Nanoemulsion of Spiramycin against Tachyzoites of Toxoplasma gondii, RH Strain: Preparation, Toxicology, and Efficacy Studies
-
Saeideh Hashemi-Hafshejani
,
Amir Amani
,
Sanaz Jafarpour Azami
,
Hossien Keshavarz Valian
,
Mehdi Mohebali
,
Mahboobeh Salimi
,
Hossien Hassani Lafmejan Pour
,
Saeedeh Shojaee
Abstract
Background: Toxoplasma infection is caused by Toxoplasma gondii, which is an intracellular protozoan parasite. This infection consequently lead various congenital disabilities during pregnancy in patients. Spiramycin (Spi), a macrolide antibiotic, is typically recommended for T. gondii infection in pregnant women. We aimed to prepare the nanoemulsion of spiramycin (NE-Spi) and to evaluate the activity of this formulation in tachyzoites of T. gondii, RH strain.
Methods: This study was conducted in 2019-2021 at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. NE-Spi was prepared by spontaneous emulsification. The effects of this nanoemulsion on the viability of cultured cells were measured using MTT assay. To estimate the effects of NE-Spi on tachyzoites of T. gondii, RH strain, different concentrations of NE-Spi, S-Spi (suspension of spiramycin), and NE (nanoemulsion without any spiramycin) were added to tachyzoites and then stored for 30, 60, 90, 120 min and 24 h in 250 µg/ml concentration at room temperature. Finally, Tachyzoites mortality rates were evaluated by trypan blue staining. Of note, flow cytometry was conducted to confirm the obtained results.
Results: The final particle size of NE-Spi was calculated to be 11.3 nm by DLS and TEM. Thereafter, using MTT assay, in 62.5 µg/ml concentration of NE-Spi, the Vero cells viability was obtained as 82%. The highest mortality rates of tachyzoites of T.gondii, RH strain were observed at 250 µg/ml concentration and after 120 min of exposure, but it was not significantly different from 24 h of exposure.
Conclusion: NE-Spi has lethal efficacy on T. gondii RH strain in-vitro.
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2. Khosravi M, Rahimi M, Doroud D, Mirsa-madi ES, Mirjalali H, Zali MR (2020). In vitro Evaluation of Mannosylated Paromomycin-Loaded Solid Lipid Na-noparticles on Acute Toxoplasmosis. Front Cell Infect Microbiol, 10:33-33.
3. Shojaee S, Firouzeh N, Keshavarz H, Aza-mi SJ-P, Salimi M, Mohebali M (2019). Nanosilver Colloid Inhibits Toxoplasma gondii Tachyzoites and Bradyzoites in Vitro. Iran J Parasitol, 14:362-67.
4. Medawar-Aguilar V, Jofre CF, Fernández-Baldo MA, et al (2019). Serological diag-nosis of Toxoplasmosis disease using a fluorescent immunosensor with chi-tosan-ZnO-nanoparticles. Anal Biochem, 564-565:116-122.
5. Shammaa AM, Powell TG, Benmerzouga I (2021). Adverse outcomes associated with the treatment of Toxoplasma infec-tions. Sci Rep, 11:1-8.
6. Zhang D, Ren L, Zhao M, et al (2019). Role of Tim-3 in Decidual Macrophage Func-tional Polarization during Abnormal Pregnancy with Toxoplasma gondii Infec-tion. Front Immunol, 10:1550.
7. Selseleh MM, Keshavarz H, Mohebali M, et al (2012). Production and evaluation of Toxoplasma gondii recombinant surface antigen 1 (SAG1) for serodiagnosis of acute and chronic Toxoplasma infection in human sera. Iran J Parasitol, 7(3):1-9.
8. Ali-Heydari S, Keshavarz H, Shojaee S, Mohebali M (2013). Diagnosis of anti-genic markers of acute toxoplasmosis by IgG avidity immunoblotting. Parasite, 20:18.
