身痛逐瘀汤疏通淋巴管瘀滞促进骨修复的实验研究

目的 观察身痛逐瘀汤对胫骨骨折小鼠的骨修复作用。方法 将6~8周龄雄性C57BL/6小鼠随机分为生理盐水组和身痛逐瘀汤组,每组20只。当两组小鼠成功建立右侧胫骨骨折复合髓内固定模型半小时后,身痛逐瘀汤组给予浓度1.375 g/mL身痛逐瘀汤药液0.2 mL/d,生理盐水组给予同体积0.9 %NaCl溶液,两组均连续灌胃5 d。造模后,活体监测小鼠的后肢淋巴管回流功能、骨折断端软组织肿胀度和足底热敏痛阈;造模14 d后取材,检测骨痂形态计量学;造模后28 d后取材,检测骨折愈合生物力学性能。结果 ①造模后第1天（P＜0.001）和第7天（P＜0.001）,身痛逐瘀汤组的淋巴管清除率优于生理盐水组。②造模后第2~7天,身痛逐瘀汤组后肢软组织肿胀度明显低于生理盐水组。③造模后第1天,身痛逐瘀汤组的足底热敏痛阈明显低于生理盐水组（P＜0.01）。④胫骨Micro-CT重建显示,身痛逐瘀汤组小鼠比生理盐水组的骨痂量更多,骨小梁更致密,骨折线更模糊;定量分析发现身痛逐瘀汤组骨痂的BV/TV（P＜0.01）和Tb.N（P＜0.05）优于生理盐水组。⑤身痛逐瘀汤最大应力（P＜0.001）和屈服应力（P＜0.001）高于生理盐水组。结论 身痛逐瘀汤能促进淋巴管回流,减轻骨折后肿痛症状,促进骨折愈合。     

脑源性神经营养因子在大鼠胫骨骨折愈合中的作用研究

目的通过研究脑源性神经营养因子(BDNF)对骨折愈合的影响,探讨神经营养因子在骨组织工程和临床治疗中的应用前景。方法使用SD大鼠胫骨横行骨折髓内针内固定模型,术后处理组每隔一天皮下注射BDNF,对照组注射生理盐水,分别在第2、3、4、5周取胫骨对骨折愈合行大体比较、力学测试和扫描电镜观察。结果术后2周两组骨折均为结缔组织连接;3周处理组已有编织骨愈合,对照组仍有明显的骨折端活动;4周两组均呈骨性愈合,但处理组的骨痂、成角畸形较小;5周时处理组骨折塑型好,对照组骨折端仍有较大骨痂。处理组骨折矢状面成角在第3、4、5周小于对照组(P<0.05)。各阶段处理组抗折应力均优于对照组(P<0.05)。结论早期应用BDNF对骨折愈合的各阶段均可能有促进作用。     

人工合成成骨生长肽促进兔胫骨骨折愈合的实验研究

目的观察人工合成的成骨生长肽(sOGP)经静脉应用对兔胫骨骨折愈合的影响。方法52只新西兰大白兔制成胫骨中段截骨、髓内钉固定骨折模型后,随机分为两组。实验组从术后第1d开始每天注射sOGP0.5μg·kg-1至取材前1d,对照组注射同样剂量的生理盐水。分别于术后第1、2、3、4、6周每组宰杀3～5只,测定血清ALP、BGP;同时取材测定骨折端骨密度、摄X线片后,再行组织学检查和形态分析,观察骨痂的大小及其组成成分的变化;其中术后6周的标本还进行了生物力学测定。结果实验组血清ALP、BGP水平高于对照组。X线片显示两组骨折均愈合,第3、4周实验组外骨痂平均面积比对照组大,3周时为265.44mm2、209.95mm2,4周时为233.10mm2、209.21mm2。第3、4、6周的骨密度实验组均大于对照组,其中第4周的骨密度经F检验两组的差异具有显著性(P<0.05)。组织学形态分析显示第2、3、4周的外骨痂平均面积实验组比对照组大;所有实验组外骨痂中小梁骨占骨痂总面积的百分比均高于对照组,其中第2、4周经F检验两组的差异具有显著性(P<0.05)。第6周骨折端平均最大破坏载荷及最大位移实验组比对照组高,但差异没有显著性(P>0.05)。结论全身应用sOGP对不稳定固定的兔胫骨横断骨折的愈合具有一定的促进作用。     

