Evaluation of expression of BMP2 protein and mineralization in the medium containing mesenchymal stem cells derived from warthon jell and mir 424
Abstract
Introduction and Objective: Restoration of the jaw bones in congenital diseases of the cleft palate, bone injuries resulted from trauma and accidents, defects caused by malignant or benign bone lesions, which sometimes lead to the removal of a large part of the jaw is always controversial. In recent years, bone formation regimens have been reported by miRNAs that suggests the role of miRNA 424 in proliferation and osteogenic differentiation of mesenchymal stem cells. The aim of this study was to investigate the effect of miRNA 424 in laboratory on the amount of osteoporosis of Wharton Jelly Stem Cells by extracting relevant proteins and staining the mineral tissue.
Methods: Vector containing MIR424 and BMP2 genes and GFP marker was designed and synthesized on the basis of lentivirus. After confirmation of the vector, HECKT293 cells were used for packaging. The medium was collected on cells at 24, 48 and 72 hours and placed on the Wharton jelly cells. The amount of BMP2 expression through PCR and the amount of BMP2 secretion in the environment via ELISA and mineral sedimentation rate were investigated by Alizarin red staining.
Results: Based on the results of this study, the amount of BMP2 secreted in the environment in the presence of microRNA424 (case group) was increased by 8.04348 compared to its absence (control group). The amount of BMP2 composition with its specific antibody in the case group was 32 and in the control group it was 9 that after statistical analysis, the difference was significant between the two groups with P <0.05. The amount of color deposition in staining with Alizarin red was 0.7 in the treatment group and in the control group, it was 0.2 at 405 nm. This color change was statistically significant (P <0.05).
Discussion: According to the results of this study, the BMP2 gene expression rate in the vicinity of MIR424 is constantly increasing. As a result, this technique can be used to reconstruct bone in animal models, so that the results of these studies provide the basis for gene therapy in bone reconstruction.