Synergistic Therapeutic Potential of Resveratrol–Quercetin Loaded Selenium Nanoparticles: Synthesis, Characterization, and In Vitro Pharmacological Evaluation

Synergistic Therapeutic Potential of Resveratrol–Quercetin Loaded Selenium Nanoparticles: Synthesis, Characterization, and In Vitro Pharmacological Evaluation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Synergistic Therapeutic Potential of Resveratrol–Quercetin Loaded Selenium Nanoparticles: Synthesis, Characterization, and In Vitro Pharmacological Evaluation Thirumalaikumaran Rathinam, Vandhana Vijayakumar, Abida Haripriya Rammohan, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8261754/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract Chronic diseases such as diabetes, inflammation, and oxidative stress continue to pose major global health challenges, while conventional therapies remain limited by poor bioavailability and side effects. Natural polyphenols like resveratrol and quercetin are well known for their antioxidant, anti-inflammatory, and antidiabetic properties, yet their clinical potential is restricted due to rapid metabolism and low solubility. Nanotechnology-based approaches offer a promising solution by enhancing stability, solubility, and targeted delivery of such bioactive compounds. In this study, resveratrol–quercetin functionalized selenium nanoparticles (QR-SeNPs) were synthesized using a green reduction method, with the phytochemicals serving as both reducing and stabilizing agents. The nanoparticles were characterized by UV–Vis spectroscopy, SEM, FTIR, DLS, zeta potential, and XRD, confirming spherical morphology, nanoscale size (200–300 nm), good colloidal stability (–30 to − 40 mV), and predominantly amorphous structure. Biological evaluations revealed that QR-SeNPs exhibited strong anti-inflammatory activity, achieving ~ 80% inhibition of protein denaturation at 50 µg/mL, alongside potent antioxidant activity with ~ 90% scavenging in DPPH and H₂O₂ assays. In antidiabetic assays, they demonstrated significant enzyme inhibition (81% for α-amylase and 75% for β-glucosidase), comparable to standard drugs. Cytotoxicity assessment showed minimal lethality at therapeutic concentrations, with moderate effects observed only at higher doses. These findings indicate that green-synthesized QR-SeNPs represent a multifunctional and biocompatible nanoplatform with strong potential for managing oxidative stress, inflammation, and diabetes, warranting further in vivo and mechanistic investigations. Resveratrol Quercetin Selenium nanoparticles Green synthesis Polyphenols Nanomedicine Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 1. Introduction Nanotechnology has emerged as a transformative field, revolutionizing medicine, agriculture, electronics, energy, and environmental sciences.[ 1 ] At the nanoscale, materials exhibit unique properties such as high surface-to-volume ratios, tunable morphologies, and distinct electronic characteristics, which enable applications in catalysis, diagnostics, and therapeutics.[ 2 , 3 ] Nanomaterials are classified into four generations: (i) passive nanostructures with intrinsic nanoscale properties, (ii) active nanostructures engineered to respond to stimuli such as pH, heat, or light, (iii) nanosystems integrating multiple components for targeted applications like drug delivery, and (iv) molecular nanosystems designed with precise molecular-level control for advanced biomedical and technological uses.[ 4 ] The field has also shifted from traditional “top-down” manufacturing to “bottom-up” approaches, where atoms and molecules self-assemble into complex structures, offering greater precision, efficiency, and sustainability.[ 5 ] Despite these advancements, the burden of chronic diseases such as diabetes, inflammation, and oxidative stress continues to rise globally.[ 6 ] Conventional therapies for these conditions are often limited by low bioavailability, poor target specificity, systemic toxicity, and suboptimal patient compliance. This highlights the urgent need for innovative therapeutic strategies that are both safe and effective.[ 7 , 8 ] Natural polyphenolic compounds, particularly quercetin and resveratrol, have gained significant attention for their antioxidant, anti-inflammatory, cardioprotective, antidiabetic, and anticancer properties.[ 9 ] Quercetin, found in onions, apples, and green tea, reduces oxidative stress and offers protection against metabolic and neurodegenerative disorders, while resveratrol, present in grapes, peanuts, and red wine, regulates key pathways such as SIRT1 and NF-κB, influencing inflammation, metabolism, and cancer progression.[ 10 ] However, the clinical translation of these phytochemicals is hindered by poor solubility, rapid metabolism, and limited bioavailability.[ 11 ] Nanotechnology-based delivery systems provide a promising solution to these challenges. Encapsulating bioactive compounds in nanoparticles improves solubility, enhances stability, allows for controlled and sustained release, and enables targeted drug delivery with reduced side effects.[ 12 , 13 ] Among various nanocarriers, selenium nanoparticles (SeNPs) stand out due to their potent antioxidant activity, biocompatibility, and lower toxicity compared to conventional selenium compounds.[ 14 ] Green synthesis methods, employing phytochemicals such as resveratrol and quercetin as reducing and stabilizing agents, further enhance the eco-friendliness and therapeutic potential of these nanocarriers.[ 15 ] The convergence of nanotechnology and natural compound pharmacology presents a powerful therapeutic strategy.[ 16 , 17 ] Selenium nanoparticles loaded with resveratrol and quercetin may synergistically enhance bioavailability, stability, and pharmacological efficacy, offering a multifunctional nanoplatform against oxidative stress, inflammation, and diabetes. This study aims to synthesize, characterize, and evaluate the in vitro pharmacological activities of resveratrol–quercetin functionalized selenium nanoparticles[QR-SeNPs], with a focus on their anti-inflammatory, antioxidant, antidiabetic, and cytotoxic properties. 2. Materials and Methods Resveratrol and quercetin were purchased from Yucca Enterprises, Mumbai. Ethanol, distilled water, and selenium sulphate were procured from Saveetha Medical College. All chemicals used were of analytical grade and used without further purification. 2.1 Preparation of QR-SeNPs Resveratrol was dissolved in a solvent mixture of 3 mL ethanol and 7 mL distilled water, while quercetin was prepared separately in a 1:1 mixture of ethanol and water (5 mL each). Both solutions were agitated for 10 minutes to achieve uniform dispersion.[ 18 ] A 30 mM selenium sulfate solution was freshly prepared and used as the precursor. To initiate nanoparticle formation, 80 mL of the selenium solution was combined with 10 mL each of the resveratrol and quercetin solutions in a conical flask. The reaction mixture was kept under continuous shaking for 12 hours to facilitate initial reduction and interaction, followed by magnetic stirring for an additional 25 hours to enhance homogeneity and ensure complete reaction.[ 19 , 20 ] The resulting suspension was then centrifuged at 8000 rpm for 10 minutes to pellet the synthesized nanoparticles, while unreacted components remained in the supernatant.[ 20 ] 2.2 Characterization Techniques The formation of selenium nanoparticles (SeNPs) was initially confirmed using UV–Vis spectroscopy (Shimadzu UV-1800) by recording absorption spectra in the range of 200–800 nm. Scanning electron microscopy (SEM, FEI QUANTA 250 FEG) coupled with energy-dispersive X-ray spectroscopy (EDS) was used to evaluate the morphology, surface structure, and elemental composition of the nanoparticles. Fourier transform infrared (FTIR) spectra were obtained using a PerkinElmer Spectrum IR spectrometer in the range of 4000–400 cm⁻¹ to identify the functional groups involved in nanoparticle stabilization by resveratrol and quercetin. Particle size distribution and zeta potential were analyzed using a Malvern Zetasizer (ZS XPLORER) at 25°C to assess hydrodynamic diameter, polydispersity index (PDI), and colloidal stability. The crystalline nature and structural characteristics of the synthesized nanoparticles were further investigated by X-ray diffraction (XRD, Bruker AXS D8 Advance) using Cu Kα radiation over a 2θ range of 10°–80°.