مقایسه‌ی تأثیر دو عامل Transforming Growth Factor Beta1 (TGF-ß1) و پیاسکلیدین بر بیان ژن‌های کلاژن II، X و اگریکان در روند کندروژنز سلول‌های بنیادی مشتق از چربی انسان در داربست کامپوزیتی فیبرین آلژینات

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

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

چکیده

مقدمه: آسیب‌های بافت غضروف در کشورهای پیشرفته، مهم‌ترین علت ناتوانی در سالمندان است. علاوه بر آن، غضروف مفصلی توانایی محدودی در ترمیم دارد. روش‌های درمانی رایج قادر به ترمیم این آسیب‌ها نمی‌باشد؛ چرا که منجر به ایجاد بافت فیبروزی در غضروف می‌شوند. سلول درمانی، یکی از روش‌های درمان است که در آن، سلول‌های بنیادی با کمک مهندسی بافت می‌توانند به کندروسیت تمایز یابند و جهت دستیابی به این هدف، از عوامل رشد و داربست‌ها استفاده می‌شود. به دلیل هایپرتروفه شدن بافت غضروف و عدم پایداری داربست‌ها، ضرورت دستیابی به عوامل القا کننده و داربست مناسب احساس می‌گردد. بر اساس مطالعات، فیبرین آلژینات از لحاظ پایداری و کشسانی (Elasticity) مناسب است و پیاسکلیدین، باعث افزایش بیان ژن‌های مخصوص غضروف می‌گردد. از این رو، در تحقیق حاضر، تأثیر پیاسکلیدین و Transforming growth factor beta1 (TGF-β1) بر القای کندروژنز سلول‌های بنیادی در داربست فیبرین آلژینات مورد مقایسه قرار گرفت.روش‌ها: Adipose derived stem cells (ADSCs) از بافت چربی سه بیمار استخراج و تکثیر داده شد. سپس به مدت 21 روز در داربست فیبرین آلژینات تحت تأثیر مدیوم کندروژنیک کشت داده شدند. میزان تکثیر و بقای سلول‌ها به 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 polymerase chain reaction (Real-time PCR) مورد ارزیابی قرار گرفت.یافته‌ها: میزان تکثیر و بقا در داربست فیبرین آلژینات، در گروه حاوی پیاسکلیدین نسبت به سایر گروه‌ها، افزایش داشت؛ اما این افزایش به صورت معنی‌دار نبود (050/0 < P). همچنین، پیاسکلیدین باعث افزایش میزان بیان ژن کلاژن II (001/0 > P) و کاهش میزان بیان ژن کلاژن X در مقایسه با TGF-β1 گردید.نتیجه‌گیری: احتمال می‌رود پیاسکلیدین در روند القای کندروژنز ADSCs در داربست فیبرین آلژینات مؤثر بوده و بر افزایش بیان ژن‌های ویژه‌ی غضروف تأثیر داشته باشد

کلیدواژه‌ها

عنوان مقاله [English]

Comparing the Effects of Transforming Growth Factor Beta1 (TGF-ß1) and Piascledine on the Expression of Collagen II, X and Aggrecan Genes in Chondrogenesis of human Adipose-Derived Stem Cells in Fibrin Alginate Composite Scaffold

نویسندگان [English]

  • Hadi Didehvar 1
  • Farhad Golshan-Iranpoor 2
  • Ali Valiani 2
  • Batool Hashemibeni 3
  • Mojtaba Esmaeeli 1
1 MSc Student, Department of Anatomical Sciences, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
2 Assistant Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Cartilage injuries are the leading cause of disability in the elderly in developed countries. In addition, articular cartilage has a limited ability to repair. Current treatment methods for cartilage tissue injuries lead to fibrous tissue formation. Cell therapy is a treatment in which stem cells using tissue engineering can be differentiated into chondrocytes by using growth factors and scaffolds. Since growth factors such as transforming growth factor beta1 (TGF-β) leads to hypertrophy of cartilage chondrocytes tissue and many scaffolds are weak in terms of mechanics and stability, it is essential to achieve the appropriate scaffolds and inducing factors. Studies have shown that fibrin alginate scaffold is appropriate in terms of mechanical and stability and piascledine increases the cartilage-specific genes expression. Therefore, in this study the chondrogenic effect of TGF-β1 and piascledine on adipose derived stem cells in fibrin alginate scaffold was compared and evaluated.Methods: Fat samples were obtained from three persons. Adipose derived stem cells (ADSCs) was extracted from adipose tissue and proliferated. Then the cells were transferred to the fibrin alginate scaffold and the cells were cultured for 21 days under the influence of the induction medium. The rate of proliferation and survival of cells was evaluated by [3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide] MTT method and the rate of gene expression of Aggrecan and Collagen II and X was evaluated with Real-time polymerase chain reaction (Real-Time PCR) method.Findings: The results showed that proliferation rate and survival of cells in a fibrin alginate scaffold in the group containing Piascledine increased compared to the other groups, but this increase is not significant (P> 0.050). Also, Piascledine increased collagen II gene expression (P < 0.001) and reduced collagen X gene expression when compared to TGF-β1.Conclusion: Piascledine was found as a proper effective inducer in chondrogenic differentiation of human adipose derived stem cells cultured in fibrin alginate scaffold.

کلیدواژه‌ها [English]

  • Adipose derived stem cell
  • Piascledine
  • Chondrogenesis
  • Transforming growth factor beta1

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مجله دانشکده پزشکی اصفهان
دوره 34، شماره 373 - شماره پیاپی 373
فروردین و اردیبهشت 1395
صفحه 157-165

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سابقه مقاله

  • تاریخ دریافت: 22 تیر 1394
  • تاریخ بازنگری: 31 فروردین 1403
  • تاریخ پذیرش: 17 فروردین 1401

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