Iranian Journal of Basic Medical Sciences

Iranian Journal of Basic Medical Sciences

Development, characterization, and molecular dynamics simulation of resveratrol-loaded nanostructured lipid carriers for enhanced anti-oxidant and antityrosinase activity

Articles in Press

Document Type : Original Article

Authors
Hoda Atapour-Mashhad 1, 2
Mohhamad Delkhosh 3
Mojgan Nejabat 4
Shiva Golmohammadzadeh 1, 2
Farzin Hadizadeh 4, 5
Samaneh Rahamouz-Haghighi 6
Vahideh Mohammadzadeh 1
Zahra Tayarani-Najaran 7
1 Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran
2 Department of Pharmaceutics, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
3 Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
4 Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
5 Biotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran
6 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
7 Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran
10.22038/ijbms.2026.97463.20959
Abstract
Objective(s): This study aimed to develop and characterize resveratrol-loaded nanostructured lipid carriers (Res-NLCs) to assess their preliminary potential as topical delivery systems, with a focus on antioxidant and antityrosinase properties.
Materials and Methods: Ten Res-NLC formulations (F1–F10) were prepared by high-shear homogenization combined with ultrasound. The formulations underwent comprehensive evaluation of morphology, particle size, long-term stability, and entrapment efficiency (EE%). Experimental findings were correlated with molecular dynamics (MD) simulations. In vitro performance was assessed by drug-release profiling (dialysis-bag diffusion), cytotoxicity and protective effects (MTT assay), reactive oxygen species (ROS) reduction (DCFH-DA), and anti-tyrosinase activity (L-DOPA assay).
Results: Formulation F10 exhibited excellent surface characteristics, including a z-average particle size of 144.0 nm, a narrow polydispersity index (PDI) of 0.117, a high negative zeta potential of -22.6 mV, favorable stability at lower temperatures, and optimal EE%. MD simulations confirmed superior molecular stability for F10 through compact cluster formation, consistent with its sustained long-term stability at 4 °C and 25 °C. In contrast, structural fragmentation observed in F2 accounted for its large particle size (>500 nm). The NLC systems exhibited a sustained-release profile, delivering 84% of the resveratrol over 48 hr. Furthermore, F10 significantly inhibited tyrosinase activity and effectively reduced intracellular ROS levels following hydrogen peroxide-induced stress.
Conclusion: The newly developed Res-NLC (F10) formulation serves as a promising, stable carrier for the sustained topical delivery of resveratrol, demonstrating notable antioxidant and antityrosinase performance, which warrants subsequent biological validation.
Keywords
Anti-oxidant
Nanostructured lipid carriers
Resveratrol
Reactive oxygen species inhibition
Tyrosinase inhibitor
Subjects
1. Solomon S, Ivy DJ, Kinnison D, Mills MJ, Neely RR, 3rd, Schmidt A. Emergence of healing in the Antarctic ozone layer. Science 2016; 353:269–274.
2. Wang PW, Hung YC, Lin TY, Fang JY, Yang PM, Chen MH, et al. Comparison of the biological impact of UVA and UVB upon the skin with functional proteomics and immunohistochemistry. Antioxidants (Basel) 2019; 8:569.
3. Papaccio F, A DA, Caputo S, Bellei B. Focus on the contribution of oxidative stress in skin aging. Antioxidants (Basel) 2022; 11:1121.
4. Widowati W, Onggowidjaja P, Vosough M, Triharsiwin DN, Distya AP, Nurani LW, et al. Hepatoprotective effect of hUC-MSC secretome in LPS-induced HepG2 cells. Iran J Basic Med Sci 2026; 29: 901-908.
5. Rostahi H, Aramshahi SMA, Vosoughi N, Shamlou S, Shahmahmoudi Z, Milan P, et al. Enhanced effects of Wharton’s jelly-derived MSC secretome and microbial exopolysaccharide on cellular responses and tissue repair in diabetic wound healing. Iran J Basic Med Sci 2026; 29: 1747-1758.
6. Nasiri N, Hemmati P, Tabaie SM. Conditioned medium from mesenchymal stem cells: Towards skin tissue engineering and wound healing. Iran J Basic Med Sci 2026; 29: 181-190.
7. Dall’Olmo L, Papa N, Surdo NC, Marigo I, Mocellin S. Alpha-melanocyte stimulating hormone (α-MSH): Biology, clinical relevance and implication in melanoma. J Transl Med 2023; 21:562.
