ارزیابی تجربی و درون‌سیلیکویی (In Silico) یک فیلم کامپوزیتی نوین ZnO/Alhagi maurorum/PVA علیه سویه‌های مقاوم به چند دارو از Pseudomonas aeruginosa حامل ژن‌های blaTEM و blaIMP

مقالات آماده انتشار

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست‌شناسی، دانشکده علوم برای بانوان، دانشگاه بابل، عراق.

2 گروه زیست‌شناسی، دانشکده علوم برای بانوان، دانشگاه بابل، عراق

3 گروه زیست‌شناسی، دانشکده علوم، دانشگاه سبز القاسم، بابل 51013، عراق

4 مرکز تحقیقات DNA، دانشگاه بابل، عراق

5 دانشگاه فنی میانه، مؤسسه فنی کوت، عراق

10.22103/jab.2026.27227.1909

چکیده

هدف: باکتری سودوموناس آئروژینوزا یکی از عوامل اصلی عفونت‌های ناشی از سوختگی است که به دلیل بیماری‌زایی بالا و مقاومت دارویی شناخته می‌شود. این مطالعه با هدف تهیه و مشخصه‌یابی فیلم نانوکامپوزیتی ZnO/PVA حاوی عصاره گیاه Alhagi maurorum برای مقابله با سویه‌های مقاوم به چند دارو از P. aeruginosa جداشده از عفونت‌های پوستی ناشی از سوختگی انجام شد. همچنین شناسایی مولکولی و تعیین توالی ژن‌های blaTEM و blaIMP و به دنبال آن تحلیل اتصال مولکولی درون‌سیلیکویی به منظور بررسی مکانیسم ضدباکتریایی انجام گرفت.
مواد و روش‌ها: سرشاخه‌های گیاه Alhagi maurorum جمع‌آوری و نگهداری شدند. آنالیز GC-MS بر روی عصاره گیاهی انجام شد. نانوذرات اکسید روی در ماتریس پلیمری به کار گرفته شدند. ارزیابی‌ برای تأیید به‌دام‌افتادن موفق و توزیع یکنواخت نانوترکیبات زیست‌فعال در ماتریس پلیمری فیلم کامپوزیتی ساخته‌شده انجام شد. تعداد 50 جدایه باکتریایی از سواب‌های بیماران بستری به دست آمد. حساسیت ضد‌میکروبی جدایه‌های P. aeruginosa (20 جدایه) تعیین شد. DNA ژنومی از جدایه‌های باکتریایی استخراج و PCR انجام گردید. توالی‌های نوکلئوتیدی ژن blaTEM مربوط به P. aeruginosa از پایگاه NCBI استخراج شد. برای تجزیه و تحلیل آماری از نرم‌افزار SPSS استفاده گردید.
نتایج: نتایج GC-MS نشان داد که طیف جرمی و ساختار دوازده ترکیب شیمیایی استخراج‌شده از گیاه Alhagi maurorum با استفاده از متانول در داده‌های GC-MS قابل مشاهده است. در میان این ترکیبات، اسید 9،12-اکتادکادی‌انوئیک (Z,Z) (اسید لینولئیک) از ویژگی خاصی برخوردار بود. با ترکیب این مواد فیتوشیمیایی در ماتریس پلیمری PVA همراه با نانوذرات روی، فیلمی زیست‌تخریب‌پذیر تولید شد. این فیلم در برابر جدایه‌های بالینی Pseudomonas aeruginosa که دارای مقاومت دارویی بودند، مورد آزمایش قرار گرفت. از میان 50 جدایه باکتریایی، 20 جدایه به عنوان P. aeruginosa شناسایی شدند. نتایج PCR وجود ژن‌های blaTEM و blaIMP را در درصد قابل توجهی از جدایه‌ها نشان داد. نتایج داکینگ مولکولی حاکی از آن بود که نانوساختار ZnO در مقایسه با اسید لینولئیک و PVA بیشترین توانایی اتصال به پروتئین را دارد. میل اتصال قوی و ثابت مهاری پایین ZnO نقش بالقوه آن را به عنوان یک متصل‌شونده یا مهارکننده پایدار نشان می‌دهد، در حالی که اسید لینولئیک و PVA کارایی کمتری داشتند.
نتیجه‌گیری: خواص ضدباکتریایی قوی این فیلم کامپوزیتی احتمالاً حاصل اثر هم‌افزای پلیمر PVA، نانوذرات روی و عصاره گیاهی است. فیلم کامپوزیتی ZnO/Alhagi/PVA فعالیت‌های ضدباکتریایی و آنتی‌اکسیدانی امیدبخشی از خود نشان داد و می‌تواند به عنوان یک ماده زیست‌تخریب‌پذیر بالقوه برای پانسمان زخم در مدیریت عفونت‌های ناشی از P. aeruginosa مقاوم به چند دارو مورد استفاده قرار گیرد.

