动物造模 z-bio 2022-07-20 278

　　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 formation of periosteum-derived stem cells in SD rats. Bone differentiation.

　　Methods: Periosteum-derived stem cells (PDSCs) from SD rats were extracted, identified by flow cytometry, and induced by three lineages of osteogenic, chondrogenic and adipogenic, and were subjected to Alizarin Red, Toluidine Blue, Alician Blue, and Oil Red. O staining and immunofluorescence experiments. Four groups of materials of FGF-2/PELA/BMP-2, FGF-2/PELA, PELA/BMP-2 and PELA were prepared, and the surface of the microcapsules was observed by scanning electron microscope (SEM), and the controlled release curve of the two factors was prepared by ELISA experiment. The extracts of the four groups of materials were co-cultured with the third-generation periosteum-derived stem cells, and alkaline phosphatase (AKP) activity was detected on 7 and 14 days, respectively, and qRT-PCR osteogenic genes were performed on 7, 14, and 21 days, respectively. Expression detection, observation and comparison of the osteogenic differentiation ability of PDSCs in each group, and statistical analysis after data collection.

　　RESULTS: Flow cytometry surface marker identification showed that PDSCs expressed mesenchymal stem cell surface markers, and the three-lineage induced differentiation staining results showed that surface PDSCs had osteogenic, chondrogenic, adipogenic and other multi-directional differentiation abilities. The results of AKP activity showed that the FGF-2/PELA/BMP-2 group had the highest AKP activity on 7 d and 14 d of PDSC culture, and the difference was significant. The qRT-PCR results of the FGF-2/PELA/BMP-2 group showed that the expression levels of RunX-2 and OCN were higher than those of the other groups, and the expression level of RunX-2 reached the peak on the 14th day, and the OCN continued to increase.

　　Conclusion: The cytokine activity in FGF-2/PELA/BMP-2 biomimetic controlled-release microcapsule scaffolds can be retained, and compared with other experimental groups, it has a higher ability to promote the osteogenic differentiation of PDSCs.