大鼠颅骨矢状缝牵引成骨体外探究.docVIP

大鼠颅骨矢状缝牵引成骨体外探究[摘要]目的：探讨建立幼年SD大鼠颅骨器官体外培养的方法，研究外源性张力对颅骨矢状缝成骨活动的影响。方法：取19日龄的SD大鼠颅顶骨矢状缝组织块，试验组加0.4 g力，对照组不加力。经器官培养后进行组织学观察和免疫组织化学检查，并检查胰岛素样生长因子-1（insulin-like growth factor-1, IGF-1）等mRNA的变化情况。结果：试验组骨缝内成骨细胞活跃，IGF-1阳性染色位于颅缝内成骨细胞，实验组IGF-1 mRNA 表达量高于对照组。 结论：大鼠颅骨骨缝器官可在体外培养中成功存活并生长。外源机械张力可以扩宽骨缝，刺激成骨活动，可显著增加IGF-1的合成。 [关键词]颅缝；器官培养技术；胰岛素样生长因子-1；机械张力 [中图分类号]R785[文献标识码]A[文章编号]1008-6455（2008）07-1015-04 Osteogenic effect of tensile force to the cranial sagittal suture of rats in vitro WU Di1,WANG Xin-gang2,ZHAO Zhen-min1 (1.Department of Cleft Lip and Palate, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, Beijing 100144；2.Department of Oral and Maxillofacial Surgery, Beijing Children Hospital, Capital Medical University, Beijing 100045,China) Abstract：ObjectiveTo develop an in vitro model for cranial suture of neonatal SD rat, and explore the osteogenic effect of exogenous tensile force.MethodsThe parietal bones with the sagittal suture were removed from SD rats (19-days old) for organ culture. In the experimental group, tensile force(0.4g) were applied by helical springs, whereas no tension (0g) were set in the control group. We dissected sagittal suture for RT-PCR. The changes of IGF-1 mRNA was investigated. Then the explants cultured for 24 hours were removed for histological observation and immunohistochemistry examination.ResultsIn the experimental group, osteoblasts and capillary vessels proliferated actively in the suture. IGF-1 mRNA were increased significantly. After immunohistochemistry staining, IGF-1、IGF-1R and PCNA were positive in osteoblasts and fibroblasts.ConclusionIn vitro model of cranial suture can be cultured successively. Exogenous tensile force has osteogenic effect in cranial suture, which companied by significant increase of IGF-1 level. Key words: cranial suture; organ culture techniques; insulin-like growth factor； tensile force 骨缝是颅面骨骼生长的主要部位，被硬脑膜等组织包绕，位于坚硬的骨骼之间。体外实验是将离体骨缝组织置于培养基中进行牵引，这不仅避免了体内综合因素的影响，还可单独对缝组织施加某种条