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XLGB-B prevents bone loss in OVX mice by inhibiting bone remodeling
Abstracts / Bone 47 (2010) S385–S458
AKT signaling pathway during osteogenic differentiation of MSCs. Taken together, our findings suggest that a combination of BMP9 and IGF2 may be explored as an effective bone regeneration agent to treat large segmental bony defects, non-union fracture, and/or osteoporatic fracture. doi:10.1016/j.bone.2010.09.267
308 Increase rate of cellular osteogenic mineralization by low-intensity ultrasound in a stimulated microgravity Y.-X. Qin, S.Z. Uddin, S. Zhang, M. Hu, Y. Huang, J. Cheng Orthopaedic Bioengineering Research Laboratory, Department of Biomedical Engineering Stony Brook University, NY, USA Objective: Osteoporosis and disuse osteopenia are increased health problems that increase the risk of fracture. Microgravity causes bone loss in the skeleton potentially due to decrease in bone mineralization by osteoblast cells. In vitro studies using stimulated microgravity have shown significant decrease in expression of osteoblastic transcription factors and low alkaline phosphatase activity (ALP). Low-intensity pulse ultrasound (LIPUS) has shown capability in promoting bony ingrowth and accelerating rate of mineralization in osteoblast cells by up regulating expression of different osteoblastic transcription factors, matrix proteins and ALP activity. Ultrasound has been known to increase calcium deposition in the osteoblast cells. The objective of this study was to study the effects of LIPUS on osteoblast mineralization in stimulated microgravity. Methods: Human fetal osteoblast (hfob 1.19) was cultured in DMEM:F12 media supplemented with 15% FBS and 0.3 mg/ml G418 with initial seeding density of 50,000 cell per Opticell cartilage. Cells were allowed to grow to confluency, and stimulations were started at day 7 of post confluency. Cells were distributed into four groups (n = 4 each), gravity, gravity + LIPUS, simulated microgravity and microgravity + LIPUS. Stimulated microgravity environment was created by putting Opticells into an in vitro rotary system with rotation set at 15 RPM. LIPUS groups were stimulated daily for total of 14 days with 30 mW/cm2 for 20 min. Cells were kept at 37 °C, 5% CO2 environment during experiment and medium was changed every second day. After 14 days of stimulations, cells were analyzed for calcification by staining with Alizarin red stain and cytoskeleton changes with antibodies against actin and microtubules. Only the area stimulated with ultrasound was used for analyses. Results: Alizarin red (AZ) staining showed significant increase in calcification in microgravity+LIPUS group (2.70±0.03 mM AZ/106 cells) compared to microgravity group (2.16±0.24 mM AZ/106 cells, p=0.023). Significant decrease of calcification was also observed (47%) from gravity (3.18± 0.21 mM AZ/106 cells) group to the microgravity group (2.16±0.24 mM AZ/106 cells, p=0.009). Higher mineralization was observed in the gravity+LIPUS group (3.69±0.31 mM AZ/106 cells) compared to gravity group (3.18±0.21 mM AZ/106 cells, p=0.08). It was also observed that microgravity samples had fewer cells. Conclusion: These results imply that LIPUS increases mineralization in osteoblast cells in a microgravity environment. It suggests that ultrasound may serve as a noninvasive modality to provide localized treatment for bone loss at critical skeletal sites. The system may also provide a novel way to simulate microgravity conditions for application of space medicine. Acknowledgment This study was supported by NIH. doi:10.1016/j.bone.2010.09.268
S433
309 XLGB-B prevents bone loss in OVX mice by inhibiting bone remodeling Xin-luan Wang1, Ge Zhang1, Yi-xin He1, Li-zhen Zheng1, Feng-juan Tu2, Xin-sheng Yao2, Ling Qin1,3 1 Musculoskeletal Research Laboratory, Department of Orthopaedics & Traumatology, The Chinese University of Hong Kong, Hong Kong, China 2 Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, Jinan University, Guangzhou, China 3 Translational Medicine R&D Center, Institute of Biomedical & Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China Objectives: “Xian Ling Gu Bao capsule” (XLGB) is the only traditional Chinese medicine for treatment of osteoporosis in the health insurance list in China. Randomized, multicenter, double-blind, placebo-controlled clinical trial demonstrated XLGB statistically significantly increased bone mineral density (BMD) in the lumbar spine at 6 months. Three fractions (XLGB-A, B, and C) were prepared for seeking the key fraction (s) for preventing osteoporosis. In this study the effect of XLGB-B on preventing osteoporosis was investigated. Material and methods: Thirty, 4-month-old female C57/BL6J mice were either sham operated (Sham Group) or OVX. After recovering for 3 days, the OVX mice were randomly divided into two groups: OVX Group and XLGB-B Group (31 mg/kg/day). Treatments were given orally for 1 month. 