Thymoquinone loaded on chitosan nanoparticles alleviated the consequences of cryptosporidiosis infection in a murine model: Evidence from parasitological, histopathological, immunohistochemical, and immunological studies

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Ali A.A.; El-Wakil E.S.; Al-Attar A.-S.R.; Fawzy M.; Samy A.; Eldin Z.E.; Al-Megrin W.A.I.; Nazeer J.T.

Journal: PLOS ONE

Publisher: Public Library of Science

Publication Date: 30 May 2025

Volume / Issue: Volume 20 / Issue 2026-05-05 00:00:00

Article No.: e0325077

ISSN: 19326203

DOI: 10.1371/journal.pone.0325077

Scopus: View on Scopus

PubMed: 40446176

Document Type: Article

Access: All Open Access; Gold Open Access; Green Open Access

Authors and Affiliations

Ali A.A., Pathology Department, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt; El-Wakil E.S., Department of Parasitology, Theodor Bilharz Research Institute, Giza, Egypt; Al-Attar A.-S.R., Pathology Department, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt; Fawzy M., Pathology Department, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt; Samy A., Pathology Department, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt; Eldin Z.E., Materials Science and Nanotechnology Department, Faculty of Postgraduate Studies for Advanced Sciences (PSAS), Beni-Suef University, Beni-Suef, Egypt; Al-Megrin W.A.I., Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia; Nazeer J.T., Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Cairo, Egypt

Abstract

Background Cryptosporidiosis, a parasitic zoonosis caused by the genus Cryptosporidium (C.), currently lacks a vaccine or fully effective treatment. Nitazoxanide (NTZ), the only medication approved by the US Food and Drug Administration for treating cryptosporidiosis, exhibits limited efficacy in immunosuppressed hosts. Thymoquinone (THQ), the active component of Nigella sativa, possesses immunomodulatory, antitumor, hepatoprotective, antioxidant, antimicrobial, and antiprotozoal properties. This study evaluated the therapeutic effects of THQ alone or loaded onto chitosan nanoparticles (CsNPs) against Cryptosporidium parvum infection compared to NTZ. Methods Chitosan nanoparticles were synthesized and characterized using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), zeta potential analysis, and scanning electron microscopy. The cytotoxicity of CsNPs, THQ/CsNPs, and NTZ/ CsNPs was evaluated on HT-29 cells. Mice were divided into seven groups to assess treatment efficacy through parasitological analysis of oocyst shedding, histopathological examination of intestinal, hepatic, and splenic tissues, immunohistochemical analysis using cyclin D1 staining of intestinal tissue, and immunological analysis measuring IFN-γ and IL-10 cytokine levels. Additionally, pharmacokinetic profiles of THQ and NTZ in free and nanoparticle-loaded forms were analyzed. Results XRD confirmed changes in peak position, shape, and intensity following the loading of THQ and NTZ into CsNPs. FTIR spectra demonstrated distinct differences in peak patterns between loaded nanoparticles and individual components, confirming successful drug encapsulation. Moreover, cytotoxicity studies showed dose-dependent effects on cell viability, with NTZ/CsNPs exhibiting the highest cytotoxicity. Regarding oocyst shedding reduction, THQ demonstrated greater efficacy than NTZ (77% vs. 54%), which was further enhanced when loaded onto CsNPs (89% for THQ/CsNPs vs. 78% for NTZ/CsNPs). Histopathological analysis revealed the restoration of structural alterations in intestinal, hepatic, and splenic tissues in treated groups. Cyclin D1 immunohistochemical staining showed a significant reduction in immunoreactivity in the THQ/CsNP-treated group compared to other treatments. Furthermore, immune responses were modulated by nanoparticle therapies, with significantly lower IFN-γ levels and higher IL-10 levels in treated groups. Pharmacokinetic analysis demonstrated that CsNP formulations significantly improved drug bioavailability by achieving higher peak plasma concentrations (Cmax), earlier time to peak concentration (Tmax), and prolonged half-life (t1/2) compared to free drugs. Conclusion Thymoquinone demonstrated significant potential as an anti-cryptosporidiosis therapeutic agent, with enhanced efficacy when loaded onto chitosan nanoparticles. Chitosan-based nanoparticle formulations improved the pharmacokinetic profiles of both THQ and NTZ, offering a promising strategy for enhancing drug bioavailability and retention while reducing parasitic burden and modulating immune responses effectively. © 2025 Ali et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Keywords

Animals; Benzoquinones; Chitosan; Cryptosporidiosis; Cryptosporidium parvum; Disease Models, Animal; Female; HT29 Cells; Humans; Mice; Nanoparticles; chitosan nanoparticle; cyclin D1; dexamethasone; dexazone; doxorubicin; gamma interferon; interleukin 10; isoflurane; nitazoxanide; thymoquinone; xylene; benzoquinone derivative; nanoparticle; anesthesia; animal cell; animal experiment; animal model; animal tissue; Article; atrophy; body weight; cell viability; controlled study; cytotoxicity; dispersity; drug bioavailability; edema; erythrocyte; field emission scanning electron microscopy; Fourier transform infrared spectroscopy; hepatocyte atrophy; histopathology; HT-29 cell line; human; human cell; hyperplasia; immunohistochemistry; inflammatory cell; Kupffer cell; male; megakaryocyte; microsteatosis; mouse; MTT assay; necrosis; nonhuman; oocyst; photon correlation spectroscopy; rat; scanning electron microscopy; submucosal edema; X ray diffraction; animal; chemistry; disease model; drug effect; drug therapy; immunology; parasitology; pathology

