Iranian Journal of Basic Medical Sciences

Iranian Journal of Basic Medical Sciences

Evaluation of excretory–secretory products of Lucilia sericata larvae loaded onto chitosan nanoparticles as a drug candidate against Leishmania major

Articles in Press

Document Type : Original Article

Authors
Farzaneh Sadeghi 1
Mohammad Saaid Dayer 2
Fatemeh Ghaffarifar 3
Amrollah Azarm 1
1 Department of Medical Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Medical Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, ran
3 Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
10.22038/ijbms.2026.95158.20522
Abstract
Objective(s): Zoonotic cutaneous leishmaniasis is an infectious disease caused by the protozoan parasite Leishmania major and characterized by chronic skin lesions. Treatment is often prolonged and associated with adverse effects. Chitosan, due to its favorable biomedical properties, including biocompatibility, biodegradability, non-toxicity, and wound-healing potential, has attracted considerable attention for therapeutic applications. This study aimed to evaluate the anti-leishmanial effects of excretory–secretory (ES) products of Lucilia sericata larvae loaded onto chitosan nanoparticles against L. major cells.
Materials and Methods: The ES was collected, filter-sterilized, and loaded onto chitosan nanoparticles (CN). The ES-loaded CN was applied at different concentrations under both in vitro and in vivo conditions to leishmanial cells, and its cytotoxicity was compared with that of the standalone CN, crude ES, and Glucantime as the first-line drug against cutaneous leishmaniasis caused by L. major.
Results: ES-loaded chitosan nanoparticles were more effective in healing leishmanial lesions than the crude ES and chitosan nanoparticles applied separately. The ES-loaded CN exhibited stronger inhibitory effects against both intracellular and extracellular forms of the parasite than its components did when used alone.
Conclusion: The results indicated that ES- loaded chitosan nanoparticles exerted antileishmanial and immunomodulatory effects on lesions. The formulation may therefore represent a promising alternative remedy for treating leishmanial lesions. Due to their biocompatibility, antimicrobial activity, and controlled-release capability, ES-loaded CN showed effective performance in wound healing and treatment.
Keywords
Chitosan
Excretory-secretory products
Leishmaniasis
Leishmania major
Lucilia sericata
Subjects
1. Firooz A, Mortazavi H, Khamesipour A, Ghiasi M, Abedini R, Balighi K, et al. Old world cutaneous leishmaniasis in Iran: Clinical variants and treatments. J Dermatolog Treat 2021;32:673-683.
2. Holakouie-Naieni K, Mostafavi E, Boloorani AD, Mohebali M, Pakzad R. Spatial modelling of cutaneous leishmaniasis in Iran from 1983 to 2013. Acta Tropica 2017;166:67-73.
3. Mahmoudzadeh‐Niknam H, Ajdary S, Riazi‐Rad F, Mirzadegan E, Rezaeian A, Khaze V, et al. Molecular epidemiology of cutaneous leishmaniasis and heterogeneity of Leishmania major strains in Iran. Trop Med Int Health 2012;17:1335-1344.
4. Jafari R, Abdoli H, Arandian MH, Shareghi N, Ghanei M, Jalali-Zand N, et al. Emerging of cutaneous leishmaniasis due to Leishmania major in a new focus in Esfahan Province, Central Iran. J Arthropod Borne Dis 2020;14:134.
5. Norouzinezhad F, Ghaffari F, Norouzinejad A, Kaveh F, Gouya MM. Cutaneous leishmaniasis in Iran: Results from an epidemiological study in urban and rural provinces. Asian Pac J Trop Biomed 2016;6:614-619.
6. Khamesipour A, Mohammadi A, Jaafari M, Eskandari S, Tasbihi M, Javadi A, et al. Pilot study of safety and efficacy of topical liposomal amphotericin B for cutaneous leishmaniasis caused by Leishmania major in the Islamic Republic of Iran. East Mediterr Health J 2022;28:658-663.
7. Reithinger R, Dujardin J-C, Louzir H, Pirmez C, Alexander B, Brooker S. Cutaneous leishmaniasis. Lancet Infect Dis 2007;7:581-596.
