Inhibitory effect of Sema7A siRNA on osteoclast activation induced by titanium particles / 中国组织工程研究

ABSTRACT

BACKGROUND:Sema7A is a kind of cel surface protein, which can promote the fusion of osteoclasts and the migration of osteoblast at the same time, affecting the dynamic balance of bone. OBJECTIVE:To investigate whether Sema7A siRNA has ainhibitory effect on the osteoclast activation in the process of osteolysis which induced by titanium particles. METHODS:The precursor osteoclasts with the concentration of 4×109 RESULTS AND CONCLUSION:At 7 days of culture, the expression levels of interleukin-1, interleukin-1β, tumor necrosis factor α, matrix metaloproteinase-9 and the receptor activator of nuclear factor-κB in the positive control, /L were seeded on 96-wel plates containing glass cover slips, and divided into four groups blank control, positive control, experiment and negative control groups. The cel culture medium was added into the control group. 20 μL un-transfected siRNA supernatant was added into the positive control group. 20 μL transfected Sema7A siRNA supernatant was added into the experiment group. 20 μL transfected control siRNA supernatant was added into the negative control group. The supernatant was obtained through the co-culture between titanium particles solution and monocyte-macrophage cel line RAW264.7of mouse for 24 hours. siRNA was transfected into mononuclear macrophage cel lines RAW264.7 of mice. negative control and experiment groups were higher than those in the control group (P < 0.05). The expression level of each factor in the experiment group was lower than that in the positive control and negative control groups (P < 0.05). At 8 days of culture, the proliferation activity of osteoclasts and the number of positive cels stained by tartrate-resistant acid phosphatase in the positive control, negative control and experiment groups were higher than those in the control group (P < 0.05). The proliferation activity of osteoclasts and the number of positive cels stained by tartrate-resistant acid phosphatase in the experiment group were lower than those in the control and negative groups (P < 0.05). These results demonstrate that Sema7A siRNA has a certain inhibitory effect on the osteoclast activation induced by titanium particles.