[ 21 , 22 ] 2.3 Biological activities The biological activities of QR-SeNPs were evaluated through standard in vitro pharmacological assays, including anti-inflammatory, antioxidant, antidiabetic, and cytotoxicity studies. 2.3.1. Anti-inflammatory activity Protein Denaturation Assay: Egg albumin was mixed with phosphate buffer saline (PBS) and treated with varying concentrations of QR-SeNPs (10–50 µg/mL). The mixtures were incubated at 70°C for 5 min to induce protein denaturation, cooled to room temperature, and absorbance was measured at 660 nm. Diclofenac served as the reference drug. Percentage inhibition of protein denaturation was calculated relative to control.[ 23 ] Membrane Stabilization Assay: A 10% human red blood cell (RBC) suspension was prepared and mixed with different concentrations of QR-SeNPs (10–50 µg/mL). The mixtures were incubated at 56°C for 30 min, cooled, and centrifuged at 2500 rpm for 5 min. Absorbance of the supernatant was recorded at 560 nm to assess hemolysis. The extent of membrane stabilization was expressed as percent inhibition compared with the standard diclofenac.[ 24 ] 2.3.2. Antioxidant activity DPPH Radical Scavenging Assay: A 0.1 mM solution of DPPH in ethanol was prepared, and equal volumes were mixed with QR-SeNPs at concentrations ranging from 10–50 µg/mL. After incubation in the dark for 30 min, absorbance was measured at 517 nm. Ascorbic acid was used as the standard. The percentage of radical scavenging was calculated using control and sample absorbance values.[ 25 ] Hydrogen Peroxide (H₂O₂) Scavenging Assay: A solution of H₂O₂ (40 mM) in phosphate buffer (pH 7.4) was prepared. QR-SeNPs at different concentrations (10–50 µg/mL) were added and incubated for 10 min in the dark. Absorbance was measured at 530 nm, and scavenging activity was calculated relative to control.[ 26 ] 2.3.3. Antidiabetic activity α-Amylase Inhibition Assay: A mixture of α-amylase enzyme solution and QR-SeNPs (10–50 µg/mL) was pre-incubated at room temperature for 30 min. Starch solution (1%) was added as substrate and incubated for 10 min. The reaction was terminated with dinitrosalicylic acid (DNSA) reagent, boiled for 5 min, cooled, and absorbance was measured at 540 nm. Acarbose was used as the standard.[ 27 ] β-Glucosidase Inhibition Assay: QR-SeNPs (10–50 µg/mL) were incubated with β-glucosidase enzyme solution for 30 min, followed by the addition of p-nitrophenyl-β-D-glucopyranoside (pNPG) as substrate. The mixture was incubated at 37°C for 15–30 min, and the reaction was stopped with 1 M sodium carbonate. Absorbance was measured at 405 nm, and percentage inhibition was calculated.[ 28 ] 2.3.4. Cytotoxicity assay The cytotoxic potential of QR-SeNPs was evaluated using a brine shrimp lethality assay. Brine shrimp eggs were hatched in artificial seawater under aeration and light for 48 h. Nauplii were collected and transferred into vials containing seawater with QR-SeNPs at different concentrations (5–80 µg/mL). After 24–48 h of exposure, surviving nauplii were counted, and mortality percentage was determined relative to the control group.[ 29 ] 3. Result 3.1 Synthesis and Visual Observation The successful synthesis of quercetin–resveratrol-loaded selenium nanoparticles (QR-SeNPs) was confirmed by the observable color change of the reaction mixture from colorless to reddish-orange, indicating the reduction of Se⁴⁺ ions into elemental selenium as shown in Fig. 1 . This aligns with previous green synthesis reports, where phytochemicals serve as both reducing and stabilizing agents. The nanoparticles remained stable at room temperature for several weeks, suggesting effective capping by polyphenolic compounds. 3.2 UV–Visible Spectroscopy The UV–Vis spectra of the synthesized nanoparticles displayed a distinct absorption peak at ~ 265 nm, characteristic of selenium nanoparticles. The Fig. 2 shows the UV spectrum of SeNPs. This peak corresponds to the surface plasmon resonance (SPR) band and is within the reported range of 250–270 nm for SeNPs. The presence of a sharp, well-defined peak supports the successful synthesis, while its position suggests nanoscale dimensions and good colloidal stability. 3.3 Scanning Electron Microscopy (SEM) SEM analysis revealed spherical nanoparticles with relatively uniform size distribution as shown in the Fig. 3 . The absence of significant aggregation indicates efficient stabilization by resveratrol and quercetin. Spherical morphology is advantageous for biomedical applications, as it enhances cellular uptake and interaction. 3.4 Fourier Transform Infrared (FTIR) Analysis The Fig. 4 FTIR spectrum confirmed the presence of functional groups associated with polyphenolic capping agents. Peaks at 3201 cm⁻¹ (O–H stretching), 1603 cm⁻¹ (C = C aromatic stretching), and 1063–985 cm⁻¹ (C–O/C–N stretching) demonstrate the involvement of hydroxyl and carbonyl groups in nanoparticle stabilization. Se–O stretching vibrations observed below 600 cm⁻¹ further indicate selenium–oxygen interactions. These findings are consistent with previous green synthesis studies, suggesting that phytochemicals act as stabilizers, preventing aggregation. 3.5 Particle Size and Zeta Potential Dynamic light scattering (DLS) analysis showed two populations of nanoparticles, with a major peak between 200–300 nm and a minor peak at larger sizes, indicating partial aggregation as shown in the Fig. 5 . Zeta potential values between − 30 and − 40 mV confirmed good colloidal stability due to electrostatic repulsion illustrated by Fig. 6 . Stable zeta values support their suitability for biological applications. 3.6 X-Ray Diffraction (XRD) XRD analysis (Fig. 7 ) indicates that the QR-SeNPs were predominantly amorphous (81.4%) with minor crystalline phases (18.6%). The broad halo observed between 15°–35° confirmed amorphous nature, while sharp peaks at 2θ values of 16.8°, 22.6°, and 24.4° indicated partial crystallinity. Such mixed-phase structures can influence solubility, reactivity, and biological activity. 3.7 Biological activity 3.7.1 Anti-Inflammatory Activity The Figs. 8 & 9 illustrate the anti-inflammatory potential of QR-SeNPs was assessed via BSA and membrane stabilization assays. Both assays showed concentration-dependent inhibition of protein denaturation, with QR-SeNPs demonstrating ~ 80% inhibition at 50 µg/mL, comparable to the standard drug diclofenac. These findings highlight their potential as natural, nanocarrier-based anti-inflammatory agents with reduced side-effect profiles compared to conventional drugs. 3.7.2 Antioxidant Activity In both DPPH and H₂O₂ assays, QR-SeNPs showed strong radical scavenging ability (Figs. 10 & 11 ). At 50 µg/mL, inhibition values reached ~ 90%, nearly matching the standard. Compared with crude plant extracts and gold nanoparticle formulations from earlier reports, the QR-SeNPs displayed superior antioxidant potential, likely due to synergistic interactions between selenium and polyphenolic compounds. 3.7.3 Antidiabetic Activity Enzyme inhibition assays revealed significant α-amylase and β-glucosidase inhibition by QR-SeNPs in a dose-dependent manner (Figs. 12 & 13 ). At 50 µg/mL, inhibition values approached 81% and 75%, respectively, closely comparable to the standard. These findings suggest QR-SeNPs can effectively reduce postprandial hyperglycemia, making them promising candidates for diabetes management. 3.7.4 Cytotoxic Evaluation Cytotoxicity was evaluated using the brine shrimp lethality assay. The Fig. 14 shows that no significant lethality was observed at lower concentrations (5–20 µg/mL), but higher doses (40–80 µg/mL) showed moderate toxicity over 48 hours. These results suggest that QR-SeNPs are relatively safe at therapeutic concentrations, but dose optimization is essential for clinical translation. 