Evaluating the Glucantime Concentration for the ex vivo Glial Cell Model of Antimony-resistant Leishmania tropica Amastigotes/Antimon Dirençli Leishmania tropica Amastigotlarinin ex vivo Glial Hücre Modeli için Glukantim Konsantrasyonunun Degerlendirilmesi

• Published in: Türkiye parazitolojii dergisi 2021-12, Vol.45 (4), p.237
• Main Authors: Zorbozan, Orcun, Evren, Vedat, Harman, Mehmet, Ozbilgin, Ahmet, Yilmaz, Ozlem Alkan, Turgay, Nevin
• Format: Article
• Language: English
• Subjects: Analysis; Leishmaniasis
• ISSN: 1300-6320
• DOI: 10.4274/tpd.galenos.2021.57966

Objective: Because the protocols used in the treatment of leishmaniasis can be toxic and have many limitations, such as the development of resistance against such protocols, new treatment options are needed, especially against resistant patients. Ex vivo models may be a good source for evaluating new drug options for patients with antimony-resistant parasites. This study aimed to evaluate the Glucantime concentration for our ex vivo glial cell amastigote model we had defined in previous work. Methods: We prepared the astroglial cell culture from brains of 2 to 3 day old neonatal Sprague-Dawley rats under sterile conditions by modifying McCarthy's method. Four plates of cells were infected with antimony-resistant Leishmania tropica promastigotes. After 24 h of incubation, we added Glucantime to 3 plates with different concentrations. After 72 h, we removed the supernatant and then dried, fixed, and stained the plates with Giemsa to count the amastigotes in the glial cells. Results: We observed the amastigotes in glial cells in the control flask. Glial cells were ruined in flasks, which include 75 [micro]g/mL and 37.5 [micro]g/mL Glucantime. The number of amastigotes per 100 glial cells was 116 for the flask with 7.5 [micro]g/mL Glucantime concentration, while 487 for the control flask. Conclusion: We found that while high concentrations of Glucantime were toxic for glial cells, 7.5 [micro]g/mL Glucantime concentration managed to reduce the number of Leishmania tropica amastigotes in glial cells. Keywords: Leishmania tropica, ex vivo, glial cell, amastigote, antimony resistance Amaç: Leishmaniasis tedavisinde kullanilan ilaçlar toksiktir ve pratikte uygulanan tedavi protokollerine karsi direnç gelisimi gibi birçok kisitliligi vardir. Özellikle dirençli hastalar için yeni tedavi seçeneklerine ihtiyaç vardir. Ex vivo modeller, antimon direnci olan hastalarda yeni ilaç seçeneklerini degerlendirmek için iyi bir kaynak olabilir. Bu çalismada, daha önce tanimladigimiz ex vivo glial hücre amastigot modelimizde kullanilabilecek Glukantim konsantrasyonunu degerlendirmeyi amaçladik. Yöntemler: Astroglial hücre kültürü, McCarthy'nin yöntemi degistirilerek steril kosullar altinda 2-3 günlük yenidogan Sprague-Dawley siçan beyinlerinden hazirlandi. Dört hücre plagi antimon dirençli Leishmania tropica promastigotlari ile enfekte edildi. Yirmi dört saatlik inkübasyondan sonra Glukantim farkli konsantrasyonlarda üç plakaya eklendi. Yetmis iki saat sonra üst sivi çikarildi, p