[Effects of Schwann cells promoting nitric oxide secretion of bone marrow mesenchymal stem cells derived endothelial cells].

ABSTRACT

OBJECTIVE: To study the effect of Schwann cells (SCs) promoting the function of nitric oxide (NO) secretion of bone marrow mesenchymal stem cells (BMSCs) derived endothelial cells so as to lay the experimental foundation for research of the effect of nerves on vessels during the process of tissue engineering bone formation. METHODS: SCs were collected from 1-day-old Sprague Dawley (SD) rats, and identified through S100 immunohistochemistry (IHC). BMSCs were collected from 2-week-old SD rats and induced into endothelial cells (IECs), which were identified through von Willebrand factor (vWF) and CD31 immunofluorescence (IF). Transwell system was used for co-culture of SCs and IECs without contact as the experimental group, and simple culture of IECs served as the control group. The NO concentration in the medium was measured at 1, 3, 5, and 7 days after culture; the mRNA expressions of nitric oxide synthetase 2 (NOS2) and NOS3 were detected by real-time fluorescence quantitative PCR (RT -qPCR) at 1, 3, 7, and 10 days. RESULTS: SCs and IECs were identified through morphology and immunology indexes of S100 IHC, vWF and CD31 IF. Significant differences were found in the NO concentration among different time points in 2 groups (P < 0.05); the NO concentration of the experimental group was significantly higher than that of the control group at the other time points (P < 0.05) except at 3 days. NOS2 mRNA expression of the experimental group was significantly higher than that of the control group (P < 0.05); difference was significant in the NOS2 mRNA expression among different time points in 2 groups (P < 0.05). NOS3 mRNA expression of the experimental group was significantly higher than that of the control group at the other time points (P < 0.05) except at 10 days. No significant difference was found in NOS3 mRNA expression among different time points in the experimental group (F = 6.673, P = 0.062), but it showed significant differences in the control group (F = 36.581, P = 0.000). CONCLUSION: SCs can promote NO secretion of BMSCs derived endothelial cells, which is due to promoting the activity of NOS.