4. Discussion The present study demonstrates the successful green synthesis of SeNPs using resveratrol and quercetin as natural reducing and stabilizing agents. The observed color change during synthesis and the characteristic UV–Vis absorption peak at ~ 265 nm confirmed nanoparticle formation, consistent with earlier reports on SeNPs synthesized via phytochemicals. SEM images revealed predominantly spherical nanoparticles with uniform morphology, while FTIR spectra confirmed the involvement of hydroxyl, carbonyl, and aromatic groups from resveratrol and quercetin in stabilization. DLS analysis showed particle sizes primarily in the 200–300 nm range, with zeta potential values between − 30 and − 40 mV, indicative of stable colloidal dispersions. XRD analysis revealed a predominantly amorphous structure with minor crystalline phases, a feature that has been reported to improve biological reactivity of selenium-based nanomaterials. Biological evaluation of the nanoparticles revealed promising pharmacological activities. In anti-inflammatory assays, QR-SeNPs exhibited significant inhibition of protein denaturation and membrane stabilization, reaching ~ 80% at 50 µg/mL, values closely comparable to diclofenac. These findings suggest that the synergistic action of selenium with polyphenolic compounds enhances the ability to suppress inflammatory processes, potentially with reduced side effects compared to synthetic drugs. Similarly, QR-SeNPs demonstrated strong antioxidant activity in both DPPH and H₂O₂ assays, with inhibition levels approaching 90% at higher concentrations. Compared to crude plant extracts and previously reported metal nanoparticles, the QR-SeNPs showed superior radical scavenging activity, likely due to their nanoscale size, high surface area, and the synergistic effects of selenium with resveratrol and quercetin. Antidiabetic activity was confirmed through α-amylase and β-glucosidase inhibition assays, where QR-SeNPs exhibited dose-dependent inhibition (up to ~ 81% and 75% at 50 µg/mL, respectively). These findings are in agreement with prior studies reporting that selenium nanoparticles can modulate carbohydrate metabolism and support glycemic control. The observed effects are further enhanced by the intrinsic antidiabetic properties of resveratrol and quercetin, which are known to improve insulin sensitivity and inhibit glucose absorption. Cytotoxicity studies revealed minimal toxicity at lower concentrations, but higher concentrations (≥ 40 µg/mL) showed moderate lethality in the brine shrimp assay after 48 h. This highlights the relative safety of QR-SeNPs at therapeutic doses, while emphasizing the importance of dose optimization in clinical applications. Collectively, these results align with previous nanoparticle-based studies but also highlight the novelty of combining resveratrol and quercetin in SeNPs. The synergistic effects of these polyphenols with selenium appear to enhance anti-inflammatory, antioxidant, and antidiabetic properties, supporting their potential as multifunctional therapeutic agents. 5. Conclusion This study successfully synthesized selenium nanoparticles using resveratrol and quercetin via a green synthesis approach, yielding stable, spherical nanoparticles with favorable physicochemical properties. Characterization confirmed effective capping by phytochemicals, amorphous structural features, and good colloidal stability. Biological evaluation demonstrated that QR-SeNPs possess significant anti-inflammatory, antioxidant, and antidiabetic activities, with efficacy comparable to standard drugs, and exhibit low cytotoxicity at therapeutic concentrations. These findings highlight the potential of resveratrol–quercetin-loaded selenium nanoparticles as a multifunctional nanoplatform for the management of oxidative stress, inflammation, and diabetes. Further in vivo studies and mechanistic investigations are warranted to validate their clinical applicability and establish safe dosage ranges. Declarations CrediT authorship contribution statement Thirumalaikumaran Rathinam: Writing – review & editing. Vandhana Vijayakumar: Writing – orginal draft. Abida Haripriya Rammohan, Dharshini Sagadevan, Udhayakumar Thangavelu: Methodology. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding Not applicable Author Contribution Vandhana Vijayakumar was involved in manuscript writing and characterization. Dr. Thirumalaikumaran Rathinam helped with manuscript corrections and valuable suggestions.Thirumalaikumaran Rathinam: Writing – review & editing. Vandhana Vijayakumar: Writing – orginal draft. Abida Haripriya Rammohan, Dharshini Sagadevan, Udhayakumar Thangavelu: Methodology. Data Availability Data will be made available on request. References P. Boisseau, B. Loubaton, Nanomedicine, nanotechnology in medicine. C R Phys. 12 (7) (2011). 10.1016/j.crhy.2011.06.001 M.F. Bertino, Introduction to Nanotechnology .; 2022. 10.1142/12142 J. Nandhini, E. Karthikeyan, S. Rajeshkumar, Nanomaterials for wound healing: Current status and futuristic frontier. Biomedical Technol. 6 (2024). 10.1016/j.bmt.2023.10.001 S. Malik, K. Muhammad, Y. Waheed, Nanotechnology, A Revolution in Modern Industry. Molecules. 28 (2) (2023). 10.3390/molecules28020661 Y. Li, J. Yao, C. Han et al., Quercetin, inflammation and immunity. Nutrients. 8 (3) (2016). 10.3390/nu8030167 G.S. Kelly, Quercetin, Altern. Med. Rev. 2011; 16 (2) S.S. Baghel, N. Shrivastava, R. Singh, B.P. Agrawal, S. Rajput, a Review of Quercetin: Antioxidant and Anticancer Properties. World J. Pharm. Pharm. Sci. 2012; 1 (1) K. Elumalai, S. Srinivasan, A. Shanmugam, Review of the efficacy of nanoparticle-based drug delivery systems for cancer treatment. Biomedical Technol. 5 (2024). 10.1016/j.bmt.2023.09.001 K.P.L. Bhat, J.W. Kosmeder, J.M. Pezzuto, Biological effects of resveratrol. Antioxid. Redox Signal. 3 (6) (2001). 10.1089/152308601317203567 R. Mikstacka, A.M. Rimando, E. Ignatowicz, Antioxidant effect of trans-Resveratrol, pterostilbene, quercetin and their combinations in human erythrocytes In vitro. Plant Foods Hum. Nutr. 65 (1) (2010). 10.1007/s11130-010-0154-8 G.M.F. Calixto, J. Bernegossi, De L.M. Freitas, C.R. Fontana, M. Chorilli, A.M. Grumezescu, Nanotechnology-based drug delivery systems for photodynamic therapy of cancer: A review. Molecules. 21 (3) (2016). 10.3390/molecules21030342 O.C. Farokhzad, R. Langer, Impact of nanotechnology on drug delivery. ACS Nano. 3 (1) (2009). 10.1021/nn900002m M. Devaraji, P.V. Thanikachalam, Phytoconstituents as emerging therapeutics for breast cancer: Mechanistic insights and clinical implications. Cancer Pathogenesis Therapy Published online Febr. 28 (2025). 10.1016/J.CPT.2025.02.006 M.J. Saadh, R.K. Jadullah, Nanotechnology in Drug Delivery. Pharmacologyonline. 3 (2021). 10.17148/ijarcce.2016.55255 K.K. Karthik, B.V. Cheriyan, S. Rajeshkumar, M. Gopalakrishnan, A review on selenium nanoparticles and their biomedical applications. Biomedical Technol. 6 (2024). 10.1016/j.bmt.2023.12.001 R. Kanwar, J. Rathee, D.B. Salunke, S.K. Mehta, Green nanotechnology-driven drug delivery assemblies. ACS Omega. 4 (5) (2019). 10.1021/acsomega.9b00304 K.K. Karunakar, P.V. Thanikachalam, S.M. Dhanalakshmi, P. Kesharwani, B.V. Cheriyan, Hinokitiol attenuates gentamicin-induced nephrotoxicity by reversing oxidative stress and inflammation. Pharmacol. Res. - Mod. Chin. Med. 10 (2024). 10.1016/j.prmcm.2024.100410 X. Xiao, H. Deng, X. Lin et al., Selenium nanoparticles: Properties, preparation methods, and therapeutic applications. Chem. Biol. Interact. 378 (2023). 10.1016/j.cbi.2023.110483 R.J. White, R. Luque, V.L. Budarin, J.H. Clark, D.J. Macquarrie, Supported metal nanoparticles on porous materials. Methods and applications. Chem. Soc. Rev. 38 (2) (2009). 10.1039/b802654h S. Kanimozhi, R. Durga, M. Sabithasree et al., Biogenic synthesis of silver nanoparticle using Cissus quadrangularis extract and its invitro study. J. King Saud Univ. Sci. 34 (4) (2022). 10.1016/j.jksus.2022.101930 M. Mutakin, R. Fauziati, F.N. Fadhilah, A. Zuhrotun, R. Amalia, Y.E. Hadisaputri, Pharmacological Activities of Soursop (Annona muricata Lin). Molecules. 27 (4) (2022). 10.3390/molecules27041201 M. Devaraji, P.V. Thanikachalam, S.R. AS R, Microwave-assisted synthesis of copper oxide nanoparticles using an Andrographis paniculata leaf extract: Characterization and multifunctional biological activities. Nano-Structures Nano-Objects. 