PTH_(1-34)对去势雌性大鼠种植体周围骨结合影响的实验研究
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摘要
目的：
通过去除Wistar雌性大鼠的双侧卵巢来建立骨质疏松动物模型,应用骨计量学方法观察甲状旁腺激素(PTHH1-34)对去势后雌性大鼠种植体骨结合的影响,从而为临床上应用甲状旁腺激素(PTH1-34)治疗更年期妇女骨质疏松患者进行口腔种植时,如何增加种植体的骨结合、提高种植成功率提供实验基础依据。
方法：
选用纯种3-4月龄健康雌性Wistar大鼠48只,随机分为A(18只)、B(15只)、C(15只)三组。其中A组去除卵巢附近与卵巢等量的脂肪组织(假手术组),B、C组摘除双侧卵巢(去势组)。去势术后8周：称量各组体重；抽取各组静脉血检测各组血清碱性磷酸酶(ALP)、雌激素(E2)、甲状旁腺激素(PTH)浓度的值；随机处死假手术组和去势组各6只(A组6只,B、C组各3只),取其胫骨和上颌骨,用双能X线测量仪测量胫骨和上颌骨的骨密度,然后制作常规骨组织石蜡切片,HE染色,进行骨组织形态学的大体观察,确定骨质疏松动物模型建立成功。骨质疏松模型建立成功后,在3组剩余大鼠右侧胫骨近干骺端分别植入纯钛螺纹柱状种植体。从种植手术后第1天开始用药：A组和B组大鼠按照lml/kg体重腹腔注射0.9%生理盐水。C组大鼠按照20μg/kg体重腹腔注射甲状旁腺激素(冻干粉由生理盐水配成20μ g/ml混悬液,用前摇匀)。剂量按体重每周校对一次,隔日一次,用药时间为8周。种植术后用药4周、8周后：取血检测各组血清碱性磷酸酶(ALP)、雌激素(E2)、甲状旁腺激素(PTH)浓度的值；处死各组半数大鼠,利用双能X线骨密度测量仪测量上颌骨、左侧胫骨骨密度；取右侧带种植体的胫骨制作不脱钙硬组织磨片,甲苯胺蓝染色,行骨组织形态学观察和骨计量学研究。
结果：
1、大鼠去势手术进展顺利。去势术后8周,去势组(B、C组)较假手术组(A组)体重显著增加,雌激素水平明显下降,两者的差异具有统计学意义(P<0.05)；运用双能X线骨密度测量仪检测,去势组(B、C组)大鼠胫骨和牙槽骨的骨密度和假手术组(A组)大鼠相比,均显著降低,差异有统计学意义(P<0.05)；同时组织学观察表明,去势组与假手术组比较,其颌骨和胫骨干骺端骨小梁数目减少,变细、稀疏而不连续,进一步证实了骨质疏松状态。以上说明该实验动物的骨质疏松模型成功建立。
2、实验大鼠用药4周、8周后,各组大鼠体重较去势8周时均增加：C、B组相比,差异无统计学意义(P>0.05),但C组体重增加值明显低于B组,差异有统计学意义(P<0.05)；C组和A组相比,差异有统计学意义(P<0.05),但C组与A组体重增加值相差不明显,差异无统计学意义(P>0.05)。
3、种植术后用药4周、8周后：C组和B组比较,血清ALP活性增高(P<0.05),ALP在种植4周时即显示出差异,而在8周显示出更大的差异；血清E2水平,虽然C组比B组下降稍缓慢,但二者在4周、8周时相比较,差异均无统计学意义(P>0.05)；血清PTH,虽然C组略有下降,B组略有升高,但均变化缓慢,二者在4周、8周时相比较,差异均无统计学意义(P>0.05)。C组和A组比较,血清ALP活性明显升高,E2水平仍然较低,二组在4周、8周时相比较,差异均有统计学意义(P<0.05)；血清PTH,C组略有下降,A组升高,两组在4周、8周时相比较,差异均无统计学意义(P>0.05)。
4、实验大鼠用药4周、8周后,利用双能X线骨密度测量仪测量各组大鼠分离的左侧胫骨和上颌牙槽骨的骨密度,发现C组大鼠胫骨和颌骨骨密度在4周时较B组虽有所增加,但增加不明显(P>0.05),但8周时两组差别具有统计学意义(P<0.05)；C组与A组比较,4周时C组大鼠胫骨和颌骨骨密度虽有增加,但仍明显低于A组,两组差别具有统计学意义(P<0.05),8周时C组胫骨及颌骨密度虽略低于A组,但差异已无统计学意义(P>0.05)。
5、种植术后用药4周、8周后,三组大鼠在骨组织形态计量学上比较：(1)种植4周时,C组的骨结合率、骨小梁平均宽度和结合骨板宽度均明显大于B组(P<0.05),而C组的骨皮质厚度、松质骨区骨量和B组相比,无显著性差异(P>0.05)；C组的松质骨区骨量、骨小梁平均宽度、结合骨板宽度均明显小于A组(P<0.05),而C组的骨皮质厚度、骨结合率与A组相比,无显著性差异(P>0.05)。(2)种植8周时,C组的骨结合率、松质骨区骨量、骨小梁平均宽度、结合骨板宽度均明显大于B组(P<0.01),而C组的骨皮质厚度和B组相比,无显著性差异(P>0.05)；A、C组除骨皮质厚度无明显改变外,各项比4周时均有升高,C组的骨结合率、骨皮质厚度、松质骨区骨量、骨小梁平均宽度和结合骨板的宽度均与A组相比,无显著性差异(P>0.05)。以上说明PTH可以提高种植体骨结合的质量。
结论：
1、去除大鼠双侧卵巢组织建立的骨质疏松模型,与绝经后女性所形成的骨质疏松类型相似,都属于高转换型骨质疏松,二者在病因和病理上表现有一定的相似性。故该模型用于研究PTH对骨质疏松状态下种植体骨结合的影响具有一定的科学性。
2、间歇性、小剂量注射PTH1-34能增加骨质疏松大鼠骨密度并改善骨结构,使种植体骨结合率增加,结合骨板宽度增加,松质骨区单位骨量明显增加,骨小梁平均宽度增加,促进骨改建,从而提高种植体骨结合的质量。
Aims:
In order to investigate the roles of parathyroid hormone (PTH1-34) on alveolar bone formation of female rats by the method of bone measurement, the models of osteoporosis animals were built by removing bilateral ovarian tissue of female Wistar rats. This research could provide basic experimental data to explain how to increase the implant osseointegration and how to improve planting success rate in dental implants, thus further to provide a theoretical basis for the clinical application of PTH1-34to treat postmenopausal women who suffer from osteoporosis.
Methods:
