شیمى کاربردى روز

شیمى کاربردى روز

نانوکپسول‌های چندمنظوره مبتنی بر PEG-PEI و عامل دار شده با فولیک اسید وگلوکز جهت انتقال هدفمند قطعات ژنی به سلول‌های سرطانی Hs-578T

نوع مقاله : مقاله علمی پژوهشی

نویسندگان
گروه زیست شناسی، واحد اردبیل، دانشگاه آزاد اسلامی، اردبیل، ایران
چکیده
مقدمه: ژن‌رسانی هدفمند می‌تواند محدودیت‌های شیمی‌درمانی را کاهش دهد. در این مطالعه، نانوکپسول‌های مبتنی بر PEI/PEG با هدف‌گیری دوگانه فولیک‌اسید و گلوکز طراحی شدند تا کارایی انتقال ژن به سلول‌های سرطان سینه Hs-578T افزایش یابد.
مواد و روش‌ها: نانوکپسول‌ها با استفاده از 1H-NMR،FTIR و TGA شناسایی شدند. اندازه، مورفولوژی و پتانسیل زتا به‌ترتیب با TEMو DLS بررسی شد. آزمون‌هایMTT، الکتروفورز ژل، فلوسیتومتری و میکروسکوپ فلورسانسی برای ارزیابی سمیت، اتصال و حفاظت DNA و کارایی انتقال توالی هدف (pEGFP-N1) انجام گرفت.
نتایج: نتایج DLS نشان داد نانوکپسول‌های PPF، PPG و PPGF دارای اندازه متوسط 174 تا 301 nm بودند و نانوکپسول‌های حاوی فولیک‌اسید اندازه بزرگ‌تری نسبت به نمونه‌های اصلاح‌شده با گلوکز داشتند. با افزایش نسبت نانوذره به DNA، اندازه کاهش یافته و بار سطحی از 7/4− به 6/2+ mV تغییر کرد. بررسی رهایش DNA نشان داد در 5/5 pH=رهایش از نانوکپسول‌های PPG،PPF و PPGF به‌طور معنی‌داری بیشتر از 4/7 pH=بود و در حضور دکستران (µg.mL-1 100) به 96/64 درصد رسید. هم چنین مشخص شد که رهایشDNA از نانوکپسول‌های پوشش داده شده توسط فولیک‌اسید نسبت به گلوکز افزایشی بود. از سویی آزمون MTT نشان داد که نانوکپسول‌های PEI/DNA در غلظت‌های بالاتر از µg/mL 30 بیش از 50 درصد سمیت ایجاد می‌کند، در حالی‌که نانوکپسول‌های اصلاح‌شده زیست‌سازگاری بالاتری داشتند. نتایج ژل الکتروفورز، فلوسیتومتری و میکروسکوپ فلورسانس بیانگر اتصال مؤثر، محافظت آنزیمی مناسب و انتقال کارآمد DNA بودند و استفاده هم‌زمان از فولیک‌اسید و گلوکز بیشترین بازدهی انتقال را نشان داد.
نتیجه‌گیری: هدف‌گیری دوگانه فولیک‌اسید و گلوکز بازدهی ژن‌رسانی را افزایش می‌دهد.
کلیدواژه‌ها
موضوعات

عنوان مقاله English

Folic Acid and Glucose Functionalized PEG-PEI-Based Multifunctional Nanocapsules for Targeted Gene Delivery to Hs-578T Cancer Cells

نویسندگان English

Hashem Yaghoubi
Afsun Bayrami
Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
چکیده English

Introduction: Targeted gene delivery can overcome limitations of conventional chemotherapy. In this study, PEI/PEG-based multifunctional nanocapsules with dual targeting were designed by simultaneous modification with folic acid and glucose to enhance gene transfer efficiency into Hs-578T breast cancer cells.
Materials and Methods: The synthesized nanocapsules were characterized using 1H-NMR, FTIR, and TGA. Particle size, morphology, and zeta potential were evaluated by DLS and TEM. Cytotoxicity, DNA binding and protection, and transfection efficiency of the target sequence (pEGFP-N1) were assessed using MTT assay, gel electrophoresis, flow cytometry, and fluorescence microscopy.
Results: DLS analysis showed that PPF, PPG, and PPGF nanocapsules had average sizes ranging from 174 to 301 nm, with folic acid–modified nanocapsules exhibiting larger sizes than glucose-modified ones. Increasing the nanoparticle/DNA ratio reduced particle size and shifted the surface charge from −7.4 to +6.2 mV. DNA release studies demonstrated significantly higher release from PPG, PPF, and PPGF nanocapsules at pH=5.5 compared to pH=7.4, reaching 64.96% in the presence of dextran (100 µg.mL-1). Moreover, folic acid modification enhanced DNA release compared to glucose. MTT results indicated that PEI/DNA complexes induced over 50% cytotoxicity at concentrations above 30 µg/mL, whereas modified nanocapsules showed improved biocompatibility. Gel electrophoresis, flow cytometry, and fluorescence microscopy confirmed effective DNA binding, enzymatic protection, and efficient gene delivery, with dual-ligand modification achieving the highest transfection efficiency.
Conclusion: Dual targeting with folic acid and glucose significantly enhances targeted gene delivery efficiency.

کلیدواژه‌ها English

Cationic Nanoparticles
Gene Delivery
Hs-578T
Nanocapsules
PEG
and PEI
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