骨折合并颅脑损伤大鼠血小板衍生生长因子表达的研究北大核心CSCD

目的观察胫骨骨折合并颅脑损伤大鼠血清及骨痂组织内血小板衍生生长因子（PDGF）的表达变化,探讨颅脑损伤对骨折愈合的影响及作用机制。方法选择144只SD大鼠随机分为4组（n=36只）：正常对照组（N组）、颅脑损伤组（TBI组）、单纯骨折组（F组）和骨折合并颅脑损伤组（TBI＋F组）,并制作相应组别的损伤模型。造模后第3天、1周、2周、3周、4周处死大鼠,4组均采用酶联免疫吸附试验法检测大鼠血清PDGF的表达;F组和TBI＋F组大鼠通过x线片观察右胫骨骨折处骨痂生长情况,通过苏木精．伊红染色观察骨痂生长及组织形态,免疫组织化学法检测骨痂组织PDGF的表达,逆转录聚合酶链反应法检测骨痂组织PDGFmRNA的水平,并进行比较。结果x线片示TBI＋F组大鼠骨折愈合速度比F组快。TBI＋F组大鼠血清PDGF表达量较其他3组高,且峰值时间提前,差异均有统计学意义（P〈0．05）。TBI＋F组大鼠骨折端局部成骨活动较F组强。TBI＋F组大鼠第3天、1周时局部骨痂组织中PDGF阳性细胞数较F组明显增多升高,且峰值时间提前,差异均有统计学意义（P〈0．05）。TBI＋F组大鼠第3天、1周时局部骨痂组织中PDGFmRNA表达水平较F组显著增高,差异均有统计学意义（P〈0．05）。结论颅脑损伤可以加速大鼠骨折愈合的进程,这可能与颅脑损伤后大鼠体内PDGF的表达增高有关。     

一氧化氮调节骨折愈合的实验研究

[目的] 探讨一氧化氮在骨折愈合中的可能作用。[方法] Wistar大鼠60只随机分为实验组和对照组。两组动物的饮用水分别含L-硝基-精氨酸甲基脂（L-NAME）和D-硝基-精氨酸甲基脂（D-NAME），建立股骨开放骨折模型，骨折后第3d、1、2和4周，取每组6只大鼠骨折处骨痂行HE染色观察。另6只大鼠于骨折后4周行骨痂的面积、骨密度和骨折的生物力学评价。[结果] 骨折后两组大鼠体重增加无差别。HE染色光镜下见骨折后不同时期L-NAME组骨折处骨痂生成少，骨痂塑形慢，未成熟细胞比例较对照组多（P〈0．05），加入L-NAME后使骨折愈合反应减小，实验组骨痂的大小、骨密度和愈合强度均明显低于对照组，有明显的统计学意义。[结论] 在骨折愈合过程中加入L-NAME会使骨折愈合减慢，一氧化氮可能促进骨折的愈合过程。     

抑制Wnt/β-catenin信号通路对骨折愈合的影响

目的 探讨Wnt/β-catenin信号通路在骨折愈合中所起的作用. 方法采用8周龄Col2al-ICAT转基因小鼠为实验组(转基因在软骨细胞中特异性表达后可竞争性阻断β-catenin信号)和同窝出生WT小鼠(对照组)作为实验动物,分别制备右下肢胫骨中段截断骨折模型.于骨折后7、9、14、21、28 d取材进行分析,通过X线片、组织学观察和组织形态计量学分析,比较两组软骨痂和骨性骨痂在骨折愈合不同时期所占的比例. 结果通过X线片观察发现,骨折后第21天,WT小鼠骨折线已消失,而Col2al-ICAT转基因小鼠骨折线仍可见.组织学观察发现,与WT小鼠相比,Col2al-ICAT转基因小鼠软骨痂出现延迟,软骨内骨化受阻,骨折的塑形改造延迟.组织形态计量学结果显示:Col2al-ICAT转基因小鼠软骨痂较晚出现高峰期,骨折后第14和第21天时骨性骨痂占总骨痂的比例明显低于WT小鼠,差异有统计学意义(P<0.05).结论 抑制Wnt/β-catenin信号通路将抑制骨折愈合的软骨内骨化过程,最终导致骨折愈合延迟.     