48Female Wistar rats which were healthy and3～4months old were choosed and randomly divided into three groups. Group A included18rats, group B included15rats and group C included15rats. Bilateral ovarian tissue were removed completely from the rats in group B and group C (group OVX), and equivalent adipose tissue around the ovary were removed from the rats in group A (group SHAM). Eight weeks later, weights as well as the contents of alkaline phosphatase (ALP), estrogen (E2), and parathyroid hormone (PTH) in serum of all rats were measured. Then,6rats from group A,3rats from group B and3rats from group C were randomly choosed to be killed. The killed rats were taken bilateral maxilla and tibia near metaphysic, followed by the measurement of the mineral density of bone tissues using dual energy X-ray absorptiometry. And then slices of bone tissues were maken to identify whether the osteoporotic models are successfully built through histopathologic study. After the models were successfully built, we implanted titanium threaded cylindrical implant in the right tibia near metaphysis in the remaining rats. After implanting,0.9%physiological saline was injected to the enterocoelia of the rats in group A and group B with the dose of1ml/kg, and parathyroid hormone was injected to enterocoelia of the rats in group C with the dose of20μ g/kg (freeze-dried powder was maked into suspension with20μ g/ml by physiological saline and the suspension was shaken up before using) every other day. The dose was proofreaded according to the weight of rats once a week. At4weeks and8weeks after implanta-tion surgery, the contents of alkaline phosphatase (ALP), estrogen (E2) and parathyroid hormone (PTH) in serum of rats in each group were measured. At the same time, half the rats in every group were killed. The mineral density of the maxilla and the left tibia near metaphysic were measured by using dual energy X-ray absorptiometry. Then, undecalcified sections were prepared with toluidine blue dyeing to examine histologically and histomorphometrically
Results:
1、Castration operation of the rats was sucessful.8weeks after castration, the weight of the rats in group OVX(group B and group C) significantly increased.compared to the rats in group SHAM(group A). it was statistical significance (P<0.05) for the discrepancy between group OVX and SHAM; Compared with the rats in group SHAM, the mineral density of tibia and maxillary of rats in group OVX were significantly lower through the dual energy X-ray absorptiometry, and it was also statistical significance (P<0.05) for the discrepancy between group OVX and SHAM. The histological observation showed that compared with SHAM group, the number of bone trabecula reduced and they got thin, sparse and not continuous in the the maxilla and the tibia near metaphysis of OVX group, which further confirmed the osteoporosis. What mentioned above proved that the experimental animal model of osteoporosis was built successfully.2、4weeks、8weeks after implantation, the weight of rats in each group increased:it was not statistical significance (P>0.05) for the discrepancy of weight between group C and B,but it wa it was statistical significance (P<0.05) for the discrepancy of weight added between group C and B; it was statistical significance (P<0.05) for the discrepancy of weight between group C and A,but it was not statistical significance (P>0.05) for the discrepancy of weight added between group C and A.
