BMPs Differential Expression Profile In Mouse Intervertebral Disc During Development And The Effect To Intervertebral Disc Cells

Adolescent idiopathic scoliosis （AIS) is a serious developmentdeformity of an unknown etiology, especially in newly diagnosed ofidiopathic scoliosis patients at the age of10~18years. Because it is notclear of its pathogenesis, the treatment mainly focus on its symptom, andthe therapeutic effects are not very perfect.Bone morphogenetic proteins(BMPs) is a multifunctional osteogenicfacto, It plays an extremely important role in the embryonic and postnatalbone formation of osteoblasts. This suggests that we can explain somepatients with bone formation of abnormal diseases such as idiopathicscoliosis(IS）from the perspective of BMPs.This study will explore the expression of BMPs in the growth processof spinal disc in mouse and the effect in the intervertebral bone andcartilage cells information.It provides important research basis to explainthe development of adolescent idiopathic scoliosis from the perspective ofabnormal skeletal development. Part oneThe expression of BMPs in the intervertebral disc of postnatalmiceObjective: To investigate the expression of BMP-2,-3,-4,-6,-7and-9in the intervertebral disc of mice of different ages, and to establish animportant foundation for the BMP signaling pathway among thedevelopment of intervertebral disc.Methods: The intervertebral disc and thoracolumbar vertebra wereobtained from the mice aged1day,3days,1week,1month,3months and6months by dissection. RT-PCR was used to detect the expression ofBMP-2,-3,-4,-6,-7and-9in the intervertebral disc of the mice withdifferent ages. Western blotting and immmunohistochemistry was appliedto validate BMP-3and BMP-6that showed different expressions inRT-PCR.Results: RT-PCR suggested that the expressions of BMP-2,-7and-9maintained at a certain level without significant difference, while BMP-3and BMP-6decreased with aging. BMP-4did not express in theintervertebral disc of the mouse. Western blotting andimmunohistochemistry showed similar results as RT-PCR.Conclusion: The expression of BMPs are not complete same duringthe development of postnatal mouse intervertebral disc, and BMP-3andBMP-6may paly some different role. Part TwoInvestigation of osteogenesis about prenatal mouseintervertabral disc cells of BMP-2and-3overexpressionObjective： To investigate the osteogenesis of prenatal mouseintervertabral disc cells which are infected with mouse recombinantadenovirus BMP-2and-3，and provide some evidences for therapy aboutthe spectrum of disc disease with BMPs.Methods： The intervertabral disc cells were infected with mouserecombinant adenovirus BMP-2and BMP-3for5-7days, the expression ofOC、OPG and OPN mRNA were detected with RT-PCR. The cartilagematrix was staining with toluidine blue staining. The alkalinephosphatase(ALP) activity was detected with ALP reading and ALPstaining.Results： The BMP-2and-3overexpression cann’t enhanceosteogenesis in AF cells. But they can promote osteogenesis in NP cells.We observe the expression of OC mRNA increasing drastic, moreover thereis only BMP-2can incite OPG and OPN mRNA increasing. The ALPactivity could be observed significant increasing in NP cells which wereinfected with adenovirus BMP-2and-3, but this change was not found inAF cells.Conclusion： BMP-2and-3can promote NP cell and AF cellosteogenic differentiation.