牵张成骨术整复猕猴腭裂后胰岛素样生长因子-1与碱性磷酸酶表达研究
Expression of insulin-like growth factor-1 and alkaline phosphatase in the reconstruction of cleft palate with distraction osteogenesis in rhesus
摘要目的 以牵张成骨术整复猕猴腭裂骨缺损,定量分析不同时段新生成骨胰岛素样生长因子-1(insulin.1ike growth factor-I,IGF-1)与碱性磷酸酶(alkaline phosphatase,ALP)表达水平,探讨其成骨调控机制.方法 用猕猴建立腭裂动物模型.实验组动物21只以牵张成骨术整复其腭部软硬组织缺损,关闭裂隙后固定.于牵张成骨术后第1、2,4…6 8 12及24周分别取材,各3只动物.采用实时定量PCR法分析比较IGF-I与ALP的mRNA表达水平,并以酶联免疫吸附试验法(ELISA)定量分析其IGF-I与ALP含量,结果与实验对照及健康对照组(动物各2只)进行比较.结果 固定期第1~2周为成骨早期,第1周时IGF-1和AIJP的mRNA表达明显上调,分别为3.67±0.35和3.30±0.21,第2周时达最高峰,分别为7.55±0.32和5.91±0.21,随后逐渐下降,至第12周与健康对照组无明显差异(P>0.05).ELISA结果显示:固定期第1~2周,IGF-1和ALP表达均增强.IGF-1含量于第2周表达最高,为(2.0±0.06)ng/mg,第4周为(1.46±0.08)ns/mg,第6周为(0.84±0.11)ng/mg,其表达逐渐下降;同期ALP则呈高水平表达,第4周为(25.34±0.44)U/mg,第6周为(26.21±0.82)U/mg.第8~12周,IGF-1和ALP的表达均下降至接近健康对照组.结论 腭骨牵张成骨区域,新骨的原位增量生成明确,增殖过程正常,最终以膜内成骨的方式形成新骨整复了腭裂骨切开牵张间隙区域.
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abstractsObjective To investigate the osteogenesis mechanism by analysis of the expression of insulin-like growth factor-I (IGF-1)and alkaline phesphatas (ALP)in the reconstruction of cleft palate(CP) with distraction osteogenesis (DO) in rhesus. Methods The CP animal models were established surgically. 21 rhesus in experimental group underwent DO to close the soft and bony defect, followed by consolidations. Every 3 animals were killed and the specimen were taken out after consolidation of 1, 2, 4, 6, 8, 12, 24 weeks. The mRNA of IGF-1 and ALP were detected with Real-time BT-PCB technique. The expression of IGF-1 and ALP was quantitatively analyzed by ELISA. The results were compared with those in control and sham groups (each of 2 animals), respectively. Results Since consolidation, the mRNA of IGF-1 and ALP increased significantly at one week and reached the peak at two weeks, but decrease to control level after 12 weeks of consolidation. The expression of IGF-1 also increased to peak level afiert two weeks of consolidation. The expression of ALT increased significantly since consolidation and reach the peak value after six weeks. They all decreased to nearly control level after 8 ~ 12 weeks. Conclusions The palate cleft can be successfully closed with new formed bone after DO. The mechanism of bone consolidation is intramembranons bone formation.
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