9. Teimouri A, Mohtasebi S, Kazemirad E, Keshavarz H (2020). Role of Toxoplasma gondii IgG avidity testing in discriminat-ing between acute and chronic toxo-plasmosis in pregnancy. J Clin Microbiol, 58 (9):e00505-20.
10. Plants M (2014). A brief insight on anti-Toxoplasma gondii activity of some medic-inal plants Aperito J Bacteriol Virol Parasi-tol, 1:107.
11. Tavares H, Cardoso J, Almeida T, et al (2021). Spiramyin-loaded PLGA im-plants for the treatment of ocular toxo-plasmosis: development, characteriza-tion, biocompatibility, and anti-Toxoplasma activity. Pharmazie, 76:68-76.
12. Omar M, Abaza BE, Mousa E, Ibrahim SM, Rashed HE, Farag TI (2021). Effect of spiramycin versus aminoguanidine and their combined use in experimental toxoplasmosis. J Parasit Dis, 45(4):1014-1025.
13. Abaza SM (2016). Applications of nano-medicine in parasitic diseases. Parasitol United J, 9(1):1.
14. Hagras NA-e, Allam AF, Farag HF, et al (2019). Successful treatment of acute ex-perimental toxoplasmosis by spiramy-cin-loaded chitosan nanoparticles. Exp Parasitol, 204:107717.
15. Kumar M, Bishnoi RS, Shukla AK, Jain CP (2019). Techniques for formulation of nanoemulsion drug delivery system: a review. Prev Nutr Food Sci, 24(3):225.
16. Hagras NA-e, Mogahed NMFH, Sheta E, et al (2022). The powerful synergistic effect of spiramycin/propolis loaded chi-tosan/alginate nanoparticles on acute murine toxoplasmosis. PLoS Negl Trop Dis, 16(3):e0010268.
17. Jafarpour Azami S, Teimouri A, Keshavarz H, et al (2018). Curcumin nanoemulsion as a novel chemical for the treatment of acute and chronic toxoplasmosis in mice. Int J Nanomedicine, 13:7363-74.
18. Assolini JP, Concato VM, Gonçalves MD, Carloto ACM, Conchon-Costa I, Pa-vanelli WR, Melanda FN, Costa IN (2017). Nanomedicine advances in tox-oplasmosis: diagnostic, treatment, and vaccine applications. J Parasitol Res, 116:1603-1615.
19. Nasr Me, Abd EL Hamid AH, et al (2020). Efficacy of azithromycin on experi-mental toxoplasmosis infected mice. J Egypt Soc Parasitol, 50(2):293-299.
20. Cheraghipour K, Masoori L, Ezzatkhah F, et al (2020). Effect of chitosan on Toxo-plasma gondii infection: A systematic re-view. Parasite Epidemiol Control, 11:e00189.
21. Mandelbrot L, Kieffer F, Sitta R, et al (2018). Prenatal therapy with py-rimethamine + sulfadiazine vs spiramy-cin to reduce placental transmission of toxoplasmosis: a multicenter, random-ized trial. Am J Obstet Gynecol, 219(4):386.e1-386.e9.
22. Montazeri M, Sharif M, Sarvi S, Mehrzadi S, Ahmadpour E, Daryani A (2017). A sys-tematic review of in vitro and in vivo activities of anti-Toxoplasma drugs and compounds (2006–2016). Front Microbiol, 8:25.
23. Yao K, Gao S, Wu Y, Zhao Z, Wang W, Mao Q (2018). Influence of dextrins on the production of spiramycin and impu-rity components by Streptomyces ambo-faciens. Folia Microbiol, 63:105-113.
24. Zhang X, Wu X, Xie F, Wang Z, Zhang X, Jiang L (2017). Physicochemical proper-ties and in vitro dissolution of Spiramy-cin microparticles using the homoge-nate-Antisolvent precipitation process. Applied Sciences, 7(1):10.