40 , 101376 (2024). 10.1016/J.NANOSO.2024.101376 S.V. Ghumre, Assessment of In-vitro Anti-Inflammatory Activity of Cynodon Dactylon and Acyclovir Showing Synergistic Effect by Albumin Denaturation and Membrane Stabilization Assay. Mod. Approaches Drug Designing. 1 (2) (2017). 10.31031/madd.2017.01.000506 C.S. Kumari, N. Yasmin, M.R. Hussain, M. Babuselvam, Invitro anti-inflammatory and anti-artheritic property of rhizopora mucronata leaves. Int. J. Pharma Sci. Res. 2015; 6 (3) S. Chatterjee, R J, S R. Green Synthesis of Zinc Oxide Nanoparticles Using Chamomile and Green Tea Extracts and Evaluation of Their Anti-inflammatory and Antioxidant Activity: An In Vitro Study. Cureus . Published online 2023. 10.7759/cureus.46088 İ. Gulcin, S.H. Alwasel, DPPH Radical Scavenging Assay. Processes. 11 (8) (2023). 10.3390/pr11082248 A.L. Hauvermale, R.S. Parveen, T.J. Harris et al., Streamlined alpha-amylase assays for wheat preharvest sprouting and late maturity alpha-amylase detection. Agrosystems Geosci. Environ. 6 (1) (2023). 10.1002/agg2.20327 S. Yin, Y. Yang, W. Xu et al., Research progress of β-glucosidase assay and its application. Food Ferment. Industries. 48 (13) (2022). 10.13995/j.cnki.11-1802/ts.031312 M.A.R. Shah, R.A. Khan, M. Ahmed, Phytochemical analysis, cytotoxic, antioxidant and anti-diabetic activities of the aerial parts of Sorghum halepense. Bangladesh J. Pharmacol. 14 (3) (2019). 10.3329/bjp.v14i3.40292 Additional Declarations No competing interests reported. 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13","display":"","copyAsset":false,"role":"figure","size":67094,"visible":true,"origin":"","legend":"\u003cp\u003eβ- Glucosidase Assay of QR-SeNPs\u003c/p\u003e","description":"","filename":"image13.png","url":"https://assets-eu.researchsquare.com/files/rs-8261754/v1/7606ac028e1cb31a399d766e.png"},{"id":97691941,"identity":"fb96dae2-a844-4894-849b-3334d8173582","added_by":"auto","created_at":"2025-12-08 11:08:24","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":30672,"visible":true,"origin":"","legend":"\u003cp\u003eCytotoxic evaluation of QR-SeNPs\u003c/p\u003e","description":"","filename":"image14.png","url":"https://assets-eu.researchsquare.com/files/rs-8261754/v1/5a2695120a43ea47e29337d2.png"},{"id":97902456,"identity":"2a6f3271-9786-4acd-8a8d-d3b85e2d05b1","added_by":"auto","created_at":"2025-12-10 15:52:25","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1797447,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8261754/v1/a4122270-13a8-40bd-931c-2e8254dbfac8.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Synergistic Therapeutic Potential of Resveratrol–Quercetin Loaded Selenium Nanoparticles: Synthesis, Characterization, and In Vitro Pharmacological Evaluation","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNanotechnology has emerged as a transformative field, revolutionizing medicine, agriculture, electronics, energy, and environmental sciences.[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e] At the nanoscale, materials exhibit unique properties such as high surface-to-volume ratios, tunable morphologies, and distinct electronic characteristics, which enable applications in catalysis, diagnostics, and therapeutics.[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e] Nanomaterials are classified into four generations: (i) passive nanostructures with intrinsic nanoscale properties, (ii) active nanostructures engineered to respond to stimuli such as pH, heat, or light, (iii) nanosystems integrating multiple components for targeted applications like drug delivery, and (iv) molecular nanosystems designed with precise molecular-level control for advanced biomedical and technological uses.[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e] The field has also shifted from traditional \u0026ldquo;top-down\u0026rdquo; manufacturing to \u0026ldquo;bottom-up\u0026rdquo; approaches, where atoms and molecules self-assemble into complex structures, offering greater precision, efficiency, and sustainability.[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/p\u003e\u003cp\u003eDespite these advancements, the burden of chronic diseases such as diabetes, inflammation, and oxidative stress continues to rise globally.[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e] Conventional therapies for these conditions are often limited by low bioavailability, poor target specificity, systemic toxicity, and suboptimal patient compliance. This highlights the urgent need for innovative therapeutic strategies that are both safe and effective.[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u003c/p\u003e\u003cp\u003eNatural polyphenolic compounds, particularly quercetin and resveratrol, have gained significant attention for their antioxidant, anti-inflammatory, cardioprotective, antidiabetic, and anticancer properties.[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] Quercetin, found in onions, apples, and green tea, reduces oxidative stress and offers protection against metabolic and neurodegenerative disorders, while resveratrol, present in grapes, peanuts, and red wine, regulates key pathways such as SIRT1 and NF-κB, influencing inflammation, metabolism, and cancer progression.[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] However, the clinical translation of these phytochemicals is hindered by poor solubility, rapid metabolism, and limited bioavailability.[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/p\u003e\u003cp\u003eNanotechnology-based delivery systems provide a promising solution to these challenges. Encapsulating bioactive compounds in nanoparticles improves solubility, enhances stability, allows for controlled and sustained release, and enables targeted drug delivery with reduced side effects.[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] Among various nanocarriers, selenium nanoparticles (SeNPs) stand out due to their potent antioxidant activity, biocompatibility, and lower toxicity compared to conventional selenium compounds.[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] Green synthesis methods, employing phytochemicals such as resveratrol and quercetin as reducing and stabilizing agents, further enhance the eco-friendliness and therapeutic potential of these nanocarriers.[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] The convergence of nanotechnology and natural compound pharmacology presents a powerful therapeutic strategy.[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] Selenium nanoparticles loaded with resveratrol and quercetin may synergistically enhance bioavailability, stability, and pharmacological efficacy, offering a multifunctional nanoplatform against oxidative stress, inflammation, and diabetes. This study aims to synthesize, characterize, and evaluate the in vitro pharmacological activities of resveratrol\u0026ndash;quercetin functionalized selenium nanoparticles[QR-SeNPs], with a focus on their anti-inflammatory, antioxidant, antidiabetic, and cytotoxic properties.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cp\u003eResveratrol and quercetin were purchased from Yucca Enterprises, Mumbai. Ethanol, distilled water, and selenium sulphate were procured from Saveetha Medical College. All chemicals used were of analytical grade and used without further purification.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Preparation of QR-SeNPs\u003c/h2\u003e\u003cp\u003eResveratrol was dissolved in a solvent mixture of 3 mL ethanol and 7 mL distilled water, while quercetin was prepared separately in a 1:1 mixture of ethanol and water (5 mL each). Both solutions were agitated for 10 minutes to achieve uniform dispersion.[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] A 30 mM selenium sulfate solution was freshly prepared and used as the precursor. To initiate nanoparticle formation, 80 mL of the selenium solution was combined with 10 mL each of the resveratrol and quercetin solutions in a conical flask. The reaction mixture was kept under continuous shaking for 12 hours to facilitate initial reduction and interaction, followed by magnetic stirring for an additional 25 hours to enhance homogeneity and ensure complete reaction.