卵巢去势对骨折愈合超微结构和BMP-4基因表达的影响

目的:探索雌激素对骨折愈合超微结构和骨形态发生蛋白-4(BMP-4)基因的定位分布的影响.方法:选用健康SD大鼠,随机分为卵巢切除(OVX)组和假手术对照(S)组.制成胫骨骨折愈合模型并分别于骨折后不同时间点处死取材.通过电镜观察超微结构变化;采用原位杂交方法研究卵巢去势对BMP-4基因表达影响.结果:(1)OVX组在骨痂形态及成骨细胞和破骨细胞活性与S组比较有明显差异;(2)骨折后1～3d BMP-4m RNA表达强度:S组＜OVX组.结论:雌激素缺乏时,骨转换加快,从而BMP-4表达增多.雌激素在骨折愈合过程中起重要作用.     

降钙素对卵巢切除大鼠股骨骨折愈合的影响

目的观察降钙素对卵巢切除大鼠股骨骨折愈合的作用，为临床上治疗骨质疏松性骨折提供实验依据。方法50只雌性、14周龄SD大鼠共分成5组，每组10只。分成假手术组（Sham，G1），双侧卵巢切除术组（OVX，G2），假手术＋骨折组（Sham＋F，G3），卵巢切除术＋骨折组（OVX＋F，G4），卵巢切除＋骨折＋降钙素药物组（OVX＋F＋CT，G5），骨折组大鼠均采用右股骨中段横行骨折，髓内针固定；降钙素采用皮下给药，隔日1次（16IU·kg^-1）。所有大鼠于术后4周杀死，取右侧股骨标本。然后，分别进行CR摄片、组织形态学观察，并应用双能X线骨密度仪（DEXA）测量右股骨整体、远段和中段骨密度以及BMP-2免疫组化观察，并应用病理图像分析仪对BMP-2免疫组化进行光密度测量。结果（1）OVX组与Sham组比较，BMD显著下降。（2）OVX＋F＋CT组与OVX＋F组比较：骨痂mBMD显著增高；BMP-2的表达无显著性差异。结论降钙素对OVX大鼠股骨骨折具有明显促进骨折愈合的作用，加速编织骨向板层骨的演变过程。     

骨形成蛋白2基因修复兔桡骨缺损的实验研究

目的观察携带人骨形成蛋白2（bone morphogenetic protein 2，BMP-2）基因的腺病毒载体（adenovirus carrying BMP-2 gene，Ad—BMP2），通过纤维蛋白凝胶与牛松质骨支架（bovine cancellous bone，BCB）复合，修复骨缺损的效果。方法将60只新西兰大耳白兔随机分为4组，每组15只。制成双侧桡骨中段1．5cm骨缺损模型，采用4种材料植入修复。A组：Ad-BMP-2＋BCB；B组：重组BMP-2＋BCB；C组：携带D-半乳糖酐酶基因的腺病毒对照载体（adenovirus carrying β—galgene，Ad—Lacz）＋BCB；D组：单纯BCB支架。修复术后各组于4、8和12周各处死动物5只取材，行X线片、组织学、生物力学和免疫组织化学染色检查。结果A、B两组骨缺损均得到了修复，但术后各时间点，A组在成骨活跃程度、新生骨量、力学强度及BMP-2表达等方面均明显优于B组；C、D两组均无新骨形成。结论BMP-2直接基冈治疗，操作简便、骨诱导能力强，是修复节段性骨缺损的有效方法。     

骨折端微动对骨形态发生蛋白-2表达的影响

目的 研究骨折端微动时轴向应力对骨形态发生蛋白-2（BMP-2）表达的影响。方法 72只新西兰大白兔分为微动组与固定组，利用免疫组化及原位杂交方法检测两组动物骨折端BMP-2蛋白及mRNA表达的不同。结果 BMP-2蛋白及mRNA表达于间质细胞、成骨细胞、成软骨细胞、骨细胞及骨基质。在骨折愈合过程早期高度表达，峰值位于骨折后14d，其后表达水平降低。BMP-2蛋白及mRNA染色阳性指数术后14、21、28d微动组大于固定组，差异有统计学意义（P〈0．05）。结论 骨折端微动时BMP- 2蛋白及mRNA表达在特定时间增强。     