3、4week、8weeks after implantation, compared with group B, the activity of ALP in serum increased obviously in group C (P<0.05), In the serum, it was not statistical significance (P<0.05) for the discrepancy of E2, PTH between the two groups (P>0.05). Comparing with the rats in group A, activity of ALP increased and E2level was still lower in group C. It was statistical significance (P<0.05) for the discrepancy between the two groups. In the serum, PTH decreased slightly in group C, while it increased in group A. However, it was not statistical significance (P>0.05) for the discrepancy between the two groups.
4、4weeks、8weeks after implantation, the bone density of the freshly isolated left tibia and the maxillaby was measured by using dual energy X-ray absorptiometry. It was found that the density of tibia and mandible bone in group C was higher than that in group B：it was not statistical significance (P>0.05) for the discrepancy between the two groups after4weeks;But it was statistical significance(P<0.05) after8weeks.Compared with A group, the bone densityis in group C was lower than that in group A in, However, it was not statistical significance (P>0.05) for the discrepancy between the two groups after8weeks.
5、4weeks、8weeks after implantation, they were examed histologically and histomorphometrically.4weeks later, it was found that implant bone contact rate，，trabercular width and combinded bone lamella width of rats were much higher in groop C than that in group B (P<0.05);However, in terms of thickness of cortical, trabercular area, there was no difference between group C and group B.It was found that trabercular area,trabercular width and combinded bone lamella width of rats were much lower in groop C than that in group A (P<0.05);However, in terms of implant bone contact rate,thickness of cortical bone, there was no difference between group C and group A.8weeks later,it was found that implant bone contact rate, trabercular area,trabercular width and combinded bone lamella width of rats were much higher in groop C than that in group B (P<0.05);However, in terms of thickness of cortical bone, there was no difference between group C and group B. There was no difference between group C and group A in all of the terms, although every term grew higher except the term of thickness of cortical bone. What mentioned above proved that PTH could improve the quality of osseointegration of the rats.
Conclusion:
1、The osteoporosis model of the rats which is built by removing bilateral ovarian tissues is similar to the osteoporosis of post-menopausal women. Both of them belong to the high turnover osteoporosis, and have certain etiological and pathological similarity. The model is built scientifically and it can be used to investigate the influence of PTH1-34on osseointegration in the condition of osteoporosis.
