• Journal of Korean Neurosurgical Society
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• v.63 no.6
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• pp.794-805
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• 2020

Objective : Teriparatide is known as an effective anabolic agent not only for severe osteoporosis but also for bone healing and union. We explored the possibility of teriparatide as an alternative treatment option for osteoporotic thoracolumbar (TL) burst fracture. Methods : This retrospective study enrolled 35 female patients with mean age of 73.77±6.71 years (61-88) diagnosed as osteoporotic TL burst fracture with ≥4 of thoracolumbar injury classification and severity (TLICS) score and no neurological deficits. All patients were treated by teriparatide only (12 of group A), teriparatide plus vertebroplasty (12 of group B), or surgical fixation with fusion (11 of group C), and followed up for 12 months. Radiological outcomes were evaluated using radiological parameters including kyphotic angle (KA), segmental vertebral kyphotic angle (SVKA), compression ratio (CR), and vertebral body height (anterior [AH], middle [MH], posterior [PH]). Functional outcomes were evaluated using visual analog scale (VAS) and Macnab classification (MC). Results : There were no statistical significant differences in age, bone mineral density (-3.36±0.73), and TLICS score (4.34±0.48) among the three groups (p>0.05). Teriparatide was administered during 8.63±2.32 months in group A and B. In 12-month radiological outcomes, there were significant restoration in SVKA, CR, AH, and MH of group B and KA, SVKA, CR, AH, and MH of group C compared to group A with no radiological changes (p<0.05). All groups showed similar significant improvements in 12-month functional outcomes, although group B and C showed a better 1-month VAS, 1-month MC, 3-month MC compared to group A (p<0.05). Conclusion : Non-surgical treatment with teriparatide showed similar 12-month functional outcomes compared to surgical fixation with fusion. The additional vertebroplasty to teriparatide and surgical fixation with fusion were more helpful to improve short-term functional outcomes with structural restoration compared to teriparatide only.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1262-1269
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• 2008

Low intensity pulsed ultrasound(LIPUS) is known to accelerate bone regeneration, but the precise cellular signaling mechanism is still unclear. The purpose if this study was to determine the effect of LIPUS on the signaling mechanism of rat chondrocyte. In the explant culture condition, there was inhibition effect of 1 $W/cm^2$ intensity LIPUS on chondrocytes proliferation but chondrocytes proliferation was increased at 0.25 $W/cm^2$ intensity. In addition, western blot analysis of MAPKs showed that LIPUS increased ERK1/2 activity from the 10 min treatment of LIPUS. Hydrogen peroxide($H_2O_2$), resulted in a time- and dose-dependent cell proliferation, which was largely attributed to apoptosis. $H_2O_2$ treatment caused marked sustained nucleus condensation in Hoechst stain. LIPUS and $H_2O_2$ activates phosphorylation of p-ERK1/2 and PD 98059($10^{-5}M$) blocked the effect of LIPUS and $H_2O_2$. Moreover, the synergistic phosphorylation of p44/42 MAPK by $H_2O_2$, LIPUS was selectively inhibited by PD 98059, ERK1/2 inhibitor. In order to determine whether the increase in cell proliferation caused by $H_2O_2$ and LIPUS could be explained by changes in the level of the prostaglandin $E_2$. Our study demonstrated that LIPUS stimulate the cell proliferation via activated phosphorylation of ERK1/2 in condrocyte. LIPUS has anabolic effects on rat cartilage in explant cultures, indicating a potential important method for the treatment of osteoarthritic cartilarge.

• Journal of the korean academy of Pediatric Dentistry
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• v.25 no.3
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• pp.635-648
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• 1998

The clinical use of fluoride with a well known osteogenic action in osteoporotic patients is rational, because this condition is characterized by impaired bone formation. However, its anabolic effect has not been demonstrated well in vitro. The purpose of this study was to investigate the effects of sodium fluoride on the physiological role of osteoblastic cell. Osteoblastic cells were isolated from fetal rat calvaria. The results were as follows : 1. Mineralized nodules were shown in osteoblastic cell cultures, which had been maintained in the presence of ascorbic acid and ${\beta}-glycerophosphate$ up to 21 days. When cultures were treated with pulses of 48 hr duration before apparent mineralization was occurring, 2-fold increased in their number was detected. 2. Alkaline phosphatase activity of osteoblastic cells was inhibited by sodium fluoride in dose dependent manner. 3. The effect of sodium fluoride on the osteoblastic cell proliferation was measured by the incorporation of $[^3H]$-thymidine into DNA. As a result, sodium fluoride at $1{\sim}100{\mu}M$ increased the $[^3H]$-thymidine incorporation into DNA in a dose dependent manner. 4. The signaling mechanism activated by sodium fluoride dose-dependently enhanced the tyrosine phosphorylation of the adaptor molecule $Shc^{p66}$ and their association with Grb2, one of earlier events in a MAP kinase activation pathway cascade used by a significant subset of G protein-coupled receptors. 5. The phosphorylation of CREB(cAMP response element binding protein)was inhibited by the sodium fluoride in MC3T3E1 cells. In conclusion, the results of this study suggested that the mitogenic effect of the sodium fluoride in MC3T3E1 cell was stimulated in a dose-dependent manner and suggested "an important role for the interaction between She and Grb2" in controlling the proliferation of osteoblasts.

