海水浸泡股骨开放性骨折的早期救治

目的利用犬动物模型研究海水浸泡股骨开放性骨折的伤情特点及早期救治方法。方法20只华北成年犬,随机分为对照与海水浸泡两组。于股骨中段截骨制备开放性骨折的动物模型,实验组犬浸泡于海水中3h,对照组于笼内放置3h,两组均于3h后行伤口清创、钢板螺钉内固定术。结果①对照组和海水浸泡组分别有1只和3只动物伤口感染(P>0.05);②病理结果显示海水浸泡组软骨骨痂较多;③生物力学测试两组骨愈合刚度分别为(451.49±183.00)N和(150.15±20.16)N(P<0.05);④影像学观察两组骨痂形成及骨愈合无明显差别。结论①海水浸泡使开放骨折感染率有增高趋势。②海水浸泡使骨折愈合质量下降,骨折愈合过程有延迟倾向。③生物力学测定示海水浸泡开放骨折愈合强度明显低于对照组。④内固定有增加伤口感染的风险,但在股骨这一特定部位可选择性应用。     

血管内皮生长因子在骨折愈合中的自分泌作用

目的探讨血管内皮生长因子（VEGF）在骨折愈合中的自分泌作用。方法利用鼠股骨闭合性骨折模型，应用组织学和逆转录聚合酶链反应（RT-PCR）方法，观察骨折不同的愈合修复阶段中骨痂的组织学变化，检测各种相应骨痂组织中VEGF及其受体VEGFR1（Flt-1）和VEG-FR2（KDR／Flk-1）的mRNA的表达。结果骨折愈合是一个高度有序的组织学变化过程，骨痂内的膜内化骨，软骨形成，软骨内化骨等过程可同时或连续出现。vEGF（251bp）及受体Flt-1（272bp）和KDR／Flk-1（252bp）在骨折后第7、14天的软骨性骨痂（软骨组织）和骨性骨痂（小梁骨-膜内化骨和软骨内化骨）中均同时清晰表达。表达信号均匀，强度较大。结论骨折愈合过程中的软骨性骨痂和骨性骨痂中的软骨-骨细胞系统共表达VEGF及其两种受体，提示VEGF自分泌作用参与骨折愈合过程。     

海水浸泡开放性骨折不同救治方法的实验研究

目的观察两种骨折固定方法救治海水浸泡开放性骨折的大体形态、组织病理学、影像学变化。方法取成年新西兰兔50只，随机分为三组：对照组10只、A组20只、B组20只。于胫骨中段截骨制成开放性骨折动物模型。对照组伤口自然旷置3h，A组和B组海水浸泡伤口3h。随后对照组和A组行伤口清创、钢板螺钉内固定术；B组行伤口清创、外固定架固定、伤口开放换药、二期缝合。观察各组的伤口感染情况、骨折愈合的组织学、影像学变化和测定骨折断端骨痂的平均比灰度值。结果a）对照组感染1只（10％），A组感染15只（75％），B组感染5只（25％）。各组间结果有统计学差异（P〈0．05）。b）术岳45d时各组间骨折断端组织愈合等级有统计学差异（P〈0．05）。c）术后45d时各组间骨折部位骨痂的生长情况在影像学上表现为对照组大于B组大于A组。各组间骨愈合率对照组为100％，A组为66．7％，B组为93．3％。d）术后45d时骨折断端间骨痂的平均比灰度值对照组为8．1149±1．2043，A组为6．2268±1．4000，B组为6．5138±1．3045，各组间有统计学差异（P〈0．05）。结论海水浸泡使开放性骨折伤口感染率增高；海水浸泡使开放性骨折断端骨痂形成不良率增高；骨外固定架与钢板内固定比较救治成功率提高。     

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.     