8. Minodier P, Parola P. Cutaneous leishmaniasis treatment. Travel Med Infect Dis 2007;5:150-158.
9. Singh S, Sivakumar R. Challenges and discoveries in the treatment of leishmaniasis. J Infect Chemother 2004;10:307-315.
10. Gazi U, Taylan‐Ozkan A, Mumcuoglu KY. The effect of Lucilia sericata larval excretion/secretion (ES) products on cellular responses in wound healing. Med Vet Entomol 2021;35:257-266.
11. Bexfield A, Bond A, Morgan C, Wagstaff J, Newton R, Ratcliffe N, et al. Amino acid derivatives from Lucilia sericata excretions/secretions may contribute to the beneficial effects of maggot therapy via increased angiogenesis. Br J Dermatol 2010;162:554-562.
12. Nigam Y, Dudley E, Bexfield A, Bond AE, Evans J, James J. The physiology of wound healing by the medicinal maggot, Lucilia sericata. Adv Insect Physiol 2010;39:39-81.
13. Tombulturk FK, Kanigur‐Sultuybek G. A molecular approach to maggot debridement therapy with Lucilia sericata and its excretions/secretions in wound healing. Wound Repair Regen 2021;29:1051-1061.
14. Nezakati E, Hasani MH, Zolfaghari P, Rashidan M, Sohrabi MB. Effects of Lucilia sericata maggot therapy in chronic wound treatment: A randomised clinical trial. Chronic Wound Care Manag Res 2020:11-17.
15. Wang D, Tang X, Ruan J, Zhu Z, Wang R, Weng Y, et al. HSP90AB1 as the druggable target of maggot extract, reverses cisplatin resistance in ovarian cancer. Oxid Med Cell Longev 2023;2023:9335440.
16. Alipour H, Shahriari-Namadi M, Ebrahimi S, Moemenbellah-Fard MD. Wound healing potential: Evaluation of molecular profiling and amplification of Lucilia sericata angiopoietin-1 mRNA mid-part. BMC Res Notes 2020;13:308.
17. Valachova I, Bohova J, Kozanek M, Takac P, Majtan J. Lucilia sericata medicinal maggots: A new source of antimicrobial compounds. Microbial Pathogens and Strategies for Combating Them: Science, Technology and Education. Badajoz, Spain: Formatex Research Center. 2013:1745-1753.
18. Sim S, Wong NK. Nanotechnology and its use in imaging and drug delivery. Biomed Rep 2021;14:42.
19. Park K. Nanotechnology: What it can do for drug delivery. J Control Release 2008;120:1.
20. Saeedi M, Vahidi O, Moghbeli MR, Ahmadi S, Asadnia M, Akhavan O, et al. Customizing nano-chitosan for sustainable drug delivery. J Control Release 2022;350:175-192.
21. Divya K, Jisha M. Chitosan nanoparticles preparation and applications. Environ Chem Lett 2018;16:101-112.
22. Aibani N, Rai R, Patel P, Cuddihy G, Wasan EK. Chitosan nanoparticles at the biological interface: Implications for drug delivery. Pharmaceutics 2021;13:1686.
23. Nagpal K, Singh SK, Mishra DN. Chitosan nanoparticles: A promising system in novel drug delivery. Chem Pharm Bull (Tokyo)2010;58:1423-1430.
24. Landriscina A, Rosen J, Friedman AJ. Biodegradable chitosan nanoparticles in drug delivery for infectious disease. Nanomedicine 2015;10:1609-1619.
25. Divakar DD, Jastaniyah NT, Altamimi HG, Alnakhli YO, Alkheraif AA, Haleem S. Enhanced antimicrobial activity of naturally derived bioactive molecule chitosan conjugated silver nanoparticle against dental implant pathogens. Int J Biol Macromol 2018;108:790-797.
26. Sherafati J, Dayer MS, Ghaffarifar F, Akbarzadeh K, Pirestani M. Evaluating leishmanicidal effects of Lucilia sericata products in combination with Apis mellifera honey using an in vitro model. PLoS One 2023;18:e0283355.
27. Sherafati J, Dayer MS, Ghaffarifar F. Therapeutic effects of Lucilia sericata larval excretion/secretion products on Leishmania major under in vitro and in vivo conditions. Parasit Vectors 2022;15:212.
28. Firoozfar F, Moosa-Kazemi SH, Rafinejad J, Saghafipour A, Baniardalani M, Ahmed Yusuf M, et al. Environmental factors influencing the growth of Lucilia Sericata larvae used for maggot therapy under laboratory condition. J Kerman Univ Med Sci 2018;25:118-127.
29. Bagheri M, Alipour H, Shahriari-Namadi M, Malekpour SH, Raz A. Improving maggot therapy: Mass rearing and molecular identification of Lucilia Sericata (Meigen, 1826) Larvae In Maggotarium, School of Health, Shiraz. 2021.
