Objective To investigate the effect of 1,25(OH)2VD3 on differentiation of embryonic stem cells (ESCs) into osteoblasts. Methods Osteoblasts were isolated and cultured from calvarium of 2-day-old Kunming white mice, embryoid bodies (EBs) were prepared with modified zur Nieden method. EBs were divided into 4 groups according to different mediums: group A, as the control group, in which EBs medium contained no leukemia inhibitory factor; group B, in which EBs medium contained supplements of Vitamin C (VC, 50 μg/mL) and β-glycerophosphate (β-GP, 50 mmol/L); group C, inwhich EBs medium was the same as that of group B and 5 × 104 osteoblasts of 3rd passage were seeded into each well; group D, in which the medium contained supplements of VC (50 μg/mL), β-GP (50 mmol/L) and 1,25(OH)2VD(4 × 10-9 mol/L), and 5 × 104 osteoblasts of 3rd passage were seeded into each well. The ALP activity was determined by ALP reagent kit every 5 days. The RQ-PCR was performed to measure the mRNA expressions of osteocalcin (OCN). Al izarin red S staining was performed to count the bone nodules. Results The expression of ALP witnessed no obvious change in each group within 5 days after adherence of EBs, but increased gradually after 5 days. The expression of ALP in group D reached the peak at 20 days. Red nodules with clear outl ine and different sizes were evident by microscope. Al izarin red S staining testified the number of bone noudles in groups A, B, C and D was 20 ± 8, 18 ± 5, 31 ± 1 and 50 ± 1, respectively, indicating significant differences between groups C, D and groups A, B (P lt; 0.05), no significant difference between group A and group B (P gt; 0.05), and a significant difference between group C and group D (P lt; 0.05). The result of RQ-PCR showed that the mRNA expressions of OCN in groups A, B, C and D was 10.18 ± 1.17, 20.29 ± 1.03, 18.84 ± 4.07 and 32.15 ± 5.23, respectively, indicating significant differences between groups C, D and groups A, B (P lt; 0.05), no significant difference between group A and group B (P gt; 0.05), and a significant difference between group C and group D (P lt; 0.05). Conclusion The combined action of 1,25(OH)2VD(4 × 10-9 mol/L), VC, and β-GP can effectively promote the differentiation of the ESCs-derived osteoblasts.
The hemodynamic parameters in arteries are difficult to measure non-invasively, and the analysis and prediction of hemodynamic parameters based on computational fluid dynamics (CFD) has become one of the important research hotspots in biomechanics. This article establishes 15 idealized left coronary artery bifurcation models with concomitant stenosis and aneurysm lesions, and uses CFD method to numerically simulate them, exploring the effects of left anterior descending branch (LAD) stenosis rate and curvature radius on the hemodynamics inside the aneurysm. This study compared models with different stenosis rates and curvature radii and found that as the stenosis rate increased, the oscillatory shear index (OSI) and relative residence time (RRT) showed a trend of increase; In addition, the decrease in curvature radius led to an increase in the degree of vascular curvature and an increased risk of vascular aneurysm rupture. Among them, when the stenosis rate was less than 60%, the impact of stenosis rate on aneurysm rupture was greater, and when the stenosis rate was greater than 60%, the impact of curvature radius was more significant. Based on the research results of this article, it can be concluded that by comprehensively considering the effects of stenosis rate and curvature radius on hemodynamic parameters, the risk of aneurysm rupture can be analyzed and predicted. This article uses CFD methods to deeply explore the effects of stenosis rate and curvature radius on the hemodynamics of aneurysms, providing new theoretical basis and prediction methods for the assessment of aneurysm rupture risk, which has important academic value and practical guidance significance.