Sadraei Fariba, Ghollasi Marzieh, Khakpai Fatemeh, Halabian Raheleh, Jalali Tehrani Hora
Cognitive and Neuroscience Research Center (CNRC), Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
Regen Ther. 2022 Aug 27;21:263-270. doi: 10.1016/j.reth.2022.07.015. eCollection 2022 Dec.
Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are undifferentiated cells with self-renewing ability and multi-lineage differentiation beneficial for regenerative medicine. Nano scaffolds are novel materials employed in bone repair and regeneration. Nisin is a prebiotic that can increase stem cells' lifespan and proliferation. This study attempted to provide a proper strategy for bone marrow mesenchymal stem cells differentiation into the Osteocytes on a Poly-L-lactic-acid (PLLA) scaffold after pretreating with Nisin.
MSC osteogenic differentiation was evaluated by measuring Calcium, Alkaline phosphatase, and quantitative tests such as Real-Time PCR, Acridine Orange, Alizarin Red, Von Kossa, and others.
The result of the MTT test showed that the optimal dose of Nisin prebiotic for the MSCs' preconditioning was 200 IU/mL on the 1st, 3rd, and 5th days of culture. Real-time PCR data indicated that the expression rate of ALP, Osteonectin, Osteocalcin, and Collagen I have increased in the presence of Nisin, while the RUNX-2 gene expression has decreased. Furthermore, the results of Alizarin Red and Von Kossa tests, as well as Scanning electron microscopy (SEM), revealed that the cell proliferation in the preconditioned samples with Nisin increased significantly.
The study concluded that the cell proliferation and differentiation increased in samples pretreated with Nisin on the PLLA Nano scaffolds.
人骨髓间充质干细胞(hBM-MSCs)是具有自我更新能力和多向分化能力的未分化细胞,对再生医学有益。纳米支架是用于骨修复和再生的新型材料。乳酸链球菌素是一种益生元,可延长干细胞寿命并促进其增殖。本研究试图提供一种合适的策略,使经乳酸链球菌素预处理后的骨髓间充质干细胞在聚L-乳酸(PLLA)支架上分化为骨细胞。
通过测量钙、碱性磷酸酶以及实时定量PCR、吖啶橙、茜素红、冯库萨染色等定量测试来评估间充质干细胞的成骨分化。
MTT试验结果表明,在培养的第1、3和5天,用于间充质干细胞预处理的乳酸链球菌素益生元的最佳剂量为200 IU/mL。实时定量PCR数据表明,在乳酸链球菌素存在的情况下,碱性磷酸酶、骨连接蛋白、骨钙素和I型胶原蛋白的表达率有所增加,而RUNX-2基因的表达则有所下降。此外,茜素红和冯库萨染色试验以及扫描电子显微镜(SEM)的结果表明,经乳酸链球菌素预处理的样品中的细胞增殖显著增加。
该研究得出结论,在PLLA纳米支架上,经乳酸链球菌素预处理的样品中细胞增殖和分化增加。