Thesis (Ph. D.)--West Virginia University, 2001. / Title from document title page. Document formatted into pages; contains xi, 211 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 171-186).

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

<p>Uvod: Osteoporoza je sistemsko oboljenje skeleta koje se karakteri&scaron;e smanjenjem mase kosti i promenama u ko&scaron;tanoj strukturi, &scaron;to sve ima za posledicu povećanu sklonost ko&scaron;tanog tkiva ka prelomima. Prema preporuci Svetske zdravstvene organizacije, dijagnoza osteoporoze postavlja se ukoliko je T-score -2,5 SD i ispod te vrednosti, a normalan nalaz ako je vrednost T-score -1,0 SD i iznad te vrednosti. Danas se smatra da je zlatni standard u dijagnostici osteoporoze primena dvostruke X apsorpcione denzitometrije lumbalne kičme i kuka putem koje se dobiju vrednosti ko&scaron;tane mase Bone mineral density i T-score. Međutim, poznato je da postoje faktori rizika koji utiču na redukciju mase kosti na taj način &scaron;to smanjuju maksimum mase kosti koji se stiče do 35. godine života i / ili ubrzavaju inače normalan proces postepenog i blagog smanjenja mase kosti koji počinje posle 35. godine života i na taj način povećavaju rizik za frakture. Takođe, poznato je da neki od faktora rizika i njihova udruženost može dovesti do povećanog rizika za frakture i nezavisno od ko&scaron;tane mase i T-score. Ciljevi istraživanja : 1. Utvrditi ko&scaron;tanu masu u postmenopauznih žena primenom dvostruke X apsorpcione denzitometrije. 2. Analizirati distribuciju faktora rizika u pacijentkinja sa T-score ispod -2.5 SD u poređenju sa pacijentkinjama sa T-score iznad -1.0 SD. 3. Utvrditi odnos između statističkog prostora koji čine pojedinačni i udruženi faktori rizika (sa karakteristikama svakih od njih) i mase kosti određene denzitometrijski. Materijal i metode rada: Istraživanje je koncipirano delom kao prospektivna, a delom kao retrospektivna studija koja je sprovedena kod pacijentkinja u postmenopauznom periodu života, životne dobi od 50 do 80 godina. Nakon urađene dvostruke X apsorpcione denzitometrije lumbalne kičme i kuka ispitivane pacijentkinje su same popunjavale upitnik uz pomoć medicinske sestre ili lekara. Nakon dobijenih podataka pacijentkinje su podeljene u dve grupe: sa osteoporozom i bez osteoporoze. U grupi sa osteoporozom je bilo 270 pacijentkinja, a u grupi bez osteoporoze 250 pacijentkinja. Potom je sprovedena statistička obrada podataka. Nakon sveobuhvatne analize dobijenih rezultata istraživanja izvedeni su sledeći zaključci: 1.Ustanovljeno je da 60% postmenopauznih žena prosečne životne dobi od 67.0 &plusmn; 7.0 godina ima osteoporozu odnosno vrednost T-score &le; -2.5 SD. 2. Postoji statistička značajna povezanost između ko&scaron;tane mase i sledećih faktora rizika: pozitivna porodična anamneza na osteoporozu i frakture, telesna težina, telesna visina, ranije frakture, česti padovi i smanjenje u visini vi&scaron;e od 3 cm. 3. Analizom faktora rizika se dobijaju karakteristike osoba sa osteoporozom: pozitivna porodična anamneza na osteoporozu i frakture, manja telesna težina i telesna visina, smanjenje u visini vi&scaron;e od 3 cm, česti padovi i ranije frakture. 4. Hipertireoidizam i hiperparatireoidizam, reumatoidni artritis, primena kortikosteroidne terapije su faktori rizika koji su vi&scaron;e zastupljeni kod ispitivanih pacijentkinja sa osteoporozom. 5. Pu&scaron;enje, rana menopauza, alergija na mleko bez adekvatne supstitucije sa kalcijumom i nedovoljan boravak na suncu bez adekvatne supstitucije sa vitaminom D su faktori rizika koji su vi&scaron;e zastupljeni kod ispitivanih pacijentkinja sa osteoporozom. 6. Najveći doprinos celini daje pozitivna porodična anamneza na osteoporozu i frakture (20.99%), zatim slede telesna težina, telesna visina, Index telesne mase (19.03%), ranije frakture, česti padovi, smanjenje u visini vi&scaron;e od 3 cm (18.41%), pu&scaron;enje i nedovoljna fizička aktivnost (12.75%), alergija na mleko i nedovoljan boravak na suncu (12.14%), rana menopauza (8.72%), hipertireoidizam, hiperparatireoidizam, reumatoidni artritis (7.93%). 