鹿瓜多肽对去卵巢大鼠骨密度、股骨生物力学及松质骨中BMP2表达的影响

目的鹿瓜多肽（松梅乐）注射液肌注对去卵巢大鼠骨密度、股骨生物力学及松质骨中BMP2表达的影响。方法将8月龄未经产雌性二级SD大鼠30只，随机分为假手术组（SHAM）组、去势组（OVX）、去势＋鹿瓜多肽（OVX＋CCP）组。OVX＋CCP组大鼠术后第1天开始给药，术后20周处死各组大鼠．应用双能X线吸收仪（DEXA）测量各组大鼠股骨粗隆、股骨干中点及第3～5腰椎中点的骨密度（BMD）；应用INSTRON1195电子拉伸试验机检测股骨生物力学性能；利用免疫组织化学染色及图像分析方法对各组大鼠松质骨切片图像进行灰度分析，观察鹿瓜多肽注射液对OVX大鼠松质骨中BMP2表达的影响。结果（1）OVX与SHAM组比较，灰度值降低明显（P〈0．01），而OVX＋CCP组灰度值较去势组降低（P〈0．01），表明OVX大鼠松质骨中骨小梁周围及髓腔中BMP2表达阳性细胞数明显多于SHAM组；而OVX＋CCP组骨小梁周围及髓腔内BMP2表达阳性细胞数多于去势组，且染色加深。（2）与SHAM组相比，OVX组股骨近端、股骨干、腰椎BMD明显降低（P〈0．01）；OVX＋CCP各部位BMD高于去势组（P〈0．01），但未达到SHAM组水平（P〈0．01）；（3）与SHAM比较，去势组大鼠股骨最大载荷、桡度、最大应力、弹性模量均明显下降（P〈0．01），治疗组上述指标高于去势组（P〈0．01），接近对照组水平（P〉0．05）。结论鹿瓜多肽注射液可以抑制和延缓骨质疏松的形成，对雌激素缺乏引起的骨质疏松具有预防作用。     

雌激素对单侧输尿管梗阻大鼠肾间质纤维化的保护作用

目的探讨雌激素对单侧输尿管梗阻（UUO）大鼠肾间质纤维化的作用。方法雌性SD大鼠30只，随机分为4组：Ⅰ组对照组；Ⅱ组生理雌激素组；Ⅲ组低雌激素组；Ⅳ组高雌激素组。UUO术后21d处死各组大鼠，光镜观察梗阻肾组织病理变化，并分别用免疫组化和RT-PCR方法检测各组肾组织α-平滑肌肌动蛋白（α-SMA）和金属蛋白酶1组织抑制剂（TIMP-1）的表达。结果低雌激素组间质纤维化病变最明显，高雌激素组病变显著减轻（P〈0．01）。与生理雌激素组相比，低雌激素组α—SMA和TIMP-1蛋白和基因的表达增加（P〈0．05）；高雌激素组上述物质表达则减少（P〈0．05）。结论雌激素可能通过抑制α-SMA和TIMP-1的表达进而减少细胞外基质的沉积而发挥肾保护作用。     

脊柱后外侧融合过程中BMP-2、BMP-4基因的动态表达

目的：研究脊柱后外侧融合过程中骨形态发生蛋白(BMP)的表达情况，探讨BMP对融合过程的作用。方法：36只成年雄性新西兰白兔，制作L4、L5双侧横突间自体骨植骨模型，按术后处死时间(0d、2d、4d、1周、2周、3周、4周、5周、6周、10周、6个月、10个月)随机平均分为12组。将融合组织平均分为3等份，与横突交界的上、下区域定为边缘区，中间区域定为中央区。以RT-PCR法检测不同时间段、不同融合区域BMP-2 mRNA、BMP-4 mRNA的表达水平。结果：在脊柱融合术后1～6周，BMP-2和BMP-4表达量明显增高，并各自出现相应的峰值。中央区BMP表达水平的增幅及峰值明显低于边缘区，其开始增高的时间及峰值出现的时间滞后于边缘区1～3周，表现出明显的时间和空间差别：结论：脊柱融合中央区BMP低表达及时间滞后可能是不融合发生的重要原因。在融合早期补充外源性BMP可能有助于提高融合率。     