8. Netcharoensirisuk P, Umehara K, De-Eknamkul W, Chaotham C. Cajanin suppresses melanin synthesis through modulating MITF in human melanin-producing cells. Molecules 2021; 26:6040.
9. Solano F. Photoprotection and skin pigmentation: Melanin-related molecules and some other new agents obtained from natural sources. Molecules 2020; 25:1537.
10. Fernandes MS, Kerkar S. Microorganisms as a source of tyrosinase inhibitors: A review. Annals of Microbiol 2017; 67:343–358.
11. Panzella L, Napolitano A. Natural and bioinspired phenolic compounds as tyrosinase inhibitors for the treatment of skin hyperpigmentation: Recent advances. Cosmetics 2019; 6:57.
12. Song H, Hwang YJ, Ha JW, Boo YC. Screening of an epigenetic drug library identifies 4-((hydroxyamino) carbonyl)-N-(2-hydroxyethyl)-N-phenyl-benzeneacetamide that reduces melanin synthesis by inhibiting tyrosinase activity independently of epigenetic mechanisms. Int J Mol Sci 2020; 21:4589.
13. Saewan N, Jimtaisong A. Photoprotection of natural flavonoids. J Appl Pharm Sci 2013; 3:129–141.
14. Jatana S, Palmer BC, Phelan SJ, Gelein R, DeLouise LA. In vivo quantification of quantum dot systemic transport in C57BL/6 hairless mice following skin application post-ultraviolet radiation. Part Fibre Toxicol 2017; 14:12.
15. Sharma B, Iqbal B, Kumar S, Ali J, Baboota S. Resveratrol-loaded nanoemulsion gel system to ameliorate UV-induced oxidative skin damage: From in vitro to in vivo investigation of anti-oxidant activity enhancement. Arch Dermatol Res 2019; 311:773–793.
16. Ungvari Z, Bagi Z, Feher A, Recchia FA, Sonntag WE, Pearson K, et al. Resveratrol confers endothelial protection via activation of the anti-oxidant transcription factor Nrf2. Am J Physiol Heart Circ Physiol 2010; 299:H18–H24.
17. Alavi M, Farkhondeh T, Aschner M, Samarghandian S. Resveratrol mediates its anti-cancer effects by Nrf2 signaling pathway activation. Cancer Cell International 2021; 21:1–9.
18. Marko M, Pawliczak R. Resveratrol and its derivatives in inflammatory skin disorders—atopic dermatitis and psoriasis: A review. Antioxidants (Basel) 2023; 12:1954.
19. Mascarenhas-Melo F, Araújo ARTS, Rodrigues M, Mathur A, Gonçalves MBS, Tanwar K, et al. Dermatological bioactivities of resveratrol and nanotechnology strategies to boost its efficacy—An updated review. Cosmetics 2023; 10:68.
20. Hecker A, Schellnegger M, Hofmann E, Luze H, Nischwitz SP, Kamolz LP, et al. The impact of resveratrol on skin wound healing, scarring, and aging. Int Wound J 2022; 19:9–28.
21. Gokce EH, Korkmaz E, Dellera E, Sandri G, Bonferoni MC, Ozer O. Resveratrol-loaded solid lipid nanoparticles versus nanostructured lipid carriers: evaluation of anti-oxidant potential for dermal applications. Int J Nanomedicine 2012; 7:1841–1850.
22. Shimojo AAM, Fernandes ARV, Ferreira NRE, Sanchez-Lopez E, Santana MHA, Souto EB. Evaluation of the influence of process parameters on the properties of resveratrol-loaded NLC using 22 full factorial design. Antioxidants 2019; 8:272.
23. Freire TB, Pinto CASdO, Gonçalves MIdA, Serra CHdR, Dario MF, Baby AR, et al. Impact of ferulic acid and resveratrol on the effectiveness and safety of sunscreen. Braz J Pharm Sci 2023; 59:e23357.
24. Atapour-Mashhad H, Nejabat M, Hadizadeh F, Hoseinsalari A, Golmohammadzadeh S. Preparation, characterization, and molecular dynamic simulation of novel coenzyme Q10 loaded nanostructured lipid carriers. Curr Pharm Des 2023; 29:2177–2190.
25. Vaziri MS, Tayarani-Najaran Z, Kabiri H, Nasirizadeh S, Golmohammadzadeh S, Kamali H. Preparation and characterization of Undecylenoyl Phenylalanine loaded-nanostructure lipid carriers (NLCs) as a new α-MSH Antagonist and antityrosinase agent. Adv Pharm Bull 2022; 13:290.