2、Intermittent application of parathyroid hormone (PTH1-34) with small doses can increase the density of alveolar bone mineral, improve their bone structure, increase implant bone contact rate, trabercular area, trabercular width and combinded bone lamella width, further to improve the osseointegration quality of the rats.
通过去除Wistar雌性大鼠的双侧卵巢来建立骨质疏松动物模型,应用骨计量学方法观察甲状旁腺激素(PTHH1-34)对去势后雌性大鼠种植体骨结合的影响,从而为临床上应用甲状旁腺激素(PTH1-34)治疗更年期妇女骨质疏松患者进行口腔种植时,如何增加种植体的骨结合、提高种植成功率提供实验基础依据。
方法：
选用纯种3-4月龄健康雌性Wistar大鼠48只,随机分为A(18只)、B(15只)、C(15只)三组。其中A组去除卵巢附近与卵巢等量的脂肪组织(假手术组),B、C组摘除双侧卵巢(去势组)。去势术后8周：称量各组体重；抽取各组静脉血检测各组血清碱性磷酸酶(ALP)、雌激素(E2)、甲状旁腺激素(PTH)浓度的值；随机处死假手术组和去势组各6只(A组6只,B、C组各3只),取其胫骨和上颌骨,用双能X线测量仪测量胫骨和上颌骨的骨密度,然后制作常规骨组织石蜡切片,HE染色,进行骨组织形态学的大体观察,确定骨质疏松动物模型建立成功。骨质疏松模型建立成功后,在3组剩余大鼠右侧胫骨近干骺端分别植入纯钛螺纹柱状种植体。从种植手术后第1天开始用药：A组和B组大鼠按照lml/kg体重腹腔注射0.9%生理盐水。C组大鼠按照20μg/kg体重腹腔注射甲状旁腺激素(冻干粉由生理盐水配成20μ g/ml混悬液,用前摇匀)。剂量按体重每周校对一次,隔日一次,用药时间为8周。种植术后用药4周、8周后：取血检测各组血清碱性磷酸酶(ALP)、雌激素(E2)、甲状旁腺激素(PTH)浓度的值；处死各组半数大鼠,利用双能X线骨密度测量仪测量上颌骨、左侧胫骨骨密度；取右侧带种植体的胫骨制作不脱钙硬组织磨片,甲苯胺蓝染色,行骨组织形态学观察和骨计量学研究。
结果：
1、大鼠去势手术进展顺利。去势术后8周,去势组(B、C组)较假手术组(A组)体重显著增加,雌激素水平明显下降,两者的差异具有统计学意义(P<0.05)；运用双能X线骨密度测量仪检测,去势组(B、C组)大鼠胫骨和牙槽骨的骨密度和假手术组(A组)大鼠相比,均显著降低,差异有统计学意义(P<0.05)；同时组织学观察表明,去势组与假手术组比较,其颌骨和胫骨干骺端骨小梁数目减少,变细、稀疏而不连续,进一步证实了骨质疏松状态。以上说明该实验动物的骨质疏松模型成功建立。
2、实验大鼠用药4周、8周后,各组大鼠体重较去势8周时均增加：C、B组相比,差异无统计学意义(P>0.05),但C组体重增加值明显低于B组,差异有统计学意义(P<0.05)；C组和A组相比,差异有统计学意义(P<0.05),但C组与A组体重增加值相差不明显,差异无统计学意义(P>0.05)。
3、种植术后用药4周、8周后：C组和B组比较,血清ALP活性增高(P<0.05),ALP在种植4周时即显示出差异,而在8周显示出更大的差异；血清E2水平,虽然C组比B组下降稍缓慢,但二者在4周、8周时相比较,差异均无统计学意义(P>0.05)；血清PTH,虽然C组略有下降,B组略有升高,但均变化缓慢,二者在4周、8周时相比较,差异均无统计学意义(P>0.05)。C组和A组比较,血清ALP活性明显升高,E2水平仍然较低,二组在4周、8周时相比较,差异均有统计学意义(P<0.05)；血清PTH,C组略有下降,A组升高,两组在4周、8周时相比较,差异均无统计学意义(P>0.05)。
4、实验大鼠用药4周、8周后,利用双能X线骨密度测量仪测量各组大鼠分离的左侧胫骨和上颌牙槽骨的骨密度,发现C组大鼠胫骨和颌骨骨密度在4周时较B组虽有所增加,但增加不明显(P>0.05),但8周时两组差别具有统计学意义(P<0.05)；C组与A组比较,4周时C组大鼠胫骨和颌骨骨密度虽有增加,但仍明显低于A组,两组差别具有统计学意义(P<0.05),8周时C组胫骨及颌骨密度虽略低于A组,但差异已无统计学意义(P>0.05)。
5、种植术后用药4周、8周后,三组大鼠在骨组织形态计量学上比较：(1)种植4周时,C组的骨结合率、骨小梁平均宽度和结合骨板宽度均明显大于B组(P<0.05),而C组的骨皮质厚度、松质骨区骨量和B组相比,无显著性差异(P>0.05)；C组的松质骨区骨量、骨小梁平均宽度、结合骨板宽度均明显小于A组(P<0.05),而C组的骨皮质厚度、骨结合率与A组相比,无显著性差异(P>0.05)。(2)种植8周时,C组的骨结合率、松质骨区骨量、骨小梁平均宽度、结合骨板宽度均明显大于B组(P<0.01),而C组的骨皮质厚度和B组相比,无显著性差异(P>0.05)；A、C组除骨皮质厚度无明显改变外,各项比4周时均有升高,C组的骨结合率、骨皮质厚度、松质骨区骨量、骨小梁平均宽度和结合骨板的宽度均与A组相比,无显著性差异(P>0.05)。以上说明PTH可以提高种植体骨结合的质量。
结论：
1、去除大鼠双侧卵巢组织建立的骨质疏松模型,与绝经后女性所形成的骨质疏松类型相似,都属于高转换型骨质疏松,二者在病因和病理上表现有一定的相似性。故该模型用于研究PTH对骨质疏松状态下种植体骨结合的影响具有一定的科学性。
2、间歇性、小剂量注射PTH1-34能增加骨质疏松大鼠骨密度并改善骨结构,使种植体骨结合率增加,结合骨板宽度增加,松质骨区单位骨量明显增加,骨小梁平均宽度增加,促进骨改建,从而提高种植体骨结合的质量。
Aims:
In order to investigate the roles of parathyroid hormone (PTH1-34) on alveolar bone formation of female rats by the method of bone measurement, the models of osteoporosis animals were built by removing bilateral ovarian tissue of female Wistar rats. This research could provide basic experimental data to explain how to increase the implant osseointegration and how to improve planting success rate in dental implants, thus further to provide a theoretical basis for the clinical application of PTH1-34to treat postmenopausal women who suffer from osteoporosis.