麝香乌龙丸对卵巢切除大鼠股骨骨折愈合的实验研究

目的研究麝香乌龙丸对卵巢切除大鼠股骨干骨折愈合的作用。方法将30只雌性、12周龄Sprague-Dawley大鼠分成3组假手术组(Sham F V)、卵巢切除 骨折 生理盐水对照组(OVX F V)、卵巢切除 骨折 麝香乌龙丸给药组(OVX F M),每组10只大鼠。所有需制造骨折的大鼠均采用右股骨中段横行骨折,髓内针固定;麝香乌龙丸给药组采用大鼠灌胃给药(1.2g.kg-1.d-1),于术后4周杀死,取大鼠右侧股骨标本;分别进行CR摄片、组织形态学染色,并应用双能X线骨密度仪(DEXA)测量右股骨整体骨密度(tBMD)、远段骨密度(dBMD)和中段骨密度(mBMD)以及BMP-2免疫组化观察,并应用病理图像分析仪对BMP-2免疫组化进行光密度测定。结果麝香乌龙丸给药组与OVX F V比较,前者骨痂mBMD和BMP-2的表达显著增高,骨小梁增宽、排列较整齐,板层骨形成,软骨组织可见。结论麝香乌龙丸对OVX大鼠股骨骨折有明显促进骨折愈合的作用,并加快编织骨向板层骨的演变过程,这与BMP-2的表达有明显相关性。     

前交叉韧带切断对卵巢切除大鼠股骨骨折愈合的影响

目的观察前交叉韧带切断(OA动物模型)对双侧卵巢切除大鼠(OVX绝经后骨质疏松模型)股骨骨折愈合的影响。方法12周龄SD大鼠共70只，分成基础对照组(Basal)、假手术组(Sham)、双侧卵巢切除术组(OVX)、卵巢切除 前交叉韧带切断术组(OVX OA)、假手术 骨折组(Sham F)、卵巢切除术 骨折组(OVX F)、卵巢切除 前交叉韧带切断 骨折组(OA OVX F)，每组10只大鼠。所有受试大鼠在处死前第10d天和第4d天分别皮下注射盐酸四环素和钙黄绿素行双荧光标记。基础对照组在实验开始时杀死，其余6组在术后6W杀死，取大鼠右侧股骨标本。然后，分别进行CR摄片、组织形态学染色，以及应用Norland-XR36双能X线骨密度仪(DXA)测量右股骨远段骨密度和中段骨密度，并将股骨远段及骨折段骨痂进行硬组织包埋、切片，作骨组织形态计量学测量。结果①OVX OA组与OVX组比较，股骨远段。BMD和BWTV显著增加；②OVX F组与Sham F组比较，骨痂(股骨中段)BMD和BV／TV显著降低：③OVX OA F组与OVX F组比较，骨痂(股骨中段)BMD利BV／TV无统计学差异。结论①骨质疏松不仅延缓骨折愈合过程，而且降低骨折愈合质量；②在此动物模型中，骨性关节炎延缓股骨骨质疏松的发生，但是，对骨质疏松性骨折的愈合没有确切的促进作用。     

雷洛昔芬对OVX大鼠骨质疏松性骨折愈合的影响

目的 探讨骨质疏松性骨折愈合的特点以及雷诺昔芬对骨质疏松性骨折愈合的影响.方法 健康12 周龄雌性SD大鼠54只,随机分成假手术组(SHAM),卵巢切除+生理盐水组(OVX),卵巢切除+雷诺昔芬组(OVX+RAL),每组18只大鼠.卵巢切除术后8周,骨密度检测确认大鼠骨质疏松模型成功.选择开放性右侧股骨中段骨折模型,以克氏针行髓内固定股骨;骨折后8周,CR片记录大鼠骨痂的连续性以及骨折愈合情况,三点弯曲试验检测股骨的最大弯曲应力和最大弯曲载荷.取骨折术后4周和8周时骨痂,行HE染色,并记录骨小梁或小梁状骨所占体积比(BV/TV)表示.免疫组化染色检测骨痂中血管内皮生长因子(VEGF)表达.结果 CR摄片显示骨折后8周各组大鼠骨折连续性都较好,骨折对位对线好.大鼠骨折8周时最大弯曲应力和最大弯曲载荷,SHAM组和OVX+RAL组都明显优于OVX组,差异显著,有统计学意义.骨折4周时骨痂的HE染色中,OVX组原始小梁状骨中新生软骨细胞较多.OVX+RAL组和SHAM组8周时骨痂中BV/TV值明显高于OVX组.骨折4周时骨痂VEGF染色中,各组大鼠的骨痂内软骨细胞或骨细胞上存在一定量的VEGF表达,SHAM组和OVX+RAL组的细胞上VEGF表达量略高于OVX组;骨折8周时各组骨痂中的VEGF几乎无明显表达.结论 OVX大鼠骨痂成熟缓慢和早期骨痂内VEGF低表达,可能是骨折愈合能力下降的重要因素.雷诺昔芬可以促进骨痂成熟和早期骨痂中VEGF的表达,有利于OVX大鼠骨折愈合.     

