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Title: The in vitro Experimental Study on Osteogenic and Angiogenic Properties of miR-378a Modified BMMSCs Membranes
Abstract:
Bone marrow mesenchymal stem cells (BMMSCs) have been extensively studied for their potential in bone and vascular tissue engineering. However, enhancing the osteogenic and angiogenic properties of BMMSCs remains a critical challenge. MicroRNAs have emerged as powerful regulators of gene expression and have shown promising effects in modulating cellular functions. In this study, we aimed to investigate the in vitro effects of miR-378a modification on BMMSCs membranes, specifically examining their osteogenic and angiogenic properties.
Introduction:
Bone regeneration and angiogenesis are essential processes in tissue engineering for repairing bone defects. BMMSCs have attracted much attention due to their multilineage differentiation potential. However, enhancing their osteogenic and angiogenic potentials is necessary to improve their therapeutic efficacy. MicroRNAs, such as miR-378a, have been found to play critical roles in regulating cellular functions, including osteogenesis and angiogenesis. Therefore, we hypothesized that the modification of BMMSCs membranes with miR-378a could enhance their osteogenic and angiogenic properties.
Materials and Methods:
1. Isolation and culture of BMMSCs: BMMSCs were obtained from bone marrow aspirates and expanded in culture.
2. Confirmation of stem cell characteristics: Immunophenotyping was performed to confirm the stem cell characteristics of BMMSCs.
3. miR-378a modification: BMMSCs were transfected with miR-378a mimics to upregulate miR-378a expression.
4. Evaluation of osteogenic properties:
a. Alkaline phosphatase (ALP) activity assay: ALP activity was measured to assess early osteogenic differentiation.
b. Alizarin Red S staining: Mineralization capacity was evaluated by Alizarin Red S staining.
c. Expression of osteogenic markers: The mRNA expression of osteogenic markers, such as Runx2, OCN, and OPN, was analyzed by quantitative real-time PCR.
5. Evaluation of angiogenic properties:
a. Endothelial cell tube formation assay: BMMSCs modified with miR-378a were co-cultured with endothelial cells to evaluate their angiogenic potential.
b. Expression of angiogenic markers: The mRNA expression of angiogenic markers, including VEGF, HIF-1α, and Ang-1, was assessed by quantitative real-time PCR.
Results:
1. Confirmation of miR-378a modification: Transfection of BMMSCs with miR-378a mimics successfully upregulated miR-378a expression.
2. Enhanced osteogenic properties: BMMSCs modified with miR-378a showed significantly increased ALP activity, mineralization capacity, and upregulated expression of osteogenic markers compared to control groups.
3. Enhanced angiogenic properties: BMMSCs modified with miR-378a demonstrated increased endothelial cell tube formation and upregulated expression of angiogenic markers compared to control groups.
Discussion:
The present study revealed that miR-378a modification could enhance the osteogenic and angiogenic properties of BMMSCs. The upregulation of miR-378a resulted in increased ALP activity, mineralization capacity, and expression of essential osteogenic markers. Additionally, the modified BMMSCs showed enhanced angiogenic potential, as demonstrated by increased endothelial cell tube formation and upregulated expression of angiogenic markers. The findings suggest that miR-378a modification could be a promising strategy to enhance the therapeutic efficacy of BMMSCs for bone and vascular tissue engineering applications.
Conclusion:
In this study, we successfully demonstrated that miR-378a modification enhances the osteogenic and angiogenic properties of BMMSCs in vitro. These findings provide valuable insights into the therapeutic potential of miRNA modification for improving the regenerative properties of BMMSCs. Further investigations are warranted to evaluate the in vivo efficacy and safety of miR-378a modified BMMSCs membranes, and their potential application in tissue engineering approaches for bone and vascular regeneration.
Keywords: miR-378a, bone marrow mesenchymal stem cells (BMMSCs), osteogenesis, angiogenesis, tissue engineering.