Methods:
48Female Wistar rats which were healthy and3～4months old were choosed and randomly divided into three groups. Group A included18rats, group B included15rats and group C included15rats. Bilateral ovarian tissue were removed completely from the rats in group B and group C (group OVX), and equivalent adipose tissue around the ovary were removed from the rats in group A (group SHAM). Eight weeks later, weights as well as the contents of alkaline phosphatase (ALP), estrogen (E2), and parathyroid hormone (PTH) in serum of all rats were measured. Then,6rats from group A,3rats from group B and3rats from group C were randomly choosed to be killed. The killed rats were taken bilateral maxilla and tibia near metaphysic, followed by the measurement of the mineral density of bone tissues using dual energy X-ray absorptiometry. And then slices of bone tissues were maken to identify whether the osteoporotic models are successfully built through histopathologic study. After the models were successfully built, we implanted titanium threaded cylindrical implant in the right tibia near metaphysis in the remaining rats. After implanting,0.9%physiological saline was injected to the enterocoelia of the rats in group A and group B with the dose of1ml/kg, and parathyroid hormone was injected to enterocoelia of the rats in group C with the dose of20μ g/kg (freeze-dried powder was maked into suspension with20μ g/ml by physiological saline and the suspension was shaken up before using) every other day. The dose was proofreaded according to the weight of rats once a week. At4weeks and8weeks after implanta-tion surgery, the contents of alkaline phosphatase (ALP), estrogen (E2) and parathyroid hormone (PTH) in serum of rats in each group were measured. At the same time, half the rats in every group were killed. The mineral density of the maxilla and the left tibia near metaphysic were measured by using dual energy X-ray absorptiometry. Then, undecalcified sections were prepared with toluidine blue dyeing to examine histologically and histomorphometrically
Results:
1、Castration operation of the rats was sucessful.8weeks after castration, the weight of the rats in group OVX(group B and group C) significantly increased.compared to the rats in group SHAM(group A). it was statistical significance (P<0.05) for the discrepancy between group OVX and SHAM; Compared with the rats in group SHAM, the mineral density of tibia and maxillary of rats in group OVX were significantly lower through the dual energy X-ray absorptiometry, and it was also statistical significance (P<0.05) for the discrepancy between group OVX and SHAM. The histological observation showed that compared with SHAM group, the number of bone trabecula reduced and they got thin, sparse and not continuous in the the maxilla and the tibia near metaphysis of OVX group, which further confirmed the osteoporosis. What mentioned above proved that the experimental animal model of osteoporosis was built successfully.2、4weeks、8weeks after implantation, the weight of rats in each group increased:it was not statistical significance (P>0.05) for the discrepancy of weight between group C and B,but it wa it was statistical significance (P<0.05) for the discrepancy of weight added between group C and B; it was statistical significance (P<0.05) for the discrepancy of weight between group C and A,but it was not statistical significance (P>0.05) for the discrepancy of weight added between group C and A.
3、4week、8weeks after implantation, compared with group B, the activity of ALP in serum increased obviously in group C (P<0.05), In the serum, it was not statistical significance (P<0.05) for the discrepancy of E2, PTH between the two groups (P>0.05). Comparing with the rats in group A, activity of ALP increased and E2level was still lower in group C. It was statistical significance (P<0.05) for the discrepancy between the two groups. In the serum, PTH decreased slightly in group C, while it increased in group A. However, it was not statistical significance (P>0.05) for the discrepancy between the two groups.
4、4weeks、8weeks after implantation, the bone density of the freshly isolated left tibia and the maxillaby was measured by using dual energy X-ray absorptiometry. It was found that the density of tibia and mandible bone in group C was higher than that in group B：it was not statistical significance (P>0.05) for the discrepancy between the two groups after4weeks;But it was statistical significance(P<0.05) after8weeks.Compared with A group, the bone densityis in group C was lower than that in group A in, However, it was not statistical significance (P>0.05) for the discrepancy between the two groups after8weeks.
5、4weeks、8weeks after implantation, they were examed histologically and histomorphometrically.4weeks later, it was found that implant bone contact rate，，trabercular width and combinded bone lamella width of rats were much higher in groop C than that in group B (P<0.05);However, in terms of thickness of cortical, trabercular area, there was no difference between group C and group B.It was found that trabercular area,trabercular width and combinded bone lamella width of rats were much lower in groop C than that in group A (P<0.05);However, in terms of implant bone contact rate,thickness of cortical bone, there was no difference between group C and group A.8weeks later,it was found that implant bone contact rate, trabercular area,trabercular width and combinded bone lamella width of rats were much higher in groop C than that in group B (P<0.05);However, in terms of thickness of cortical bone, there was no difference between group C and group B. There was no difference between group C and group A in all of the terms, although every term grew higher except the term of thickness of cortical bone. What mentioned above proved that PTH could improve the quality of osseointegration of the rats.
Conclusion:
1、The osteoporosis model of the rats which is built by removing bilateral ovarian tissues is similar to the osteoporosis of post-menopausal women. Both of them belong to the high turnover osteoporosis, and have certain etiological and pathological similarity. The model is built scientifically and it can be used to investigate the influence of PTH1-34on osseointegration in the condition of osteoporosis.