Early period of fracture healing in ovariectomized rats

Objective. To evaluate the effect of osteoporosis on fracture healing through observing the hlstomorphological changes, bone mineral density of callus and expression and distribution of transforming growth factor beta 1 (TGF-β1 ), basic fibroblast growth factor (bFGF)and bone morphogenetic protein.2 (BMP-2) in ovariectomized rats. Methods. Sixty female Sprague-Dawley rats ( aged 12 weeks and weighing 235 g on average ) were randomly divided into an ovariectomized (OVX) group (n =30) anda sham-operated (SO) group ( n = 30). Ovariectomy was performed in the OVX rats and same incision was made in the SO rats. Three months later, fracture of femoral shaft was made on all the rats. Then they were killed at different time points. Callus formation was observed with histological and imethods. Results: A reduction in callus and bone mineral density in the healing femur and a decrease of osteoblasts expressing TGF-β1 near the bone trabecula were observed in the OVX rats 3-4 weeks after fracture.Histomorphological analysis revealed a higher content of soft callus in the OVX rats than that in the SO rats.Immunohistochemistry results showed that no remarkable difference in expression and distribution of BMP-2 and bFGF between the OVX and SO groups was found. Conclusions: Osteoporosis influences the quantity and quality of callus during the early period of fracture healing. The effect of osteoporosis on fracture healing has no relationship with the expression of BMP-2 or bFGF. The decreased expression of TGF-I31 in osteoblasts may cause a decrease in quality of facture healing after osteoporosis.     

雌激素对实验性骨质疏松症骨折愈合的影响

目的 研究不同剂量的雌激素对卵巢除后发生骨质疏检习股骨骨折愈合的影响。方法 成年ｂａｌｂ／ｃ小鼠卵巢除后３个月开始制作右股骨中段闭合骨折模型,同一天开始皮下注射高低两种剂量雌激素,小鼠于折骨后５、１０、１５、２０、３０天分批处死,采用Ｘ射线、骨痂称重、骨痂中钙盐沉积率测定、组织学检查、血清生化检测等方法研究雌激素对骨折愈合的影响,并和卵巢除组、假手术对照组进行比较。结果 卵巢除组骨痂较小,     

补正续骨丸对去卵巢大鼠骨量及其相关骨折的作用机制研究

目的探索中药复方补正续骨丸对去卵巢大鼠骨量及相关骨折的影响。方法雌性大鼠卵巢切除法(OVX)造PMOP模型,分组给药6周后DXA行大鼠股骨BMD检测; HE染色测量股骨骨小梁密度; ELISA检测法检测大鼠血清中P1NP、β-CTX; Western blot法检测大鼠股骨中4EBP-1、p70S6K蛋白表达;同时比较不同处理组大鼠骨折愈合时间及相同时间骨痂密度,观察补正续骨丸对OVX大鼠骨折愈合时间及骨痂密度的影响。结果补正续骨丸可增加OVX大鼠股骨BMD、减少骨小梁断裂、降低去卵巢大鼠血清中β-CTX水平、抑制mTOR1信号通路中p70S6K、4EBP1的蛋白表达,补正续骨丸可促进OVX大鼠骨折处骨痂密度增加及减少骨折愈合时间。结论中药复方补正续骨丸通过抑制mTOR1信号通路中p70S6K、4EBP1蛋白的表达抑制骨吸收、促进OVX大鼠骨折的愈合。     