25. Shafiq S, Shakeel F, Talegaonkar S, Ahmad FJ, Khar RK, Ali M (2007). Development and bioavailability assessment of rami-pril nanoemulsion formulation. Eur J Pharm Biopharm, 66(2):227-243.
26. Sutradhar KB, Amin ML (2013). Nanoemul-sions: increasing possibilities in drug de-livery. Eur J Nanomed, 5(2):97-110.
27. Jafarpour Azami S, Rahimi HM, Mirjalali H, Zali MR (2021). Unravelling Toxoplas-ma treatment: conventional drugs toward nanomedicine. World J Microbiol Biotechnol, 37:1-9.
28. Teimouri A, Azami SJ, Keshavarz H, et al (2018). Anti-Toxoplasma activity of various molecular weights and concentrations of chitosan nanoparticles on tachyzoites of RH strain. Int J Nanomedicine, 13:1341-51.
29. Keyhani A, Shakibaie M, Mahmoudvand H, et al (2020). Prophylactic activity of bio-genic selenium nanoparticles against chronic Toxoplasma gondii infection. Re-cent Pat Antiinfect Drug Discov, 15(1):75-84.
30. FarahatAllam A, Shehab AY, Fawzy Hus-sein Mogahed NM, Farag HF, Elsayed Y, Abd El-Latif NF (2020). Effect of nita-zoxanide and spiramycin metronidazole combination in acute experimental tox-oplasmosis. Heliyon, 6(4):e03661.
31. Saadatmand M, Al-Awsi GRL, Alanazi AD, et al (2021). Green synthesis of zinc na-noparticles using Lavandula angustifolia Vera. Extract by microwave method and its prophylactic effects on Toxoplasma gondii infection. Saudi J Biol Sci, 28(11):6454-6460.
32. Machado LF, Sanfelice RA, Bosqui LR, et al (2020). Biogenic silver nanoparticles re-duce adherence, infection, and prolifer-ation of Toxoplasma gondii RH strain in HeLa cells without inflammatory media-tors induction. Exp Parasitol, 211:107853.
33. Schöler N, Krause K, Kayser O, et al (2001). Atovaquone nanosuspensions show excellent therapeutic effect in a new murine model of reactivated toxo-plasmosis. Antimicrob Agents Chemother, 45(6):1771-1779.
34. Hamad HK, Ramadan NF, Mohamed SH, Aly I, Zalat R (2020). Study the synergis-tic Effect between Nanoparticles and Spiramycin on Immunological Re-sponse against Toxoplasmosis. IOP Conference Series: Materials Science and Engineering, IOP Publishing, pp. 022091.
2. Khosravi M, Rahimi M, Doroud D, Mirsa-madi ES, Mirjalali H, Zali MR (2020). In vitro Evaluation of Mannosylated Paromomycin-Loaded Solid Lipid Na-noparticles on Acute Toxoplasmosis. Front Cell Infect Microbiol, 10:33-33.
3. Shojaee S, Firouzeh N, Keshavarz H, Aza-mi SJ-P, Salimi M, Mohebali M (2019). Nanosilver Colloid Inhibits Toxoplasma gondii Tachyzoites and Bradyzoites in Vitro. Iran J Parasitol, 14:362-67.
4. Medawar-Aguilar V, Jofre CF, Fernández-Baldo MA, et al (2019). Serological diag-nosis of Toxoplasmosis disease using a fluorescent immunosensor with chi-tosan-ZnO-nanoparticles. Anal Biochem, 564-565:116-122.
5. Shammaa AM, Powell TG, Benmerzouga I (2021). Adverse outcomes associated with the treatment of Toxoplasma infec-tions. Sci Rep, 11:1-8.
6. Zhang D, Ren L, Zhao M, et al (2019). Role of Tim-3 in Decidual Macrophage Func-tional Polarization during Abnormal Pregnancy with Toxoplasma gondii Infec-tion. Front Immunol, 10:1550.
