Karim Shamsasenjan
1* 
, Younes Beygi Khosrowshahi
2, Mahsa Mahmoodi
1 , Parvin Akbarzadehlaleh
3, Nesrin Gareayaghi
4, Babak Nejati
5 1 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Stem Cell and Tissue Engineering Research Laboratory, Azerbaijan Shahid Madani University, Tabriz, Iran.
3 Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Istanbul Sisli Hamidiye Etfal Training and Research Hospital, Blood Center, University of Health Science, Istanbul, Turkey.
5 Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract
Purpose: Insoluble fibronectin as an extracellular matrix (ECM) protein has the potential topromote proliferation, differentiation, and migration of mesenchymal stem cells (MSCs).However, there is limited information about the effects of fibronectin various concentrations onbone marrow-derived MSCs (BMMSCs) function and differentiationMethods: In this experimental study, using a gel injection device, BMMSCs were encapsulatedin sodium alginate microcapsules containing 1.25% alginate, 1% gelatin, and fibronectin (0.01,0.05, 0.1, and 0.2 μg/ml). MTT assay was used to examine the proliferation of BMMSCs. Also,BMMSCs apoptosis were analyzed using Annexin-V/PI staining and fluorescence activated cellsorting (FACS). Alkaline phosphatase (ALP) test was conducted to assess BMMSCs osteogenicdifferentiation potential. Finally, mRNA expression levels of the SP7, osteocalcin (OCN), TwistFamily BHLH Transcription Factor 1 (Twist1), Peroxisome proliferator‐activated receptor γ2(PPARγ2), Cyclin-dependent kinase 1 (CDK1), and Zinc Finger and BTB Domain Containing 16(ZBTB16), following exposure with fibronectin 0.1 μg/ml.Results: According to results, fibronectin had the potential to promote proliferation rates of theBMMSCs, in particular at 0.1 and 0.2 μg/ml concentrations. we showed that the fibronectinwas not able to modify apoptosis rates of the BMMSCs. ALP test results approved the notablepotential of the fibronectin, to trigger osteogenic differentiation of the BMMSCs. Also, RT-PCRresults indicated that fibronectin 0.1 μg/ml could augment osteogenic differentiation of culturedBMMSCs through targeting of OCN, SP7, Twist1, CDK1, and ZBTB16, strongly or slightly.Conclusion: Results showed that fibronectin can improve proliferation and osteogenicdifferentiation of BMMSCs without any effect on these cells’ survival.