7. Analizom tri grupe obeležja koja daju najveći doprinos celini ustanovljeno je da pozitivna porodična anamneza na frakture (37.7%) i telesna težina (31.3%) predstavljaju major faktore rizika za osteoporozu. 8. Matematičkom obradom dolazi se do formule pomoću koje bi sa verovatnoćom od 64.0 % mogla predvideti osteoporoza, a sa verovatnoćom 73.2 % odsustvo osteoporoze, čime se između ostalog u na&scaron;em istraživanju donekle relativizuje neophodnost određivanja ko&scaron;tane mase u proceni rizika za prelome i u proceni potrebe za uvođenje antiosteoporotične terapije. Formula je +.214 O +.562 F +.202 R +.223 P +.335 S +.493 T +.057 V +.020 9. Potrebno je testirati dobijenu formulu na ispitivanim pacijentkinjama i nastaviti istraživanje na većem uzorku na faktore rizika koji nisu pokazali statističku značajnost.</p> / <p>Introduction: Osteoporosis is a systematic disease of skeleton characterized by the reduction of bone mass and changes in bone structure which result in the increased aptitude of bone tissue to fractures. According to the suggestion of the World Health Organization, the diagnosis for osteoporosis is set if the T-score is -2.5 SD and below it and the normal report if the value of T-score is -1.0 SD and above it. Nowadays, it is considered that the golden standard in osteoporosis diagnostic is the use of double X absorption densitometry of lumbal spine and hipe which provides the values of bone mass Bone mineral density as well as T-score. However, it has been known that there are risk factors whish influence the reduction of bone mass by reducing maximum bone mass gained by the age of 35 and/or by quckening, the normal process of gradual and mild reduction of bone mass starting after 35 and in that way increase the risk toward fractures. It mas also been known that some of the risk factors and their correlation may cause the increasement of the risk factor toward fractures not having the connection with the bone mass and T-score. Researchment aims: 1. Determine bone mass in postmenopausal women using double X absorption densitometry. 2. Analyse distribution of risk factors in patients whith the T-score below -2.5 SD comparing to the patients with T-score above -1.0SD. 3. Determine the relation between statistical space made by individual and associated risk factors (with the characteristics of each of them) and the bone mass specified by densitometry. Material and methods of working: Researchment is outlined partly as prospective and partly as retrospective study which was carried out in patients in postmenopausal life period, aged 50-80. After applying double X absorption densitometry of lumbal spine and hip the examined patients did the questionnaire by themselves whith the help of nurses and doctors. After obtaining the data, patients were divided into two groups: with and without osteoporosis. There were 270 patients in the group with osteoporosis and 250 of them without it. Thereafter, the statistic data processing was carried out. After the overall analysis of obtained results of researchment, following conclusions were conducted: 1. It has been determined that 60 % of postmenopausal women of average age 67.0&plusmn;7.0 have osteoporosis, in other words, their T-score is &le; -2.5 SD. 2. There is statistically important relationship between the bone mass and following risk factors: positive family anamnesis to osteoporosis and fractures, body weight, height, previos fractures, frequent falls and reduction of height for more than 3 cm. 3. Analysing the risk factors, characteristics of persons with osteoporosis have been obtained: positive family anamnesis to osteoporosis and fractures, smaller body weight and height, the reduction in height for more than 3 cm, frequent falls and previous fractures. 4. Hyperthyroidism and hyperparathyroidism, rheumatoid arthritis and the usage of corticosteroid therapy are the risk factors more incident in the examined patients with osteoporosis. 5. Smoking, early menopause, allergy to milk with no adequate substitution of calcium and insufficient exposition to sun rays with no adequate substitution of vitamine D are the risk factors more incident in patients with osteoporosis. 