Investigation of In-vitro Osteogenic Potential of Human Adipose-Derived Stem Cells Cultured on Hydrogel Based 3D Nanocomposite Scaffold Made of PVA - VBP – nGO

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 294

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شناسه ملی سند علمی:

NSCMRMED03_269

تاریخ نمایه سازی: 30 دی 1397

چکیده مقاله:

Background and Aim: The purpose of the present study was to investigatethe osteogenic differentiation of human adipose-derived stem cells(hASCs) in three-dimensional (3D) constructs in vitro. Previously, wecreate two groups of 3D composite scaffolds made of polyvinyl alcohol(PVA), vinyl based polymer (VBP), and nano graphene oxide (nGO) (NP+group), and PVA and VBP (NP- group) through controlled freeze-dryingtechnique. In this study, we assessed the potential of these scaffoldscombined with hASCs for bone tissue application.Methods: human adipose-derived stem cells (hASCs) were isolated fromsubcutaneous adipose tissue of 7 healthy female donors (30-40 yearsold) that had undergone liposuction through the enzymatic procedure.In passages 3, cell surface markers (CD73, CD90, CD105, CD34, CD45)were identified by flow cytometry. After sterilization of the two groupsof the scaffold, 5×106 hASCs were seeded on each scaffold and culturedin osteogenic media (OM). The presence of the hASCs on the scaffoldwere emphasized by SEM, DAPI staining, and hematoxylin-eosin (H&E)staining on the 1st, 7th,14th, 21th day of culture. Then, osteogenicdifferentiation of seeded cells was evaluated by alkaline phosphataseactivity and Alizarin Reds in 14th, 21th day of culture. Moreover, realtimePCR, Western blot, and immunocytochemistry (ICC) were done forspecific osteogenic markers including Runx2, col Iα, Osteocalcin, andOsteonectin.Results: The positive expression of CD73, CD90, and CD105 andnegative expression of CD34, CD45 of hASCs were identified by flowcytometry. Results from SEM, DAPI staining, and H&E staining showedthat the presence of most of the cells on the two groups of scaffolds(NP+,NP-) after the 1st, 7th, 14th, 21th day of culture. In vitro osteogenicinduction of the 3D construct showed that the expression levels ofRunx2, col I, Osteocalcin and Osteonectin in NP+ scaffolds group weresignificantly higher than NP- scaffolds group on days 7 and 14 aftercultured in osteogenic medium (OM). Western blotting and ICC showedthat the expression of osteogenesis-related proteins was significantlyhigher in cells seeded on NP+ scaffolds group than NP- scaffolds group.Conclusion: These results indicated that 3D nanocomposite constructs(NP+) consisting of hASCs had the great ability to promote mineralizedmatrix formation and could be used for the repair of bone tissue defects.

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نویسندگان

Fatemeh SadatMaryam Sadat Khoramghah

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Simzar Hosseinzadeh

Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, ShahidBeheshti University of Medical Sciences, Tehran, Iran

Javad Ranjbari

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Fatemeh Sadat Tabatabaei Mirakabad

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran