Title
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Fibronectin within Sodium Alginate Microcapsules Improved
Osteogenic Differentiation of BMMSCs in Dose Dependent Manner
by Targeting SP7, OCN, CDK1, ZBTB16, and Twist1 Expression
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Keywords
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Bone marrow-derived MSCs(BMMSCs), Alginate, Fibronectin, Microcapsules, Osteogenic differentiation
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Abstract
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Insoluble fibronectin as an extracellular matrix (ECM) protein has the potential to
promote proliferation, differentiation, and migration of mesenchymal stem cells (MSCs).
However, there is limited information about the effects of fibronectin various concentrations on
bone marrow-derived MSCs (BMMSCs) function and differentiation
Methods: In this experimental study, using a gel injection device, BMMSCs were encapsulated
in 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 cell
sorting (FACS). Alkaline phosphatase (ALP) test was conducted to assess BMMSCs osteogenic differentiation potential. Finally, mRNA expression levels of the SP7, osteocalcin (OCN), Twist Family 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 the BMMSCs, in particular at 0.1 and 0.2 μg/ml concentrations. we showed that the fibronectin was not able to modify apoptosis rates of the BMMSCs. ALP test results approved the notable potential of the fibronectin, to trigger osteogenic differentiation of the BMMSCs. Also, RT-PCR results indicated that fibronectin 0.1 μg/ml could augment osteogenic differentiation of cultured BMMSCs through targeting of OCN, SP7, Twist1, CDK1, and ZBTB16, strongly or slightly. Conclusion: Results showed that fibronectin can improve proliferation and osteogenic differentiation of BMMSCs without any effect on these cells’ survival.
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