去势延迟雄性小鼠骨折愈合

目的观察去势雄性小鼠骨折愈合的情况,探讨男性骨质疏松骨折愈合过程。方法40只7周龄C57BL/6J雄性小 鼠,其中8只作为基线组,32只随机分为对照组与去势组,小鼠去势后1周建立右股骨中段骨折模型。术后2周和4周,测量 血清睾酮浓度,骨折端骨密度（BMD)。骨折愈合情况通过显微CT和组织学切片显示。结果去势后小鼠血清睾酮浓度明显 下降;术后4周,与对照组相比,去势组BMD明显降低。三维重建图像显示去势组骨痂改建差于对照组。病理切片可见去势 组初级骨小梁少于对照组,骨痂重建差于对照组。结论去势延迟的骨折的愈合,表现在骨折区骨量减少以及骨改建延迟, 本研究为男性骨质疏松骨折的研究提供了动物模型。     

连续碳纤维聚烯烃髓内钉治疗兔胫骨骨折

目的 观察连续碳纤维增强聚烯烃复合材料髓内钉对兔胫骨骨折愈合的影响.方法 雄性新西兰兔39只,制作兔双侧胫骨中断骨折髓内钉固定模型,左侧实验侧以碳纤维复合材料髓内钉固定,右侧以不锈钢髓内钉做对照.24只动物术后2、4、8、12周取材行大体观察、影像学观察和组织学检查.另15只术后4、8、12周取材做愈合骨强度的生物力学分析.结果 术后4周及8周,试验组矿化沉积率及平均类骨质宽度均优于对照组.愈合8周与12周,实验组骨愈合负载优于对照组25～30%(P＜0.05),弯曲强度实验组高于对照组15%(P＜0.05).结论 与不锈钢髓内钉比较,连续碳纤维增强聚烯烃复合材料髓内钉更有利于兔胫骨骨折愈合.     

高能震波对骨痂中骨形成蛋白表达的影响

目的 观察高能震波（ＨＥＳＷ）对骨痂中骨形成蛋白（ＢＭＰ）表达的影响，探讨其促进骨折愈合的机理。方法 手术造成家兔双侧胫骨中段标准骨折模型，对一侧胫骨骨折部施以ＨＥＳＷ，另一侧为对照组，术后每周进行Ｘ线及组织学检查，并取骨痂组织切片用ＢＭＰ抗体进行免疫组化染色（ＳＡＢＣ法（）。结果 实验组骨痂中ＢＭＰ表达强度在第２－４周时明在于对照组（Ｐ〈０．０１），且Ｘ线摄片及组织学观察均显示实验组骨折愈合时间     

植入型复方丹参缓释剂对骨折愈合影响的实验研究

目的:观察丹参缓释剂植入骨折局部对骨折愈合的影响.方法:42只新西兰健康白兔随机分为两组,均于右前肢桡骨中段制成3mm骨缺损的骨折模型,同时实验组放置丹参缓释剂60mg,对照组放置等量赋形剂.分别于术后第2、4、5、6周取右桡骨行X线片及组织学检查;第2、4、6周行电镜及骨密度检查;第5、6周进行生物力学测试.结果:X线片显示实验组较对照组骨折愈合提前.组织学观察实验组纤维性骨痂、软骨性骨痂、骨小梁之间过渡加快,毛细血管增生明显.电镜检查实验组示骨折修复细胞形态学变化均有利于修复骨折.术后第2、4、6周时骨密度值实验组均较对照组高,其中第4周时两者差异具有显著性(P＜0.05).术后第5、6周实验组抗折力均较对照组高(P＜0.05).结论:在骨折局部使用丹参缓释剂对骨折愈合具有一定的促进作用.     

Hyperbaric oxygen therapy mitigates the adverse effect of cigarette smoking on the bone healing of tibial lengthening: an experimental study on rabbits.