6. The largest contribution to the total makes positive family anamnesis to osteoporosis and fractures (20.99%), followed by body weight, height, Body mass index (19.03%), previos fractures, frequent falls and reduction in height for more than 3 cm (18.41%), smoking and insufficient physical activity (12.75%), allergy to milk and insufficient exposition to the sun (12.14%), early menopause (8.72%), hyperthyroidism and hyperparathyroidism, rheumatoid arthritis (7.93%). 7. By the analysis of all three goups of features giving the largest cintribution to the total, it has been determined that positive family anamnesis to fractures (37.7%), and body weight (31.3%), present the major risk factors for osteoporosis. 8. By mathematical processing we obtain the formula which can with the probability of 64.0% predict osteoporosis, and with the probability of 73.2% the absence of osteoporosis, which can, among other things in our research to some extent, require relative necessity for introduction of antiosteoporotic therapy. The formula is +.214 O +.562 F +.202 R +.223 P +.335 S +.493 T +.057 V +.020. 9. It is necessary to test the formula obtained in examined patients and continue the reseachment, on larger sample, of risk factors which have not shown statistic importance.</p>

A pilot clinical trial was conducted after the in-vivo and in-vitro studies: Eight subjects fulfilled the inclusion criteria were recruited. However, the liver function tests of three subjects out of eight were found to be abnormal with elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) level, which was not reported in previous toxicity test. The trial was suspended immediately and a follow-up test showed that the elevated AST and ALT level had reverted back to normal within one month after termination of OPR intake. Although we could not accomplish a RCT, the pilot study revealed potential hepatotoxicity of OPR on human beings and it would raise the safety awareness of investigators on the use of herbal remedies in future clinical studies. / After the in-vivo and in-vitro studies, a double-blinded, randomized, placebo-controlled clinical trial (RCT) was planned. Due to a lack of a Chinese version of instrument to measure osteoporosis-specific quality of life, an English version of the Osteoporosis-Targeted Quality of Life Questionnaire (OPTQoL) was translated into Chinese and linguistically validated according to the standard guideline. The newly formed Chinese OPTQoL can be used to assess the impact of new interventions on quality of life among Chinese osteoporosis patients. / Association of the incidence of postmenopausal osteoporosis and Kidney-Vacuity Syndromes in TCM was investigated with the aid of a Kidney-Vacuity Syndromes scoring questionnaire. In the study, postmenopausal women, who suffered from deficiency of kidney "qi" and kidney "essence", had a significantly higher incidence of osteoporosis. These findings strongly supported that replenishing kidney qi and kidney essence was a logical therapeutic principle in the formulation of OPR. / In conclusion, this study investigated the use of TCM on the treatment of postmenopausal osteoporosis in a systematic manner. It started from herbal formulation, basic science studies to clinical trial. It revealed beneficial effects of OPR on bones through in-vivo and in-vitro studies and demonstrated certain possible mechanism behind. On the other hand, the hepatotoxicity of OPR on human beings was also exposed and had not been reported in previous toxicity tests. The study provided valuable clinical data for other investigators on the potential hazards of herbal remedies although they had been validated as safe and effective in pre-clinical stage. / In search for safe, effective and low-priced medicine, the public have turned their attention to Traditional Chinese Medicine (TCM). Extensive experience has been accumulated in TCM regarding the diagnosis and treatment of osteoporosis, which often involves the prescription of kidney-tonifying herbs. Therefore, the aim of the study, firstly, was to explore the association of the incidence of postmenopausal osteoporosis and Kidney-Vacuity Syndromes in TCM, so as to formulate a rational kidney-tonifying herbal formula for