ObjectiveTo investigate the regulation of human bone marrow mesenchymal stem cells (hBMSCs) osteogenic and adipogenic differentiations mediated by Wnt10b adenoviral vector in vitro. MethodsThe hBMSCs from ilial bone tissue in adults at passage 4 were infected by Wnt10b gene expression adenoviral vector (group A), Wnt10b-shRNA adenoviral vector (group B), and empty vector (group C), and non-transfected hBMSCs served as the blank control group. Then the cells were cultured separately in the circumstance of osteogenic induction, adipogenic induction, and non-induction. The alkaline phosphatase (ALP) staining, alizarin red staining, and oil red O staining were used to detect the osteogenic and adipogenic differentiations; real-time fluorescent quantitative PCR and Western blot were used to analyze the expressions of osteoblast and adipocyte genes and proteins. ResultsThe results of ALP staining were positive after osteogenic induction, group A showed strong staining, and group B showed the weakest staining. The results of alizarin red staining showed that there were a lot of patchy confluent brown mineralized nodules in group A; a few punctate brown mineralized nodules were seen in group B; and many punctuate brown mineralized nodules were found in groups C and D. The results of oil red O staining showed strong staining in groups B, C, and D after adipogenic induction, especially in group B; scattered or small clustered staining was observed in group A. The expressions of osteoblast genes and proteins were significantly higher in group A than groups B, C, and D, and in groups C and D than group B by real-time fluorescent quantitative PCR and Western blot test; however, the expressions of adipocyte genes and proteins showed a contrary tendency. ConclusionThe high level expression of Wnt10b can enhance osteogenic differentiation of hBMSCs, and the low level expression of Wnt10b can increase adipogenic differentiation of hBMSCs.
ObjectiveTo analyze the correlation between the trabecular microstructure and the clinical imaging parameters in the fracture region of osteoporotic hip so as to provide a simple method to evaluate the trabecular microstructure by a non-invasive way. MethodsBetween June 2012 and January 2013, 16 elderly patients with femoral neck fracture underwent hip arthroplasty were selected as the trial group; 5 young patients with pelvic fracture were selected as the control group. The hip CT examination was done, and cancellous bone volume/marrow cavity volume (CV/MV) was analyzed with Mimics 10.01 software in the control group. The CT scan and bone mineral density (BMD) measurement were performed on normal hips of the trial group, and cuboid specimens were gained from the femoral necks at the place of the tensional trabeculae to evaluate the trabecular microstructure parameters by Micro-CT, including bone volume fraction (BV/TV), trabecular number (Tb. N), trabecular spacing (Tb.Sp), trabecular thickness (Tb.Th), connect density (Conn.D), and structure model index (SMI). The correlation between imaging parameters and microstructure parameters was analyzed. ResultsIn the trial group, the BMD value was 0.491-0.698 g/cm2 (mean, 0.601 g/cm2); according to World Health Organization (WHO) standard, 10 cases were diagnosed as having osteoporosis, and 6 cases as having osteopenia. The CV/MV of the trial group (0.670 1±0.102 0) was significantly lower than that of the control group (0.885 0±0.089 1) (t=-4.567, P=0.000). In the trial group, CV/MV had correlation with BV/TV, Tb.Th, and SMI (P<0.05); however, CV/MV had no correlation with Tb.N, Tb.Sp, or Conn.D (P>0.05). BV/TV had correlation with Tb.Th, Tb.N, Tb.Sp, and SMI (P<0.05), but it had no correlation with Conn.D (P=0.075). There was no correlation between BMD and microstructure parameters (P>0.05). ConclusionCV/MV obviously decreases in the osteoporotic hip, and there is a correlation between CV/MV and the microstructure parameters of BV/TV, Tb.Th, and SMI, to some extent, which can reflect the variety of the microstructure of the trabeculae. There is no correlation between BMD of femoral neck and microstructure parameters.