• YAKHAK HOEJI
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• v.58 no.5
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• pp.307-313
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• 2014

Rumex crispus (curled dock), which is a perennial wild plant, has long been used as a laxative, astringent, and medicine to treat blood and skin diseases. We recently reported that the roots of R. crispus are an effective nutraceutical for bone. This study prepared ethanol extracts of the leaves and roots of R. crispus, and analyzed the major constituents using liquid chromatography and mass spectrometry. In addition, their effects on the proliferation and differentiation of human osteoblast-like MG-63 cells, such as cell viability, alkaline phosphatase (ALP) activity, collagen content, and mineralization, were compared. The chromatograms of the chemical constituents of the two extracts exhibited quite different profiles: quercetin and quercitrin were identified as major peaks in the leaf extract, whereas cinnamtannin B1 and procyanidin isomers were the major peaks for the root extract. Neither extract was cytotoxic at concentrations of < $25{\mu}g/ml$. ALP activity and collagen synthesis-which are markers of the early stage of osteogenesis-in MG-63 cells were significantly increased upon the addition of the root extract compared with the addition of the leaf extract. In contrast, the leaf extract had a more stimulatory effect on mineralization-which is marker of the late stage of osteogenesis-in MG-63 cells than did the root extract. In conclusion, extracts of both leaves and roots of R. crispus stimulated the bone-forming activity of osteoblasts; in particular, the root extract was more effective in the early stage of osteoblast differentiation, while the leaf extract was more effective in the late stage. This difference in anabolic activity may be due to differences in the constituents of the leaves and roots. The leaves and roots of R. crispus appear to complement each other as stimulators of bone formation.

• Molecules and Cells
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• v.41 no.3
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• pp.234-243
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• 2018

In recent years, the interest towards the relationship between incretins and bone has been increasing. Previous studies have suggested that glucagon-like peptide-1 (GLP-1) and its receptor agonists exert beneficial anabolic influence on skeletal metabolism, such as promoting proliferation and differentiation of osteoblasts via entero-osseous-axis. However, little is known regarding the effects of GLP-1 on osteoblast apoptosis and the underlying mechanisms involved. Thus, in the present study, we investigated the effects of liraglutide, a glucagon-like peptide-1 receptor agonist, on apoptosis of murine MC3T3-E1 osteoblastic cells. We confirmed the presence of GLP-1 receptor (GLP-1R) in MC3T3-E1 cells. Our data demonstrated that liraglutide inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as detected by Annexin V/PI and Hoechst 33258 staining and ELISA assays. Moreover, liraglutide upregulated Bcl-2 expression and downregulated Bax expression and caspase-3 activity at intermediate concentration (100 nM) for maximum effect. Further study suggested that liraglutide stimulated the phosphorylation of AKT and enhanced cAMP level, along with decreased phosphorylation of $GSK3{\beta}$, increased ${\beta}-catenin$ phosphorylation at Ser675 site and upregulated nuclear ${\beta}-catenin$ content and transcriptional activity. Pretreatment of cells with the PI3K inhibitor LY294002, PKA inhibitor H89, and siRNAs GLP-1R, ${\beta}-catenin$ abrogated the liraglutide-induced activation of cAMP, AKT, ${\beta}-catenin$, respectively. In conclusion, these findings illustrate that activation of GLP-1 receptor by liraglutide inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation through $cAMP/PKA/{\beta}-catenin$ and $PI3K/Akt/GSK3{\beta}$ signaling pathways.