2、Intermittent application of parathyroid hormone (PTH1-34) with small doses can increase the density of alveolar bone mineral, improve their bone structure, increase implant bone contact rate, trabercular area, trabercular width and combinded bone lamella width, further to improve the osseointegration quality of the rats.
引文
[1]Ekelund JA, Lindquist LW, Carlsson GE, Jemt T. Implant treatment in the edentulous mandible:a prospective study on Branemark system implants over more than 20 years.[J]Prosthodont,2003,16:602-608.
[2]Curran D, Maravic M, Kiefer P, Tochon V, Fardellone P. Epidemiology of osteoporosis-related fractures in France:A literature review. [J] Joint Bone Spine, 2010,77:546-551.
[3]Stanford CM, Schneider G B. Functional behaviour of bone around dental implants. [J] Gerodontology,2004,21:71-77.
[4]Tsolaki IN, Madianos PN, Vrotsos JA. Outcomes of dental implants in osteoporotic patients. A literature review. [J] Prosthodont,2009,18:309-323.
[5]Madrid C, Sanz M. What impact do systemically administrated bisphosphonates have on oral implant therapy? A systematic review. [J] Clin Oral Implants,2009,20(4): 87-95.
[6]Gulsahi A, Paksoy CS, Yazlcloglu N, Arpak N, Kucuk NO, Terzioglu H. Assessment of bone density differences between conventional and bone-condensing techniques using dual energy x-ray absorptiometry and radiography. [J] Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology,2007,104: 692-698.
[7]褚娴骨质疏松症的药物治疗进展[J]天津药学,2004,16(3)：49-51.
[8]蒋利华骨质疏松症药物治疗进展[J]中国药业,2006,15(4)：3-5.
[9]李颖骨质疏松症的药物治疗[J]上海铁道大学学报,2000,21(11)：76-79.
[10]Pham JW, Radhakrishnan I, Sontheimer EJ. Thermodynamic and structural characterization of 2'2 nitrogen2 modified RNA dup lexes. [J]N ucleic Acids Res, 2004,32 (11):3446-3455.
[11]宋庆明,盛正妍,刘皋林特立帕肽治疗骨质疏松症的应用进展[J]国际药学研究杂志,2008,35(6)：415-418,424.
[12]Jee WS, Ma Y. Animal models of immobilization osteopenia. [J] Morphologic, 1999,83:25-34.
[13]邓红文,刘耀中骨质疏松学前沿[M]北京：高等教育出版社,2005：245.
[14]赵和平,陈列等骨质疏松动物实验模型的建立和评价[J]中国中医骨伤科杂志,2001,9(3)：55-57.
[15]程群,朱汉民建立骨质疏松动物模型的标准化问题[J]老年医学与保健,2003,9(2)：122-124.
[16]陈守平,周正炎,陆卫青去势法兔骨质疏松模型建立[J]口腔颌面外科杂志,2001,11(3)：221-224.
[17]刘忠厚骨质疏松学[M]北京：高等教育出版社,1998：287,507.
[18]Thompson DD,Simmons HA,et al. FAD guidelines and animal models for osteoporsis.[J] Bone,1995,17(4):125-133.
[19]骆凯,闫福华,等绝经后骨质疏松大鼠实验性牙周炎动物模型研究[J]Journal of Oral Science Research,2006,4(22):130-132.
[20]崔燎,吴铁,刘晓青,等,低剂量雌激素与中药联合用药有效防治大鼠去卵巢所致骨丢失[J]中国骨质疏松杂志,2003,9(3)：200-204.
[21]Motohashi M, Shirota T, Tokugawa Y,et al. Bone reactions around hydroxyap-atitecoated implants in ovariectomized rats.[J] Oral Surg Oral Med Oral Pathol Ordiol Endod,1999,87(2):145-152.
[22]Yamazaki M, Shirota T, Tokugawa Y,et al.Bone reactions to titanium screw implants in ovariectomized animals. [J] Oral Surg Oral Med Oral Pathol Ordiol Endod,1999,87(4):411-418.
[23]Pan J,Shirota T,Ohno K,et al. Effect of ovariectomy on bone remodeling adjacent to hydroxyapatitecoated implants in the tibia of mature rats.[J] Oral Maxillofac Surg,2000,58(8):817-882.
[24]Riggs BL. The mechanisms of estrogen regulation of bone resorption.[J] Clin Invest,2000,106:1203-1204.
[25]Jimi E, Shuto T, Koga T. Macrophage colony-stimulating factor and interleukin-1 alpha maintain the survival of osteoelast-like cells.[J] Endocrinology, 1995,136(2):808-811.