OBJECT: We investigated whether -intermittent hyperbaric oxygen (HBO) therapy can mitigate the adverse effects of cigarette smoking on the bone healing of tibial lengthening by using a previously validated rabbit model. METHODS: Eighteen male rabbits were randomly divided into three groups of six animals each. Group 1 (smoking plus HBO) went through intermittent cigarette smoke inhalation and hyperbaric oxygen therapy, group 2 (control) did not go through intermittent cigarette smoke inhalation or hyperbaric oxygen therapy and group 3 (smoking) went through intermittent cigarette smoke inhalation. Each animal's right tibia was lengthened 5 mm by using an uniplanar lengthening device. Bone mineral density (BMD) study was performed for all the animals at 1 day before operation and 3, 4, 5, and 6 weeks after operation. All of the animals were killed at 6 weeks postoperatively for biomechanical testing. RESULTS: By using the preoperative BMD as an internal control, we found that the BMD of group 1 (smoking plus HBO)and group 2 (control) was superior to that of group 3 (smoking). The mean %BMD at 3, 4, 5, and 6 weeks were 58.6%, 66.6%, 73.7%, and 83.8%, respectively, in group 1, whereas the mean %BMD were 52.0%, 64.3%, 70.1%, and 76.2%, respectively, in group 2, and the mean %BMD were 46.2%, 54.0%, 64.9%, and 69.4%, respectively, in group 3 (two-tailed t test, p > 0.05, p > 0.05, p > 0.05, and p < 0.05 at 3, 4, 5, and 6 week respectively between group 1 and group 2, p < 0.01,p < 0.01,p < 0.01, and p < 0.01 at 3, 4, 5, and 6 week, respectively, between group 1 and group 3 and p < 0.05, p < 0.05, p < 0.05, and p < 0.05 at 3, 4, 5, and 6 week respectively between group 2 and group 3). By using the contralateral nonoperated tibia as an internal control, we found that the torsional strength of group 1 (smoking plus HBO) and group 2 (control) was superior to that of group 3 (smoking). The mean percentage of maximum torque was 80.9% in group 1 (smoking plus HBO) and was 78.0% in group 2 (control), whereas the mean percentage of maximum torque was 59.6 % in group 3 (smoking) (two-tailed t test, p < 0.05 between groups land 3 and between groups 2 and 3, whereas p > 0.05 between groups 1 and 2). CONCLUSION: This study suggests that smoke inhalation delays the bone healing in tibial lengthening; however, HBO mitigates the delayed healing effect of smoke inhalation and, thus, helps the smoking animal in achieving an expeditious bone healing in tibial lengthening.     

鞘注神经生长因子促进大鼠胫骨骨折愈合的实验研究

目的 研究鞘注神经生长因子(NGF)对大鼠胫骨骨折愈合的影响,以及相应脊髓背根神经节(DRG)与胫骨骨痂中的降钙素基因相关肽、P物质的表达变化. 方法 成年雄性SD大鼠48只随机分为两组.实验组予NGF 1μg/d连续鞘注14 d,对照组予等量生理盐水.术后7、14、21、28 d分批处死动物,行骨痂X线评分、骨痂/骨干比值测量.免疫组织化学法测定相应节段DRG与胫骨骨痂中的CGRP、SP表达.结果 术后21 d,NGF组X线评分低于对照组,21 d、28 d骨痂/骨干比值低于对照组.HE染色显示各时期NGF组软骨内成骨过程增强,骨痂成熟度高于对照组,骨痂改建过程提前且更为完全.CGRP、SP在DRG及胫骨骨痂中表达的平均光密度(OD)值高于同期对照组,差异有统计学意义(P＜0.05).相关性分析显示DRG神经肽表达OD值与骨痂OD值明显正相关. 结论 鞘注神经生长因子可促进DRG与骨折骨痂中的神经肽表达,促进成骨细胞增殖与软骨内骨化,减小骨痂体积并加速骨折的愈合及塑形改建过程.推测神经生长因子与DRG神经肽参与了中枢神经调控外周成骨活动的过程.     

生物热熔胶与同种异体骨板治疗胫骨骨折的实验研究

目的 观察生物热熔胶与同种异体骨板治疗犬胫骨骨折的疗效,探讨应用同种异体骨板联合生物热熔胶治疗长骨骨折的可行性.方法 将32只Labrador犬的一侧胫骨制成横行骨折模型,随机分为粘接固定组16只,用生物热熔胶与冻干同种异体骨板粘接固定;金属接骨板固定组16只,用金属接骨板与螺丝钉内固定.记录两组的手术时间和术中出血量...     

