甲状旁腺激素联合降钙素对大鼠骨质疏松性骨折骨生长因子的影响

目的探讨甲状旁腺素联合降钙素对大鼠骨质疏松性骨折骨相关生长因子及骨折愈合的影响。方法75只雌性SD大鼠随机分为:假手术组、去势组、甲状旁腺素组、降钙素组、联合用药组,15只/组,首先构建去卵巢大鼠骨质疏松性骨折动物模型。观察并评估骨折愈合,测定骨痂BMD和BMC,检测血清BMP-2、VEGF、TGF-β、IGF-1水平和骨痂蛋白表达,观察骨痂形态结构变化。结果去势组较假手术组骨折愈合评分、骨痂BMD和BMC、血清BMP-2、VEGF、TGF-β、IGF-1水平和骨痂蛋白表达均显著降低(P均<0.05),而联合用药组较去势组上述指标均显著升高(P均<0.05)。去势组骨痂骨小梁明显减少,生长稀疏,排列紊乱,大量间充质干细胞及纤维软骨细胞,成骨细胞及微血管少见;联合用药组骨痂大量骨小梁,生长旺盛,互相交错网状,排列致密有序,可见较多成熟的骨细胞及成骨细胞。结论甲状旁腺素联合降钙素通过介导提高相关骨生长因子表达,提高骨质疏松性骨折骨密度和矿物含量,改善骨组织形态学,加快骨折愈合。     

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。     

葛根素对去卵巢大鼠骨质疏松性骨折骨痂血管形成的影响

目的探讨葛根素对骨质疏松性骨折骨痂血管形成的影响及对骨折愈合的作用。方法60只雌性SD大鼠分为3组:假手术组、去势组、葛根素组,20只/组,去卵巢大鼠骨质疏松性骨折动物。观察评估骨折愈合情况,检测血清BMP-2和VEGF浓度,观察骨痂形态结构变化,检测骨痂BMP-2和VEGF表达。结果去势组较假手术组骨折愈合评分、血清BMP-2和VEGF浓度、骨痂BMP-2和VEGF表达、微血管数均显著降低(P<0.05),葛根素组较去势组上述指标均显著增高(P<0.05)。去势组骨痂组织可见少量骨小梁,生长稀疏,排列紊乱,大量纤维软骨细胞等纤维组织,成骨细胞及新生小血管少见;葛根素组骨痂可见较多骨小梁,生长较旺盛,排列有序,可见较多成熟的骨细胞,新生微小血管较多。结论葛根素通过介导去卵巢大鼠骨质疏松性骨折骨痂BMP-2和VEGF表达,促进骨痂血管形成,改善骨组织形态学,加快骨折愈合。     

仙灵骨葆胶囊对骨质疏松性骨折大鼠骨生长因子及骨折愈合的影响

目的 探讨仙灵骨葆胶囊对骨质疏松性骨折大鼠骨生长因子BMP-2、IGF-1表达及骨折愈合的影响。方法 48只雌性SD大鼠随机分为：假手术组、模型组、雌二醇组、仙灵骨葆组，12只/组，采用“双侧卵巢切除术+右侧股骨干骨折髓内固定术”构建骨质疏松性骨折大鼠模型，评估骨折愈合情况，检测股骨骨痂BMD、股骨骨生物力学指标和血清骨代谢相关指标，检测骨痂BMP-2、IGF-1蛋白表达。结果 模型组较假手术组骨折愈合评分、股骨痂BMD、股骨骨生物力学指标（最大载荷、最大应力、最大位移）、骨痂BMP-2和IGF-I阳性表达均显著降低（P<0.05），雌二醇组、仙灵骨葆组较模型组骨折愈合评分、股骨痂BMD、股骨骨生物力学指标、骨痂BMP-2和IGF-I阳性表达均显著升高（P<0.05），均以仙灵骨葆组最高。模型组较假手术组血清骨代谢指标（BGP、PICP、TRACP-5b）均显著升高（P<0.05），雌二醇组、仙灵骨葆组较模型组血清骨代谢指标均显著降低（P<0.05），以仙灵骨葆组最低。结论 仙灵骨葆胶囊可能通过介导提高骨质疏松性骨折大鼠骨生长因子BMP-2和IGF-1表达，改善骨代谢，加速骨痂形成，增加骨密度，提高骨生物力学，促进骨折愈合。     

