelial/circular muscle layers. Quantitative PCR identified pex-miR-219 as uniquely suppressed by selenium, showing progressive downregulation (0.60-, 0.47-, 0.36-, and 0.22-fold at 1 µM, 100 µM, 200 µM, and 300 µM, respectively; p < 0.001). Computational target prediction (miRanda, RNAhybrid, RNA22) and qPCR validation confirmed transcription factor Su(H) as a high-affinity target of pex-miR-219, with Su(H) mRNA upregulated 8.26-fold at 300 µM (p < 0.001). Molecular dynamics simulations demonstrated stable binding between pex-miR-219 and Su(H), evidenced by RMSD stabilization (~ 0.25 nm after 4 ns), low RMSF in helical regions, and persistent hydrogen bonds (30–35 bonds). These findings establish a selenium-mediated regulatory axis where suppression of pex-miR-219 relieves Su(H) repression, accelerating blastemal growth and tissue restoration. The study shows that selenium could be used as a treatment to influence miRNA pathways, which helps improve healing processes. By elucidating the pex-miR-219/Su(H) interaction, this work provides a molecular framework for developing selenium-based interventions in regenerative medicine, particularly for conditions requiring accelerated tissue repair, such as wound healing or post-traumatic regeneration. These insights could inform future strategies to harness trace elements for targeted gene regulation in human regenerative therapies.