ارزیابی اثرات حفاظتی نانوذرات PLGA حاوی لیپوپلی‌ساکارید سمیت‌زدایی شده باکتری اسینتوباکتر بومانی (Acinetobacter baumannii) در عفونت ریوی موش

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست شناسی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه زیست شناسی، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران

3 گروه ایمنوتراپی، انستیتو علوم دارویی، دانشگاه علوم پزشکی، تهران، ایران

چکیده

اسینتوباکتر بومانی، باکتری هوازی فرصت طلبی است که به عنوان یکی از شایعترین پاتوژن­ های میکروبی با مقاومت آنتی بیوتیکی در ایجاد عفونت تنفسی در بیماران بستری در بخش ICU شناخته می شود. ساخت واکسن می­تواند یکی از راهکارهای موثر در مقابله با این عفونت باشد. این مطالعه، به منظور ارزیابی اثرات حفاظتی نانو ذرات PLGA حاوی لیپوپلی ساکارید(LPS)  سمیت­زدایی شده باکتری اسینتوباکتربومانی به عنوان واکسن در عفونت ریوی موش انجام شد. برای تکثیر انبوه باکتری از محیط کشت مولر هینتون براث استفاده شد.LPS  باکتری به روش آب-فنل داغ استخراج و به کمک NaOH 2/0 مولار سمیت­زدایی شد. انکپسوله کردن LPS سمیت زدایی شده در ذرات  PLGA به روش تبخیر حلال – امولسیون دوتایی آب - در روغن - در آب انجام شد. ذرات تهیه شده بین ۱۵۰ تا ۲۰۰ نانومتر قطر داشتند و دارای بار سطحی منفی بودند. 40 سر موش Balb/C به طور تصادفی به  4 گروه 10 تایی (کنترل، گروه دریافت کننده PLGA، گروه دریافت کننده  D-LPS و گروه دریافت کننده           PLGA-D-LPS ) تقسیم و تمامی گروه ها سه بار به فاصله 14 روز با واکسن تیمار شدند. در روز 35 باکتری های زنده از طریق ریه به گروه ها عرضه شد و پس از 48 ساعت ریه موش­ها جهت مطالعات باکتری شناسی و هیستوپاتولوژی خارج گردید. کشت عصاره همگن شده بافت ریه، تفاوت معنی داری را بین گروه 4 با سایر گروه ها نشان داد (05/0 p <). مطالعه بافت شناسی نیز اثرحفاظتی نانوذرات PLGA حاوی LPS سم زدایی شده را کاملا نشان داد. این مطالعه نشان داد که ذرات PLGA حاوی LPS سمیت زدایی شده اسینتوباکتر بومانی در تحریک سیستم ایمنی موش موفق بوده و می تواند به عنوان واکسن استفاده شود.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of Protective Effects of PLGA Nanoparticles Containing Detoxified Lipopolysaccharide (LPS) Derived from Acinetobacter Baumannii in Mouse Lung Infection

نویسندگان [English]

  • Afshin Gholizadeh 1
  • Reza Shapoori 2
  • Parviz Pakzad 1
  • Mehdi Mahdavi 3
1 Department of Biology, North Tehran Branch, Islamic Azad University, Tehran, Iran
2 Department of Biology, Zanjan Branch, Islamic Azad University, Zanjan, Iran
3 Department of Immunotherapy, Institute of Pharmaceutical Sciences, University of Medical Sciences, Tehran, Iran
چکیده [English]

common microbial pathogens with antibiotic resistance in causing respiratory infections in patients admitted to the ICU. Making a vaccine can be one of the effective ways to combat this infection. This study was performed to evaluate the protective effects of PLGA nanoparticles containing detoxified lipopolysaccharide (D-LPS) of Acinetobacter baumannii as a vaccine in mouse lung infection. Müller Hinton Broth culture medium was used for mass propagation of bacteria. Bacterial LPS was extracted by hot water-phenol method and detoxified with 0.2M NaOH. Encapsulation of detoxified LPS in PLGA particles was performed by Double emulsion solvent evaporation (Water/Oil/Water emulsion). The prepared particles were between 150 and 200 nm in diameter with a negative surface charge. Forty Balb/C mice were randomly divided into four groups of 10 (control, PLGA-receiving group, D-LPS-receiving group, and PLGA-D-LPS-receiving group). All groups were vaccinated three times at intervals of 14 days. On day 35, live bacteria were delivered to the groups through the lungs, and after 48 hours, the mice’s lungs were removed for bacteriological and histopathological studies. Culture of homogenized extract of lung tissue showed a significant difference between group 4 and other groups. (P <0.05) Histological study also showed the protective effect of PLGA nanoparticles containing detoxified LPS. This study showed that PLGA particles containing detoxified LPS of Acinetobacter baumannii were successful in stimulating the immune system of mice and could be used as a vaccine..

کلیدواژه‌ها [English]

  • Acinetobacter baumannii
  • PLGA
  • LPS
  • nanoparticles
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