Figure 4
Osteogenesis & angiogenesis. Line 1. Calcein-Tetracycline double labeling of new bone formation. Representative images of the central femurs from SD rats by fluorescence microscopy. The distance between CAL and TCY strips (yellow inside and green outside, respectively) of PDTC group were much wider than model group, and generally close or wider with that of control group. Bars show that the fluorescence intensity comparisons demonstrated a similar result. Line 2&3. Immunohistochemical staining of OPG, and Masson staining. The administration of LPS + MPS significantly suppressed the expressions of OPG and collagen in the model group. However, PDTC treatment effectively reversed the degradation, even superior to the control group. Bar graphs represent the mean optical density of OPG, collagen in bone marrow. Line 4&5. Immunohistochemical staining of CD31 and Ink artery infusion of the femoral head. Images of immunohistochemical staining of CD 31 and ink artery infusion. Few blood vessels and less ink perfusion area were found in the model group, however, more stained blood vessels and much more perfusion ratio were detected in the PDTC group. These results even superior in the PDTC group to control group. Bar graphs show the numeber of blood vessels and ratio of perfusion in the femoral head. Line 6. Expression levels of Runx2 and VEGF by RT-qPCR. The mRNA expression levels of Runx2 and VEGF activations were further confirmed by RT-qPCR. Compared with the control group, the expressions of Runx2 and VEGF were significantly decreased in the model group. While, PDTC treatment efficiently elevated the expressions of VEGF and Runx2. (Above staining quantitative analysis parameters were based on at least 10 fields <n = 10>. Data from RT-qPCR illustrated are from three separate experiments. Bars show relative values of three groups. *p < 0.01 v.s. the control; ▲P < 0.05 vs. model & control; #p < 0.05 vs. model, P < 0.01 vs. control. Further tested by Dunnett-t).
