ارزیابی ویژگی های داربست کلاژنی سنتز شده به روش آنزیمی و کارایی آن جهت لود کردن سلول‌های فیبروبلاست‌ نئوناتال

نوع مقاله : مقاله پژوهشی

نویسندگان

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

2 مرکز تحقیقات پوست و سلول‌های بنیادی، دانشگاه علوم پزشکی تهران، تهران، ایران

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

4 گروه زیست شناسی، دانشکده علوم پایه، واحد تهران شرق، دانشگاه آزاد اسلامی ، تهران، ایران

5 مرکز تحقیقات بیوتکنولوژی پزشکی، واحد اشکذر، دانشگاه آزاد اسلامی، اشکذر، یزد، ایران

چکیده

مقدمه و هدف: داربست‌های کلاژنی یکی از داربست‌های مناسب جهت لود شدن سلول‌های بنیادی می باشند، اما میزان زنده‌مانی سلولهای فیبروبلاست لود شده بر روی داربستهای کلاژنی از مسایل مهم می باشد. بر‌این اساس مطالعه حاضر به بررسی میزان زنده‌مانی سلولهای فیبروبلاست لود شده بر  داربست کلاژنی پرداخته است.
مواد و روش ها: در این مطالعه تجربی آزمایشگاهی، نمونه پوست انسان طی عمل جراحی ختنه از نوزادان دریافت  شد و کلاژن گاوی به صورت آماده تهیه شد. سلول‌های فیبروبلاست پس از جدا شدن از قطعات بافتی به وسیله لام هموسیتومتر شمارش شدند. زنده‌مانی سلول‌ها با استفاده از روش‌های فلوسایتومتری بررسی شدند. به منظور بررسی هویت سلول‌های فیبروبلاست از مارکر ویمنتین استفاده شد. سلول‌های فیبروبلاست بر روی هیدروژل کشت شدند. به منظور بررسی اثر سیتوتوکسیک هیدروژل بر سلول‌ها از روش MTT استفاده شد. داده‌ها با استفاده از آزمون تی تست مستقل آنالیز شدند.
نتایج: زنده‌مانی سلول‌های فیبروبلاست جدا شده از پوست ختنه گاه در اولین روز پس از جدا سازی برابر 89 درصد بود.  نتایج حاصل از بررسی ایمنوسیتوشیمی هویت سلول‌های فیبروبلاست را تایید کرد. نتایج MTT نشان دادند که هیدروژل کلاژن اثر سمیت معناداری بر سلول‌های فیبروبلاست لود شده نداشت. نتایج بررسی میکروسکوپی فلورسنت نشانگر آن بود که زنده مانی سلول‌های لود شده بر هیدروژل برابر 96 درصد بود.
نتیجه‌گیری: نتایج حاصل از این پژوهش نشان دادند که داربست‌های کلاژنی جهت لود کردن سلول‌های فیبروبلاست جدا شده از پوست ختنه گاه مناسب بوده و می توانند در مهندسی بافت و انتقال سلول‌های فیبروبلاست به نواحی آسیب دیده کاربرد داشته باشند.

کلیدواژه‌ها

عنوان مقاله [English]

Evaluation of collagen scaffold synthetized by enzymatic method and its efficacy for loading neonatal fibroblast cells

نویسندگان [English]

  • Roqieh Shalbafian 1
  • Sona Zare 2
  • Rahim Ahmadi 3
  • Sanaz Sahraee 4
  • Maryam Hassan Nasab 5
1 Department of Biology, Faculty of Basic Sciences, Hamedan Branch, Islamic Azad University, Hamedan, Iran
2 Skin and Stem Cell Research Center, Tehran University of Medical Sciences, Tehran, Iran
3 Department of Biology, Avicenna International College, Budapest, Hungary
4 Department of Biology, Faculty of Basic Sciences, East Tehran Branch, Islamic Azad University, Tehran, Iran
5 Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran
چکیده [English]

Background and Objective: Collagen scaffolds are one of the suitable scaffolds for stem cell loading, but the viability of fibroblast cells loaded on collagen scaffolds is one of the important issues. Therefore, the present study investigated the viability of fibroblasts loaded on collagen scaffold.
Materials and Methods: In this in vitro experimental study, human skin samples were obtained from neonates during circumcision surgery and bovine collagen was prepared. Fibroblast cells were counted using a hemocytometer after isolation from tissue fragments. Cell viability was evaluated using flow cytometry. Vimentin marker were used to identify fibroblast cells. Fibroblast cells were cultured on hydrogels. MTT method was used to evaluate the cytotoxic effect of hydrogels on cells. Data were analyzed using independent t-test.
Results: The viability of fibroblast cells isolated from the foreskin was 89% on the first day after isolation. The results of immunocytochemical examination confirmed the identity of fibroblast cells. MTT results showed that collagen hydrogel had no significant toxicity on loaded fibroblasts. The results of fluorescent microscopy showed that the viability of cells loaded on hydrogel was 96%.
Conclusion: The results of this study showed that collagen scaffolds are suitable for loading fibroblast cells isolated from the foreskin and can be used in tissue engineering and transfer of fibroblast cells to damaged areas.

کلیدواژه‌ها [English]

  • Foreskin
  • Fibroblast cells
  • Collagen scaffold
  • Viability

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دانشور پزشکی
دوره 30، شماره 2 - شماره پیاپی 159
خرداد و تیر 1401
صفحه 1-11

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