Citation Information

Scopus Citations: 3

For comprehensive information about the Theodor Bilharz Research Institute (TBRI), its institutional activities, scientific and research achievements, clinical and hospital services, and the diverse expertise offered through its 22 specialized research and clinical departments, as well as opportunities for professional training, specialized workshops, and scientific conferences, readers are invited to visit the Institute’s official website.

The website provides regularly updated information on the Institute’s latest news, research activities, scientific initiatives, clinical services, institutional programs, and academic and professional opportunities.

English Website: https://www.tbri.sci.eg/en/

Arabic Website: https://www.tbri.sci.eg/ar/

Prepared and Uploaded by:

Abdalla F. Abdalla

Electronic Portal Unit

Electronic Portal Unit

Popular Posts

Transcriptome analysis for Schistosoma vector snail’s Oncomelania hupensis exposed to silver nanoparticles

Image
Bibliographic Information Authors: Zayed K.M.; Zhou X.-N. Journal: Journal of Parasitic Diseases Publisher: Springer Publication Date: 3 September 2026 ISSN: 9717196 DOI: 10.1007/s12639-026-01986-5 Scopus: View on Scopus Document Type: Article Authors and Affiliations Zayed K.M., Medical Malacology Department, Theodor Bilharz Research Institute, Giza, 12411, Egypt, National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention, Shanghai, 200025, China, NHC Key Laboratory of Parasite and Vector Biology, WHO Collaborating Centre for Tropical Diseases, National Center for International Research On Tropical Diseases, Ministry of Science and Technology, Shanghai, 200025, China; Zhou X.-N., National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention, Shanghai, 200025, China, NHC Key Laboratory of Parasite and Vector Biology, WHO Collaborating Centre for Tropical Diseases, National Center for International Res...
Read more

Interpretation of liver stiffness measurement in patients with mixed liver disease etiologies

Image
Bibliographic Information Authors: El Ray A.; El Ghannam M.; Rautou P.E. Journal: Arab Journal of Gastroenterology Publisher: Elsevier Ltd Publication Date: August 2026 ISSN: 16871979 DOI: 10.1016/j.ajg.2026.05.002 Scopus: View on Scopus PubMed: 42362465 Document Type: Letter Authors and Affiliations El Ray A., Theodor Bilharz Research Institute, Hepatogastroenterology Department, Giza, Egypt, Faculty of Medicine, October 6 University, Egypt; El Ghannam M., Theodor Bilharz Research Institute, Hepatogastroenterology Department, Giza, Egypt; Rautou P.E., Université Paris-Cité, Inserm, Centre de recherche sur l’inflammation, UMR 1149, Paris, France, AP-HP, Hôpital Beaujon, Service d’Hépatologie, DMU DIGEST, Centre de Référence des Maladies Vasculaires du Foie, FILFOIE, ERN RARE-LIVER, Clichy, France Abstract Vibration-controlled transient elastography (VCTE)–based liver stiffness measurement (LSM) is widely used for non-invasive assessment of liver fibrosis and p...
Read more

Multifunctional polycaprolactone-based composite fibers with icariin and glutathione for effective repair of critical-sized bone defects

Image
Bibliographic Information Authors: Yessuf A.M.; Zhang Y.; Sharma B.P.; Wang Y.; Kassa T.S.; Qadir A.; Jinqiu Y.; Mohideen M.M.; Salama A.M.; Wang W.; Zhang Q.; Liu Y. Journal: BMEMat Publisher: John Wiley and Sons Inc Publication Date: 30 July 2026 ISSN: 27517438 DOI: 10.1002/bmm2.70115 Scopus: View on Scopus Document Type: Article Access: All Open Access; Gold Open Access Authors and Affiliations Yessuf A.M., Key Laboratory of Advanced Functional Polymer Composites, College of Materials Science and Engineering, Beijing University of Chemical Technology, Beijing, China; Zhang Y., Henan Provincial Orthopedic Hospital, Henan, Zhengzhou, China; Sharma B.P., Key Laboratory of Electrochemical Process and Technology of Materials, College of Materials Science and Engineering, Beijing University of Chemical Technology, Beijing, China; Wang Y., Beijing University of Chinese Medicine, Beijing, China; Kassa T.S., Key Laboratory of Advanced Functional Polymer Composites, Col...
Read more

Holothuria arenicola Extract-Loaded Polycaprolactone Nanocapsules Attenuate Bile Duct Ligation-Induced Acute Liver Injury