[26]刘忠厚.骨质疏松学[M]北京：科学出版社,1998：67-78.
[27]严鹏霄,崔维顶,等阿仑膦酸钠、辛伐他汀联合应用治疗大鼠骨质疏松的作用[J]南京医科大学学报(自然科学版),2009,29(8)：1099-1102.
[28]赵勇,何冀川活血补肾方对去卵巢大鼠骨密度及骨力学性能的影响[J]中国中医药信息杂志,2010,17(4)：31-32.
[29]伍贤平 双能X线吸收法测定大鼠骨量的评价及去卵巢骨丢失敏感区的选择[J]中华内分泌代谢杂志,2000,16(4)：212-215.
[30]Whitfield JF. Osteoporosis'treating parathyroid hormonepeptides:What are they?What do they do?How mightthey do it? [J] Curr Opin Investig Drugs,2006, 7(4):349-359.
[31]Whitfield JF. Growing Bone. [M] Georgetown:Landes Bio-science,2005: 113-116.
[32]Keller H, Kneissel M. SOST is a target gene for PTH in bone. [J] Bone, 2005,37(2):148-159.
[33]Li X, Zhang Y, Kang H, et al. Sclerostin binds to LRP5/6 and antagonizes canonical Wnt signaling.[J] J Biol Chem,2005,280(20):19883-19887.
[34]Dobnig H, Turner RT. Evidence that intermittent treatment with parathyroid hormone increases bone formation in adult rats by activation of bone lining cells. [J] Endocrinology,1995,136(8):3632-3638.
[35]Hodsman AB, Steer BM. Early histomorphometric changes in response to parathyroid hormone therapy in osteoporosis:Evidence for de novo bone formation on quiescent cancellous surfaces. [J] Bone,1993,14(3):523-527.
[36]冯坤,刘越桂,张灵菊等高转换型骨质疏松模型的生化特点[J]中国骨质疏松杂志,1997,3：25-30.
[37]Mundy GR.Inflammatory mediators and the destruction of bone.[J] Periontal Res,1991,26:213-217.
[38]邱蔚六.口腔颌面外科学[M]北京：人民卫生出版社,2007：325-327.
[39]Rozalia D,Eleftherios T,Peter V,et al.Current concepts of molecular,aspects of bone healing. [J] Injury,2005,36(12):1392-1404.
[40]Jung RE,Cochran D L,Domken O,et al.The effect of matrix bound parathyroid hormone on bone regeneration. [J] Clin Oral Implants Res,2007,18(3):319-325.
[41]Jung R E,H mmerle C H,Kokovic V,et al.Bone regeneration using asynthetic matrix containing a parathyroid hormone peptide combined witha grafting material.[J]Oral Maxillofac Implants,2007,22(2):258-66.
[2]Curran D, Maravic M, Kiefer P, Tochon V, Fardellone P. Epidemiology of osteoporosis-related fractures in France:A literature review. [J] Joint Bone Spine, 2010,77:546-551.
[3]Stanford CM, Schneider G B. Functional behaviour of bone around dental implants. [J] Gerodontology,2004,21:71-77.
[4]Tsolaki IN, Madianos PN, Vrotsos JA. Outcomes of dental implants in osteoporotic patients. A literature review. [J] Prosthodont,2009,18:309-323.
[5]Madrid C, Sanz M. What impact do systemically administrated bisphosphonates have on oral implant therapy? A systematic review. [J] Clin Oral Implants,2009,20(4): 87-95.
[6]Gulsahi A, Paksoy CS, Yazlcloglu N, Arpak N, Kucuk NO, Terzioglu H. Assessment of bone density differences between conventional and bone-condensing techniques using dual energy x-ray absorptiometry and radiography. [J] Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology,2007,104: 692-698.
[7]褚娴骨质疏松症的药物治疗进展[J]天津药学,2004,16(3)：49-51.
[8]蒋利华骨质疏松症药物治疗进展[J]中国药业,2006,15(4)：3-5.
[9]李颖骨质疏松症的药物治疗[J]上海铁道大学学报,2000,21(11)：76-79.
[10]Pham JW, Radhakrishnan I, Sontheimer EJ. Thermodynamic and structural characterization of 2'2 nitrogen2 modified RNA dup lexes. [J]N ucleic Acids Res, 2004,32 (11):3446-3455.
[11]宋庆明,盛正妍,刘皋林特立帕肽治疗骨质疏松症的应用进展[J]国际药学研究杂志,2008,35(6)：415-418,424.
[12]Jee WS, Ma Y. Animal models of immobilization osteopenia. [J] Morphologic, 1999,83:25-34.
[13]邓红文,刘耀中骨质疏松学前沿[M]北京：高等教育出版社,2005：245.
[14]赵和平,陈列等骨质疏松动物实验模型的建立和评价[J]中国中医骨伤科杂志,2001,9(3)：55-57.