osteoporosis research (OPR). Secondly, the effect of the formula was evaluated by in-vitro and in-vivo studies. Thirdly, the Osteoporosis-Targeted Quality of Life Questionnaire was linguistically validated from English to Chinese, which was expected to be one of the outcome measurement tools in future clinical trials. Lastly, a pilot clinical study was performed, which revealed some potential hazards of the formula on human beings which have not been shown in previous works. / Osteoporosis is a skeletal disorder which leads to an increased risk of bone fracture, disability or even death. It has become a major public health threat and the worldwide incidence of osteoporotic fracture is projected to increase two fold within the next 50 years. Postmenopausal women, being affected by a lack of estrogen, face a much higher risk of the disease. This study would therefore focus on type I osteoporosis (i.e. postmenopausal osteoporosis). Although current medications can slow down the bone deterioration process, their side effects and high cost had impaired patients' compliance with long term treatment. / The effect of OPR for the treatment of postmenopausal osteoporosis was then evaluated by in-vitro and in-vivo studies. In the in-vivo study, an osteoporosis model was established by performing ovariectomy on the four-week-old C57BL/6 mice. A high bone turnover rate was induced and OPR successfully slowed down the high turnover rate of bones by decreasing bone formation and resorption process without increasing the uterine linings. However, its beneficial effect on bones could not be detected on bone mineral density measurement. / The potential mechanism of action of OPR on bones was explored by in-vitro study. OPR was shown to induce cell proliferation and differentiation of osteoblast-like UMR 106 cells. Furthermore, the estrogenic activity of OPR was detected by MCF-7 cell line, which has been stably transfected with estrogen responsive elements (ERE). OPR was shown to possess an estrogenic activity in a dose dependent manner and was comparable to the positive control at a concentration of 200 and 1000 mug/ml. The induced estrogenic activity by OPR may be associated with the presence of phytoestrogen within the herbal formula. These findings suggested that the beneficial effect of OPR on bones might relate to its direct positive effect on osteoblast and its estrogenic-like activity. / Liong, Ching. / Adviser: Chun-tao Che. / Source: Dissertation Abstracts International, Volume: 73-03, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves ). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

成人骨量的更新与维持通过骨重塑来实现。骨重塑包括骨吸收与骨形成两个偶联的过程，其中成骨细胞介导骨形成，破骨细胞介导骨吸收，当偶联的骨吸收超过骨吸收就会导致骨量丢失，进而导致发生骨质疏松症的发生。目前，临床治疗骨质疏松的药物如阿仑膦酸盐、雌激素受体调节剂、活性维生素D、雌激素替代治疗、降钙素、骨化三醇等都是基于针对破骨细胞的调控来抑制骨吸收，但是对于已经丢失的骨量无法恢复。唯一被美国FDA批准用来通过刺激新骨形成来恢复丢失的骨量的治疗药物就是甲状旁腺激素。然而，这种药物在刺激新骨形成的同时也刺激了骨吸收，即：在使用18个月后有明显促进骨吸收的副作用。 / 酪蛋白激酶相互作用蛋白-1（CKIP-1）基因是一个新发现的骨形成的负调控基因，CKIP-1基因敲除小鼠在骨发育和正常骨代谢过程中均未发现激活骨吸收。CKIP-1敲除导致小鼠胫骨近端骨量与胫骨皮质骨形成速率显著高于野生型，且这一差异随着小鼠的增龄而显著，而骨外器官没有发现异常表型，提示CKIP-1是潜在相对安全的治疗骨质疏松的靶向基因。特别是我们最近研发的一种天门冬氨酸－丝氨酸－丝氨酸重复三肽修饰的脂质体递送（(Asp-Ser-Ser)₆-liposome）系统能够实现靶向骨形成表面的小干扰核酸的递送，并明显减少了小干扰核酸在非骨组织的分布。因此，提出本课题的研究假设：特异性静默骨内CKIP-1可以促进骨形成而不刺激骨吸收，从而为骨质疏松的临床治疗提供安全有效的治疗手段。 / 为了确定CKIP-1基因表达在老年绝经后妇女骨骼中与骨形成内在联系，首先，我们通过对发生骨折的老年绝经后妇女的骨痂标本中CKIP-1 mRNA和蛋白表达的测定，发现CKIP-1基因mRNA和蛋白表达水平与骨形成能力负相关。并且，这种相关性在骨质疏松动物模型中进一步得到证实。其次，针对我们研究假设，从一组针对大鼠、小鼠、猴和人类的成骨样细胞的CKIP-1 mRNA的跨种属siRNA序列中筛选出体外静默效率最高CKIP-1小干扰核酸序列si-3。接着，体内外实验证实si-3序列在健康动物体内的静默效率和促进成骨的功能。同时，确定尾静脉注射（Asp-Ser-Ser）₆-liposome 包裹的CKIP-1小干扰核酸在 大鼠和小鼠为的最佳剂量分别为3.75mg/kg和7.5mg/kg以及注射周期为每两周一次。最后，为了检验CKIP-1 小干扰核酸是否可通过促进骨形成从而逆转绝经后骨质疏松症中的骨丢失，我们分别以绝经后骨质疏松大鼠和小鼠为实验动物模型，通过测定骨形态计量学参数、骨量和骨结构等来评价骨靶向递送系统（(Asp-Ser-Ser)₆-liposome）递送的CKIP-1 siRNA对老年绝经后骨质疏松症的治疗效果。动态活体CT分析结果表明，与0周未治疗的基础值相比，经6周治疗骨密度（BMD）, 相对骨体积分数（BV/TV）和骨小梁厚度（Tb.Th）在小干扰核酸治疗组显著增加。此外，在治疗6周后小干扰核酸治疗组骨密度，相对骨体积和骨小梁厚度显示较高于模型对照组。0周与其它检测时间点之间的对比分析较显示，小干扰核酸治疗组的新生骨显著高于模型组或假手术组。组织形态学分析结果表明在治疗6周后，无论是股骨远端或中段的矿化沉积率（MAR）、骨形成速率（BFR） 和组的骨形成表面（Ob.S/ BS）在OVX组和siRNA组均显著高于模型对照组，而模型对照组和小干扰核酸治疗组的骨吸收表面（Oc.S/ BS）之间无显著性差异。 / 结论：CKIP-1基因小核酸干扰治疗在老年绝经后骨质疏松中能够显著促进骨形成并不会加剧骨吸收，该药物具有显著逆转骨丢失的作用。 / Osteoporosis is characterized by an imbalance between bone formation and bone resorption. Therefore, promoting bone formation and inhibiting bone resorption are the two major therapeutic strategies in the treatment of osteoporosis. Currently, the only Food and Drug Administration (FDA)-approved anabolic agent capable of stimulating bone formation is parathyroid hormone (PTH). However, dominant bone resorption after 18-month treatment with PTH is a great concern (Rubin and Bilezikian 2003). Thus, development of alternative bone anabolic agents is highly desirable. / Casein kinase-2 interacting protein-1 (CKIP-1), which is encoded by Plekho1, and thus also known as Plekho1, is a newly discovered negative regulator of bone formation during bone development and subsequent bone maintenance that does not activate bone resorption (Lu, Yin et al. 2008). Specifically, CKIP-1 protein functions as the auxiliary factor of ubiquitin ligase Smad ubiquitylation regulatory factor 1 (Smurf1) to interrupt the bone anabolic BMP-signalling pathway, which has been demonstrated to be a specific suppressor of bone formation (Yamashita, Ying et al. 2005). In a previous study, we found that CKIP-1 expression in female rat bone increases with aging, whereas bone formation decreases with aging (Guo, Zhang et al. 2010). Systemic examination of the tissue distribution of CKIP-1 expression has revealed that is abundantly expressed in the musculoskeletal system but sparingly expressed in the liver, lungs, kidneys, pancreas, and other organs (Zhang, Tang et al. 2007). In addition, an abnormal tissue phenotype in heart, liver, spleen, lung, and kidney tissue has not been observed in CKIP-1 gene knockout mice (KO), even at an advanced age (Lu, Yin et al. 2008). Thus, CKIP-1 gene silencing might be a potential strategy for promoting bone anabolic action in reversing bone loss. / RNA interference (RNAi), a natural cellular process that regulates gene expression by a highly precise mechanism of sequence-directed gene silencing at the stage of translation by degrading specific messenger RNA and then blocking translation of the specific gene, has been employed for gene silencing in vivo (Frank-Kamenetsky, Grefhorst et al. 2008). Accordingly, RNAi should be an appropriate target for CKIP-1 gene silencing in vivo. / We raised the hypothesis that therapeutic RNAi targeting of CKIP-1 might promote bone formation for reversing postmenopausal bone loss. To test the hypothesis, we performed several studies to achieve the following specific aims: (1) To explore the relationship between CKIP-1 expression and bone formation in aged postmenopausal osteoporosis; (2) To Identify a cross-species CKIP-1 siRNA sequence with high knockdown efficiency; (3) To validate of the identified CKIP-1 siRNA in healthy rodents in vivo; (4) To examine the anabolic effect of the identified CKIP-1 siRNA on bone in osteoporotic animal models. / The relationship between CKIP-1 gene expression and bone formation in bone specimens from aged postmenopausal women: To explore the association between CKIP-1 gene expression and bone formation in bone specimens from aged postmenopausal women, the gene expression of CKIP-1 and ALP in the bone specimens from aged female patients were examined. We found the protein expression of CKIP-1 increased during aging and negatively correlate to bone formation as indicated by the mRNA expression of ALP (Guo., Zhang. et al. 2011). Further, we also found the decreased bone formation during aging was partly rescued in Ckip-1 KO mice during aging. / A cross-species CKIP-1 siRNA sequence: Recently, we identified a specific CKIP-1 siRNA sequence (CKIP-1 siRNA si-3) with high knockdown efficiency across rat, mouse, rhesus, and human osteoblast-like cells that does not induce immunostimulatory activity and promotes osteoblast differentiation across the species in vitro and bone formation in rats in vivo (Guo, Zheng et al. 2012). / Validation of the CKIP-1 siRNA si-3 capsulated by bone-targeted siRNA delivery system in healthy rodents in vivo: We developed a bone-targeting siRNA delivery system (tripeptide aspartate-serine-serine linked with liposome, i.e. (Asp-Ser-Ser)₆-liposome) that can remarkably reduce the exposure