摘要
目的探讨"手风琴"技术对大鼠牵张区骨再生的作用、刺激时机以及可能机制。方法取12周龄雄性SD大鼠54只,制备右侧胫骨牵张成骨模型。术后经5 d潜伏期、7 d牵张期后进入为期6周的矿化期。根据在矿化期实施"手风琴"操作的时间不同,将大鼠随机分为4组:对照组(n=18)为观察组,不行"手风琴"操作;早期组(n=18)、中期组(n=12)及晚期组(n=6)分别于矿化第1、3、5周行"手风琴"操作,操作时间为7 d,其中3.5 d压缩、3.5 d牵张。对照组和早期组分别于矿化第2、4、6周结束时,中期组于矿化第4、6周结束时,晚期组于矿化第6周结束时,各取6只大鼠处死,取双侧胫骨进行相关观测。矿化期间每周对各组大鼠右侧胫骨摄正侧位X线片,观察骨再生与矿化进展;各组每次取材时对右侧胫骨行Micro-CT扫描三维重建观察,评估牵张区新生骨结构改变,并计算矿化第6周各组158～211、211～1 000及158～1 000阈值范围的骨体积(bone volume,BV)和组织体积(tissue volume,TV),计算两者比值BV/TV,同时计算骨密度(bone mineral density,BMD)。矿化第6周,对所取胫骨行四点弯曲生物力学检测最大压力、弹性模量、折断能量;行Von Kossa、番红O、HE染色及成骨相关转录因子(osterix,OSX)、骨钙蛋白(osteocalcin,OCN)、VEGF免疫组织化学染色观察。结果 X线片显示牵张区骨痂在矿化中期组行"手风琴"刺激后矿化速度最快。Micro-CT三维重建可见中期组骨重建最快,牵张后6周皮质获得连续。矿化第6周,中期组BMD及158～1 000、211～1 000阈值范围的BV/TV均高于其余各组。生物力学检测示,中期组胫骨标本的最大压力、弹性模量、折断能量均显著高于其余3组(P<0.05)。组织学染色示中期组骨重建速度最快,且至牵张后第6周髓腔基本获得再通。免疫组织化学染色结果进一步提示,中期组新生骨组织内成骨指标(OCN、OSX)及成血管指标(VEGF)随着骨重建的进程明�
Objective To investigate the effect, right timing, and mechanism of ＂accordion＂ technique on bone regeneration in rat distraction osteogenesis model. Methods Fifty-four 12-week-old male Sprague Dawley rats underwent right tibial distraction osteogenesis procedure. After a 5-day latency, the distraction was performed for 7 days followed by 6-week consolidation. All animals were randomly divided into 4 groups based on different periods of＂accordion＂ maneuvers in consolidation phase： control group（n=18） with no manipulation, and three experimental groups including early-phase group（n=18）, mid-phase group（n=12）, and late-phase group（n=6） with ＂accordion＂maneuvers applied at 1, 3, and 5 weeks, respectively. The duration of the ＂accordion＂ maneuver was 7 days consisting of a 3.5-day compression and 3.5-day distraction. Rats in control group and early-phase group were sacrificed at 2, 4, and6 weeks of the consolidation phase; rats in mid-phase group were sacrificed at 4 and 6 weeks of the consolidation phase;and rats in late-phase group were sacrificed at 6 weeks of the consolidation phase. Bilateral tibias from 6 rats in each groupat each time point were obtained. Callus formation was monitored by X-ray radiography every week; new bone was reconstructed by Micro-CT three-dimensional reconstruction. The change of bone structure was evaluated, and parameters containing bone volume（BV）/tissue volume（TV） ratio（BV/TV） and bone mineral density（BMD） in three thresholds（158-211, 211-1 000, 158-1 000） were recorded and calculated at 6 weeks. Mechanical test consisting of ultimate load, modulus of elasticity, and energy to failure was performed. Histological analysis, such as Von Kossa staining,Safranin O staining, and HE staining, was done. Immunohistochemical staining using markers of osterix（OSX）,osteocalcin（OCN）, and vascular endothelial growth factor（VEGF） was analyzed. Results Images of X-ray showed that callus formation increased significantly in the mid-ph
作者
沈洁
叶晓健
SHEN Jie;YE Xiaojian(Department of Orthopedics, Changzheng Hospital, the Second Military Medical University, Shanghai, 200003, P.R.Chin)
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2018年第5期558-567,共10页
Chinese Journal of Reparative and Reconstructive Surgery
关键词
“手风琴”技术
牵张成骨
骨矿化
大鼠
＂Accordion＂ technique
distraction osteogenesis
bone consolidation
rat