[15]程群,朱汉民建立骨质疏松动物模型的标准化问题[J]老年医学与保健,2003,9(2)：122-124.
[16]陈守平,周正炎,陆卫青去势法兔骨质疏松模型建立[J]口腔颌面外科杂志,2001,11(3)：221-224.
[17]刘忠厚骨质疏松学[M]北京：高等教育出版社,1998：287,507.
[18]Thompson DD,Simmons HA,et al. FAD guidelines and animal models for osteoporsis.[J] Bone,1995,17(4):125-133.
[19]骆凯,闫福华,等绝经后骨质疏松大鼠实验性牙周炎动物模型研究[J]Journal of Oral Science Research,2006,4(22):130-132.
[20]崔燎,吴铁,刘晓青,等,低剂量雌激素与中药联合用药有效防治大鼠去卵巢所致骨丢失[J]中国骨质疏松杂志,2003,9(3)：200-204.
[21]Motohashi M, Shirota T, Tokugawa Y,et al. Bone reactions around hydroxyap-atitecoated implants in ovariectomized rats.[J] Oral Surg Oral Med Oral Pathol Ordiol Endod,1999,87(2):145-152.
[22]Yamazaki M, Shirota T, Tokugawa Y,et al.Bone reactions to titanium screw implants in ovariectomized animals. [J] Oral Surg Oral Med Oral Pathol Ordiol Endod,1999,87(4):411-418.
[23]Pan J,Shirota T,Ohno K,et al. Effect of ovariectomy on bone remodeling adjacent to hydroxyapatitecoated implants in the tibia of mature rats.[J] Oral Maxillofac Surg,2000,58(8):817-882.
[24]Riggs BL. The mechanisms of estrogen regulation of bone resorption.[J] Clin Invest,2000,106:1203-1204.
[25]Jimi E, Shuto T, Koga T. Macrophage colony-stimulating factor and interleukin-1 alpha maintain the survival of osteoelast-like cells.[J] Endocrinology, 1995,136(2):808-811.
[26]刘忠厚.骨质疏松学[M]北京：科学出版社,1998：67-78.
[27]严鹏霄,崔维顶,等阿仑膦酸钠、辛伐他汀联合应用治疗大鼠骨质疏松的作用[J]南京医科大学学报(自然科学版),2009,29(8)：1099-1102.
[28]赵勇,何冀川活血补肾方对去卵巢大鼠骨密度及骨力学性能的影响[J]中国中医药信息杂志,2010,17(4)：31-32.
[29]伍贤平 双能X线吸收法测定大鼠骨量的评价及去卵巢骨丢失敏感区的选择[J]中华内分泌代谢杂志,2000,16(4)：212-215.
[30]Whitfield JF. Osteoporosis'treating parathyroid hormonepeptides:What are they?What do they do?How mightthey do it? [J] Curr Opin Investig Drugs,2006, 7(4):349-359.
[31]Whitfield JF. Growing Bone. [M] Georgetown:Landes Bio-science,2005: 113-116.
[32]Keller H, Kneissel M. SOST is a target gene for PTH in bone. [J] Bone, 2005,37(2):148-159.
[33]Li X, Zhang Y, Kang H, et al. Sclerostin binds to LRP5/6 and antagonizes canonical Wnt signaling.[J] J Biol Chem,2005,280(20):19883-19887.
[34]Dobnig H, Turner RT. Evidence that intermittent treatment with parathyroid hormone increases bone formation in adult rats by activation of bone lining cells. [J] Endocrinology,1995,136(8):3632-3638.
[35]Hodsman AB, Steer BM. Early histomorphometric changes in response to parathyroid hormone therapy in osteoporosis:Evidence for de novo bone formation on quiescent cancellous surfaces. [J] Bone,1993,14(3):523-527.
[36]冯坤,刘越桂,张灵菊等高转换型骨质疏松模型的生化特点[J]中国骨质疏松杂志,1997,3：25-30.
[37]Mundy GR.Inflammatory mediators and the destruction of bone.[J] Periontal Res,1991,26:213-217.
[38]邱蔚六.口腔颌面外科学[M]北京：人民卫生出版社,2007：325-327.
[39]Rozalia D,Eleftherios T,Peter V,et al.Current concepts of molecular,aspects of bone healing. [J] Injury,2005,36(12):1392-1404.
[40]Jung RE,Cochran D L,Domken O,et al.The effect of matrix bound parathyroid hormone on bone regeneration. [J] Clin Oral Implants Res,2007,18(3):319-325.
[41]Jung R E,H mmerle C H,Kokovic V,et al.Bone regeneration using asynthetic matrix containing a parathyroid hormone peptide combined witha grafting material.[J]Oral Maxillofac Implants,2007,22(2):258-66.