7. Selseleh MM, Keshavarz H, Mohebali M, et al (2012). Production and evaluation of Toxoplasma gondii recombinant surface antigen 1 (SAG1) for serodiagnosis of acute and chronic Toxoplasma infection in human sera. Iran J Parasitol, 7(3):1-9.
8. Ali-Heydari S, Keshavarz H, Shojaee S, Mohebali M (2013). Diagnosis of anti-genic markers of acute toxoplasmosis by IgG avidity immunoblotting. Parasite, 20:18.
9. Teimouri A, Mohtasebi S, Kazemirad E, Keshavarz H (2020). Role of Toxoplasma gondii IgG avidity testing in discriminat-ing between acute and chronic toxo-plasmosis in pregnancy. J Clin Microbiol, 58 (9):e00505-20.
10. Plants M (2014). A brief insight on anti-Toxoplasma gondii activity of some medic-inal plants Aperito J Bacteriol Virol Parasi-tol, 1:107.
11. Tavares H, Cardoso J, Almeida T, et al (2021). Spiramyin-loaded PLGA im-plants for the treatment of ocular toxo-plasmosis: development, characteriza-tion, biocompatibility, and anti-Toxoplasma activity. Pharmazie, 76:68-76.
12. Omar M, Abaza BE, Mousa E, Ibrahim SM, Rashed HE, Farag TI (2021). Effect of spiramycin versus aminoguanidine and their combined use in experimental toxoplasmosis. J Parasit Dis, 45(4):1014-1025.
13. Abaza SM (2016). Applications of nano-medicine in parasitic diseases. Parasitol United J, 9(1):1.
14. Hagras NA-e, Allam AF, Farag HF, et al (2019). Successful treatment of acute ex-perimental toxoplasmosis by spiramy-cin-loaded chitosan nanoparticles. Exp Parasitol, 204:107717.
15. Kumar M, Bishnoi RS, Shukla AK, Jain CP (2019). Techniques for formulation of nanoemulsion drug delivery system: a review. Prev Nutr Food Sci, 24(3):225.
16. Hagras NA-e, Mogahed NMFH, Sheta E, et al (2022). The powerful synergistic effect of spiramycin/propolis loaded chi-tosan/alginate nanoparticles on acute murine toxoplasmosis. PLoS Negl Trop Dis, 16(3):e0010268.
17. Jafarpour Azami S, Teimouri A, Keshavarz H, et al (2018). Curcumin nanoemulsion as a novel chemical for the treatment of acute and chronic toxoplasmosis in mice. Int J Nanomedicine, 13:7363-74.
18. Assolini JP, Concato VM, Gonçalves MD, Carloto ACM, Conchon-Costa I, Pa-vanelli WR, Melanda FN, Costa IN (2017). Nanomedicine advances in tox-oplasmosis: diagnostic, treatment, and vaccine applications. J Parasitol Res, 116:1603-1615.
19. Nasr Me, Abd EL Hamid AH, et al (2020). Efficacy of azithromycin on experi-mental toxoplasmosis infected mice. J Egypt Soc Parasitol, 50(2):293-299.
20. Cheraghipour K, Masoori L, Ezzatkhah F, et al (2020). Effect of chitosan on Toxo-plasma gondii infection: A systematic re-view. Parasite Epidemiol Control, 11:e00189.
21. Mandelbrot L, Kieffer F, Sitta R, et al (2018). Prenatal therapy with py-rimethamine + sulfadiazine vs spiramy-cin to reduce placental transmission of toxoplasmosis: a multicenter, random-ized trial. Am J Obstet Gynecol, 219(4):386.e1-386.e9.
22. Montazeri M, Sharif M, Sarvi S, Mehrzadi S, Ahmadpour E, Daryani A (2017). A sys-tematic review of in vitro and in vivo activities of anti-Toxoplasma drugs and compounds (2006–2016). Front Microbiol, 8:25.