of non-bone tissue to CKIP-1 siRNA (Zhang, Guo et al. 2012). To validate the identified CKIP-1 siRNA in healthy rodents in vivo, the established continuous CKIP-1 gene silencing protocol is optimized in adult rats and mice in vivo by hydrodynamic tail vein injection of 3.75mg/kg for rats and 7.5 mg/kg for mice every 2 weeks (Guo, Zhang et al. 2010). The osteogenic effects of CKIP-1 siRNA in both rats and mice were further validated in vivo. / Anabolic effect of CKIP-1 siRNA si-3 on bone in aged postmenopausal osteoporosis: For evaluation of the anabolic effect of CKIP-1 siRNA si-3 on reversing bone loss due to osteoporosis in an animal model, we intravenously injected ovariectomized (OVX) rats and mice with CKIP-1 siRNA delivered by the (Asp-Ser-Ser)₆-liposome, a liposome linked with six repeated aspartate-serine-serine moiety, every 2 weeks for 6 weeks. In vivo and ex vivo microCT analysis demonstrated a change over time in the variables examined and different change patterns over time among the groups examined after administration. We found that the siRNA group had experienced a significant increase in bone mineral density (BMD), relative bone volume (BV/TV), and trabecular thickness (Tb.Th) between weeks 0 and 6; had a higher BMD, BV/TV, and Tb.Th compared to the OVX group at week 6; and had a similar Tb.Th to that of the SHAM group at week 6. Registration analysis between week 0 and other time points revealed that the siRNA had a greater number of newly formed bone than the OVX and SHAM groups. Histomorphometric analysis showed that the siRNA group had a significantly higher mineralization rate (MAR), a significantly higher bone-formation rate (BFR), a significantly larger osteoblast surface (Ob.S/BS) at both the distal and mid-shaft femur compared to the OVX group after 6 weeks of treatment but not a significantly different Oc.S/BS. / Significance: Confirmation of our hypothesis by our results helps establish CKIP-1’s role as a pivotal negative regulator of bone formation in the aging skeleton and provides evidence that inhibiting CKIP-1 is a novel anabolic treatment for osteoporosis, indicating great potential for the use of therapeutic RNAi in orthopaedics and traumatology. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Guo, Baosheng. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves [132-150]). / Abstract also in Chinese. / Declaration --- p.i / Acknowledgements --- p.ii / Abstract --- p.iii / 论文摘要 --- p.vii / Table of Content --- p.ix / Abbreviations --- p.xvii / List of Figures --- p.xix / List of Tables --- p.xxii / Chapter CHAPTER 1 --- Review of recent anabolic therapy for osteoporosis --- p.1 / Chapter 1.1. --- Epidemiology of postmenopausal osteoporosis --- p.1 / Chapter 1.1.1. --- Definition of osteoporosis --- p.1 / Chapter 1.1.2. --- Epidemiology and health challenge of postmenopausal osteoporosis --- p.2 / Chapter 1.2. --- General pathophysiological understanding of osteoporosis and current challenge for osteoporosis treatment --- p.3 / Chapter 1.2.1. --- Bone modeling and remodeling --- p.3 / Chapter 1.2.2. --- Pathophysiological process of osteoporosis --- p.4 / Chapter 1.2.3. --- Systemic risk factors in the pathophysiology of osteoporosis --- p.5 / Chapter 1.2.4. --- Local risk factors in the osteoporosis pathophysiology --- p.6 / Chapter 1.2.5. --- Two therapeutic strategies for osteoporosis treatment --- p.7 / Chapter 1.3. --- Current and potential anabolic agents for osteoporosis treatment --- p.8 / Chapter 1.3.1. --- PTH analogues --- p.8 / Chapter 1.3.2. --- Potential concerns regarding PTH administration --- p.9 / Chapter 1.3.3. --- Potential PTH alternatives --- p.10 / Chapter 1.3.4. --- Modulation of Wnt/β-cateinin pathway --- p.10 / Chapter 1.3.5. --- Aptamer-based technology in osteoporosis treatment --- p.14 / Chapter 1.4. --- CKIP-1: A novel negative regulator of bone formation --- p.15 / Chapter 1.4.1. --- TGF-β/BMP signaling pathways involved in regulating bone formation --- p.15 / Chapter 1.4.2. --- CKIP-1 interrupts BMP signaling pathway --- p.16 / Chapter 1.4.3. --- CKIP-1 negatively regulates bone formation without activating bone resorption --- p.17 / Chapter 1.5. --- RNA