Comparative in vitro anti-leishmanial activity of Thymus vulgaris-mediated silver nanoparticles against Leishmania tropica and L. infantum
Main Article Content
Abstract
Background: Leishmaniasis remains a significant global health concern, with current treatments often limited by toxicity, cost, and emerging drug resistance. We aimed to evaluate the antileishmanial potential of eco-friendly silver nanoparticles synthesized using Thymus vulgaris extract (TSNPs) against Leishmania tropica and L. infantum in vitro.
Methods: The antileishmanial effects were evaluated against the promastigote stages of L. tropica and L. infantum using cell counting, MTT assay, and flow cytometry techniques. Cytotoxic effects on RAW 264.7 macrophages were determined by the MTT assay, and the half-maximal inhibitory concentration (IC₅₀) values were calculated for both species.
Results: TSNPs exhibited a characteristic plasmon resonance peak at 440 nm and spherical morphology with an average size of 18 nm. Treatment with TSNPs significantly reduced promastigote viability in a dose- and time-dependent manner. The IC₅₀ values after 72 h were 2.27 ppm for L. tropica and 2.04 ppm for L. infantum. Flow cytometry revealed induction of apoptosis in promastigotes, with 56.78% and 59.27% apoptotic cells for L. tropica and L. infantum, respectively, at 25 ppm. Macrophage cytotoxicity was observed only at higher concentrations (≥25 ppm).
Conclusion: Thyme-derived silver nanoparticles demonstrate potent and selective antileishmanial activity against both L. tropica and L. infantum, primarily through the induction of apoptosis. These findings highlight TSNPs as a promising green therapeutic candidate for leishmaniasis, warranting further in vivo investigation.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Tiuman TS, Santos AO, Ueda-Nakamura T, Dias Filho BP, Nakamura CV. Recent advances in leishmaniasis treatment. Int J Infect Dis. 2011;15(8):e525–e532.doi:10.1016/j.ijid.2011.03.021
Mohammadi M, Zaki L, KarimiPourSaryazdi A, Tavakoli P, Tavajjohi A, Poursalehi R, et al. Effica-cy of green synthesized silver nanoparticles via gin-ger rhizome extract against Leishmania major in vitro. PLoS One. 2021;16(8):e0255571. doi:10.1371/journal.pone.0255571
Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 2012;7(5):e35671. doi:10.1371/journal.pone.0035671
Ready PD. Epidemiology of visceral leishmaniasis. Clin Epidemiol. 2014;6:147–154. doi:10.2147/CLEP.S44267
Coura-Vital W, Marques MJ, Veloso VM, Roatt BM, Aguiar-Soares RDO, Reis LE, et al. Prevalence and factors associated with Leishmania infantum in-fection of dogs from an urban area of Brazil as iden-tified by molecular methods. PLoS Negl Trop Dis. 2011;5(8):e1291. doi:10.1371/journal.pntd.0001291
Jamil KM, Haque R, Rahman R, Faiz MA, Bhuiyan AT, Hossain M, et al. Effectiveness study of paromomycin IM injection (PMIM) for the treatment of visceral leishmaniasis (VL) in Bangla-desh. PLoS Negl Trop Dis. 2015;9(10):e0004118. doi:10.1371/journal.pntd.0004118
Reithinger R, Dujardin JC, Louzir H, Pirmez C, Alexander B, Brooker S. Cutaneous leishmaniasis. Lancet Infect Dis. 2007;7(9):581–596. doi:10.1016/S1473-3099(07)70256-8
Singh N, Kumar M, Singh RK. Leishmaniasis: current status of available drugs and new potential drug targets. Asian Pac J Trop Med. 2012;5(6):485–497. doi:10.1016/S1995-7645(12)60084-4
