动物实验,干细胞 z-bo 2020-08-28 477

　　Objective: To observe the effect of fibroblast growth factor 2/polylactic acid-polyethylene glycol-polylactic acid/bone morphogenetic protein 2 (FGF-2/PELA/BMP-2) microcapsule scaffold on the growth of periosteum stem cells in SD rats.

　　Method: Extract SD rat periosteum-derived stem cells (PDSC), identify surface markers by flow cytometry, and induce the three lines of bone formation, cartilage and adipogenesis, alizarin red, toluidine blue and Alsin. Blue, oil red O staining and immunofluorescence experiments. Prepare four groups of materials, FGF-2/PELA/BMP-2, FGF-2/PELA, PELA/BMP-2, PELA, and observe the surface of the microcapsules with a scanning electron microscope (SEM). The ELISA experiment generated a controlled release curve of these two factors. The four groups of material extracts were co-cultured with stem cells derived from the third generation of periosteum. The alkaline phosphatase (AKP) activity was tested on the 7th and 14th days, and the qRT-PCR was performed on the 7, 14 and 21 days. Osteogenic gene expression. The day after the test. Observe and compare the osteogenic differentiation potential of each group of PDSC, and perform statistical analysis after collecting the data.

　　Result: Through the identification of the surface markers of flow cytometry, it was found that PDSC expressed the surface markers of mesenchymal stem cells, and through 3-line induced differentiation staining, PDSC showed that PDSC showed bone formation, cartilage, fat formation, etc. Facts have proved that it has the ability of multi-directional differentiation. The result of AKP activity showed that the FGF-2/PELA/BMP-2 group showed the highest AKP activity on the 7th and 14th day of PDSC culture, and the difference was significant. The qRT-PCR results of the FGF-2/PELA/BMP-2 group showed that the expression of RunX-2 and OCN was higher than that of other groups, and the expression of RunX-2 reached a peak on the 14th day. OCN has continued to grow.

　　Conclusion: The FGF-2/PELA/BMP-2 biomimetic controlled-release microcapsule scaffold retains cytokine activity than other experimental groups and has a higher ability to promote osteogenic differentiation of PDSC.