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BACKGROUND: Diabetes mel itus is one of the most common systemic diseases, which often leads to the changes of the jaw and other bone structure, as wel as the abnormal changes of mineral metabolism. OBJECTIVE: To observe the three-dimensional structure and histopathological changes of the mandible in type 1 diabetes mel itus mice. METHODS: The mice were randomly divided into control group and diabetes mel itus group. The diabetes mel itus group received intraperitoneal injection of 50 mg/kg streptozotocin for 5 days to establish a type 1 diabetes mel itus model, and the control group received intraperitoneal injection of citrate buffer. RESULTS AND CONCLUSION: At 3 weeks after modeling, the micro-CT technique was used to observe the three-dimensional structure of the mandibles in the two groups. The quantitative analysis on the microstructure of cancel ous bone and cortical bone showed that the bone mineral density, bone volume fraction, trabecular number and trabecular thickness of cancel ous bone in the interest region in the mandible of type 1 diabetes mel itus mice were significantly decreased when compared with that in the control group (P < 0.01, P < 0.05), while the structure model index was increased significantly (P < 0.05); the mineral density and area of cortical bone were decreased in the diabetes mel itus group (P < 0.05). Hematoxylin-eosin staining showed that the number and volume of mandibular trabeculae of type 1 diabetes mel itus mice were decreased. The results suggest that the three-dimensional structure of the cancel ous bone and cortical bone in the streptozotocin-induced type 1 diabetes mel itus mice are changed significantly, and the microstructure change of the cancel ous bone is more obvious.
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BACKGROUND: The transcription factor Runx2 is the key factor that regulates osteogenic differention and bone development. It has been reported that the C2C12 mesenchymal cells can be induced to differentiate into osteoblasts by Runx2 overexpression, but the molecular mechanism of induction is stil largely unclear. OBJECTIVE: To investigate the role of the members of the miR-376 family during Runx2-induced osteogenic differentiation in C2C12 cells. METHODS: The expression of the members of the miR-376 family was detected by real-time quantitative PCR at different time points using C2C12/Runx2Dox sub-line with conditional Runx2 expression. In miR-376b-3p-transfected C2C12/Runx2Dox cells, the expression of osteoblast markers, such as alkaline phosphatase and osteocalcin, was detected by real-time quantitative PCR, and the alkaline phosphatase activity was also examined by alkaline phosphatase staining. The putative miR-376b-3p targets were commonly predicted by online tools (miRanda, miRWalk and TargetScan). The functional classification of these putative targets was performed by DAVID Bioinformatics Resources database. RESULTS AND CONCLUSION: The expression of miR-376b-3p was significantly increased during Runx2-induced osteogenic differentiation of C2C12 cells, but the expression of other members was not changed. Transfection of miR-376b-3p mimic upregulated alkaline phosphatase expression, but had no effect on osteocalcin expression. The alkaline phosphatase activity was also increased by transfection of miR-376b-3p. The functional classification of miR-376b-3p putative targets showed that miR-376b-3p is involved in the skeleton development, indicating the role of miR-376b-3p in osteoblast differentiation. Taken together, these results suggest that Runx2 promotes early osteogenic differentiation in C2C12 cells by regulating the expression of genes related to osteogenic differentiation through upregulation of miR-376b-3p.
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BACKGROUND: Nitric oxide plays an important role in bone metabolism. However, the effects of different doses of L-arginine, nitric oxide substrate, on the healing of osteoporotic fractures remain unclear. OBJECTIVE: To investigate the influence of different doses of L-arginine on the healing of osteoporotic fractures and blood biochemistry in rats. METHODS: A total of 50 female Wistar rats, aged 6-month-old, were randomly divided into sham-surgery (n=10) and osteoporosis (bilateral ovariectomy, n=40). After osteoporosis model was established, left middle femoral fractures were made in al 50 rats, and the osteoporosis group was further divided into four groups, low, middle and high dose of L-arginine, and control groups. The L-arginine groups were intraperitoneal y injected with 1, 3, and 5 mg/kg L-arginine at the second day fol owing surgery, while the control and sham-surgery groups were injected with the same volume of normal saline. At 8 weeks, the lumbar and cal us bone mineral density, serum nitric oxide, and nitric oxide synthase were detected. Meanwhile, the bone cal us histological examination was conducted. RESULTS AND CONCLUSION: During fracture healing, osteoporosis rats showed a low level of serum nitric oxide and nitric oxide synthase compared with normal fractured rats (P < 0.05). L-arginine can improve the concentration of serum nitric oxide and nitric oxide synthase in osteoporosis rats. Moreover, middle dose of L-arginine can increase the cal us and lumbar spine bone mineral density of osteoporosis rats, so the number and continuity of bone trabecula were enhanced. These findings suggest that middle dose of L-arginine (3 mg/kg) can promote healing process of osteoporotic fractures and improve osteoporosis.