کلیدواژه‌ها

عنوان مقاله [English]

Experimental and In Silico evaluation of a novel ZnO/Alhagi maurorum/PVA composite film against multidrug-resistant Pseudomonas aeruginosa harboring blaTEM and blaIMP genes

نویسندگان [English]

  • Nisreen Kaddim Radi 1
  • Shireen Sabah Kamil 2
  • Safa Hasan Radhi 3
  • Hussein J. Hussein 2
  • Ali H. Al-Marzoqi 2
  • Lubna Abdulazeem 4
  • Israa A. Ibraheam 2
  • Ali Salam Kashkool 5
1 Department of Biology, College of Science for Women, University of Babylon, Iraq.
2 Department of Biology, College of science for women, University of Babylon, Iraq.
3 Biology Department, College of science, Al-Qasim Green University, Babylon 51013, Iraq
4 DNA Research center, University of Babylon, Iraq.
5 Middle Technical University, Kut Technical Institute, Iraq
چکیده [English]

Objective
A major contributor to burn infections, Pseudomonas aeruginosa is known for its aggressiveness and medication resistance. This study was carried out to prepare and characterize ZnONPs/PVA (nanoparticles) film loaded with Alhagi maurorum extract for multidrug-resistant Pseudomonas aeruginosa isolated from skin burn infections, as well as to molecularly detect and sequence blaTEM and blaIMP genes followed by an in-silico docking analysis to investigate the antibacterial mechanism.

Materials and methods
Alhagi maurorum shoots were collected. Voucher specimens were retained. The GC-MS analysis was performed. Zinc oxide nanoparticles (ZnO-NPs) were used in the polymer matrix. Comprehensive assessments were conducted to demonstrate the successful entrapment and homogeneous distribution of bioactivating nanocomponents in the polymeric matrix of the fabricated composite film. Fifty bacterial isolates were obtained from burn swabs that had been isolated from the burn unit at AL-Sadiq Teaching Hospital, Babylon, Iraq. The antimicrobial susceptibility of P. aeruginosa isolates (n = 20) was determined. Genomic DNA from bacterial isolates was isolated and PCR was performed. The nucleotide sequences of the blaTEM gene from P. aeruginosa were acquired from the National Center for Biotechnology Information (NCBI). SPSS software was used for statistical analysis.
Results
GC-MS revealed that the mass spectrum and structure of twelve chemical components extracted from Alhagi maurorum using methanol are displayed in the GC-MS data. Among them, 9,12-Octadecadienoic acid (Z,Z)-(Linoleic) was unique. By incorporating these phytochemicals into a PVA polymer matrix with zinc nanoparticles, we created a film that decomposes naturally. The film was tested against medical isolates of Pseudomonas aeruginosa, which are drug-resistant bacteria. Among 50 bacterial isolates, 20 were identified as P. aeruginosa. PCR analysis revealed the presence of blaTEM and blaIMP genes in a substantial proportion of the isolates. Molecular docking results suggest that the ZnO nanostructure has the highest binding potential toward the 4b7q protein compared to linoleic acid and PVA. The strong binding affinity and low inhibition constant of ZnO highlight its potential role as a stable binder or inhibitor, whereas linoleic acid and PVA are less effective.
Conclusion: The strong antibacterial properties of the composite film are probably the result of the PVA polymer, zinc nanoparticles, and plant extract working in concert. The ZnO/Alhagi/PVA composite film demonstrated promising antibacterial and antioxidant activities and may represent a potential biodegradable wound-dressing material for managing MDR P. aeruginosa infections.

کلیدواژه‌ها [English]

  • Alhagi marorum
  • antibacterial films
  • beta-lactamase
  • polyvinyl alcohol
  • zinc nanoparticles

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