扩髓治疗骨折不愈合中的血管分布研究

目的 研究扩髓治疗骨折不愈合过程中血管生成和血管内皮细胞生长因子(VEGF)表达的时间和空间特点以及相互联系.探讨扩髓并更换粗直径髓内钉治疗骨折不愈合的生物学机理,为临床运用扩髓治疗长骨骨折延迟愈合及不愈合提供理论依据. 方法 3个月龄雄性SD大鼠200只.对大鼠一侧股骨干骨折用不稳定固定方法制备肥大性骨折不愈合模型,随机选取40只作为模型鉴定,剩下160只随机分为试验组扩髓治疗和对照组不扩髓治疗,每组80只进行随机配对研究.术后第1、3、5、7、14、21、28、42天取材.用血管性血友病因子(VWF)标记血管,免疫组化方法检测骨折处VWF和VEGF的表达,来研究骨折处血管生成的时空特点以及与VEGF表达的关系. 结果 实验组的骨折愈合率显著高于对照组.时空分布上,在早期(1～7 d),实验组血管生成主要在骨外膜和软骨痂中.对照组主要在骨内膜;中期(7～14 d)实验组血管随着肉芽组织长入骨折间隙,对照组近骨内膜处血管增加,但是骨折间隙没有血管;后期(14～42 d)实验组骨内膜处出现血管,骨折间隙血管大量增加,对照组血管仍然局限在肉芽组织和骨内膜处,骨折间隙没有血管.实验组血管总数始终高于对照组.VEGF出现的时间和空间顺序与血管生成顺序一致,实验组VEGF表达普遍高于对照组. 结论 在骨折不愈合的治疗中,扩髓可使骨外膜和骨痂血供代偿性增加以及刺激VEGF表达增加,血管总数要高于不扩髓组.VEGF表达时间和分布与血管的生成关系密切,提示治疗骨折不愈合中,VEGF可能起着至关重要的作用,决定着血管形成的时间、部位与数量.     

血管内皮生长因子对骨折愈合相关因子表达的调控

目的探讨应用和拮抗血管内皮生长因子(VEGF）对骨折愈骨相关因子表达的影响，并观察骨折愈台过程中的病理改变。方法用105只天上I=白兔制作骨折模型．随机分为对照组、应用VECF组和拮抗vEGF组，分别于伤后8、24、72b和1．3、5、8周测定各组动物骨折端相关因子的表达变化、同时取骨折端标本脱钙进行兜镜观察。结果应甩VEGF使骨折端埘BMP的表达时同提前和延良，拈抗VEGF抑制rBMP的表达，应用VEGF组伤后3J嗣时骨折端充填有纤维性骨痴，软骨骨痂和骨性骨痂．5周时新生骨以编织骨为主，8周时骨折正常愈合；括抗VH押导受伤后初期骨细胞坏死增多，1～5周各时相点骨折端均有灶性坏死出现，3同时骨折部位血管生成明显减少。结论骨折愈台过程需要多因子参与、协调，VEGF有可能是作为骨折愈台过程中的一种重要因子在发挥作用     

仙灵骨葆胶囊联合阿仑膦酸钠对骨质疏松骨折大鼠骨痂血管形成的影响

目的探讨仙灵骨葆胶囊联合阿仑膦酸钠对骨质疏松性骨折大鼠骨痂血管形成及VEGF、BMP-2表达的影响。方法50只雌性SD大鼠随机分为假手术组(SHAM组)、模型组(MODEL组)、仙灵骨葆组(XLGB组)、阿仑膦酸钠组(ALLSN组)、联合药物组(LHYW组),10只/组,构建骨质疏松性骨折大鼠模型,放射性X线观察评估骨折愈合,双能X线检测骨密度,Mirco-CT检测骨结构形态学参数,番红O固绿染色观察骨痂组织形态学,免疫组化检测骨痂VEGF和BMP-2蛋白表达。结果所有实验大鼠均进入结果分析。与SHAM组比较,MODEL组大鼠骨折愈合评分、骨密度、骨组织形态学参数、骨痂VEGF和BMP-2表达均显著降低(P0.05),与MODEL组比较,XLGB组、ALLSN组和LHYY组骨折愈合评分、骨密度、骨组织形态学参数、骨痂VEGF和BMP-2表达均显著升高(P0.05),尤以LHYY组最高。SHAM组骨小梁结构正常,几乎均为骨性骨痂。MODEL组骨小梁明显稀疏、断裂,未见明显骨性骨痂。XLGB组和ALLSN组骨小梁增多,排列稍紊乱,大部分为骨性骨痂。LHYW组骨小梁明显增多,排列密集整齐,大量骨性骨痂。结论仙灵骨葆胶囊联合阿仑膦酸钠可能通过介导提高骨质疏松性骨折大鼠骨生长因子VEGF和BMP-2表达,促进骨痂血管形成,加速骨痂形成,增加骨密度,改善骨结构形态,促进骨折愈合。     

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.     

雄性激素减少对骨折愈合过程中骨痂显微结构变化的影响

目的探讨雄性激素减少对骨折愈合过程中骨痂显微结构变化的影响。方法清洁级SD大鼠40只,6月龄,随机分为骨折组（A组）及睾丸切除＋骨折组（B组）,建立骨折模型,造模后第3、7、14、21d,每组随机选取5只大鼠,切取骨痂标本,在显微镜下观察骨痂显微结构并拍片观察。结果A组毛细血管侵入时间、成纤维细胞、成软骨细胞、成骨细胞出现时间早于B组,且功能较B组活跃。结论睾丸切除后雄性激素下降对骨折愈合的影响,主要发生在骨折中后期,表现为软骨骨痂向骨性骨痂演变缓慢和骨改建期间骨吸收加剧。     