30. Nguyen TA, Nguyen TH. Study on fabrication of antibacterial low molecular weight nanochitosan using sodium tripolyphosphate and hydrogen peroxide. J Nanotechnol 2022;2022:8368431.
31. Simin A, Ghaffarifar F, Delavari H, Dayer MS, Hamidianfar N, Baghkhani F. In vitro and in vivo effects of ethanolic extract of Fumaria parviflora Lam. embedded in chitosan nanoparticles against Leishmania major. Acta Parasitol 2024;69:628-638.
32. Van Bavel N, Issler T, Pang L, Anikovskiy M, Prenner EJ. A simple method for synthesis of chitosan nanoparticles with ionic gelation and homogenization. Molecules 2023;28:4328.
33. Soltani S, Mojiri-Forushani H, Soltani S, Kahvaz MS, Foroutan M. Evaluation of antileishmanial activity employing conventional and solid lipid nanoparticles of amphotericin B on Leishmania major in vitro and in vivo. Infect Disord Drug Targets 2020;20:822-827.
34. Maleki F, Zarebavani M, Mohebali M, Dayer MS, Hajialiani F, Tabatabaie F. In vitro and in vivo susceptibility of Leishmania major to some medicinal plants. Asian Pac J Trop Biomed 2017;7:37-42.
35. Azizi K, Shahidi-Hakak F, Asgari Q, Hatam GR, Fakoorziba MR, Miri R, Moemenbellah-Fard MD. In vitro efficacy of ethanolic extract of Artemisia absinthium (Asteraceae) against Leishmania major L. using cell sensitivity and flow cytometry assays. J Parasit Dis 2016;40:735-740.
36. Zarandi HZ-A, Shirani-Bidabadi L, Aghaei-Afshar A, Eghbalian M, Zolala J, Mirtadzadini M, et al. In vitro evaluation of hydroalcoholic extracts of Capparis spinosa, Ricinus communis, and Solanum luteum on Leishmania major (MRHO/IR/75/ER) Promastigotes. Jundishapur J Nat Pharm Prod 2021;17:e115306.
37. Basmaciyan L, Azas N, Casanova M. Different apoptosis pathways in Leishmania parasites cell death discovery. 2018;4:90.
38. Khamesipour F, Khamesipour A, Hejazi SH, Ghanadian M. In vitro assessment of Dracocephalum lindbergii for growth inhibition, apoptosis induction, and cytokine modulation against Leishmania major. Heliyon 2024;10:e38331.
39. Feiz Haddad MH, Rezvan H, Ghasemi M, Mohseni E, Dalvand R, Hamoonnavard S, Habibpour H. Cellular and histopathological changes in BALB/c mice infected with live attenuated Leishmania major parasite. Archives of Razi Institute. 2026; 81:353-364.
40. Carrion J, Nieto A, Iborra S, Iniesta V, Soto M, Folgueira C, et al. Immunohistological features of visceral leishmaniasis in BALB/c mice. Parasite Immunol 2006;28:173-183.
41. Zahirnia AH, Bordbar A, Ebrahimi S, Spotin A, Mohammadi S, Ghafari SM, et al. Predominance of Leishmania major and rare occurrence of Leishmania tropica with haplotype variability at the center of Iran. Braz J Infect Dis 2018;22:278-287.
42. Lopes ME, Dos Santos LM, Sacks D, Vieira LQ, Carneiro MB. Resistance against Leishmania major infection depends on microbiota-guided macrophage activation. Front Immunol 2021;12:730437.
43. Rahimi S, Khamesipour A, Akhavan AA, Rafinejad J, Ahmadkhaniha R, Bakhtiyari M, et al. The leishmanicidal effect of Lucilia sericata larval saliva and hemolymph on in vitro Leishmania tropica. Parasit Vectors 2021;14:40.
44. Sanei-Dehkordi A, Khamesipour A, Akbarzadeh K, Akhavan AA, Mohammadi AMA, Mohammadi Y, et al. Anti Leishmania activity of Lucilia sericata and Calliphora vicina maggots in laboratory models. Exp Parasitol2016;170:59-65.
45. Akküçük Ş, Kaya ÖM, Yaman M. What is the best animal model for leishmaniasis studies? Turkish J Vet Res 2018;2:28-31.
46. Song M, Bai X, Wang D, Wang Q, Pan L, He P, et al. Combined application of moist exposed burn ointment and maggot therapy in wound healing. J Wound Care2022;31:S41-S52.
47. Benbow M. Modern wound therapies: Maureen Benbow gives an overview of just some of the vast range of modern wound therapies available to community nurses. J Community Nurs 2008;22:20-25.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 31 August 2026