نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست شناسی، دانشکده علوم پایه، واحد همدان، دانشگاه آزاد اسلامی، همدان، ایران

2 گروه بیولوژی، کالج بین المللی اویسینا، بوداپست، مجارستان

3 گروه زست شناسی، دانشکده علوم پایه، واحد کرج، دانشگاه خوارزمی، ایران

4 رکز تحقیقات بیوتکنولوژی پزشکی، واحد اشکذر، دانشگاه آزاد اسلامی، اشکذر، یزد، ایران

10.30495/rkctc.2022.67131.1043

چکیده

زمینه و هدف: مطالعات بسیاری نشان داده‌اند که داربست‌های کیتوزان می توانند جهت لود کردن سلولهای بنیادی مورد استفاده قرار گیرند اما نتایج هنوز هم در این مورد چالش برانگیزند. براین اساس مطالعه حاضر به ارزیابی ویژگی‌های داربست کیتوزانی سنتز شده به روش آنزیمی و کارایی آن جهت لود کردن سلول‌های فیبروبلاست نئوناتال پرداخته است.
روش بررسی: طی این تحقیق تجربی-زمایشگاهی پوست ختنه‌گاه نوزاد انسان تهیه و سلول‌های فیبروبلاست لایه درم آن جداسازی شده و کشت شدند. میزان زنده‌مانی سلول‌ها  به روش فلوسایتومتری و تعیین هویت سلول‌ها با استفاده از مارکر وایمنتین صورت گرفت. پس از تهیه هیدروژل کیتوزان، سلول های فیبروبلاست بر روی آن لود شدند و اثر سمیت هیدروژل بر سلول‌ها با استفاده از روشMTT ارزیابی شد. داده‌ها با استفاده از آزمون تی - تست مورد تجزیه و تحلیل قرار گرفت.
نتایج: شمارش سلول‌ها نشان داد که هر فلاسک T75 متراکم، حدود 2 میلیون سلول داشت. سلول‌های جداسازی شده مارکر ویمنتین را در سطح بالایی بیان کردند. فیبروبلاست‌ها با چسبندگی مناسب و توزیع یکنواخت برروی هیدروژل کیتوسان مشاهده شدند. طبق نتایج تستMTT ، هیدروژل تهیه شده اثر سمیت معناداری بر سلول‌های فیبروبلاست نداشت.
نتیجه‌گیری: نتایج این تحقیق نشان دادند که داربست کیتوزان می‌تواند داربست مناسبی برای لود کردن سلول‌های فیبروبلاست و انتقال این سلول‍‌ها جهت پیوند به بافتهای آسیب دیده باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Evaluation of Chitosan Scaffold Synthetized by Enzymatic Method and Its Efficacy for Loading Neonatal Fibroblast Cells

نویسندگان [English]

  • Akram Fadavi 1
  • Rahim Ahmadi 1 2
  • Sepideh Shahbaz Gahrouei 3
  • Maryam Hassan Nasab 4

1 Department of Biology, Faculty of Basic Sciences, Hamedan Branch, Islamic Azad University, Hamedan, Iran

2 Department of Biology, Faculty of Basic Sciences, Hamedan Branch, Islamic Azad University, Hamedan, Iran|Avicenna International College, Budapest, Hungary

3 Department of Biology, Faculty of Basic Sciences, Karaj Branch , Kharazmi University, Iran

4 Medical Biotechnology Research Center, AshkezarBranch, Islamic Azad University, Ashkezar, Yazd, Iran

چکیده [English]

Background and aim: Many studies have shown that chitosan scaffolds can be used to load stem cells, but the findings are still challenging. Therefore, the present study evaluated the properties of chitosan scaffolds synthesized by the enzymatic method and their efficiency for loading neonatal fibroblast cells.
Methods: During this experimental-laboratory study, the human neonatal foreskin was prepared and the fibroblast cells of the dermal layer were isolated and cultured. Cell viability was determined by flow cytometry and cell identity was determined using a Vimentin marker. After the preparation of chitosan hydrogel, fibroblast cells were loaded on it and the cytotoxic effect of hydrogel on cells was evaluated using the MTT assay. Data were analyzed using a t-test.
Results: Cell counts showed that each dense T75 flask contained about 2 million cells. Isolated cells expressed the Vimentin marker at a high level. Fibroblasts were observed with good adhesion and uniform distribution on chitosan hydrogel. According to the results of the MTT test, the prepared hydrogel had no significant toxicity effect on fibroblast cells.Conclusion: The results of this study showed that chitosan scaffolding can be a suitable scaffold for loading fibroblast cells and transporting these cells for transplantation to damaged tissues.

کلیدواژه‌ها [English]

  • Chitosan
  • Fibroblast cells
  • Viability
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