[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] The resulting suspension was then centrifuged at 8000 rpm for 10 minutes to pellet the synthesized nanoparticles, while unreacted components remained in the supernatant.[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Characterization Techniques\u003c/h2\u003e\u003cp\u003eThe formation of selenium nanoparticles (SeNPs) was initially confirmed using UV\u0026ndash;Vis spectroscopy (Shimadzu UV-1800) by recording absorption spectra in the range of 200\u0026ndash;800 nm. Scanning electron microscopy (SEM, FEI QUANTA 250 FEG) coupled with energy-dispersive X-ray spectroscopy (EDS) was used to evaluate the morphology, surface structure, and elemental composition of the nanoparticles. Fourier transform infrared (FTIR) spectra were obtained using a PerkinElmer Spectrum IR spectrometer in the range of 4000\u0026ndash;400 cm⁻\u0026sup1; to identify the functional groups involved in nanoparticle stabilization by resveratrol and quercetin. Particle size distribution and zeta potential were analyzed using a Malvern Zetasizer (ZS XPLORER) at 25\u0026deg;C to assess hydrodynamic diameter, polydispersity index (PDI), and colloidal stability. The crystalline nature and structural characteristics of the synthesized nanoparticles were further investigated by X-ray diffraction (XRD, Bruker AXS D8 Advance) using Cu Kα radiation over a 2θ range of 10\u0026deg;\u0026ndash;80\u0026deg;.[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 Biological activities\u003c/h2\u003e\u003cp\u003eThe biological activities of QR-SeNPs were evaluated through standard in vitro pharmacological assays, including anti-inflammatory, antioxidant, antidiabetic, and cytotoxicity studies.\u003c/p\u003e\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\u003ch2\u003e2.3.1. Anti-inflammatory activity\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eProtein Denaturation Assay: Egg albumin was mixed with phosphate buffer saline (PBS) and treated with varying concentrations of QR-SeNPs (10\u0026ndash;50 \u0026micro;g/mL). The mixtures were incubated at 70\u0026deg;C for 5 min to induce protein denaturation, cooled to room temperature, and absorbance was measured at 660 nm. Diclofenac served as the reference drug. Percentage inhibition of protein denaturation was calculated relative to control.[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eMembrane Stabilization Assay: A 10% human red blood cell (RBC) suspension was prepared and mixed with different concentrations of QR-SeNPs (10\u0026ndash;50 \u0026micro;g/mL). The mixtures were incubated at 56\u0026deg;C for 30 min, cooled, and centrifuged at 2500 rpm for 5 min. Absorbance of the supernatant was recorded at 560 nm to assess hemolysis. The extent of membrane stabilization was expressed as percent inhibition compared with the standard diclofenac.[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\u003ch2\u003e2.3.2. Antioxidant activity\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eDPPH Radical Scavenging Assay: A 0.1 mM solution of DPPH in ethanol was prepared, and equal volumes were mixed with QR-SeNPs at concentrations ranging from 10\u0026ndash;50 \u0026micro;g/mL. After incubation in the dark for 30 min, absorbance was measured at 517 nm. Ascorbic acid was used as the standard. The percentage of radical scavenging was calculated using control and sample absorbance values.[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eHydrogen Peroxide (H₂O₂) Scavenging Assay: A solution of H₂O₂ (40 mM) in phosphate buffer (pH 7.4) was prepared. QR-SeNPs at different concentrations (10\u0026ndash;50 \u0026micro;g/mL) were added and incubated for 10 min in the dark. Absorbance was measured at 530 nm, and scavenging activity was calculated relative to control.[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.3.3. Antidiabetic activity\u003c/h2\u003e\u003cp\u003e\u003cul\u003e\u003cli\u003e\u003cp\u003eα-Amylase Inhibition Assay: A mixture of α-amylase enzyme solution and QR-SeNPs (10\u0026ndash;50 \u0026micro;g/mL) was pre-incubated at room temperature for 30 min. Starch solution (1%) was added as substrate and incubated for 10 min. The reaction was terminated with dinitrosalicylic acid (DNSA) reagent, boiled for 5 min, cooled, and absorbance was measured at 540 nm. Acarbose was used as the standard.[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003cli\u003e\u003cp\u003eβ-Glucosidase Inhibition Assay: QR-SeNPs (10\u0026ndash;50 \u0026micro;g/mL) were incubated with β-glucosidase enzyme solution for 30 min, followed by the addition of p-nitrophenyl-β-D-glucopyranoside (pNPG) as substrate. The mixture was incubated at 37\u0026deg;C for 15\u0026ndash;30 min, and the reaction was stopped with 1 M sodium carbonate. Absorbance was measured at 405 nm, and percentage inhibition was calculated.[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]\u003c/p\u003e\u003c/li\u003e\u003c/ul\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003e2.3.4. Cytotoxicity assay\u003c/h2\u003e\u003cp\u003eThe cytotoxic potential of QR-SeNPs was evaluated using a brine shrimp lethality assay. Brine shrimp eggs were hatched in artificial seawater under aeration and light for 48 h. Nauplii were collected and transferred into vials containing seawater with QR-SeNPs at different concentrations (5\u0026ndash;80 \u0026micro;g/mL). After 24\u0026ndash;48 h of exposure, surviving nauplii were counted, and mortality percentage was determined relative to the control group.[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"3. Result","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e3.1 Synthesis and Visual Observation\u003c/h2\u003e\u003cp\u003eThe successful synthesis of quercetin\u0026ndash;resveratrol-loaded selenium nanoparticles (QR-SeNPs) was confirmed by the observable color change of the reaction mixture from colorless to reddish-orange, indicating the reduction of Se⁴⁺ ions into elemental selenium as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. This aligns with previous green synthesis reports, where phytochemicals serve as both reducing and stabilizing agents. The nanoparticles remained stable at room temperature for several weeks, suggesting effective capping by polyphenolic compounds.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e3.2 UV\u0026ndash;Visible Spectroscopy\u003c/h2\u003e\u003cp\u003eThe UV\u0026ndash;Vis spectra of the synthesized nanoparticles displayed a distinct absorption peak at ~\u0026thinsp;265 nm, characteristic of selenium nanoparticles. The Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the UV spectrum of SeNPs. This peak corresponds to the surface plasmon resonance (SPR) band and is within the reported range of 250\u0026ndash;270 nm for SeNPs. The presence of a sharp, well-defined peak supports the successful synthesis, while its position suggests nanoscale dimensions and good colloidal stability.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e3.3 Scanning Electron Microscopy (SEM)\u003c/h2\u003e\u003cp\u003eSEM analysis revealed spherical nanoparticles with relatively uniform size distribution as shown in the Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The absence of significant aggregation indicates efficient stabilization by resveratrol and quercetin. Spherical morphology is advantageous for biomedical applications, as it enhances cellular uptake and interaction.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e3.4 Fourier Transform Infrared (FTIR) Analysis\u003c/h2\u003e\u003cp\u003eThe Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e FTIR spectrum confirmed the presence of functional groups associated with polyphenolic capping agents. Peaks at 3201 cm⁻\u0026sup1; (O\u0026ndash;H stretching), 1603 cm⁻\u0026sup1; (C\u0026thinsp;=\u0026thinsp;C aromatic stretching), and 1063\u0026ndash;985 cm⁻\u0026sup1; (C\u0026ndash;O/C\u0026ndash;N stretching) demonstrate the involvement of hydroxyl and carbonyl groups in nanoparticle stabilization. Se\u0026ndash;O stretching vibrations observed below 600 cm⁻\u0026sup1; further indicate selenium\u0026ndash;oxygen interactions. These findings are consistent with previous green synthesis studies, suggesting that phytochemicals act as stabilizers, preventing aggregation.