Abstract
Cystic echinococcosis (CE), which is caused by the metacestode of Echinococcus granulosus, is one of the most important zoonoses affecting humans. Benzimidazoles (in particular albendazole) and praziquantel (PZQ) are effective against CE, but poor water solubility of these agents often leads to inadequate results. Here, we evaluate the effects of chitosan-albendazole (ChABZ) and chitosan-praziquantel (ChPZQ) nanoparticles as a new formulation on hydatid cysts both in vitro and in vivo. Developed microcysts in culture were treated with different concentrations of ChABZ and ChPZQ nanoparticles (either alone or in combination), and ABZ + PZQ suspension. The viability rate of microcysts was used to evaluate the drug efficacies. In addition, the prophylactic and therapeutic effects of the drugs were studied on infected DBA/2 mice. Transmission electron microscopy was used to observe the ultra-structural changes. The viability rate of microcysts and differences in cyst weights were compared by ANOVA, and the cyst numbers were compared using the Kruskal-Wallis test. The combination of ChABZ + ChPZQ nanoparticles was more effective than the ABZ + PZQ suspension in vitro (p < 0.05). In prophylaxy, a significant reduction was observed both in size and in number of the cysts in ChABZ + ChPZQ nanoparticle groups compared with the control group (p < 0.05). In the therapeutic stage, however, this treatment only reduced the cyst numbers. Degeneration of the microcysts treated with the drugs was evident in the ultra-structural imaging. Overall, the nanoparticulate drugs were more effective than their suspension counterparts, but further studies are recommended to evaluate the full potential of these nanoparticles in the treatment of human CE.
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References
Abulaihaiti M, Wu XW, Qiao L, Lv HL, Zhang HW, Aduwayi N, Wang YJ, Wang XC, Peng XY (2015) Efficacy of albendazole-chitosan microsphere-based treatment for alveolar echinococcosis in mice. PLoS Negl Trop Dis 9:e0003950. https://doi.org/10.1371/journal.pntd.0003950
Ahmadnia S, Moazeni M, Mohammadi-Samani S, Oryan A (2013) In vivo evaluation of the efficacy of albendazole sulfoxide and albendazole sulfoxide loaded solid lipid nanoparticles against hydatid cyst. Exp Parasitol 135:314–319. https://doi.org/10.1016/j.exppara.2013.07.017
Amidon GL, Lennernas H, Shah VP, Crison JR (1995) A theoretical basis for a biopharmaceutic drug classification: the correlation of in vitro drug product dissolution and in vivo bioavailability. Pharm Res 12:413–420
Brunetti E, Kern P, Vuitton DA, WHO-IWGE WP (2010) Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop 114:1–16. https://doi.org/10.1016/j.actatropica.2009.11.001
Casado N, PerezSerrano J, Denegri G, Rodriguez-Caabeiro F (1996) Development of a chemotherapeutic model for the in vitro screening of drugs against Echinococcus granulosus cysts: the effects of an albendazole albendazole sulphoxide combination. Int J Parasitol 26:59–65. https://doi.org/10.1016/0020-7519(95)00095-X
Casado N, Rodriguezcaabeiro F, Hernandez S (1986) In vitro survival of Echinococcus granulosus protoscolices in several media, at +4-degrees-C and +37-degrees-C. Z Parasitenkd Parasitol Res 72:273–278. https://doi.org/10.1007/Bf00931155
Casado N, Urrea-Paris MA, Moreno MJ, Rodriguez-Caabeiro F (2001) Combined praziquantel and albendazole chemoprophylaxis in experimental hydatidosis. Parasitol Res 87:787–789
Choi B, Jo DH, Anower AKMM, Islam SMS, Sohn S (2016) Chitosan as an immunomodulating adjuvant on T-cells and antigen-presenting cells in herpes simplex virus type 1 infection. Mediat Inflamm. https://doi.org/10.1155/2016/4374375
Cobo F et al (1998) Albendazole plus praziquantel versus albendazole alone as a pre-operative treatment in intra-abdominal hydatidosis caused by Echinococcus granulosus. Tropical Med Int Health 3:462–466