3D bone structure of the sixth lumber was analyzed using Micro-CT (Extreme CT, Scanco Medical, Brüttisellen, Switzerland) with an isotropic resolution of 12 μm in all three spatial dimensions. The trabecular bone volume (BV/TV, %) was measured. Concentrations of N-terminal propeptides of type I procollagen (PINP, bone formation marker) and C-propeptide of type I collagen (CTX, bone resorption marker) in serum were measured using ELISA kits (Immunodiagnostic Systems Ltd. UK). Results: 3D images of the 6th lumbar clearly demonstrated that the trabecular bone in Sham and XLGB-B group was obviously more than that in OVX group (p < 0.05). Similarly, BV/TV in Sham and XLGB-B group was significantly higher than that in the OVX group (p < 0.05). Both the concentrations of PINP and CTX in serum of the OVX group increased compared to that of the Sham group, which indicated that OVX could promote the bone remodeling, and XLGB-B significantly inhibited the increases of both PINP and CTX induced by OVX (p < 0.05). Conclusion: In conclusion, XLGB-B could prevent bone loss in OVX mice by inhibiting bone remodeling.
Acknowledgment This work was supported by Tongjitang Pharmaceutical Co. Ltd. doi:10.1016/j.bone.2010.09.269
311 Poor bone quality in old Chinese women with bound feet—A QUS study Mian Xu1, Hanchang Lao1, Yi Pan1, Vivian Wing-yin Hung2, Ming Zhang3, Ling Qin4 1 The 2nd Affiliated Hospital of Kunming Medical College, Kunming, China 2 Department of Orthopaedics & Traumatology, Bone Quality & Health Centre, The Chinese University of Hong Kong, Hong Kong, China 3 Department of Health Technology & Informatics, The Hong Kong Polytechnic University, Hong Kong, China 4 Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
AKT signaling pathway during osteogenic differentiation of MSCs. Taken together, our findings suggest that a combination of BMP9 and IGF2 may be explored as an effective bone regeneration agent to treat large segmental bony defects, non-union fracture, and/or osteoporatic fracture. doi:10.1016/j.bone.2010.09.267
308 Increase rate of cellular osteogenic mineralization by low-intensity ultrasound in a stimulated microgravity Y.-X. Qin, S.Z. Uddin, S. Zhang, M. Hu, Y. Huang, J. Cheng Orthopaedic Bioengineering Research Laboratory, Department of Biomedical Engineering Stony Brook University, NY, USA Objective: Osteoporosis and disuse osteopenia are increased health problems that increase the risk of fracture. Microgravity causes bone loss in the skeleton potentially due to decrease in bone mineralization by osteoblast cells. In vitro studies using stimulated microgravity have shown significant decrease in expression of osteoblastic transcription factors and low alkaline phosphatase activity (ALP). Low-intensity pulse ultrasound (LIPUS) has shown capability in promoting bony ingrowth and accelerating rate of mineralization in osteoblast cells by up regulating expression of different osteoblastic transcription factors, matrix proteins and ALP activity. Ultrasound has been known to increase calcium deposition in the osteoblast cells. The objective of this study was to study the effects of LIPUS on osteoblast mineralization in stimulated microgravity. Methods: Human fetal osteoblast (hfob 1.19) was cultured in DMEM:F12 media supplemented with 15% FBS and 0.3 mg/ml G418 with initial seeding density of 50,000 cell per Opticell cartilage. Cells were allowed to grow to confluency, and stimulations were started at day 7 of post confluency. Cells were distributed into four groups (n = 4 each), gravity, gravity + LIPUS, simulated microgravity and microgravity + LIPUS. Stimulated microgravity environment was created by putting Opticells into an in vitro rotary system with rotation set at 15 RPM. LIPUS groups were stimulated daily for total of 14 days with 30 mW/cm2 for 20 min. Cells were kept at 37 °C, 5% CO2 environment during experiment and