辛伐他汀联合阿仑膦酸钠对去卵巢骨质疏松大鼠骨代谢的影响

目的探讨辛伐他汀联合阿仑膦酸钠干预对去卵巢大鼠骨质疏松骨代谢的影响。方法 60只雌性SD大鼠随机平均分为5组:假手术组、去势组、辛伐他汀组、阿仑膦酸钠组、联合药物组,首先构建去卵巢大鼠骨质疏松性模型。分别检测骨代谢相关生化指标、氧化应激生化指标和骨组织骨密度(bone mineral density,BMD),HE染色观察骨组织形态学。结果去势组大鼠血清Ca、P、SOD、CAT和骨组织BMD均较假手术组显著降低(P0.05),辛伐他汀组、阿仑膦酸钠组、联合药物组上述指标均较去势组升高,以联合用药组升高最显著(P0.05)。去势组大鼠血清ALP、BGP、PICP、TRAP、GLA、ICIP和MDA较假手术组均显著增高(P0.05),辛伐他汀组、阿仑膦酸钠组、联合药物组上述指标较去势组均降低,以联合用药组降低最显著(P0.05)。去势组股骨骨小梁明显稀疏,连接不完整,大量纤维组织,髓腔内大量空泡状脂肪细胞。联合药物组骨小梁数目明显增多,结构较完整,粗细均匀致密,连接成网状。结论辛伐他汀联合阿仑膦酸钠通过调节去卵巢大鼠骨代谢,抗氧化应激,增加骨密度,改善骨组织结构,发挥抗骨质疏松作用。     

阿仑膦酸钠和强骨胶囊单用与合用对大鼠骨质疏松性骨折骨愈合影响的实验研究

目的通过骨质疏松性骨折动物模型,观察阿仑膦酸钠、强骨胶囊单用及合用对骨折愈合的影响,以探讨中西医结合治疗骨质疏松性骨折的意义。方法 70只SD大鼠给予维甲酸灌胃,制造骨质疏松性骨折模型成功后分别予以阿仑膦酸钠、强骨胶囊及两药合用,于2周、4周和6周处死5只试验鼠,进行骨痂大小的测量及组织学研究。结果 2周时合用组骨痂大小较其他组无显著差异;4周时合用组骨痂大小较阿仑膦酸钠组小,较强骨胶囊组大;6周时各组骨痂大小无异。组织学检测发现阿仑膦酸钠组破骨细胞数目最少,骨小梁成熟较其他组缓慢;强骨胶囊组成骨细胞数目最多,小梁骨成熟最快;强骨胶囊联合阿仑膦酸钠组与对照组无差别。结论强骨胶囊与阿仑膦酸钠联合使用治疗维甲酸所致大鼠骨质疏松性骨折疗效并不显著。     

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．     

甲状旁腺素联合降钙素对大鼠骨质疏松的治疗作用

目的探讨甲状旁腺素联合降钙素对大鼠骨质疏松的影响及其作用机制,为临床治疗提供理论依据。方法 100只雌性SD大鼠随机分为5组:假手术组、去势组、甲状旁腺素组、降钙素组、联合用药组,每组20只,构建去卵巢大鼠骨质疏松动物模型。骨组织骨密度(bone mineral density,BMD)和骨矿物质含量(bone mineral content,BMC)测定,血清E2检测,骨代谢相关生化指标和标志物检测,观察骨形态结构变化。结果去势组较假手术组腰椎和股骨BMD和BMC、股骨最大负荷及血清E2、Ca、P、骨保护素水平均显著降低(P0. 05),而联合用药组上述指标较去势组均显著增高(P0. 05)。去势组血清碱性磷酸酶、OC、RANK水平较假手术组均显著升高(P0. 05),而联合用药组上述指标较去势组均显著降低(P0. 05)。去势组骨小梁明显减少,排列稀疏错乱,并出现大片断裂现象,大量纤维组织,骨细胞少见;联合用药组骨小梁较多且致密,形态结构较规整,骨细胞排列较整齐,骨连续性好,骨密质较厚且均匀。结论甲状旁腺素联合降钙素通过提高雌激素水平,调节维持骨代谢,提高骨密度和矿物含量,改善生物力学性能,改善骨组织病理形态学,起到抗骨质疏松疗效和骨保护作用。     