interference strategy in anabolic therapy of osteoporosis --- p.18 / Chapter 1.5.1. --- siRNA-mediated gene silencing in osteoporosis treatment --- p.18 / Chapter 1.5.2. --- MicroRNAs as potential therapeutic targets in the anabolic treatment of osteoporosis --- p.20 / Chapter 1.5.3. --- Bone targeted RNAi-based anabolic-agents delivery --- p.23 / Chapter 1.6. --- Summary --- p.24 / Chapter CHAPTER 2 --- The relationship between CKIP-1 expression and bone formation in aged postmenopausal osteoporosis --- p.26 / Chapter 2.1 --- Introduction --- p.26 / Chapter 2.2 --- Materials and methods --- p.28 / Chapter 2.2.1 --- Bone specimen collection from aged postmenopausal women --- p.28 / Chapter 2.2.2 --- Total RNA extraction, reverse transcription and quantitative real-time PCR --- p.28 / Chapter 2.2.3 --- Total protein extraction and western blot analysis --- p.30 / Chapter 2.2.4 --- CKIP-1 expression in bone and other tissues --- p.31 / Chapter 2.2.5 --- Relationship between CKIP-1 expression and bone formation in aged ovariectomized rats --- p.31 / Chapter 2.2.6 --- Role of CKIP-1 in regulating bone formation in aged ovariectomized mice --- p.32 / Chapter 2.2.7 --- Statistics --- p.32 / Chapter 2.3 --- Results --- p.33 / Chapter 2.3.1 --- Correlation analysis between CKIP-1 expression and bone formation-related gene expression in bone specimens from agedd postmenopausal women across age --- p.33 / Chapter 2.3.2 --- CKIP-1 gene expression pattern in bone and other tissues --- p.37 / Chapter 2.3.3 --- Correlation between CKIP-1 expression and bone formation in rat bone --- p.38 / Chapter 2.3.4 --- CKIP-1 negatively regulates bone formation in aged ovariectomized mice by using CKIP-1 knockout mice --- p.39 / Chapter 2.4 --- Summary --- p.41 / Chapter CHAPTER 3 --- Identification of a cross-species CKIP-1 siRNA sequence --- p.43 / Chapter 3.1 --- Introduction --- p.43 / Chapter 3.2 --- Materials and methods --- p.44 / Chapter 3.2.1 --- Design rationale and modification for cross-species CKIP-1 siRNA --- p.44 / Chapter 3.2.2 --- In vitro screening for cross-species CKIP-1 siRNA sequences --- p.45 / Chapter 3.2.3 --- Investigation of the effects of the identified CKIP-1 siRNA on the expression of osteoblast phenotype genes --- p.47 / Chapter 3.2.4 --- Total RNA extraction, reverse transcription and quantitative real-time PCR --- p.47 / Chapter 3.2.5 --- Western blot analysis --- p.51 / Chapter 3.2.6 --- Evaluation of calcium deposition --- p.51 / Chapter 3.2.7 --- BMP-2 reporter activity assay in MC3T3-E1 cells --- p.52 / Chapter 3.2.8 --- Isolation of the primary human blood monocytes and IFN-α and TNF-α measurement --- p.53 / Chapter 3.2.9 --- Statistics --- p.54 / Chapter 3.3 --- Results --- p.54 / Chapter 3.3.1 --- Bio-informatic analysis of the designed CKIP-1 siRNA sequences --- p.54 / Chapter 3.3.2 --- Identified the cross-species CKIP-1 siRNA sequences by In vitro screening --- p.56 / Chapter 3.3.3 --- Effects of the identified CKIP-1 siRNA on the expression of osteoblast phenotype genes --- p.60 / Chapter 3.3.4 --- Effects of the identified CKIP-1 siRNA on matrix mineralization --- p.65 / Chapter 3.3.5 --- Effect of the identified CKIP-1 siRNA on BMP signaling --- p.67 / Chapter 3.3.6 --- Effects of the identified CKIP-1 siRNA on the ratio of RANKL/OPG --- p.67 / Chapter 3.3.7 --- Effects of the identified CKIP-1 siRNA on immunostimulatory activity --- p.68 / Chapter 3.4 --- Summary --- p.71 / Chapter 3.4.1 --- CKIP-1 siRNA si-3 as the identified sequence --- p.71 / Chapter 3.4.2 --- CKIP-1 siRNA si-3 promoted osteoblast differentiation in vitro --- p.72 / Chapter CHAPTER 4 --- Validation of the identified CKIP-1 siRNA in healthy rodents in vivo --- p.74 / Chapter 4.1 --- Introduction --- p.74 / Chapter 4.2 --- Materials and methods --- p.74 / Chapter 4.2.1 --- Localization of intraosseous siRNA delivered by (Asp-Ser-Ser)₆-liposome --- p.75 / Chapter 4.2.2 --- Cell-selective delivery in vivo of CKIP-1 siRNA --- p.76 / Chapter 4.2.3 --- Dose-response study of CKIP-1 siRNA --- p.77 / Chapter 4.2.4 --- Time-course study of CKIP-1 siRNA --- p.77 / Chapter 4.2.5 --- Examination of the effect of the identified siRNA on the