Effect of recombinant human osteogenic protein-1 on the healing of a freshly closed diaphyseal fracture

Osteogenic protein-1 (OP-1), or bone morphogenetic protein-7, is an osteoinductive morphogen that is involved in embryonic skeletogenesis and in bone repair. In bone defect models without spontaneous healing, local administration of recombinant human OP-1 (rhOP-1) induces complete healing. To investigate the ability of rhOP-1 to accelerate normal physiologic fracture healing, an experimental study was performed. In 40 adult female goats a closed tibial fracture was made, stabilized with an external fixator, and treated as follows: (1) no injection; (2) injection of 1 mg rhOP-1 dissolved in aqueous buffer; (3) injection of collagen matrix; and (4) injection of 1 mg rhOP-1 bound to collagen matrix. The test substances were injected in the fracture gap under fluoroscopic control. At 2 and 4 weeks, fracture healing was evaluated with radiographs, three-dimensional computed tomography (CT), dual-energy X-ray absorptiometry, biomechanical tests, and histology. At 2 weeks, callus diameter, callus volume, and bone mineral content at the fracture site were significantly increased in both rhOP-1 groups compared with the no-injection group. As signs of accelerated callus maturation, bending and torsional stiffness were higher and bony bridging of the fracture gap was observed more often in the group with rhOP-1 dissolved in aqueous buffer than in uninjected fractures. Treatment with rhOP-1 plus collagen matrix did not result in improved biomechanical properties or bony bridging of the fracture gap at 2 weeks. At 4 weeks there were no differences between groups, except for a larger callus volume in the rhOP-1 plus collagen matrix group compared with the control groups. All fractures showed an advanced stage of healing at 4 weeks. In conclusion, the healing of a closed fracture in a goat model can be accelerated by a single local administration of rhOP-1. The use of a carrier material does not seem to be crucial in this application of rhOP-1.     

原始骨血肿的骨再生潜能的实验研究

目的　探索骨折后冷藏原始骨血肿的骨再生潜能和对骨折愈合的影响。方法　在新西兰兔股骨、胫骨上制造骨缺损模型 ,3d后取出骨血肿并 4℃冷藏 ,3d后冷藏的原始骨血肿回置到实验组骨缺损区 ,定期摄X线片和取出骨痂行组织学观察。结果　实验组从 2、 4、 6周均见明显骨痂形成和骨膜反应 ,组织学观察可见有成骨样细胞形成。结论　冷藏的原始骨血肿仍有很好的骨再生潜能 ,可促进骨折、骨缺损区骨愈合。骨折行手术治疗尤其是闭合性骨折行手术治疗时 ,应考虑到原始骨血肿丢失对骨折愈合的影响 ,术中注意保存部分原始骨血肿可能有利于骨折、骨缺损的愈合     

Healing of tibial fractures is not impaired after acute hindlimb ischemia in rats

The influence of transient circulatory arrest on the healing of closed tibial fractures was investigated in rats by the use of a hindlimb tourniquet technique. Twenty-four animals were randomly divided into three groups. In all animals, the left lower leg was fractured and fixed with an intramedullary nail system. In the ischemic group, complete acute transient ischemia for 4.5 h and neurapraxia of the sciatic and femoral nerves were induced prior to fracture. In the neurapraxia group, the sciatic and femoral nerves were crushed with forceps before fracture. In the control group, no other intervention than fracture was made. The rats of the control group ambulated normally 3–4 days after the operation. The animals of the ischemic and neurapraxia groups resumed normal weight-bearing after about 3 weeks. After 6 weeks, all animals were killed, and mechanical strength and bone mineral turnover of the healing tibia as well as blood flow of the bone and musculature were evaluated. The weight of the tibia and the corresponding anterior tibial muscle in the ischemic and neurapraxia animals were reduced compared with the control rats. Bone mineral turnover was found to be lower in the ischemic group. There were no differences between the groups in mechanical strength nor in blood circulation of bone and muscle. In conclusion, complete, acute hindlimb ischemia for 4.5 h in rats did not cause delayed healing of closed tibial fractures.     

