تأثیر پیاسکلیدین بر القای کندروژنز از سلول‌های بنیادی مشتق از چربی انسان در داربست فیبرین

نوع مقاله : مقاله های پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه علوم تشریح، دانشکده‌ی پزشکی و کمیته‌ی تحقیقات دانشجویی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 دانشیار، گروه علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

3 استادیار، گروه علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

4 دانشجوی کارشناسی ارشد، گروه مهندسی بافت، دانشکده‌ی مهندسی پایه، دانشگاه آزاد اسلامی، واحد نجف‌آباد، نجف‌آباد، ایران

5 دانشجوی دکتری، گروه ژنتیک، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

مقدمه: آسیب‌های بافت غضروف یکی از مشکلات موجود در کلینیک است و روش‌های درمانی رایج منجر به ترمیم کامل آسیب‌ها نمی‌شود. از این رو، مهندسی بافت در این جهت مورد توجه است. در مهندسی بافت از مواد زیستی، عوامل رشد و سلول مناسب استفاده می‌شود. از آن جایی که عوامل رشد مانند TGF-β (Transforming growth factor beta) منجر به هایپرتروفه شدن بافت غضروف تهیه شده می‌گردد، دستیابی به عوامل القا کننده‌ی مناسب‌تر ضرورت دارد. مطالعات نشان داده است که پیاسکلیدین بر کندروسیت‌های کشت یافته تأثیر مثبت داشته است. از این رو، در این تحقیق تأثیر پیاسکلیدین بر القای کندروژنز و غضروف‌سازی از سلول‌های بنیادی مورد بررسی قرار گرفت.روش‌ها: نمونه‌های چربی از سه بیمار، طی جراحی لیپوساکشن به دست آمد. سلول‌های بنیادی مشتق از چربی (Adipose tissue-derived stem cells یا ADSCs) از بافت چربی استخراج و تا پاساژ سوم تکثیر داده شد. سپس، سلول‌ها در داربست فیبرین تحت تأثیر محیط کشت القای کندروژنیک به مدت 21 روز کشت داده شد. میزان تکثیر و بقا‌ی سلول‌ها با روش MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] و میزان بیان ژن‌های اگریکان،کلاژن II و X با استفاده از تکنیک Real-time PCR ( Real-time polymerase chain reaction) مورد ارزیابی قرار گرفت.یافته‌ها: میزان تکثیر و بقای سلول‌ها در داربست فیبرین، در گروهای مختلف، تفاوت قابل توجهی نداشت (05/0 < P)؛ اما میزان بیان ژن‌های کلاژن II و اگریکان تحت پیاسکلیدین به طور چشم‌گیری در مقایسه با گروه دارای TGF-β افزایش و بیان ژن کلاژن X به طور معنی‌داری کاهش نشان داد (01/0 > P).نتیجه‌گیری: ترکیب پیاسکلیدین در روند القای غضروف‌سازی از سلول‌های بنیادی به طور کامل مؤثر است و بر کاهش هایپرتروفه شدن سلولی نیز تأثیر چشم‌گیری دارد.

کلیدواژه‌ها

عنوان مقاله [English]

Effect of Piasclidin on Induction of Chondrogenesis by Human Adipose-Derived Stem Cells in Fibrin Scaffold

نویسندگان [English]

  • Mojtaba Esmaeily 1
  • Batool Hashemibeni 2
  • Ali Valiani 3
  • Noushin Amirpour 3
  • Babak Purmollaabbasi 4
  • Mohammad Kazemi 5
1 MSC Student, Department of Anatomical Sciences, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
2 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Assistant Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
4 MSC Student, Department of Tissue Engineering, School of Basic Engineering, Najafabad Branch, Islamic Azad University, Isfahan, Iran
5 PhD Student, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Cartilage tissue damage is one of the problems in the clinic and the current treatment methods would not be lead to full repair of damage; therefore, tissue engineering in this field is considered. In tissue engineering, biomaterials such as growth factors and appropriate cells are used. Since the growth factors such as transforming growth factor beta (TGF-β) leads to hypertrophy of cartilage, it is essential to achieve the appropriate inducer factors. Studies have shown that piasclidine had a positive effect on cultured chondrocytes. In this study, the effect of the piasclidine on chondrogenesis of human adipose-derived stem cells was examined.Methods: Fat samples were obtained from three liposuction surgeries. Stem cells were extracted from adipose tissue and proliferated via the third passage. Then, the cells were transferred to the fibrin scaffold and  cultured for 21 days under the influence of the chondrogenic imedium in three groups. The rate of cell proliferation was evaluated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method and the rate of gene expression of aggrecan and types II and X collagen was evaluated via real-time polymerase chain reaction (PCR) method.Findings: The rate of proliferation and survival of cells in a fibrin scaffold was not different significantly between the groups (P > 0.05); but the gene expression of type II collagen and aggrecan was significantly more expression of type X collagen was significantly less in piasclidine group compared to the of TGF-β group (P < 0.01).Conclusion: Piasclidine is an effective factor in induction of chondrogenesis of stem cells and also has a significant effect on reduction of the cell hypertrophy.

کلیدواژه‌ها [English]

  • Adipose-derived stem cell
  • Chondrogenesis
  • Piasclidine

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مجله دانشکده پزشکی اصفهان
دوره 33، شماره 357 - شماره پیاپی 357
آذر و دی 1394
صفحه 1862-1870

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سابقه مقاله

  • تاریخ دریافت: 13 بهمن 1393
  • تاریخ بازنگری: 05 اردیبهشت 1403
  • تاریخ پذیرش: 17 فروردین 1401

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