针刺对实验性骨质疏松大鼠股骨骨折的预防作用

目的研究"健脾益肾强骨针法"对去势SD大鼠股骨骨质疏松性骨折的预防作用。方法 6月龄SD大鼠在去势3个月检测骨质疏松症模型造模成功后,被随机分为3个组:对照组、阿仑膦酸钠组和针刺组。针刺组针刺足三里、肾俞、大杼,每日1次,每周5次,连续治疗6个月;阿仑膦酸钠组每日皮下注射阿仑膦酸钠,每日1次,连续治疗6个月;对照组每日在其他两组治疗期间进行同样抓放。治疗6个月后,检测包括在体股骨骨折最小外力、新鲜股骨形态学、骨密度、骨力学性能等反映股骨的硬度和弹性的指标。结果针刺组、阿仑膦酸钠组的在体股骨骨折最小外力明显大于对照组;针刺组新鲜股骨近端横径明显小于阿仑膦酸钠组和对照组;针刺组、阿仑膦酸钠组骨密度明显大于对照组;针刺组、阿仑膦酸钠组离体股骨的部分力学性能指标(如压缩、拉伸)明显高于对照组。结论 1针刺足三里、肾俞、大杼6个月具有预防绝经后骨质疏松症模型大鼠股骨骨折的作用,其机制可能与针刺促进股骨的形态适应、增加骨密度、提高骨的抗外力性能密切相关。2"健脾益肾强骨针法"与阿仑膦酸钠均能提高绝经后骨质疏松大鼠在体股骨骨折最小外力,但针刺的作用机制与阿仑膦酸钠不完全相同。     

雷尼酸锶促进大鼠股骨骨质疏松性骨折愈合的研究

目的 探讨雷尼酸锶(strontium ranelate,SR)对大鼠股骨骨质疏松性骨折愈合的影响。方法 6个月龄Wistar雌性处鼠66只,切除双侧卵巢制作骨质疏松动物模型,模型建立成功后,在大鼠股骨中段横行截骨,制作大鼠股骨骨质疏松性骨折动物模型,术后死亡6只,然后将剩余60只大鼠随机分为实验组(SR干预治疗组）和空白对照组,每组30只。骨折术后第一天起,实验组给予SR 400 mg/(kg d)灌胃,对照组给予同等体积的生理盐水灌胃。通过影像学、组织学和免疫组织化学（骨形态发生蛋白-2,BMP-2)观察,骨组织形态计量学、骨密度（bone mineral density,BMD)和生物力学测量,观察骨折愈合情况。结果 实验组腰4、5椎体BMD、BMP-2阳性表达细胞数、骨小梁面积比、骨小梁平均宽度、最大负荷和最大桡度高于同时期对照组,差异有统计学意义(P <0. 01);骨小梁平均间隔小于同期对照组,差异有统计学意义(P <0. 01)。结论 SR具有抑制骨吸收和促进骨形成的双重作用,能增加BMD,加快骨痴形成,促进骨质疏松性骨折愈合,改善骨的显微结构,提高骨折愈合后骨的生物力学特性。     

Relationship among bone mineral density, collagen composition, and biomechanical properties of callus in the healing of osteoporotic fracture

Objective： To study the change and relationship among bone mineral density （BMD）, collagen composition and biomechanical properties of the callus in the healing process of osteoporotic fracture. Methods： The osteoporotic rat model and fracture model were established through bilateral ovariectomy （OV＇X） and osteotomy of the middle shaft of the right hind tibiae, respectively. Ninety female SD rats were randomly divided into OVX group and sham group. With the samples of blood and callus, roentgenoraphic and histological observation were performed for the assessment of the healing progress of the fracture, and the serum concentration of TRAP-5b, proportion of type I collagen, BMD and biomechanical properties of the callus were measured. Results： The OVX group experienced a significant delay of fracture healing. The mean serum concentration of TRAP-5b of rats in the OVX group was much higher than that in the sham group after the operation （P 〈 0.05）, but the difference at the same time point after fracture was smaller than that before fracture （P 〈 0.05）. The BMD of the callus in both groups reached the peak value at the 6 th week after fracture while the proportion of the type I collagen and the biomechanical strength reached the peak at the 8th week. Conclusions. The deficiency of estrogen after the ovariectomy could induce the up-regulation of the osteoclasts activities, whereas the potency of further activation after fracture was depressed. Although the synthesis of collagen together with its mineralization determines the biomechanical properties of new bone, the accumulation of collagen could be assessed as an index in the prediction of biomechanical strength of bones independent of the bone mineral deposition.     