Image
Bibliographic Information Authors: Mansour E.M.; Shemis M.; Fahmy S.R.; Amer M.A.; Mohamed A.S. Journal: Current Nanomedicine Publisher: Bentham Science Publishers Publication Date: September 2026 Volume / Issue: Volume 16 / Issue 5 Pages: 771–786 ISSN: 24681873 DOI: 10.2174/0124681873401484250924125945 Scopus: View on Scopus Document Type: Article Authors and Affiliations Mansour E.M., Department of Biochemistry and Molecular Biology, Theodor Bilharz Research Institute, Giza, Egypt; Shemis M., Department of Biochemistry and Molecular Biology, Theodor Bilharz Research Institute, Giza, Egypt; Fahmy S.R., Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt; Amer M.A., Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt; Mohamed A.S., Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt Abstract Introduction: Cholestatic liver diseases are characterized by jaundice and cholestasis with various conseq...
Read more

Mytilus edulis-mediated green synthesis of selenium nanoparticles with antimicrobial and molluscicidal applications

Image
Bibliographic Information Authors: Dokmak H.-A.A.; AlSedawy M.; Ramadan M.A.; Amin B.H.; Faid A.H.; Abdelmaksoud H.F.; El-Sayed S.M. Journal: Scientific Reports Publisher: Nature Research Publication Date: 7 September 2026 Volume / Issue: Volume 16 / Issue 1 Article No.: 27918 ISSN: 20452322 DOI: 10.1038/s41598-026-65860-3 Scopus: View on Scopus PubMed: 42706305 Document Type: Article Access: All Open Access; Gold Open Access; Green Open Access Authors and Affiliations Dokmak H.-A.A., Department of Medical Malacology, Theodor Bilharz Research Institute, Giza, Egypt; AlSedawy M., Microbiology and Immunology Department, Faculty of Pharmacy (For Boys), Al-Azhar University, Cairo, Egypt; Ramadan M.A., Department of Laser Application in Metrology, Photochemistry and Agriculture, National Institute of Laser Enhanced Science (NILES) Cairo University (CU), Giza, Egypt; Amin B.H., The Regional Center for Mycology and Biotechnology, Al-Azhar University, Cairo, 11787,...
Read more

Microbial levan potentiates hepatic retention and antitumor activity of a PEGylated benzimidazole–curcumin nanocomplex through TLR2–FXR/FGF15-associated immunometabolic remodeling in experimental liver cancer

Image
Bibliographic Information Authors: Shawky H.; Ibrahim N.E.; Amer M.N.; Al-Ashmawy A.A.K.; Hammam O.A.; Fayed D.B.; Maghraby A.S. Journal: Scientific Reports Publisher: Nature Research Publication Date: 12 August 2026 Volume / Issue: Volume 16 / Issue 1 Article No.: 25094 ISSN: 20452322 DOI: 10.1038/s41598-026-64576-8 Scopus: View on Scopus PubMed: 42581056 Document Type: Article Access: All Open Access; Gold Open Access; Green Open Access Authors and Affiliations Shawky H., Therapeutic Chemistry Department, Pharmaceutical and Drug Industries Research Institute, National Research Centre, Dokki, Cairo, 12622, Egypt; Ibrahim N.E., Microbial Biotechnology Department, Biotechnology Research Institute, National Research Centre, Dokki, Cairo, 12622, Egypt; Amer M.N., Chemistry of Natural and Microbial Products Department, Pharmaceutical and Drug Industries Research Institute, National Research Centre, Dokki, Cairo, 12622, Egypt; Al-Ashmawy A.A.K., Therapeutic Chem...
Read more

Variable clinical presentations of pulmonary hydatid cysts: a four-case series from a single center in United Arab Emirates, non-endemic region

Image
Bibliographic Information Authors: Yehya M.; Abdallah M.F.H.; Ajeka S.; Al-Masalmeh F.H.; Albaghdadi M.O.J. Journal: Journal of Surgical Case Reports Publisher: Oxford University Press Publication Date: 29 July 2026 Volume / Issue: Volume 2026 / Issue 7 Article No.: rjag658 ISSN: 20428812 DOI: 10.1093/jscr/rjag658 Scopus: View on Scopus Document Type: Article Access: All Open Access; Gold Open Access; Green Open Access Authors and Affiliations Yehya M., Department of Thoracic Surgery, SAQR Hospital, Al Juwais Al Kadi St, Ras Al-Khaimah, United Arab Emirates; Abdallah M.F.H., Department of Radiology, SAQR Hospital, Al Juwais Al Kadi St, Ras Al-Khaimah, United Arab Emirates, Theodor Bilharz Research Institute, Mahad Al Abhas Al bahari St, Kornish Al Nile, Warraq Al Arab, Al Warak, Governorate, Giza, 3863341, Egypt; Ajeka S., Department of General Surgery, SAQR Hospital, Al Juwais Al Kadi St, Ras Al-Khaimah, United Arab Emirates; Al-Masalmeh F.H., Department of ...
Read more