Daniel-Mwambete K, Torrado S, Cuesta-Bandera C, Ponce-Gordo F, Torrado JJ (2004) The effect of solubilization on the oral bioavailability of three benzimidazole carbamate drugs. Int J Pharm 272:29–36. https://doi.org/10.1016/j.ijpharm.2003.11.030
Diaz A, Casaravilla C, Allen JE, Sim RB, Ferreira AM (2011a) Understanding the laminated layer of larval Echinococcus II: immunology. Trends Parasitol 27:263–272. https://doi.org/10.1016/j.pt.2011.01.008
Diaz A, Casaravilla C, Irigoin F, Lin G, Previato JO, Ferreira F (2011b) Understanding the laminated layer of larval Echinococcus I: structure. Trends Parasitol 27:204–213. https://doi.org/10.1016/j.pt.2010.12.012
Elissondo MC, Dopchiz MC, Brasesco M, Denegri G (2004) Echinococcus granulosus: first report of microcysts formation from protoscoleces of cattle origin using the in vitro vesicular culture technique. Parasite 11:415–418. https://doi.org/10.1051/parasite/2004114415
Elissondo M, Ceballos L, Dopchiz M, Andresiuk V, Alvarez L, Sánchez Bruni S, Lanusse C, Denegri G (2007) In vitro and in vivo effects of flubendazole on Echinococcus granulosus metacestodes. Parasitol Res 100:1003–1009. https://doi.org/10.1007/s00436-006-0381-y
Elissondo MC, Ceballos L, Alvarez L, Bruni SS, Lanusse C, Denegri G (2009) Flubendazole and ivermectin in vitro combination therapy produces a marked effect on Echinococcus granulosus protoscoleces and metacestodes. Parasitol Res 105:835–842. https://doi.org/10.1007/s00436-009-1469-y
Fakhar M, Chabra A, Rahimi-Esboei B, Rezaei F (2013) In vitro protoscolicidal effects of fungal chitosan isolated from Penicillium waksmanii and Penicillium citrinum. J Parasit Dis 39:162–167. https://doi.org/10.1007/s12639-013-0300-y
Farhadi M, Fazaeli A, Haniloo A (2015) Genetic characterization of livestock and human hydatid cyst isolates from Northwest Iran, using the mitochondrial cox1 gene sequence. Parasitol Res 114:4363–4370. https://doi.org/10.1007/s00436-015-4673-y
Hemphill A, Muller J (2009) Alveolar and cystic echinococcosis: towards novel chemotherapeutical treatment options. J Helminthol 83:99–111. https://doi.org/10.1017/S0022149x0928936x
Horton J (2003) Albendazole for the treatment of echinococcosis. Fundam Clin Pharmacol 17:205–212
Jung H, Medina L, Garcia L, Fuentes I, Moreno-Esparza R (1998) Absorption studies of albendazole and some physicochemical properties of the drug and its metabolite albendazole sulphoxide. J Pharm Pharmacol 50:43–48
Kang BS, Lee SE, Ng CL, Kim JK, Park JS (2015) Exploring the preparation of albendazole-loaded chitosan-tripolyphosphate nanoparticles. Materials 8:486–498. https://doi.org/10.3390/ma8020486
Li X, Min M, du N, Gu Y, Hode T, Naylor M, Chen D, Nordquist RE, Chen WR (2013) Chitin, chitosan, and glycated chitosan regulate immune responses: the novel adjuvants for cancer vaccine. Clin Dev Immunol 2013:387023. https://doi.org/10.1155/2013/387023
Liang HF, Yang TF, Huang CT, Chen MC, Sung HW (2005) Preparation of nanoparticles composed of poly(gamma-glutamic acid)-poly(lactide) block copolymers and evaluation of their uptake by HepG2 cells. J Control Release 105:213–225. https://doi.org/10.1016/j.jconrel.2005.03.021
Liang W, Wang XC, Wu XW, Zhang SJ, Sun H, Ma X, Peng XY (2014) Efficacy of albendazole chitosan microspheres against Echinococcus granulosus infection in mice. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 32:188–192
Liu Y, Wang XQ, Ren WX, Chen YL, Yu Y, Zhang JK, Bawudong D, Gu JP, Xu XD, Zhang XN (2013) Novel albendazole-chitosan nanoparticles for intestinal absorption enhancement and hepatic targeting improvement in rats. J Biomed Mater Res B Appl Biomater 101B:998–1005. https://doi.org/10.1002/jbm.b.32908
Mohamed AE, Yasawy MI, Al Karawi MA (1998) Combined albendazole and praziquantel versus albendazole alone in the treatment of hydatid disease. Hepatogastroenterology 45:1690–1694