26. Moradi A, Seifati SM, Darroudi M, Golmohammadzadeh S, Ashkezari MD. Nanocrystal structure of minoxidil: A safe stimulator for hair growth factor for C57BL/6 mice. Curr Med Chem 2025; 32:4343–4355.
27. Kabiri H, Tayarani-Najaran Z, Rahmanian-devin P, Vaziri MS, Nasirizadeh S, Golmohammadzadeh S, et al. Preparation, characterization, and evaluation of anti-tyrosinase activity of solid lipid nanoparticles containing Undecylenoyl phenylalanine (Sepiwhite®). J Cosmet Dermatol 2022; 21:6061–6071.
28. Smith BC. Fundamentals of Fourier Transform Infrared Spectroscopy: CRC press; 2011.
29. He Y, Li H, Zheng X, Yuan M, Yang R, Yuan M, et al. Preparation, in vivo and in vitro release of polyethylene glycol monomethyl ether-polymandelic acid microspheres loaded panax notoginseng saponins. Molecules 2019; 24:2024.
30. Van Der Spoel D, Lindahl E, Hess B, Groenhof G, Mark AE, Berendsen HJC. GROMACS: fast, flexible, and free. J Comput Chem 2005; 26:1701–1718.
31. Hess B, Bekker H, Berendsen HJC, Fraaije JGEM. LINCS: A linear constraint solver for molecular simulations. J Comput Chem 1997; 18:1463–1472.
32. Bussi G, Donadio D, Parrinello M. Canonical sampling through velocity rescaling. J Chem Phys 2007; 126:014101.
33. Berendsen HJC, Postma JPMv, Van Gunsteren WF, DiNola A, Haak JR. Molecular dynamics with coupling to an external bath. J Chem Phys 1984; 81:3684–3690.
34. Atapour-Mashhad H, Soukhtanloo M, Golmohammadzadeh S, Chamani J, Nejabat M, Hadizadeh F. Synthesis and molecular dynamic simulation of novel cationic and non-cationic pyrimidine derivatives as potential G-quadruplex-ligands. Anticancer Agents Med Chem 2024; 24:1126–1141.
35. Hoseinsalari A, Atapour-Mashhad H, Asili J, Tayarani-Najaran Z, Mohtashami L, Khanavi M, et al. Preparation and characterization of nanostructured lipid carriers (NLCs) containing glycyrrhiza glabra extract for the treatment of skin hyperpigmentation. Curr Pharm Des 2024; 30:3213–3232.
36. Im YS, Ryu YK, Moon EY. Mouse melanoma cell migration is dependent on production of reactive oxygen species under normoxia condition. Biomol Ther (Seoul) 2012; 20:165–170.
37. Hashemi-Shahri SH, Golshan A, Mohajeri SA, Baharara J, Amini E, Salek F, et al. ROS-scavenging and Anti-tyrosinase Properties of Crocetin on B16F10 Murine Melanoma Cells. Anticancer Agents Med Chem 2018; 18:1064–1069.
38. Ghasemiyeh P, Azadi A, Daneshamouz S, Heidari R, Azarpira N, Mohammadi-Samani S. Cyproterone acetate-loaded nanostructured lipid carriers: Effect of particle size on skin penetration and follicular targeting. Pharm Dev Technol 2019; 24:812–823.
39. Elmowafy M, Al-Sanea MM. Nanostructured lipid carriers (NLCs) as drug delivery platform: Advances in formulation and delivery strategies. Saudi Pharm J 2021; 29:999–1012.
40. Kim MH, Kim KT, Sohn SY, Lee JY, Lee CH, Yang H, et al. Formulation and evaluation of nanostructured lipid carriers (NLCs) Of 20(S)-protopanaxadiol (PPD) by box-behnken design. Int J Nanomedicine 2019; 14:8509–8520.
41. Masoomzadeh S, Sadighian S, Gholikhani T, Rostamizadeh K. Potential use of nanostructured lipid carriers mediated neuronal delivery of carbamazepine. Nanomed Res J 2022; 7:330–337.
42. Mardhiah Adib Z, Ghanbarzadeh S, Kouhsoltani M, Yari Khosroshahi A, Hamishehkar H. The effect of particle size on the deposition of solid lipid nanoparticles in different skin layers: A histological study. Adv Pharm Bull 2016; 6:31–36.
43. Corrigan DO, Healy AM, Corrigan OI. Preparation and release of salbutamol from chitosan and chitosan co-spray dried compacts and multiparticulates. Eur J Pharm Biopharm 2006; 62:295–305.