medium was changed every second day. After 14 days of stimulations, cells were analyzed for calcification by staining with Alizarin red stain and cytoskeleton changes with antibodies against actin and microtubules. Only the area stimulated with ultrasound was used for analyses. Results: Alizarin red (AZ) staining showed significant increase in calcification in microgravity+LIPUS group (2.70±0.03 mM AZ/106 cells) compared to microgravity group (2.16±0.24 mM AZ/106 cells, p=0.023). Significant decrease of calcification was also observed (47%) from gravity (3.18± 0.21 mM AZ/106 cells) group to the microgravity group (2.16±0.24 mM AZ/106 cells, p=0.009). Higher mineralization was observed in the gravity+LIPUS group (3.69±0.31 mM AZ/106 cells) compared to gravity group (3.18±0.21 mM AZ/106 cells, p=0.08). It was also observed that microgravity samples had fewer cells. Conclusion: These results imply that LIPUS increases mineralization in osteoblast cells in a microgravity environment. It suggests that ultrasound may serve as a noninvasive modality to provide localized treatment for bone loss at critical skeletal sites. The system may also provide a novel way to simulate microgravity conditions for application of space medicine. Acknowledgment This study was supported by NIH. doi:10.1016/j.bone.2010.09.268
S433
309 XLGB-B prevents bone loss in OVX mice by inhibiting bone remodeling Xin-luan Wang1, Ge Zhang1, Yi-xin He1, Li-zhen Zheng1, Feng-juan Tu2, Xin-sheng Yao2, Ling Qin1,3 1 Musculoskeletal Research Laboratory, Department of Orthopaedics & Traumatology, The Chinese University of Hong Kong, Hong Kong, China 2 Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, Jinan University, Guangzhou, China 3 Translational Medicine R&D Center, Institute of Biomedical & Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China Objectives: “Xian Ling Gu Bao capsule” (XLGB) is the only traditional Chinese medicine for treatment of osteoporosis in the health insurance list in China. Randomized, multicenter, double-blind, placebo-controlled clinical trial demonstrated XLGB statistically significantly increased bone mineral density (BMD) in the lumbar spine at 6 months. Three fractions (XLGB-A, B, and C) were prepared for seeking the key fraction (s) for preventing osteoporosis. In this study the effect of XLGB-B on preventing osteoporosis was investigated. Material and methods: Thirty, 4-month-old female C57/BL6J mice were either sham operated (Sham Group) or OVX. After recovering for 3 days, the OVX mice were randomly divided into two groups: OVX Group and XLGB-B Group (31 mg/kg/day). Treatments were given orally for 1 month. 3D bone structure of the sixth lumber was analyzed using Micro-CT (Extreme CT, Scanco Medical, Brüttisellen, Switzerland) with an isotropic resolution of 12 μm in all three spatial dimensions. The trabecular bone volume (BV/TV, %) was measured. Concentrations of N-terminal propeptides of type I procollagen (PINP, bone formation marker) and C-propeptide of type I collagen (CTX, bone resorption marker) in serum were measured using ELISA kits (Immunodiagnostic Systems Ltd. UK). Results: 3D images of the 6th lumbar clearly demonstrated that the trabecular bone in Sham and XLGB-B group was obviously more than that in OVX group (p < 0.05). Similarly, BV/TV in Sham and XLGB-B group was significantly higher than that in the OVX group (p < 0.05). Both the concentrations of PINP and CTX in serum of the OVX group increased compared to that of the Sham group, which indicated that OVX could promote the bone remodeling, and XLGB-B significantly inhibited the increases of both PINP and CTX induced by OVX (p < 0.05). Conclusion: In conclusion, XLGB-B could prevent bone loss in OVX mice by inhibiting bone remodeling.
Acknowledgment This work was supported by Tongjitang Pharmaceutical Co. Ltd. doi:10.1016/j.bone.2010.09.269
311 Poor bone quality in old Chinese women with bound feet—A QUS study Mian Xu1, Hanchang Lao1, Yi Pan1, Vivian Wing-yin Hung2, Ming Zhang3, Ling Qin4 1 The 2nd Affiliated Hospital of Kunming Medical College, Kunming, China 2 Department of Orthopaedics & Traumatology, Bone Quality & Health Centre, The Chinese University of Hong Kong, Hong Kong, China 3 Department of Health Technology & Informatics, The Hong Kong Polytechnic University, Hong Kong, China 4 Translational Medicine Research & Development Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China