Fibronectin within Sodium Alginate Microcapsules Improved Osteogenic Differentiation of BMMSCs in Dose Dependent Manner by Targeting SP7, OCN, CDK1, ZBTB16, and Twist1 Expression

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.