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BACKGROUND: Vascular endothelial growth factor play an important role in promoting healing of osteoporotic fractures, but whether it can affect the bone mineral density is not clear. OBJECTIVE: To observe the correlation between serum vascular endothelial growth factor, bone mineral density and the number of osteoblasts in the ovariectomized rats. METHODS: Forty female Sprague-Dawley rats were randomly divided into ovariectomized group and control group. After 3 months, the bone mineral density of the whole body, femur and lumbar spine was measured. Rat enzyme-linked immunosorbent assay kit was used to measure the level of serum vascular endothelial growth factor. Then, the rats in two groups received femoral metaphyseal fixation, decalcified, dehydrated, embeding in paraffin, slicing and hematoxylin-eosin staining. Each slice was free to take five fields of view (10×40) in order to count the osteoblasts of femur distal metaphysis under optical microscope. RESULTS AND CONCLUSION: After ovariectomized for 3 months, the rats body mass was increased significantly (P 0.05), and the difference of the osteoblast number between ovariectomized group and control group was not significant (P > 0.05). This indicated that there was no correlation between bone mineral density and the number of osteoblasts and vascular endothelial growth factor level in the ovariectomized group and the control group. These findings suggest that the bone mineral density is reduced and the body mass is increased in the ovariectomized rats, and the reduced bone mineral density of ovariectomized rats may be irrelevant with the change of serum vascular endothelial growth factor.
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BACKGROUND: Now, dual-energy X-ray absorptiometry is international y recognized as gold standard for the diagnosis of osteoporosis, but the errors can be found in the measurement results due to the heterotopic ossification and bone hyperplasia exists in the measurement part. OBJECTIVE: To investigate the clinical significance of bone metabolic markers in the diagnosis and treatment of elderly patients with osteoporotic fractures, and to research its correlation with the changes of pathological histology and bone mineral density. METHODS: Four bone biochemical markers in 50 elderly patients with osteoporosic fractures were measured preoperatively. According to the results, 25 patients had significantly increased tartrate-resistant acid phosphatase 5b (considered as the increased tartrate-resistant acid phosphatase 5b group), and 25 patients had increased bone alkaline phosphatase (considered as the increased bone alkaline phosphatase group). During operation, the bone tissues of eight patients in each group were treated with hematoxylin-eosin staining and electron microscopy scanning in order to detect the pathological changes. After operation, the patients in the increased tartrate-resistant acid phosphatase 5b group received salmon calcitonin anti-osteoporosis treatment, and the patients in the increased bone alkaline phosphatase group received the anti-osteoporosis treatment of bone peptide injection. The bone mineral density and the four bone biochemical markers were detected again at 6 months after treatment. RESULTS AND CONCLUSION: There were no significant differences in the preoperative bone mineral density and four biomechanical markers between two groups (P > 0.05). The pathological examination results of bone tissue on the fracture site showed that the number of osteoblasts was reduced and the number of oeteoclasts was increased in the increased tartrate-resistant acid phosphatase 5b group; while in the increased bone alkaline phosphatase group, the pathological examination results showed the number of osteoblasts was reduced; the trabecular bone/bone area ratio was decreased in two groups, and there was a significant difference in the decrease degree between two groups (P < 0.05). The electron microscope scanning showed that the osteoclasts of two groups were more active than that of the normal group. The sloppy of trabecular bone in the increased tartrate-resistant acid phosphatase 5b group was more obvious than that in the increased bone alkaline phosphatase group, and the absorption vacuoles were increased. There were significant differences in the bone mineral density and four biomechanical markers between two groups before and after anti-osteoporosis treatment (P < 0.05). The detection of bone metabolic markers could help us to make it clearly that the main function of osteoblast reduce or osteoclast increase in bone tissue of patients, and guide us to use anti-osteoporosis drugs in target. Pathological histology examination can better reflect the condition of osteoblasts, osteoclasts and trabecular bone in bone tissue on the fracture site. Target application of anti-osteoporosis drugs in the osteoporosis patients can effectively improve the efficacy and reduce the relative complications.