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e3.5 Particle Size and Zeta Potential\u003c/h2\u003e\u003cp\u003eDynamic light scattering (DLS) analysis showed two populations of nanoparticles, with a major peak between 200\u0026ndash;300 nm and a minor peak at larger sizes, indicating partial aggregation as shown in the Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Zeta potential values between \u0026minus;\u0026thinsp;30 and \u0026minus;\u0026thinsp;40 mV confirmed good colloidal stability due to electrostatic repulsion illustrated by Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e. Stable zeta values support their suitability for biological applications.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e3.6 X-Ray Diffraction (XRD)\u003c/h2\u003e\u003cp\u003eXRD analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e) indicates that the QR-SeNPs were predominantly amorphous (81.4%) with minor crystalline phases (18.6%). The broad halo observed between 15\u0026deg;\u0026ndash;35\u0026deg; confirmed amorphous nature, while sharp peaks at 2θ values of 16.8\u0026deg;, 22.6\u0026deg;, and 24.4\u0026deg; indicated partial crystallinity. Such mixed-phase structures can influence solubility, reactivity, and biological activity.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e3.7 Biological activity\u003c/h2\u003e\u003cdiv id=\"Sec18\" class=\"Section3\"\u003e\u003ch2\u003e3.7.1 Anti-Inflammatory Activity\u003c/h2\u003e\u003cp\u003eThe Figs.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e \u0026amp; \u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e illustrate the anti-inflammatory potential of QR-SeNPs was assessed via BSA and membrane stabilization assays. Both assays showed concentration-dependent inhibition of protein denaturation, with QR-SeNPs demonstrating\u0026thinsp;~\u0026thinsp;80% inhibition at 50 \u0026micro;g/mL, comparable to the standard drug diclofenac. These findings highlight their potential as natural, nanocarrier-based anti-inflammatory agents with reduced side-effect profiles compared to conventional drugs.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\u003ch2\u003e3.7.2 Antioxidant Activity\u003c/h2\u003e\u003cp\u003eIn both DPPH and H₂O₂ assays, QR-SeNPs showed strong radical scavenging ability (Figs.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e \u0026amp; \u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003e). At 50 \u0026micro;g/mL, inhibition values reached\u0026thinsp;~\u0026thinsp;90%, nearly matching the standard. Compared with crude plant extracts and gold nanoparticle formulations from earlier reports, the QR-SeNPs displayed superior antioxidant potential, likely due to synergistic interactions between selenium and polyphenolic compounds.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec20\" class=\"Section3\"\u003e\u003ch2\u003e3.7.3 Antidiabetic Activity\u003c/h2\u003e\u003cp\u003eEnzyme inhibition assays revealed significant α-amylase and β-glucosidase inhibition by QR-SeNPs in a dose-dependent manner (Figs.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e12\u003c/span\u003e \u0026amp; \u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e13\u003c/span\u003e). At 50 \u0026micro;g/mL, inhibition values approached 81% and 75%, respectively, closely comparable to the standard. These findings suggest QR-SeNPs can effectively reduce postprandial hyperglycemia, making them promising candidates for diabetes management.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec21\" class=\"Section3\"\u003e\u003ch2\u003e3.7.4 Cytotoxic Evaluation\u003c/h2\u003e\u003cp\u003eCytotoxicity was evaluated using the brine shrimp lethality assay. The Fig.\u0026nbsp;\u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e14\u003c/span\u003e shows that no significant lethality was observed at lower concentrations (5\u0026ndash;20 \u0026micro;g/mL), but higher doses (40\u0026ndash;80 \u0026micro;g/mL) showed moderate toxicity over 48 hours. These results suggest that QR-SeNPs are relatively safe at therapeutic concentrations, but dose optimization is essential for clinical translation.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe present study demonstrates the successful green synthesis of SeNPs using resveratrol and quercetin as natural reducing and stabilizing agents. The observed color change during synthesis and the characteristic UV\u0026ndash;Vis absorption peak at ~\u0026thinsp;265 nm confirmed nanoparticle formation, consistent with earlier reports on SeNPs synthesized via phytochemicals. SEM images revealed predominantly spherical nanoparticles with uniform morphology, while FTIR spectra confirmed the involvement of hydroxyl, carbonyl, and aromatic groups from resveratrol and quercetin in stabilization. DLS analysis showed particle sizes primarily in the 200\u0026ndash;300 nm range, with zeta potential values between \u0026minus;\u0026thinsp;30 and \u0026minus;\u0026thinsp;40 mV, indicative of stable colloidal dispersions. XRD analysis revealed a predominantly amorphous structure with minor crystalline phases, a feature that has been reported to improve biological reactivity of selenium-based nanomaterials.\u003c/p\u003e\u003cp\u003eBiological evaluation of the nanoparticles revealed promising pharmacological activities. In anti-inflammatory assays, QR-SeNPs exhibited significant inhibition of protein denaturation and membrane stabilization, reaching\u0026thinsp;~\u0026thinsp;80% at 50 \u0026micro;g/mL, values closely comparable to diclofenac. These findings suggest that the synergistic action of selenium with polyphenolic compounds enhances the ability to suppress inflammatory processes, potentially with reduced side effects compared to synthetic drugs. Similarly, QR-SeNPs demonstrated strong antioxidant activity in both DPPH and H₂O₂ assays, with inhibition levels approaching 90% at higher concentrations. Compared to crude plant extracts and previously reported metal nanoparticles, the QR-SeNPs showed superior radical scavenging activity, likely due to their nanoscale size, high surface area, and the synergistic effects of selenium with resveratrol and quercetin.\u003c/p\u003e\u003cp\u003eAntidiabetic activity was confirmed through α-amylase and β-glucosidase inhibition assays, where QR-SeNPs exhibited dose-dependent inhibition (up to ~\u0026thinsp;81% and 75% at 50 \u0026micro;g/mL, respectively). These findings are in agreement with prior studies reporting that selenium nanoparticles can modulate carbohydrate metabolism and support glycemic control. The observed effects are further enhanced by the intrinsic antidiabetic properties of resveratrol and quercetin, which are known to improve insulin sensitivity and inhibit glucose absorption. Cytotoxicity studies revealed minimal toxicity at lower concentrations, but higher concentrations (\u0026ge;\u0026thinsp;40 \u0026micro;g/mL) showed moderate lethality in the brine shrimp assay after 48 h. This highlights the relative safety of QR-SeNPs at therapeutic doses, while emphasizing the importance of dose optimization in clinical applications.\u003c/p\u003e\u003cp\u003eCollectively, these results align with previous nanoparticle-based studies but also highlight the novelty of combining resveratrol and quercetin in SeNPs. The synergistic effects of these polyphenols with selenium appear to enhance anti-inflammatory, antioxidant, and antidiabetic properties, supporting their potential as multifunctional therapeutic agents.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study successfully synthesized selenium nanoparticles using resveratrol and quercetin via a green synthesis approach, yielding stable, spherical nanoparticles with favorable physicochemical properties. Characterization confirmed effective capping by phytochemicals, amorphous structural features, and good colloidal stability. Biological evaluation demonstrated that QR-SeNPs possess significant anti-inflammatory, antioxidant, and antidiabetic activities, with efficacy comparable to standard drugs, and exhibit low cytotoxicity at therapeutic concentrations.