BMP-7 stimulates early diaphyseal fracture healing in estrogen deficient rats

Estrogen deficiency causes postmenopausal osteoporosis. The relationship between estrogen deficiency and the high failure rate after osteoporotic fracture treatment is unclear, as is the effect of possible interventions, either with anti-resorptive agents or with anabolic agents such as bone morphogenetic proteins (BMPs). To investigate the influence of estrogen deficiency as well as the effect of early intervention, forty female wistar rats underwent ovarectomy (OVX) followed by low calcium diet. Ten rats underwent sham operations, followed by normal diet. After 6 weeks, a closed midshaft femoral fracture was induced. Ten animals received a systemic bisphosphonate injection, 10 injection of BMP-7 in the fracture, and 10 a combination. All then received a normal diet. After 2 weeks healing was evaluated using radiographs, CT, biomechanical testing, and histology. Radiography showed significant increase of bridging in groups treated with BMP-7. Callus volume was higher in these groups. Bending stiffness and strength were similar between OVX and sham, and not influenced by bisphosphonates. Significant increase was seen in groups treated with BMP-7. Histology was in accordance with other endpoints. Early fracture healing was not affected by estrogen deficiency. While no beneficiary effect of bisphosphonate treatment was found, injection of BMP-7 stimulated healing in ovarectomized rats.     

Mechanical strength of fracture callus in osteopenic bone at different phases of healing

OBJECTIVES: To quantify and compare peak bending force and stiffness of fractured femurs during healing of ovariectomized (OVX) and sham-operated (SHAM) rats. DESIGN: Temporal biomechanical animal study. SETTING: Rat femurs were fractured and surgically fixed by a qualified surgeon. The inherent instability of the fixation system employed produced delayed union of the fracture. All biomechanical assessments were performed with servohydraulic test machines (Instron Inc., Canton, MA, U.S.A.; and MTS Corp., Eden Prairie, MN, U.S.A.). INTERVENTION: OVX was performed sixteen weeks before femur fracture, and the effect of OVX on healing fractures was determined. MAIN OUTCOMES: Peak bending force and stiffness of the healing femurs at four, six, and eight weeks after fracture. RESULTS: Peak bending loads of the healing fractured femurs in the OVX and SHAM animals were not significantly different. Peak bending loads for the OVX animals at four and six weeks were significantly lower than the peak load at eight weeks (p < 0.05), whereas no difference was found in the peak load with respect to time for the SHAM animals. Both SHAM and OVX animals had greater bending stiffness of the healing fractured femur after eight weeks of healing than at four weeks (p < 0.05). CONCLUSIONS: OVX is known to reduce cancellous bone mass and strength, but the effect of OVX on healing of fractures in cortical bone is controversial. This study, using a delayed-union model, found no significant differences between OVX and SHAM animals in the breaking strength of healing fractures.     

Postsurgical administration of estradiol benzoate decreases tensile strength of healing skin wounds in ovariectomized rats

BACKGROUND: The most effective method of increasing the level of estrogen in the wounds of post-menopausal women undergoing routine surgical procedures is by long-term preoperative administration. However, in the case of acute surgery or trauma, the most effective method of increasing the level of estrogen is administration immediately pre- or postsurgery. This study, therefore, was aimed at assessing the effect of postsurgical administration of estradiol benzoate on wound healing in ovariectomized (OVX) Sprague Dawley rats. MATERIALS AND METHODS: Three months prior to the wound healing experiment, 16 rats were anesthetized and underwent ovariectomy, while the other eight rats were sham operated. Two parallel full thickness skin incisions and two round full thickness skin excisions were performed on the dorsum of each rat. Dose of 10 microg/d of estradiol benzoate was administered to eight OVX rats for 6 d postoperatively, whereas the other animals received a placebo. After 6 d, all animals were sacrificed and samples removed for biomechanical and histological evaluation. RESULTS: The mean wound tensile strength of OVX estrogen treated rats (9.54 +/- 3.24 g/mm(2)) was significantly lower compared with vehicle-treated OVX animals (14.57 +/- 4.12 g/mm(2)) as well as with control rats subjected to sham-OVX surgery (11.71 +/- 3.33 g/mm(2)). Nevertheless, the histological evaluation in OVX estrogen treated rats showed a significantly increased process of neo-angiogenesis associated with slightly decreased collagen deposition. CONCLUSION: Our results indicate that the question of the clinical significance of this type of hormone replacement therapy remains open and requires further research.     

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。