نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم دامی، دانشکده علوم کشاورزی و منابع طبیعی، دانشگاه گنبد کاووس،گنبد کاووس، گلستان، ایران

2 گروه علوم دام و طیور، پردیس ابوریحان، دانشگاه تهران، ورامین، ایران

3 گروه زیست فناوری، پردیس علوم، دانشگاه تهران، تهران، ایران

چکیده

هدف از این مطالعه بررسی امکان جداسازی سلول‌های بنیادی مزانشیمی از بافت چربی اسب و مطالعه بازده تمایز استخوانی این سلول‌ها در شرایط کشت تک بعدی (در ظرف کشت بافت پلاستیکی) و سه بعدی (روی داربست های پلی-ال-لاکتیک-اسید) بود. بافت چربی به روش بیوپسی از ناحیه قاعده دم اسب تهیه  و سلول‌های بنیادی مزانشیمی با کمک هضم مکانیکی و آنزیمی از بافت چربی جدا شد. سلول‌ها بنیادی جداشده، در دو شرایط جداگانه شامل شرایط ظرف کشت بافت پلاستیکی (گروه شاهد) و شرایط داربست پلی-ال-لاکتیک-اسید با سه تکرار، به رده استخوان تمایز داده شدند. در طول 21 روز تمایز آزمونهای آلیزارین رد، اندازه‌گیری آنزیم آلکالین‏فسفاتاز و اندازه‌گیری میزان کلسیم رسوبی برای ارزیابی راندمان هر یک از این شرایط در تمایز سلول‌ها به رده استخوان مورد استفاده قرار‌گرفت. دادهای حاصل در قالب طرح کاملاً تصادفی تجزیه شدند. رنگ‌آمیزی آلیزارین رد بهعنوان یک آزمون کیفی نشان داد در هر دو شرایط سلول‏های بنیادی مزانشیمی مشتقشده از بافت چربی اسب می‌توانند به رده استخوان تمایز یابند. با این حال سلول‌های بنیادی مزانشیمی کشت داده شده روی داربست پلی-ال-لاکتیک-اسید در مقایسه با سلول‌های کشت داده شده در شرایط ظرف کشت بافت پلاستیکی دارای فعالیت آنزیم آلکالین‏فسفاتاز و مقدار کلسیم رسوبی بیشتری بودند. یافتههای این مطالعه نشان داد استفاده از داربستهای پلی-ال-لاکتیک-اسید امکان رشد و تمایز بهینه سلول‌های بنیادی مزانشیمی مشتقشده از بافت چربی اسب را به رده استخوانی فراهم می‌سازد.

کلیدواژه‌ها

عنوان مقاله [English]

Effect of poly (L-lactide) nanofiber scaffolds on osteogenic differentiation of equine adipose tissue-derived mesenchymal stem cells

نویسندگان [English]

  • behnaz bageshlooyafshar 1
  • Reza Rahchamani 1
  • Abdollah Mohammadi-Sangcheshmeh 2
  • Ehsan Seyedjafari 3
  • Yussof Mostafaloo 1

1 2. Department of Animal Sciences, College of Agriculture and Natural Resources, University of Gonbad-e Qabus ,golestan,iran

2 Department of Livestock and Poultry Sciences, ABOURYHAN, University of Tehran,varamin,iran

3 Department of Biotechnology, College of Science, University of Tehran ,Tehran, Iran

چکیده [English]

This study was conducted to investigate the differentiation potential of equine adipose-derived mesenchymal stem cell into bone in single-dimensional culture system (in plastic tissue culture) and in three-dimensional system (on poly-l-lactic acid scaffolds; PLLA). A porous structure that allows use of three-dimensional distribution and provides optimal growth of cells is of great clinical significance in the field of tissue engineering. In current study using equine adipose-derived stem cells (ASCs), we intended to compare the osteogenic differentiation potential of PLLA nanofibrous scaffold with tissue culture plastic (TCP). Adipose tissues were collected from 3 adult horses, and ASCswere isolated by enzymatic digestion. PLLA nanofibrous scaffold was successfully prepared using a phase separation method. Viability and growth characteristics of ASCs on TCP and scaffold were investigated by tetrazolium (MTT) based colorimetric assay. Alizarin Red staining was performed for determination of calcium deposition following osteogenic differentiation. Furthermore, other common osteogenic markers such as alkaline phosphatase (ALP) activity, and calcium content were also analyzed. Our data showed that the PLLA scaffold had no detrimental effect on the cell growth rate as evaluated by MTT assay. However, ASCs that differentiated on PLLA nanofibrous scaffolds indicated higher ALP activity and more calcium content than that on TCP. Adequate proliferation rate and higher expression of osteogenic markers of stem cells cultured on PLLA nanofibrous scaffolds provide this scaffold as a suitable substrate to support proliferation and differentiation of ASCs in equine.

کلیدواژه‌ها [English]

  • adipose tissue
  • Mesenchymal stem cells
  • osteogenic differentiation
  • poly-L-lactic-acid
  • scaffolds
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