\u003c/p\u003e\u003cp\u003eThese findings highlight the potential of resveratrol\u0026ndash;quercetin-loaded selenium nanoparticles as a multifunctional nanoplatform for the management of oxidative stress, inflammation, and diabetes. Further in vivo studies and mechanistic investigations are warranted to validate their clinical applicability and establish safe dosage ranges.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eCrediT authorship contribution statement\u003c/h2\u003e\u003cp\u003eThirumalaikumaran Rathinam: Writing \u0026ndash; review \u0026amp; editing. Vandhana Vijayakumar: Writing \u0026ndash; orginal draft. Abida Haripriya Rammohan, Dharshini Sagadevan, Udhayakumar Thangavelu: Methodology.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003ch2\u003eDeclaration of Competing Interest\u003c/h2\u003e\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eNot applicable\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eVandhana Vijayakumar was involved in manuscript writing and characterization. Dr. Thirumalaikumaran Rathinam helped with manuscript corrections and valuable suggestions.Thirumalaikumaran Rathinam: Writing \u0026ndash; review \u0026amp; editing. Vandhana Vijayakumar: Writing \u0026ndash; orginal draft. Abida Haripriya Rammohan, Dharshini Sagadevan, Udhayakumar Thangavelu: Methodology.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eData will be made available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eP. Boisseau, B. Loubaton, Nanomedicine, nanotechnology in medicine. C R Phys. \u003cb\u003e12\u003c/b\u003e(7) (2011). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.crhy.2011.06.001\u003c/span\u003e\u003cspan address=\"10.1016/j.crhy.2011.06.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM.F. Bertino, \u003cem\u003eIntroduction to Nanotechnology\u003c/em\u003e.; 2022. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1142/12142\u003c/span\u003e\u003cspan address=\"10.1142/12142\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJ. Nandhini, E. Karthikeyan, S. Rajeshkumar, Nanomaterials for wound healing: Current status and futuristic frontier. Biomedical Technol. \u003cb\u003e6\u003c/b\u003e (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bmt.2023.10.001\u003c/span\u003e\u003cspan address=\"10.1016/j.bmt.2023.10.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS. Malik, K. Muhammad, Y. Waheed, Nanotechnology, A Revolution in Modern Industry. Molecules. \u003cb\u003e28\u003c/b\u003e(2) (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules28020661\u003c/span\u003e\u003cspan address=\"10.3390/molecules28020661\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eY. Li, J. Yao, C. Han et al., Quercetin, inflammation and immunity. Nutrients. \u003cb\u003e8\u003c/b\u003e(3) (2016). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/nu8030167\u003c/span\u003e\u003cspan address=\"10.3390/nu8030167\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eG.S. Kelly, Quercetin, Altern. Med. Rev. 2011;\u003cb\u003e16\u003c/b\u003e(2)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS.S. Baghel, N. Shrivastava, R. Singh, B.P. Agrawal, S. Rajput, a Review of Quercetin: Antioxidant and Anticancer Properties. World J. Pharm. Pharm. Sci. 2012;\u003cb\u003e1\u003c/b\u003e(1)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eK. Elumalai, S. Srinivasan, A. Shanmugam, Review of the efficacy of nanoparticle-based drug delivery systems for cancer treatment. Biomedical Technol. \u003cb\u003e5\u003c/b\u003e (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bmt.2023.09.001\u003c/span\u003e\u003cspan address=\"10.1016/j.bmt.2023.09.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eK.P.L. Bhat, J.W. Kosmeder, J.M. Pezzuto, Biological effects of resveratrol. Antioxid. Redox Signal. \u003cb\u003e3\u003c/b\u003e(6) (2001). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1089/152308601317203567\u003c/span\u003e\u003cspan address=\"10.1089/152308601317203567\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eR. Mikstacka, A.M. Rimando, E. Ignatowicz, Antioxidant effect of trans-Resveratrol, pterostilbene, quercetin and their combinations in human erythrocytes In vitro. Plant Foods Hum. Nutr. \u003cb\u003e65\u003c/b\u003e(1) (2010). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11130-010-0154-8\u003c/span\u003e\u003cspan address=\"10.1007/s11130-010-0154-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eG.M.F. Calixto, J. Bernegossi, De L.M. Freitas, C.R. Fontana, M. Chorilli, A.M. Grumezescu, Nanotechnology-based drug delivery systems for photodynamic therapy of cancer: A review. Molecules. \u003cb\u003e21\u003c/b\u003e(3) (2016). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules21030342\u003c/span\u003e\u003cspan address=\"10.3390/molecules21030342\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eO.C. Farokhzad, R. Langer, Impact of nanotechnology on drug delivery. ACS Nano. \u003cb\u003e3\u003c/b\u003e(1) (2009). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/nn900002m\u003c/span\u003e\u003cspan address=\"10.1021/nn900002m\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM. Devaraji, P.V. Thanikachalam, Phytoconstituents as emerging therapeutics for breast cancer: Mechanistic insights and clinical implications. Cancer Pathogenesis Therapy Published online Febr. \u003cb\u003e28\u003c/b\u003e (2025). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/J.CPT.2025.02.006\u003c/span\u003e\u003cspan address=\"10.1016/J.CPT.2025.02.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM.J. Saadh, R.K. Jadullah, Nanotechnology in Drug Delivery. Pharmacologyonline. \u003cb\u003e3\u003c/b\u003e (2021). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.17148/ijarcce.2016.55255\u003c/span\u003e\u003cspan address=\"10.17148/ijarcce.2016.55255\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eK.K. Karthik, B.V. Cheriyan, S. Rajeshkumar, M. Gopalakrishnan, A review on selenium nanoparticles and their biomedical applications. Biomedical Technol. \u003cb\u003e6\u003c/b\u003e (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bmt.2023.12.001\u003c/span\u003e\u003cspan address=\"10.1016/j.bmt.2023.12.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eR. Kanwar, J. Rathee, D.B. Salunke, S.K. Mehta, Green nanotechnology-driven drug delivery assemblies. ACS Omega. \u003cb\u003e4\u003c/b\u003e(5) (2019). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acsomega.9b00304\u003c/span\u003e\u003cspan address=\"10.1021/acsomega.9b00304\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eK.K. Karunakar, P.V. Thanikachalam, S.M. Dhanalakshmi, P. Kesharwani, B.V. Cheriyan, Hinokitiol attenuates gentamicin-induced nephrotoxicity by reversing oxidative stress and inflammation. Pharmacol. Res. - Mod. Chin. Med. \u003cb\u003e10\u003c/b\u003e (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.prmcm.2024.100410\u003c/span\u003e\u003cspan address=\"10.1016/j.prmcm.2024.100410\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eX. Xiao, H. Deng, X. Lin et al., Selenium nanoparticles: Properties, preparation methods, and therapeutic applications. Chem. Biol. Interact. \u003cb\u003e378\u003c/b\u003e (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.cbi.2023.110483\u003c/span\u003e\u003cspan address=\"10.1016/j.cbi.2023.110483\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eR.J. White, R. Luque, V.L. Budarin, J.H. Clark, D.J. Macquarrie, Supported metal nanoparticles on porous materials. Methods and applications. Chem. Soc. Rev. \u003cb\u003e38\u003c/b\u003e(2) (2009). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1039/b802654h\u003c/span\u003e\u003cspan address=\"10.1039/b802654h\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS. Kanimozhi, R. Durga, M. Sabithasree et al., Biogenic synthesis of silver nanoparticle using Cissus quadrangularis extract and its invitro study. J. King Saud Univ. Sci. \u003cb\u003e34\u003c/b\u003e(4) (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jksus.2022.101930\u003c/span\u003e\u003cspan address=\"10.1016/j.jksus.2022.101930\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM. Mutakin, R. Fauziati, F.N. Fadhilah, A. Zuhrotun, R. Amalia, Y.E. Hadisaputri, Pharmacological Activities of Soursop (Annona muricata Lin). Molecules. \u003cb\u003e27\u003c/b\u003e(4) (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules27041201\u003c/span\u003e\u003cspan address=\"10.3390/molecules27041201\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM. Devaraji, P.V. Thanikachalam, S.R. AS R, Microwave-assisted synthesis of copper oxide nanoparticles using an Andrographis paniculata leaf extract: Characterization and multifunctional biological activities. Nano-Structures Nano-Objects. \u003cb\u003e40\u003c/b\u003e, 101376 (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/J.NANOSO.2024.101376\u003c/span\u003e\u003cspan address=\"10.1016/J.NANOSO.2024.101376\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS.V. Ghumre, Assessment of In-vitro Anti-Inflammatory Activity of Cynodon Dactylon and Acyclovir Showing Synergistic Effect by Albumin Denaturation and Membrane Stabilization Assay. Mod. Approaches Drug Designing. \u003cb\u003e1\u003c/b\u003e(2) (2017). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.31031/madd.2017.01.000506\u003c/span\u003e\u003cspan address=\"10.31031/madd.2017.01.000506\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eC.S. Kumari, N. Yasmin, M.R. Hussain, M. Babuselvam, Invitro anti-inflammatory and anti-artheritic property of rhizopora mucronata leaves. Int. J. Pharma Sci. Res. 2015;\u003cb\u003e6\u003c/b\u003e(3)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS. Chatterjee, R J, S R. Green Synthesis of Zinc Oxide Nanoparticles Using Chamomile and Green Tea Extracts and Evaluation of Their Anti-inflammatory and Antioxidant Activity: An In Vitro Study. \u003cem\u003eCureus\u003c/em\u003e. Published online 2023. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.7759/cureus.46088\u003c/span\u003e\u003cspan address=\"10.7759/cureus.46088\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eİ. Gulcin, S.H. Alwasel, DPPH Radical Scavenging Assay. Processes. \u003cb\u003e11\u003c/b\u003e(8) (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/pr11082248\u003c/span\u003e\u003cspan address=\"10.3390/pr11082248\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eA.L. Hauvermale, R.S. Parveen, T.J. Harris et al., Streamlined alpha-amylase assays for wheat preharvest sprouting and late maturity alpha-amylase detection. Agrosystems Geosci. Environ. \u003cb\u003e6\u003c/b\u003e(1) (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/agg2.20327\u003c/span\u003e\u003cspan address=\"10.1002/agg2.20327\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eS. Yin, Y. Yang, W. Xu et al., Research progress of β-glucosidase assay and its application. Food Ferment. Industries. \u003cb\u003e48\u003c/b\u003e(13) (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.13995/j.cnki.11-1802/ts.031312\u003c/span\u003e\u003cspan address=\"10.13995/j.cnki.11-1802/ts.031312\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM.A.R. Shah, R.A. Khan, M. Ahmed, Phytochemical analysis, cytotoxic, antioxidant and anti-diabetic activities of the aerial parts of Sorghum halepense. Bangladesh J. Pharmacol. \u003cb\u003e14\u003c/b\u003e(3) (2019). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3329/bjp.v14i3.40292\u003c/span\u003e\u003cspan address=\"10.3329/bjp.v14i3.40292\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"biomedical-materials-and-devices","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Biomedical Materials \u0026 Devices](https://link.springer.com/journal/44174)","snPcode":"44174","submissionUrl":"https://submission.springernature.com/new-submission/44174/3","title":"Biomedical Materials \u0026 Devices","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Resveratrol, Quercetin, Selenium nanoparticles, Green synthesis, Polyphenols, Nanomedicine","lastPublishedDoi":"10.21203/rs.3.rs-8261754/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8261754/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChronic diseases such as diabetes, inflammation, and oxidative stress continue to pose major global health challenges, while conventional therapies remain limited by poor bioavailability and side effects. Natural polyphenols like resveratrol and quercetin are well known for their antioxidant, anti-inflammatory, and antidiabetic properties, yet their clinical potential is restricted due to rapid metabolism and low solubility. Nanotechnology-based approaches offer a promising solution by enhancing stability, solubility, and targeted delivery of such bioactive compounds. In this study, resveratrol\u0026ndash;quercetin functionalized selenium nanoparticles (QR-SeNPs) were synthesized using a green reduction method, with the phytochemicals serving as both reducing and stabilizing agents. The nanoparticles were characterized by UV\u0026ndash;Vis spectroscopy, SEM, FTIR, DLS, zeta potential, and XRD, confirming spherical morphology, nanoscale size (200\u0026ndash;300 nm), good colloidal stability (\u0026ndash;30 to \u0026minus;\u0026thinsp;40 mV), and predominantly amorphous structure. Biological evaluations revealed that QR-SeNPs exhibited strong anti-inflammatory activity, achieving\u0026thinsp;~\u0026thinsp;80% inhibition of protein denaturation at 50 \u0026micro;g/mL, alongside potent antioxidant activity with ~\u0026thinsp;90% scavenging in DPPH and H₂O₂ assays. In antidiabetic assays, they demonstrated significant enzyme inhibition (81% for α-amylase and 75% for β-glucosidase), comparable to standard drugs. Cytotoxicity assessment showed minimal lethality at therapeutic concentrations, with moderate effects observed only at higher doses. These findings indicate that green-synthesized QR-SeNPs represent a multifunctional and biocompatible nanoplatform with strong potential for managing oxidative stress, inflammation, and diabetes, warranting further in vivo and mechanistic investigations.\u003c/p\u003e","manuscriptTitle":"Synergistic Therapeutic Potential of Resveratrol–Quercetin Loaded Selenium Nanoparticles: Synthesis, Characterization, and In Vitro Pharmacological Evaluation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-08 11:08:10","doi":"10.21203/rs.3.rs-8261754/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-27T17:18:54+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-26T16:55:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"29258807735547380994777486397278559513","date":"2025-12-14T05:50:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"170698706146248728017340026224774355601","date":"2025-12-12T06:55:46+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-11T19:42:44+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-10T06:00:27+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"80491917885934331084229439962809592095","date":"2025-12-07T19:10:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"59474008646772236485521218839170081807","date":"2025-12-07T17:03:42+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"137139385312590709270093409994585947974","date":"2025-12-05T03:33:09+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-04T20:18:24+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-12-04T10:13:19+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-12-04T10:13:13+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biomedical Materials \u0026 Devices","date":"2025-12-02T14:27:41+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biomedical-materials-and-devices","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Biomedical Materials \u0026 Devices](https://link.springer.com/journal/44174)","snPcode":"44174","submissionUrl":"https://submission.springernature.com/new-submission/44174/3","title":"Biomedical Materials \u0026 Devices","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"820ecc16-0f8b-4484-a7b9-477ed70e3682","owner":[],"postedDate":"December 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-07T06:38:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-08 11:08:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8261754","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8261754","identity":"rs-8261754","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}