44. Danaei M, Dehghankhold M, Ataei S, Hasanzadeh Davarani F, Javanmard R, Dokhani A, et al. Impact of particle size and polydispersity index on the clinical applications of lipidic nanocarrier systems. Pharmaceutics 2018; 10:57.
45. Pizzol CD, Filippin-Monteiro FB, Restrepo JA, Pittella F, Silva AH, Alves de Souza P, et al. Influence of surfactant and lipid type on the physicochemical properties and biocompatibility of solid lipid nanoparticles. Int J Environ Res Public Health 2014; 11:8581–8596.
46. Pochapski DJ, Carvalho dos Santos C, Leite GW, Pulcinelli SH, Santilli CV. Zeta potential and colloidal stability predictions for inorganic nanoparticle dispersions: Effects of experimental conditions and electrokinetic models on the interpretation of results. Langmuir 2021; 37:13379–13389.
47. Tamjidi F, Shahedi M, Varshosaz J, Nasirpour A. Design and characterization of astaxanthin-loaded nanostructured lipid carriers. Innov Food Sci Emerg Technol 2014; 26:366–374.
48. Atapour-Mashhad H, Tayarani-Najaran Z, Golmohammadzadeh S. Preparation and characterization of novel nanostructured lipid carriers (NLC) and solid lipid nanoparticles (SLN) containing coenzyme Q10 as potent anti-oxidants and antityrosinase agents. Heliyon 2024; 10:e31429.
49. Gomes MJ, Martins S, Ferreira D, Segundo MA, Reis S. Lipid nanoparticles for topical and transdermal application for alopecia treatment: Development, physicochemical characterization, and in vitro release and penetration studies. Int J Nanomedicine 2014; 9:1231–1242.
50. Jayan H, Leena MM, Sundari SKS, Moses JA, Anandharamakrishnan C. Improvement of bioavailability for resveratrol through encapsulation in zein using electrospraying technique. J Funct Foods 2019; 57:417–424.
51. Balata GF, Essa EA, Shamardl HA, Zaidan SH, Abourehab MA. Self-emulsifying drug delivery systems as a tool to improve solubility and bioavailability of resveratrol. Drug Des Devel Ther 2016; 10:117–128.
52. Tang H, Xiang S, Li X, Zhou J, Kuang C. Preparation and in vitro performance evaluation of resveratrol for oral self-microemulsion. PLoS One 2019; 14:e0214544.
53. Yu M, Shen Z, Zhang S, Zhang Y, Zhao H, Zhang L. The active components of Erzhi wan and their anti-Alzheimer’s disease mechanisms determined by an integrative approach of network pharmacology, bioinformatics, molecular docking, and molecular dynamics simulation. Heliyon 2024; 10:e33761.
54. Chen H, Levitas VI, Popov D, Velisavljevic N. Nontrivial nanostructure, stress relaxation mechanisms, and crystallography for pressure-induced Si-I→ Si-II phase transformation. Nature Communications 2022; 13:982.
55. Yin LL, Xu JK, Wang XJ, Gao SQ, Lin YW. Molecular dynamics simulation and kinetic study of fluoride binding to V21C/V66C myoglobin with a cytoglobin-like disulfide bond. Int J Mol Sci 2020; 21:2512.
56. Jomova K, Raptova R, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, et al. Reactive oxygen species, toxicity, oxidative stress, and anti-oxidants: chronic diseases and aging. Arch Toxicol 2023; 97:2499–2574.
57. Astley C, Houacine C, Zaabalawi A, Wilkinson F, Lightfoot AP, Alexander Y, et al. Nanostructured lipid carriers deliver resveratrol, restoring attenuated dilation in small coronary arteries, via the AMPK pathway. Biomedicines 2021; 9:1852.
58. Radeva L, Stefanova D, Yordanov Y, Kamenova K, Petrov PD, Marinova MK, et al. Incorporation of resveratrol in polymeric nanogel for improvement of its protective effects on cellular and microsomal oxidative stress Models Gels 2023; 9:450.
59. Brugè F, Damiani E, Puglia C, Offerta A, Armeni T, Littarru GP, et al. Nanostructured lipid carriers loaded with CoQ10: Effect on human dermal fibroblasts under normal and UVA-mediated oxidative conditions. Int J Pharm 2013; 455:348–356.
60. Lee TH, Seo JO, Baek SH, Kim SY. Inhibitory effects of resveratrol on melanin synthesis in ultraviolet B-induced pigmentation in Guinea pig skin. Biomol Ther (Seoul) 2014; 22:35–40.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 17 October 2026