Shamsasenjan Karim, Beygi Khosrowshahi Younes, Mahmoodi Mahsa, Akbarzadehlaleh Parvin, Gareayaghi Nesrin, Nejati Babak
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Stem Cell and Tissue Engineering Research Laboratory, Azerbaijan Shahid Madani University, Tabriz, Iran.
Adv Pharm Bull. 2022 Jan;12(1):109-117. doi: 10.34172/apb.2022.012. Epub 2021 Aug 7.
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. 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 γ (PPARγ), Cyclin-dependent kinase 1 (CDK1), and Zinc Finger and BTB Domain Containing 16 (ZBTB16), following exposure with fibronectin 0.1 μg/ml. 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. Results showed that fibronectin can improve proliferation and osteogenic differentiation of BMMSCs without any effect on these cells' survival.
不溶性纤连蛋白作为一种细胞外基质（ECM）蛋白，具有促进间充质干细胞（MSCs）增殖、分化和迁移的潜力。然而，关于不同浓度纤连蛋白对骨髓来源的间充质干细胞（BMMSCs）功能和分化的影响，相关信息有限。在本实验研究中，使用凝胶注射装置，将BMMSCs封装在含有1.25%海藻酸钠、1%明胶和纤连蛋白（0.01、0.05、0.1和0.2μg/ml）的海藻酸钠微胶囊中。采用MTT法检测BMMSCs的增殖情况。此外，使用膜联蛋白V/碘化丙啶染色和荧光激活细胞分选（FACS）分析BMMSCs的凋亡情况。进行碱性磷酸酶（ALP）检测以评估BMMSCs的成骨分化潜能。最后，检测了在暴露于0.1μg/ml纤连蛋白后，SP7、骨钙素（OCN）、 Twist家族BHLH转录因子1（Twist1）、过氧化物酶体增殖物激活受体γ（PPARγ）、细胞周期蛋白依赖性激酶1（CDK1）和含锌指和BTB结构域16（ZBTB16）的mRNA表达水平。根据结果，纤连蛋白具有促进BMMSCs增殖率的潜力，特别是在0.1和0.2μg/ml浓度下。我们发现纤连蛋白不能改变BMMSCs的凋亡率。ALP检测结果证实了纤连蛋白具有显著的触发BMMSCs成骨分化的潜力。此外，逆转录-聚合酶链反应（RT-PCR）结果表明，0.1μg/ml的纤连蛋白可通过靶向OCN、SP7、Twist1、CDK1和ZBTB16，强烈或轻微地增强培养的BMMSCs的成骨分化。结果表明，纤连蛋白可以改善BMMSCs的增殖和成骨分化，而对这些细胞的存活没有任何影响。