葛根素联合阿仑膦酸钠对绝经后骨质疏松大鼠氧化应激、炎症反应及骨代谢的影响

目的观察研究葛根素联合阿仑膦酸钠对绝经后骨质疏松大鼠氧化应激、炎症因子和骨代谢的影响。方法实验分为假手术组、去卵巢组、A药组、B药组和AB药组,构建绝经后骨质疏松大鼠动物模型,分别给予不同药物干预。分别检测血清氧化应激、炎性因子、骨代谢指标及骨密度,观察骨组织形态结构。结果去卵巢组大鼠血清SOD和GSH-Px水平较假手术组均显著降低(P<0.05),AB药组较去卵巢组均显著升高(P<0.05);去卵巢组大鼠血清H2O2和MDA水平较假手术组均显著升高(P<0.05),AB药组较去卵巢组均显著降低(P<0.05)。去卵巢组大鼠血清炎性因子和骨代谢指标较假手术组均显著升高(P<0.05),AB药组较去卵巢组均显著下降(P<0.05)。去卵巢组大鼠骨组织BMD较假手术组显著降低(P<0.05),AB药组较去卵巢组显著升高(P<0.05)。去卵巢组骨皮质明显变薄,骨小梁明显稀疏,排列紊乱,髓腔扩大。AB药组骨皮质轻微改变,骨小梁明显增多,排列规则,髓腔明显变小。结论葛根素联合阿仑膦酸钠具有协同抑制绝经后骨质疏松大鼠氧化应激和炎症反应,降低氧化应激和炎性因子分泌,调节骨代谢,改善骨组织形态学结构,发挥抗骨质疏松作用。     

Effects of calcium supplements on fracture healing in a rat osteoporotic model

Fracture healing is a complex process, which is further complicated if the bone is osteoporotic. Calcium is one of the important minerals in bone and has been found to prevent osteoporosis but its role in fracture healing of osteoporotic bone is still unclear. We carried out a study on the effects of calcium supplementation on the late phase healing of fractured osteoporotic bone using an ovariectomized rat model. Twenty‐four female Sprague–Dawley rats were divided into three groups: sham‐operated (SO), ovariectomized‐control (OVXC), and ovariectomized + calcium supplements (Ca). The right femurs of all the rats were fractured at mid‐epiphysis and a K‐wire was inserted for internal fixation. After 2 months of treatment, the rats were sacrificed and the femora were dissected out for radiological and biomechanical assessment. As expected, osteoporosis resulted in impaired healing as shown by the poor radiological and biomechanical properties of the OVXC group. CT scans showed significantly lower callus volumes in the SO and Ca groups compared to the OVXC group. Radiological scoring of fracture healing and callus staging of the SO and Ca groups were better than the OVXC group. However, the biomechanical parameters of the Ca group were significantly lower than the SO group and similar to the OVXC group. Therefore, calcium supplements may appear to improve fracture healing of osteoporotic bone but failed to improve strength. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1651–1656, 2010     

Dsteoporosis influences the middle and late periods of racture healing in a rat osteoporotic model

OBJECTIVE: To evaluate the influence of osteoporosis on the middle and late periods of fracture healing process through observing the histomorphological changes, bone mineral density and biomechanical properties in ovariectomized rats. METHODS: Eighty-four female SD rats of 4 months old were randomly divided into osteoporosis group and sham operation group, 42 in each. Rats in osteoporosis group were performed ovariectomy operation while those in sham operation group were given sham operation. A midshaft tibia fracture model was established 10 weeks after ovariectomy. Tibias were harvested 2, 4, 6, 12, 18 weeks after fracture for bone mineral density, histomorphological and biomechanical evaluation. RESULTS: Compared with the sham operation group, callus bone mineral density was 12.8%, 18.0%, 17.0% lower in osteoporosis group 6, 12, 18 weeks after fracture, respectively (P<0.05); callus failure load was 24.3%, 31.5%, 26.6%, 28.8% lower in osteoporosis group, and callus failure stress was 23.9%, 33.6%, 19.1%, 24.9% lower in osteoporosis group 4, 6, 12, 18 weeks after fracture, respectively (P<0.05). In osteoporosis group, endochondral bone formation was delayed, more osteoclast cells could be seen around the trabecula, and the new bone trabecula arranged loosely and irregularly. CONCLUSIONS: Osteoporosis influences the middle and late periods of fracture healing in the rat osteoporotic model. The impairment is considered to be the result of combined effects of prolonged endochondral calcification, high activated osteoclast cell and the deceleration of the increase in bone mineral density.     

Osteoporosis influences the middle and late periods of fracture healing in a rat osteoporotic model

Osteoporosisischaracterizedbydecreasedbonemass, increasedbonefragilityinducedbydemolitionofbonemicrostructureandincreasedsusceptibilitytofracture. Currentstudiesmainlyfocusonthepreventionoffracture. However, theinfluenceofosteoporosisonthefracturehealingremainspoorlyunderstoodandcontroversial.Inourpreviousstudy, wehaveevaluatedtheeffectofosteoporosisontheearlyperiodoffracturehealing, andfoundthatosteoporosisinfluencesthequantityandqualityofcallusduringtheearlyperiodofracturehealing.1 Incurrent…