• Journal of Life Science
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• v.21 no.2
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• pp.300-308
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• 2011

Osteoporosis is a disease involving a decrease in bone mineral density and increased risk of fractures. Osteoblast and osteoclast activities are important for bone formation. The MC3T3-E1 osteoblastic cell line is a well-accepted model of osteogellsis in vitro. Hijikia fusiforme is a kind of edible brown seaweed that grows mainly in the Northwest Pacific region, including the countries of Korea, Japan and China, and it has been widely used as a medicinal and health food in Korea. In this study, by using osteoblasts, the effects of Hijikia fusiforme fractions on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and mineralization of cells were investigated. Hijikia fusiforme were subjected to fractionation by using hexane, methanol, butanol and aqueous. Proliferation of the MC3T3-E1 osteoblastic cells that were treated with Hijikia fusiforme fractions increased by approximately 120%. Regarding effects of Hijikia fusiforme fractions on ALP activity, 1 ${\mu}g$/ml butanol fraction showed the highest activity. The synthesis of collagen increased significantly in response to treatment with Hijikia fusiforme fractions, with the exception of the hexane fraction. Moreover, mineralization in the MC3T3-E1 cells that were treated with 100 ${\mu}g$/ml butanol fraction increased by 281%. Also, when 100 ${\mu}g$/ml aqueous fraction was added, mineralization increased by 240%. These results indicate that Hijikia fusiforme fractions have anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.664-670
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• 2016

Cheonggukjang (CKJ) is a Korean traditional food made of fermented soybeans. In comparison to normal intake of soybeans, Cheonggukjang has high digestibility with bioactive, antioxidant substances, and thrombolytic enzymes. Recent studies have reported anti-oxidant, anti-cancer, anti-inflammatory, anti-obesity activities as well as inhibitory activities against osteoporosis for CKJ. In this study, we identified the effects of CKJ on osteoblast differentiation by increasing the polyglutamic acid (PGA) content of CKJ. Alkaline phosphatase (ALP) activity and mineralization significantly increased in response to treatment with both natural CKJ (CKJ A) and PGA-increased CKJ (CKJ B). However, CKJ B exhibited higher ALP activity and mineralization than CKJ A. Real-time reverse transcription PCR demonstrated that mRNA expression of osteoblastic-associated genes such as type I collagen, alkaline phosphatase, osteocalcin, and osteopontin in C2C12 cells was significantly up-regulated by CKJ A or B treatment. These results indicate that treatment with CKJ has an anabolic effect on bone by increasing osteoblastic differentiation and ALP activity. Increasing PGA content in CKJ had a greater effect than CKJ A on up-regulation of osteoblastic gene expression in osteoblast cells.

• Journal of Life Science
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• v.20 no.4
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• pp.607-613
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• 2010

Osteoporosis is a disease involving a decrease in bone mineral density and an increased risk of fractures. The MC3T3-E1 pre-osteoblastic cell line is a well-accepted model of osteogenesis in vitro. Pine needles have long been used as a traditional health-promoting medicinal food in Korea. In this study, MTT assay, the alkaline phosphatase (ALP) activity and collagen synthesis of osteoblast cells were investigated to determine the effects of pine needle extracts on cell proliferation and differentiation. Pine needle extracts were prepared using hexane, ethanol and water. The effects of the pine needle extracts were examined by comparing the results with those of commercial agents, such as proanthocyanidin. The MC3T3-E1 cells exposed to proanthocyanidin showed increased proliferation in a concentration-dependent manner. The cells exposed to the hexane extract showed a similar increase in proliferation to that observed with proanthocyanidin. The hexane extract showed the highest ALP activity. Moreover, a supplement of pine needle extracts induced collagen synthesis in MC3T3-E1 cells. The pine needle extract produced the highest level of collagen synthesis at concentrations of $10{\sim}50\;{\mu}g/ml$. These results indicate that pine needle extracts have an anabolic effect on bone by promoting osteoblastic differentiation, and may be used in the treatment of common metabolic bone diseases.

• Journal of Life Science
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• v.21 no.12
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• pp.1752-1760
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• 2011

In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.4
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• pp.525-530
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• 2011

Pine needles have long been used as a traditional health-promoting medicinal food in Korea. This study was carried out to investigate the effects of pine needle extracts on the antioxidant activity, and proliferation of osteoclastic RAW 264.7 cells. Pine needle extracts were examined using hot water, ethanol, hexane, hot water-ethanol, and hot water-hexane. The effects of the pine needle extracts were examined by comparing the results with that of a commercial agents, proanthocyanidin. Analysis of each extract indicated that hot water-ethanol and ethanol extracts contained the highest total polyphenol concentrations. The hot water-ethanol and ethanol extracts also showed relatively the highest SOD-like activity. The proliferation of osteoclastic RAW 264.7 cells treated with pine needle extracts was decreased by lower than 70%. In addition, the hot water and ethanol extracts of pine needle significantly reduced the number of tartrate-resistant acid phosphatase-positive ($TRAP^+$) multinucleated cells from osteoclatic RAW 264.7 cells. These results indicate that pine needle extracts had an anabolic effect on bone through the promotion of osteoclast differentiation, suggesting that they could be used for the treatment of common metabolic bone diseases.