Moro P, Schantz PM (2009) Echinococcosis: a review. Int J Infect Dis 13:125–133. https://doi.org/10.1016/j.ijid.2008.03.037
Nazligul Y, Kucukazman M, Akbulut S (2015) Role of chemotherapeutic agents in the management of cystic echinococcosis. Int Surg 100:112–114. https://doi.org/10.9738/INTSURG-D-14-00068.1
Ogawara K, Yoshida M, Higaki K, Toshikiro Kimura, Shiraishi K, Nishikawa M, Takakura Y, Hashida M (1999) Hepatic uptake of polystyrene microspheres in rats: effect of particle size on intrahepatic distribution. J Control Release 59:15–22. https://doi.org/10.1016/S0168-3659(99)00015-2
Palomares F, Palencia G, Ambrosio JR, Ortiz A, Jung-Cook H (2006) Evaluation of the efficacy of albendazole sulphoxide and praziquantel in combination on Taenia crassiceps cysts: in vitro studies. J Antimicrob Chemother 57:482–488. https://doi.org/10.1093/jac/dki484
Pensel PE, Castro S, Allemandi D, Bruni SS, Palma SD, Elissondo MC (2014) Enhanced chemoprophylactic and clinical efficacy of albendazole formulated as solid dispersions in experimental cystic echinococcosis. Vet Parasitol 203:80–86. https://doi.org/10.1016/j.vetpar.2014.01.027
Richards KS, Morris DL, Taylor DH (1989) Echinococcus multilocularis: ultrastructural effect of in vivo albendazole and praziquantel therapy, singly and in combination. Ann Trop Med Parasitol 83:479–484
Rokni MB (2009) Echinococcosis/hydatidosis in Iran. Iran J Parasitol 4:1–16
Rudt S, Muller RH (1992) Invitro phagocytosis assay of nanoparticles and microparticles by vhemiluminescence. 1. Effect of analytical parameters, particle-size and particle concentration. J Control Release 22:263–271. https://doi.org/10.1016/0168-3659(92)90101-V
Sashiwa H, Aiba SI (2004) Chemically modified chitin and chitosan as biomaterials. Prog Polym Sci 29:887–908. https://doi.org/10.1016/j.progpolymsci.2004.04.001
Torabi N, Dobakhti F, Haniloo A (2017) Albendazole and praziquantel chitosan nanoparticles: preparation, characterization, and in vitro release study. Iran J Sci Technol Trans Sci. https://doi.org/10.1007/s40995-017-0150-z
Torgerson PR (2014) Helminth-Cestode:Echinococcus granulosus and Echinococcus multilocularis. In: Motarjemi Y (ed) Encyclopedia of food safety, vol 2. Academic press, Switzerland, pp 63–69
Urrea-Paris MA, Casado N, Moreno MJ, Rodriguez-Caabeiro F (2001) Chemoprophylactic praziquantel treatment in experimental hydatidosis. Parasitol Res 87:510–512
Urrea-Paris MA, Moreno MJ, Casado N, Rodriguez-Caabeiro F (2000) In vitro effect of praziquantel and albendazole combination therapy on the larval stage of Echinococcus granulosus. Parasitol Res 86:957–964. https://doi.org/10.1007/Pl00008526
Yasawy MI, Alkarawi MA, Mohamed ARE (1993) Combination of praziquantel and albendazole in the treatment of hydatid-disease. Trop Med Parasitol 44:192–194
Acknowledgements
The authors wish to thank Dr. M.C. Elissondo (National Scientific and technical Research Council, Argentina) for helpful and constructive comments. We are also grateful to Dr. Vahid Pirhajati (Iran University of Medical Sciences, Iran) for the technical assistance with the transmission electronic microscopy. Last but not least, we would like to thank Dr. Mehrdad Pedram (Dept. of Genetics and Molecular Medicine, ZUMS) for his science editing and English skills helping us for preparing and proofreading the manuscript.
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This study was financially supported by Zanjan University of Medical Sciences (A-12-95-7).
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Torabi, N., Dobakhti, F., Faghihzadeh, S. et al. In vitro and in vivo effects of chitosan-praziquantel and chitosan-albendazole nanoparticles on Echinococcus granulosus Metacestodes. Parasitol Res 117, 2015–2023 (2018). https://doi.org/10.1007/s00436-018-5849-z
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DOI: https://doi.org/10.1007/s00436-018-5849-z