去势对骨折早期愈合过程的影响

目的 通过对去势大鼠股骨骨折模型愈合早期骨痂的组织学、骨密度以及转化生长因子β1(Transforming growth factor beta 1.TGF-β1)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF）、骨形态发生蛋白-2(bone morphogenetic proteins-2，BMP-2)合成变化的观察．探讨骨质疏松骨折早期愈合过程的变化。方法 将60只雌性SD大鼠分为去势组和对照组，3个月后制成股骨干骨折模型．伤后不同阶段处死．分别进行组织学、骨密度以及TGF-β1,bFGF,BMP-2免疫组化染色方法观察。结果 大鼠去势后3个月全身骨密度检查证实去势组骨质疏松形成。骨折后第3d两组均开始形成原始骨痂；第4～6周，去势组骨痂比对照组少，且软骨痂比例较高．骨密度较低，免疫组化染色显示，在两组间bFGF、BMP-2的表达与分布、高峰出现与持续时间差异无显著性．去势组骨小梁附近表达TGF-β1的成骨细胞数目减少，结论 骨质疏松使大鼠骨折早期骨痂的数量与质量降低，自软骨性骨痂至骨性骨痂演变过程减缓．对骨折愈合过程的影响与BMP-2、bFGF的表达无明显关联。TGF-β1在成骨细胞中的表达减少可能是引起骨质疏松骨折愈合质量下降的因素之一．     

骨折愈合过程中血管内皮细胞生长因子表达的实验研究

目的 观察骨折愈合过程中骨痂组织内血管内皮细胞生长因子（ｖａｓｃｕｌａｒ ｅｎｄｏｔｈｅｌｉａｌ ｇｒｏｗｔｈ ｆａｃｔｏｒ,ＶＥＧＦ）的表达,进一步探讨ＶＥＧＦ对骨折愈合的影响。方法 采用免疫组织化学方法愈合的不同阶段,骨痂组织内表达ＶＥＧＦ的细胞来源不同,同时表达程度也存在差异。结论 骨折愈合的不同时期,骨痂组织内间充质细胞、软骨细胞、成骨细胞等出现程度不同的ＶＥＧＦ表达,表明ＶＥＧＦ参与调节     

Effect of fluvastatin on vascular endothelial growth factor in rats with osteoporosis in process of fracture healing

OBJECTIVE: To explore the effect of fluvastatin on vascular endothelial growth factor (VEGF) in rats with osteoporosis in the process of fracture healing. METHODS: Fractures at the intermediate piece of the femur were made on 72 Sprague Dawley (SD) rats (weighing initially 290-340 g and aged 6 months) with osteoporosis after ovariectomy for three months, then these rats were divided randomly into the medication administration group (the experimental group) and the control group, 36 rats each. In the experimental group, the rats received fluvastatin lavage (10 mg/kg per day) since the next day of operation lasting for 6 weeks, and the rats in the control group received placebo. Then the expression of VEGF and VEGF mRNA in bony callus of the two groups was measured respectively with immunohistochemistry and in situ hybridization on days of 3rd, 7th, 14th, 21st, 28th, and 42nd, and image analysis was made with real-color image analysis machine. RESULTS: No difference was found in the cellular localization of VEGF and VEGF mRNA gene expression between the experimental group and the control group in process of fracture healing and their expression modes were almost similar. On the 14th day postoperatively, the positive extent of positive cells in the experimental group was higher than that of the control group (P < 0.05). CONCLUSION: Fluvastatin can promote the VEGF level in rats with osteoporosis in process of fracture healing.     

Effects of recombinant human basic fibroblast growth factor on cell proliferation during mandibular fracture healing in rabbits

ＯＤｅｐａｒｔｍｅｎｔｏｆＯｒａｌ ＭａｘｉｌｌｏｆａｃｉａｌＳｕｒｇｅｒｙ ,ＣｏｌｌｅｇｅｏｆＳｔｏｍａｔｏｌｏｇｙ ,ＦｏｕｒｔｈＭｉｌｉｔａｒｙＭｅｄｉｃａｌＵｎｉｖｅｒｓｉｔｙ ,Ｘｉ ａｎ 710 0 32 ,Ｃｈｉｎａ (ＧｏｎｇＺＹ ,ＺｈｏｕＳＸ ,ＣａｏＪＧａｎｄＧｕＸＭ)ｎｃｅｆｒａｃｔｕｒｅｏｃｃｕｒｓ ,ｃｅｌｌｓａｒｅｎｅｅｄｅｄｆｏｒｂｏｎｅｒｅｐａｉｒｔｏａｇｇｒｅｇａｔｅａｔｔｈｅｆｒａｃｔｕｒｅｓｉｔｅｓａｎｄｉｔｉｓａｌｓｏｉｍｐｏｒｔａｎｔｆｏｒｆｒａｃｔｕｒｅｈｅａｌｉｎｇｔｏｉｎ…     