专家级髓内钉治疗胫骨远端骨折的疗效观察

目的观察专家级髓内钉治疗胫骨远端骨折的临床疗效。方法将2010年1月至2012年12月我院收治的74例胫骨远端骨折患者随机分为实验组与对照组,两组均37例。实验组接受专家级髓内钉内固定术,对照组接受解剖钢板内固定术,比较两组患者手术及术后恢复情况。结果实验组与对照组在手术时间、术中出血量、X线透视次数、术后住院时间、术后主要并发症发生率等方面无明显差异(P>0.05);实验组在待床时间、总住院时间、完全负重时间及骨性愈合时间等方面均明显优于对照组(P<0.05);实验组与对照组术后踝关节症状与功能评分及Johner&Wruhs评分优良率的差异比较无统计学意义(P>0.05)。结论胫骨专家级髓内钉治疗胫骨远端骨折疗效优良,有利于患者的早期恢复,值得推广。     

组织工程化同种异体骨移植修复骨缺损

目的解决骨缺损修复时自体骨移植存在修复材料来源有限,异体骨移植又为爬行替代,存在愈合慢、假关节率高的问题。方法以兔骨膜成骨细胞为种子细胞,经分离、体外培养、传代,再粘附于冷冻干燥表面脱钙同种异体骨,共同复合培养,制作兔胫骨缺损,分异体骨移植组(对照组)、组织工程化异体骨移植组,术后2、4、6周各处死2只兔子,大体观测骨痂大小及硬度;苏木精-伊红染色,光镜观察细胞在材料上的生长情况,了解其愈合快慢及质量。结果与对照组比较,实验组炎性明显较轻,细胞生长活跃,缺损愈合快。结论组织工程化同种异体骨移植能解决骨缺损修复时自体植骨材料来源有限,特别是在小儿及骨缺损大时修复材料的来源问题;同时,植入的异体骨又具有支架及自体成骨细胞活性,使植入的异体骨愈合加快,克服其愈合慢、假关节率高的问题。     

闭合性胫骨骨折骨膜微血管功能的动力学研究

目的通过建立闭合性胫骨骨折的动物模型,在荧光显微镜下动态观察骨折后骨膜微循环的动力学改变.方法 Sprague-Dawlay大鼠14只,随机分为对照组和骨折组.使用改良快速冲击法造成大鼠左下肢胫骨中段标准的横行骨折,显微手术解剖胫骨,充分暴露骨膜,在显微镜及计算机图像分析软件下进行骨膜微血管的动力学研究.结果闭合性胫骨骨折导致骨膜功能性毛细血管密度、毛细血管网微动、静脉内红细胞移动速率及毛细血管回流量下降(P＜0.05);此外,亦引起毛细血管网动、静脉的管径扩大,毛细血管网动、静脉渗透率增加,毛细血管白细胞及内皮细胞的交互作用增强(P＜0.01).结论骨折导致骨膜微血管损伤,而骨膜微血管损伤的程度又直接影响甚至决定骨折的愈合.如何修复与重建骨折后骨膜微循环是治疗骨折的又一关键.     

Immunosuppression with FK506 has no influence on fracture healing in the rat

Immunosuppressant drugs like cyclosporine A and FK506 are widely used for solid organ transplantation. They are accelerating bone remodeling but cause net bone loss. The aim of this study was to investigate the effect of FK506 on fracture healing in the rat. Eighty Lewis rats were divided into four groups, which received FK506 (1 mg/kg BW) or no treatment for 2 or 4 weeks, beginning after production of a closed, nondisplaced unilateral tibial fracture. Radiographic, histological, and biomechanical studies were used to evaluate fracture healing and histomorphometric analysis of the tibial metaphysis of the intact contralateral side was performed. Radiographs revealed no difference of the healing of the control fractures compared with the fractures in the FK506-treated group at 2 and 4 weeks. The mechanical parameters of the tested contralateral intact tibiae and of the fracture callus demonstrated no difference between control and immunosuppressed animals. Tibial bone histomorphometry revealed increased measures of bone formation and bone resorption, accompanied by a significant reduction of percent trabecular area. At 4 weeks, the fractures showed osseous healing with woven bone at the fracture site and only minimal amounts of cartilage. Histological grading was not different between the control and the FK506 group at both time points. We conclude that systemic application of FK506 has no biomechanical and histological effects of experimental fracture healing in the rat. However, resorption far in excess of formation leads to a net bone loss in the trabecular bone of the tibia that has no effect on the stability of the intact bone.