expression of osteoblast phenotype genes --- p.78 / Chapter 4.2.6 --- Measurement for serum PINP and urinary DPD --- p.80 / Chapter 4.2.7 --- 5’-RACE Analysis --- p.81 / Chapter 4.2.8 --- Laser captured micro-dissection (LCM) --- p.82 / Chapter 4.2.9 --- Evaluation the anabolic effect of the identified siRNA on healthy rat bone --- p.82 / Chapter 4.2.10 --- Evaluation the anabolic effect of the identified siRNA on healthy mouse bone --- p.84 / Chapter 4.2.11 --- Micro CT analysis --- p.84 / Chapter 4.2.12 --- Dynamic bone histomorphometric analysis --- p.85 / Chapter 4.2.13 --- Statistics --- p.86 / Chapter 4.3 --- Results --- p.87 / Chapter 4.3.1 --- Rationale of bone targeted delivery of CKIP-1 siRNA by (Asp-Ser-Ser)₆-liposome --- p.87 / Chapter 4.3.2 --- Intraosseous distribution of siRNA delivered by (Asp-Ser-Ser)₆-liposome --- p.89 / Chapter 4.3.3 --- Optimal dosage and duration for CKIP-1 siRNA administration in vivo --- p.92 / Chapter 4.3.4 --- Knockdown efficiency of CKIP-1 siRNA in osteoblasts by LCM in combination with Q-PCR --- p.94 / Chapter 4.3.5 --- Examination of the effect of the identified siRNA on the expression of osteoblast phenotype genes --- p.96 / Chapter 4.3.6 --- RNAi mechanism of CKIP-1 siRNA action in vivo --- p.99 / Chapter 4.3.7 --- Anabolic effect of the identified siRNA on healthy rat bone --- p.101 / Chapter 4.3.8 --- Anabolic effect of the identified siRNA on healthy mouse bone . --- p.104 / Chapter 4.4 --- Summary --- p.107 / Chapter 4.4.1 --- Intraosseous localization of CKIP-1 siRNA after systemic administration --- p.107 / Chapter 4.4.2 --- Evidence of RNAi in bone tissue from systemic administration of CKIP-I siRNA --- p.107 / Chapter 4.4.3 --- CKIP-1 siRNA si-3 promots bone formation in rats and mice in vivo --- p.108 / Chapter CHAPTER 5 --- Anabolic effect of the identified CKIP-1 siRNA on bone in postmenopausal osteoporostic animal models --- p.110 / Chapter 5.1. --- Introduction --- p.110 / Chapter 5.2. --- Materials and Methods --- p.110 / Chapter 5.2.1. --- Evaluation of anabolic effect of CKIP-1 siRNA on osteoporotic mouse bone --- p.111 / Chapter 5.2.2. --- Evaluation of anabolic effect of CKIP-1 siRNA on osteoporotic rat bone --- p.112 / Chapter 5.2.3. --- In vivo micro-CT analysis and registration of proximal tibia from osteoporotic rats --- p.112 / Chapter 5.2.4. --- Ex vivo micro-CT analysis of the distal femur and 5th lumbar vertebrae body of osteoporotic rats --- p.115 / Chapter 5.2.5. --- Ex vivo micro-CT analysis of distal femur from osteoporotic mice --- p.115 / Chapter 5.2.6. --- Bone histomorphometric analysis --- p.116 / Chapter 5.2.7. --- Mechanical testing --- p.117 / Chapter 5.2.8. --- Statistics --- p.118 / Chapter 5.3. --- Results --- p.116 / Chapter 5.3.1. --- Anabolic effect of CKIP-1 siRNA si-3 on osteoporotic mouse bone --- p.118 / Chapter 5.3.2. --- In vivo microCT data of proximal tibia from aged osteoporotic rats --- p.121 / Chapter 5.3.3. --- Ex vivo microCT data of distal femur from aged osteoporotic rats --- p.124 / Chapter 5.3.4. --- Ex vivo microCT data of 5th LV body from aged osteoporotic rats --- p.126 / Chapter 5.3.5. --- Bone histomorphometric analysis of aged osteoporotic rats --- p.129 / Chapter 5.3.6. --- Mechanical testing of the mid-shaft femur of aged osteoporotic rats --- p.132 / Chapter 5.4. --- Summary --- p.134 / Chapter CHAPTER 6 --- Discussions --- p.134 / Chapter 6.1 --- CKIP-1 siRNA design rationale and further modification --- p.135 / Chapter 6.1.1 --- Specificity design rationale of the CKIP-1 siRNA --- p.135 / Chapter 6.1.2 --- Stability enhancing modification of CKIP-1 siRNA --- p.136 / Chapter 6.1.3 --- Safety concerns with CKIP-1 siRNA therapy --- p.136 / Chapter 6.2 --- Development of bone-targeted siRNA delivery --- p.136 / Chapter 6.3 --- Prospects for and limitation of application of study findings to clinical therapeutics --- p.137 / References --- p.139 / Publications --- p.159

Diss. (sammanfattning) Umeå : Umeå universitet, 2009. / Härtill 4 uppsatser. Även tryckt utgåva.

Wong Chi Ming. / "August 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 182-215). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.