23. Yao K, Gao S, Wu Y, Zhao Z, Wang W, Mao Q (2018). Influence of dextrins on the production of spiramycin and impu-rity components by Streptomyces ambo-faciens. Folia Microbiol, 63:105-113.
24. Zhang X, Wu X, Xie F, Wang Z, Zhang X, Jiang L (2017). Physicochemical proper-ties and in vitro dissolution of Spiramy-cin microparticles using the homoge-nate-Antisolvent precipitation process. Applied Sciences, 7(1):10.
25. Shafiq S, Shakeel F, Talegaonkar S, Ahmad FJ, Khar RK, Ali M (2007). Development and bioavailability assessment of rami-pril nanoemulsion formulation. Eur J Pharm Biopharm, 66(2):227-243.
26. Sutradhar KB, Amin ML (2013). Nanoemul-sions: increasing possibilities in drug de-livery. Eur J Nanomed, 5(2):97-110.
27. Jafarpour Azami S, Rahimi HM, Mirjalali H, Zali MR (2021). Unravelling Toxoplas-ma treatment: conventional drugs toward nanomedicine. World J Microbiol Biotechnol, 37:1-9.
28. Teimouri A, Azami SJ, Keshavarz H, et al (2018). Anti-Toxoplasma activity of various molecular weights and concentrations of chitosan nanoparticles on tachyzoites of RH strain. Int J Nanomedicine, 13:1341-51.
29. Keyhani A, Shakibaie M, Mahmoudvand H, et al (2020). Prophylactic activity of bio-genic selenium nanoparticles against chronic Toxoplasma gondii infection. Re-cent Pat Antiinfect Drug Discov, 15(1):75-84.
30. FarahatAllam A, Shehab AY, Fawzy Hus-sein Mogahed NM, Farag HF, Elsayed Y, Abd El-Latif NF (2020). Effect of nita-zoxanide and spiramycin metronidazole combination in acute experimental tox-oplasmosis. Heliyon, 6(4):e03661.
31. Saadatmand M, Al-Awsi GRL, Alanazi AD, et al (2021). Green synthesis of zinc na-noparticles using Lavandula angustifolia Vera. Extract by microwave method and its prophylactic effects on Toxoplasma gondii infection. Saudi J Biol Sci, 28(11):6454-6460.
32. Machado LF, Sanfelice RA, Bosqui LR, et al (2020). Biogenic silver nanoparticles re-duce adherence, infection, and prolifer-ation of Toxoplasma gondii RH strain in HeLa cells without inflammatory media-tors induction. Exp Parasitol, 211:107853.
33. Schöler N, Krause K, Kayser O, et al (2001). Atovaquone nanosuspensions show excellent therapeutic effect in a new murine model of reactivated toxo-plasmosis. Antimicrob Agents Chemother, 45(6):1771-1779.
34. Hamad HK, Ramadan NF, Mohamed SH, Aly I, Zalat R (2020). Study the synergis-tic Effect between Nanoparticles and Spiramycin on Immunological Re-sponse against Toxoplasmosis. IOP Conference Series: Materials Science and Engineering, IOP Publishing, pp. 022091.
| Files | ||
| Issue | Vol 52 No 7 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v52i7.13252 | |
| Keywords | ||
| Nanoemulsion Spiramycin Toxoplasma Tachyzoite RH strain | ||
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How to Cite
1.
Hashemi-Hafshejani S, Amani A, Jafarpour Azami S, Keshavarz Valian H, Mohebali M, Salimi M, Hassani Lafmejan Pour H, Shojaee S. Nanoemulsion of Spiramycin against Tachyzoites of Toxoplasma gondii, RH Strain: Preparation, Toxicology, and Efficacy Studies. Iran J Public Health. 2023;52(7):1495-1503.
