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The study aimed to assess the effectiveness of pyrimethamine-loaded mPEG\u2013PCL copolymeric nanoparticles (mPEG\u2013PCL\u2013Pyr) against <jats:italic>Toxoplasma gondii<\/jats:italic> (<jats:italic>T. gondii<\/jats:italic>) tachyzoites. The nanoparticles were fabricated using a nano-precipitation method and characterized using DLS and AFM. Through MTT assay, the anti-toxoplasmic activity was gauged. Healthy and infected cell viability, infection index, proliferation index, plaque count, and size were evaluated. Results indicated a maximum drug loading capacity of 11.23% in the nanoparticles. Toxicity testing on Vero cells infected with tachyzoites revealed notable cytotoxic effects at pyrimethamine-equivalent concentrations of 20 and 40 mg\/mL in the mPEG\u2013PCL copolymeric nanoparticle formulation. However, as the results indicated, the administration of mPEG\u2013PCL\u2013Pyr resulted in the reduction of intracellular proliferation, infection index, and the number and size of plaques in comparison to uninfected treated cells (<jats:italic>P<\/jats:italic>\u2009&lt;\u20090.05). The synthesized Nano drug exhibited slightly better performance than pyrimethamine, despite the lack of statistical significance in the observed distinction (<jats:italic>p<\/jats:italic>\u2009&gt;\u20090.05). The study demonstrated that mPEG\u2013PCL copolymeric nanoparticles containing pyrimethamine effectively impede the <jats:italic>in vitro<\/jats:italic> replication of <jats:italic>T. gondii<\/jats:italic>. The study demonstrated that mPEG\u2013PCL copolymeric nanoparticles containing pyrimethamine effectively impede the in vitro replication of <jats:italic>T. gondii<\/jats:italic>. The impact of pyrimethamine resembled that of the synthesized nano-drug on <jats:italic>T. gondii<\/jats:italic> tachyzoites, suggesting the potential of these nanoparticles as a promising avenue for toxoplasmosis treatment.<\/jats:p>","DOI":"10.1186\/s13568-025-01876-8","type":"journal-article","created":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T18:17:40Z","timestamp":1746209860000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Study of anti-Toxoplasma gondii effect of mPEG\u2013PCL copolymeric loaded with pyrimethamine, in vitro"],"prefix":"10.1186","volume":"15","author":[{"given":"Mobina","family":"Gholami","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8956-7895","authenticated-orcid":false,"given":"Hossein","family":"Danafer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7607-7229","authenticated-orcid":false,"given":"Mitra","family":"Sadeghi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8571-5803","authenticated-orcid":false,"given":"Ahmad","family":"Daryani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2990-1123","authenticated-orcid":false,"given":"Seyed Abdollah","family":"Hosseini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2429-7702","authenticated-orcid":false,"given":"Shirzad","family":"Gholami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,5,2]]},"reference":[{"key":"1876_CR1","doi-asserted-by":"publisher","DOI":"10.1042\/BSR20190379","author":"RAA-MW Alajmi","year":"2019","unstructured":"Alajmi RAA-MW, Metwally D, Al-Subaie H, Altamrah N, Barakat AM, Abdel Moneim AE, Al-Otaibi TT, El-Khadragy M (2019) Anti-Toxoplasma activity of silver nanoparticles green synthesized with Phoenix dactylifera and Ziziphus spina-christi extracts which inhibits inflammation through liver regulation of cytokines in Balb\/c mice. 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