Sundar S, Chakravarty J. An update on pharma-cotherapy for leishmaniasis. Expert Opin Pharma-cother. 2015;16(2):237–252. doi:10.1517/14656566.2015.973850
Sundar S, More DK, Singh MK, Singh VP, Shar-ma S, Makharia A, et al. Failure of pentavalent an-timony in visceral leishmaniasis in India: report from the center of the Indian epidemic. Clin Infect Dis. 2000;31(4):1104–1107. doi:10.1086/318121
Karimipour-Saryazdi A, Ghaffarifar F, Tavakoli P, Karimipour-Saryazdi Y, Zaki L, Bahadory S, et al. Anti-parasitic effects of herbal extract-based silver nanoparticles on the trophozoite and cystic forms of Acanthamoeba protozoa. Int J Enteric Pathog. 2020;8(3):84–88. doi:10.34172/ijep.2020.19
Zaki L, Mohammadi M, Karimipour-Saryazdi A, Baghkhani F, Badri M, Ghaffarifar F. In vitro activi-ty of green synthesized silver nanoparticles via Thymus vulgaris extract against Leishmania major. Iran J Parasitol. 2023;18(4):494–502. doi: 10.18502/ijpa.v18i4.14258
Zazo H, Colino CI, Lanao JM. Current applications of nanoparticles in infectious diseases. J Control Re-lease. 2016; 224:86–102. doi: 10.1016/j.jconrel.2016.01.008
Ghosh SK, Pal A, Jana NR. Nanoparticles in treat-ment of parasitic diseases: a review on anti-leishmanial perspectives. J Control Release. 2012;159(2):140–153. doi:10.1016/j.jconrel.2012.01.046
Bessa IAA, D’Amato DL, Souza ABC, Levita DP, Mello CC, da Silva AFM, et al. Innovating leish-maniasis treatment: a critical chemist’s review of inorganic nanomaterials. ACS Infect Dis. 2024;10(8):2485–2506. doi:10.1021/acsinfecdis.4c00231.
Nilforoushzadeh MA, Shirani-Bidabadi L, Zolfaghari-Baghbaderani A, Saberi S, Siadat AH, Mahmoudi M, et al. Comparison of Thymus vulgar-is (Thyme), Achillea millefolium (Yarrow) and prop-olis hydroalcoholic extracts versus systemic glucan-time in the treatment of cutaneous leishmaniasis in Balb/c mice. J Vector Borne Dis. 2008;45(4):301–306.
Mohammadi M, Shahisaraee SA, Tavajjohi A, Pournoori N, Muhammadnejad S, Roodbar Mo-hammadi S, et al. Green synthesis of silver nano-particles using Zingiber officinale and Thymus vul-garis extracts: characterisation, cell cytotoxicity, and its antifungal activity against Candida albicans in comparison to fluconazole. IET Nanobiotechnol. 2019;13(2):114–119. doi:10.1049/iet-nbt.2018.5146.
Zaki L, Ghaffarifar F, Sharifi Z, Horton J, Sadraei J. Toxoplasma gondii: preventive and therapeutic ef-fects of morphine and evaluation of treatment pa-rameters of tachyzoites and infected macrophages in vitro and in a murine model. EXCLI J. 2020; 19:514–527. doi:10.17179/excli2019-1961.
Soosaraei M, Fakhar M, Teshnizi SH, Ziaei Hezar-jaribi H, Banimostafavi ES. Medicinal plants with promising antileishmanial activity in Iran: a sys-tematic review and meta-analysis. Ann Med Surg (Lond). 2017;21:63–80. doi:10.1016/j.amsu.2017.07.057
Agha MIH, Baaj R. Contribution study to the chemical composition of the volatile oil in the leaves of Thymus vulgaris L. and determination of the anti-bacterial activity and anti-leishmanial vitali-ty. J Essent Oil Res. 2018;30(1):10–16
Mohebali M, Rezayat MM, Gilani K, Sarkar S, Ak-houndi B, Esmaeili J, et al. Nanosilver in the treat-ment of localized cutaneous leishmaniasis caused by Leishmania major (MRHO/IR/75/ER): an in vitro and in vivo study. Daru. 2009;17(4):285–289.