بررسی رفتار سلول‌‌های MG63 بر روی داربست‌‌های سه‌بعدی پلی‌‌کاپرولاکتون و پلی‌‌کاپرولاکتون/کلاژن برای بازسازی استخوان

محورهای موضوعی : فصلنامه زیست شناسی جانوری

زهرا ابراهیمی ¹ , شیوا ایرانی ² , عبدالرضا اردشیری لاجیمی ³ , احسان سیدجعفری ⁴

1 - گروه زیست شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
2 - گروه زیست شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
3 - مرکز تحقیقات سلول‌‌های بنیادی در مجاری ادراری تناسلی، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران
4 - گروه زیست فناوری، پردیس علوم، دانشگاه تهران، تهران، ایران

تاریخ دریافت : 1401/10/26 تاریخ پذیرش : 1401/12/24 تاریخ انتشار : 1402/09/01

کلید واژه: مهندسی بافت استخوان, پلی‌‌کاپرولاکتون, کلاژن, سلول‌‌های بنیادی, داربست سه‌بعدی,

چکیده مقاله :

چاپ سه‌بعدی داربست‌‌ها در دمای پایین نویدبخش زیادی برای ساخت جایگزین‌‌های مصنوعی پیوند استخوان با عملکرد بیشتر نسبت به تکنیک‌‌های سنتی است. یکی از امیدوارکننده‌‌ترین استراتژی‌‌ها در مهندسی بافت استخوان روی توسعه داربست‌‌های biomimetic متمرکز شده است. داربست‌‌ها با پایه سرامیک با توانایی استئوژنیک و خواص مکانیکی، کاندیدای امیدوارکننده‌‌ای برای ترمیم استخوان هستند. هدف از این مطالعه متناسب‌‌سازی انعطاف‌‌پذیری و خاصیت القای استخوان داربست سه‌بعدی پلی‌‌کاپرولاکتون (PCL) تهیه شده با روش مدل رسوب ذوب شده (FDM)، با استفاده از تلفیق کلاژن (COL) به عنوان پلیمر طبیعی همراه با پلیمر مصنوعی و بررسی رفتار سلول‌‌های MG63بر روی آن بود. بعد از تهیه داربست، از آزمون‌‌های میکروسکوپ الکترونی روبشی (SEM)، طیف سنجی پراش انرژی ایکس (EDX) و طیف‌‌سنجی ATR-FTIR استفاده شد. پس از گذشت 1،7،14 روز، بررسی روند استخوان‌‌سازی سلول‌‌های در تیمارهای مختلف، با استفاده از رنگ‌آمیزی آلیزارین‌‌رد و فعالیت آلکالین فسفاتاز (ALP) صورت گرفت. همچنین عدم سمیت داربست‌‌ها برای اطمینان از تکثیر سلول‌‌ها توسط آزمون MTT مورد بررسی قرار گرفت. از مشاهده زیر میکروسکوپ، مشخص شد که داربست‌‌های مهندسی بافت منافذ مربعی شکل را به طور یکنواخت توزیع و بهم متصل کرده‌‌اند. داربست COL/PCL تفاوت معنی‌‌داری را از لحاظ قابلیت بقا نسبت به داربست PCL تنها در محیط تمایزی نشان داد (P ≤ 0.0001). نتایج ارزیابی فعالیت ALP در داربست PCL/COL به طور معنی‌‌داری بالاتر از داربست PCL بدون پوشش و کنترل بود (P ≤ 0.0001). نتایج بدست آمده در این تحقیق نشان داد استفاده از داربست PCL به همراه COL می‌تواند به عنوان محیط مناسبی به منظور تکثیر و تمایز سلول‌‌های MG63 در نظر گرفته شود. بنابراین، داربست‌‌ کامپوزیت PCL/COL که توسط چاپگر FDM تهیه شده‌اند، بدلیل بقای سلولی بواسطه COL است، می‌تواند کاربرد وسیع‌‌تری در مهندسی بافت استخوان داشته باشد.

چکیده انگلیسی:

3D printing of scaffolds at low temperatures is very promising for making artificial bone graft alternatives with more performance than traditional techniques. One of the most promising strategies in bone tissue engineering has focused on the development of biomimetic scaffolds. Ceramic-based scaffolds with osteogenic ability and mechanical properties are promising candidates for bone repair. The aim of this study was to adapt the flexibility and increasing the effect of inducing osteogenesis of Polycaprolactone (PCL) scaffold prepared by fused deposition modeling (FDM) method, using the combination of Collagen (COL) as a natural polymer with synthetic polymer and to investigate the behavior of MG63 cells on it. on it. After preparing the scaffold, scanning electron microscope (SEM), energy dispersive X-ray (EDX) and ATR-FTIR spectroscopy were used. After 1,7,14 days, the ossification process of MG63 cells in different treatments was performed using alizarin red staining and alkaline phosphatase activity. The non-toxicity of scaffolds was also evaluated by MTT assay to ensure cell proliferation. From under a microscope, it was found that tissue engineering scaffolds distributed and connected almost rectangular pores evenly. The PCL/COL scaffold showed a significant difference in terms of viability compared to the polycaprolactone scaffold only in the differential medium (P ≤ 0.0001). The results of evaluating ALP activity in PCL/COL scaffolds were significantly higher than uncoated PCL scaffolds and control (P ≤ 0.0001). The results of this study showed that the use of PCL/COL scaffold can be considered a suitable medium for proliferation and differentiation of MG63 cells in bone tissue engineering. Therefore, the PCL/COL composite scaffold prepared by FDM printer can be widely used in bone tissue engineering due to the cell survival by COL.

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