Effect of recombinant human basic fibroblast growth factor on angiogenesis during mandible fracture healing in rabbits

Objective: To investigate the effect of recombinant human basic fibroblast growth factor ( rhbFGF) on angiogenesis during mandible fracture healing in rabbit.Methods: Fifty adult white rabbits were used for animal model and randomly divided into a control group (25 rabbits) and an experimental group (25 rabbits). The membranous complex of rhbFGF and bovine type I collagen was prepared and implanted into the rabbit mandible fracture site under periosteum. The animals were sacrificed on 7, 14, 28, 56 and 84 days respectively after operation and the whole mandibles were harvested. The expression of factor Vm related antigen (F8-RA) in callus was examined with immimohistochemical staining.Results: The amounts of microvascular formation in calluses in the rhbFGF-treating group on days 7, 14, 28 and 56 were more than those of the control group (P < 0.01).Conclusions: The results indicated that rhbFGF could stimulate microvascular formation during mandible fracture healing in rabbits.     

整合素αv β3和VEGF在骨折愈合过程中的表达及意义

[目的]探讨整合素α,β3和VEGF在骨折愈合过程中的表达及意义.[方法]将新西兰大白兔20只,随机分为4组,每组5只,手术制作左侧兔桡骨中段骨干缺损模型,作为实验组.右侧作假手术处理,作为对照组;应用免疫组织化学链霉亲合素-生物素酶复合物(SABC)法检测术后2、4、8、12周骨痂组织中整合素αvβ3和VEGF的表达.[结果]在骨折愈合的不同时期,整合素仅αvβ3和VEGF的表达上调,对照组与实验组的表达差异有统计学意义(P＜0.05).术后2周整合素αvβ3和VEGF的表达水平最高,术后12周表达水平最低.[结论]整合素αvβ3和VEGF可能在骨折愈合的过程中起重要作用,检测整合素αvβ3和VEGF的表达有助于对骨折预后的判断.     

Systemic leptin administration alters callus VEGF levels and enhances bone fracture healing in wildtype and ob/ob mice

IntroductionLeptinʼs role in bone formation has been reported, however, its mechanism of affecting bone metabolism is remaining unclear. In this study, we aimed to test whether leptin has a positive effect on fracture healing through the possible mechanism of increasing vascular endothelial growth factor (VEGF) expression in callus tissue.MethodsStandardized femur fractures were created in leptin-deficient ob/ob and wildtype C57BL/6J mice, and recombinant mouse leptin or its vehicle (physiological saline) was administered intraperitoneally during the study. Body weight, radiological, histologic and immunoblotting analyses were performed at different stages of fracture healing.Key findingsThe results showed that leptin treatment led to lower rate of body weight change in both mice genotypes. Radiological and histological analyses showed that the experimental groups had better fracture healing at 14, 21 and 28 days compared to the control groups. Leptin-treated groups had significantly higher VEGF expression in callus compared with the control groups at 2 and 3 weeks post-fracture except normal mice at 2 weeks, and leptin-deficient mice had higher VEGF levels in calluses than normal mice at the same timepoint.ConclusionLow-dose systemically-administered leptin has a positive effect on promoting fracture healing during the latter stages in a clinically-relevant mouse bone fracture model, and increase callus VEGF levels.     

Regulation of vascular endothelial growth factor on the expression of fracture healing,related factors

Objective ： To study the effect of vascular endothelial growth factor （VEGF）and anti-VEGF on the expression of fracture healing-related factors and observe pathological changes at fractured sites. Methods： Fracture models were established in 105 New Zealand white rabbits and they were randomly divided into control group, VEGF group and anti-VEGF group. The relevant factors expression at fractured sites was assayed and pathological changes were observed in decalcified samples at 8, 24, 72 hours and 1,3,5,8 weeks after fracture. Results： After application of VGEF, the expression of BMP appeared earlier and expression time lasted longer. On the contrary, anti-VEGF completely inhibited the expression of BMP. The fractured sites were filled with fibrous callus, cartilaginous callus and bony callus at the 3rd week and woven bone was constructed at the 5th week. Fracture healing was accomplished at the 8th week in VEGF group. In anti-VEGF polyclonal antibody group, cellular necrosis increased at early period. Continuous focal necrosis was seen in the fractured sites from the 1st week to 5th week. Vascularization reduced obviously at the 3rd week. Conclusions： Fracture healing is a result of mutual regulation and coordination among many factors. VEGF may be an important factor in fracture healing.