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10.3969/j.issn.2095-4344.2013.25.010
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10.3969/j.issn.2095-4344.2013.24.002
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10.3969/j.issn.2095-4344.2013.24.004
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BACKGROUND:The reports on bone morphogenetic protein-7 as a stimulating factor to induce osteogenic are relatively rare. OBJECTIVE:To study the expression of alkaline phosphatase of periosteal cel s after induced by bone morphogenetic protein-7 in vitro. METHODS:Periosteal cel s were obtained from adult tibial periosteum, and then the periosteal cel s were cultured by routine method in vitro. The cel s were divided into experimental group and control group, and then cultured with bone morphogenetic protein-7 plus osteoblast culture adjuvants and simple osteoblast culture adjuvants, respectively. The phase contrast microscope was used to observe the morphology and ultrastructure of periosteal cel s. Each group was observed at 7, 14 and 21 days, and three samples were observed at each time point. Alkaline phosphatase kit was used to detect the expression of osteoblast-specific markers alkaline phosphatase. RESULTS AND CONCLUSION:After cultured for 7 days, the proliferation of periosteal cel s in the experimental group and the control group was increased obviously, and the expression of alkaline phosphatase was detected but less. The cel s were spindle in shape, while the expression of alkaline phosphatase in the experimental group was higher than that in the control group. After cultured for 14 days, the proliferation of periosteal cel s in the experimental group and the control group was increased obviously, the cel morphology was changed from spindle-shaped to wide spindle-shaped, and the expression of alkaline phosphatase in the experimental group was increased significantly when compared with the control group. After cultured for 21 days, the proliferation of periosteal cel s was detected in the experimental group and the control group, and the proliferation in the experimental group was more significant than that in the control group, the cel morphology was wide spindle-shaped, and the number of alkaline phosphatase in the experimental group was higher than that in the control group. Statistical analysis showed that the positive rate of osteogenic markers alkaline phosphatase of bone morphogenetic protein-7 induced periosteal cel s in the experimental group was higher than that in the control group (Posteogenic and regeneration ability, the bone morphogenetic protein-7 could induce periosteal cel s, promote the expression of alkaline phosphatase, and could induce the periosteal cel s to transform into osteoblasts.
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BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis in Fujian University of Traditional Chinese Medicine, and the previous studies mainly focus on effect to cartilage. OBJECTIVE:To observe the effect of Tougu Xiaotong capsule on the proliferation and differentiation of osteoblasts as wel as the expressions of bone remodeling correlated factors. METHODS:Rat osteoblast-like cel line ROS17/2.8 cel s were incubated with Tougu Xiaotong capsule. The ROS17/2.8 cel s were divided into blank control group and Tougu Xiaotong capsule groups with different concentrations. The cel proliferation was determined by methylthiazolyldiphenyl-tetrazolium bromide assay. Osteoblast differentiation biomarkers alkaline phosphatase activity, osteocalcin and bone mineralized nodules were measured with colorimetry, enzyme-linked immunosorbent assay and alizarin red staining, respectively. The real-time PCR and enzyme-linked immunosorbent assay were used to detect the expressions of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand. RESULTS AND CONCLUSION:Compared with the control group, the Tougu Xiaotong capsule with the concentration of 0.25-2 g/L could significantly promote the ROS17/2.8 cel proliferation (Ppercentage of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand (Pproliferation and differentiation of osteoblasts and bone remodeling.
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BACKGROUND:Sclerostin can negatively regulate the bone metabolism, and the sclerostin monoclonal antibody can antagonize the negative regulation effect, inhibit bone resorption and promote bone formation. OBJECTIVE:To explore the mechanism and application progress of sclerostin monoclonal antibody in the treatment of osteoporosis. METHODS:An online search of PubMed database, CNKI database, VIP database and Wanfang database between May 2005 and May 2013 was performed by the first author to search the related articles with the key words of“osteoporosis, antibody, sclerostin, Wnt, SOST”in both English and Chinese. Articles related to sclerostin monoclonal antibody were included. For the articles in the same field, those published earlier or in the authorized journals were preferred. RESULTS AND CONCLUSION:A total of 170 articles were obtained after initial search, and final y 54 articles related to sclerostin monoclonal antibody were included for review according to the inclusion criteria. The sclerostin can block Wnt pathway through combining with low-density lipoprotein receptor-related protein 5/6, thus inhibiting the differentiation and mineralization of osteoblasts. By specifical y binding to sclerostin, the sclerosin monoclonal antibody can indirectly promote bone formation and restrain bone absorption which has great significance in the treatment of osteoporosis. Meanwhile, compared with the other treatment method, the specific targeting of sclerostin and the binding specificity of sclerostin monoclonal antibody provide application advantages for the treatment of osteoporosis.
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BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear. OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts. METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse transcription-PCR in al groups. RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (Pcore binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.