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Manganese superoxide dismutase is required to maintain osteoclast differentiation and function under static force.

Guo T, Zhang L, Konermann A, Zhou H, Jin F, Liu W - Sci Rep (2015)

Bottom Line: Total respective protein spot numbers of 549 ± 13, 612 ± 19 and 634 ± 16 were detected in each of the gels by image analysis.The five proteins identified were plasminogen activator inhibitor 2 (PAI-2, Spot 1), peroxiredoxin-6 (PRD-6, Spot 3), manganese superoxide dismutase (SOD2, Spot 6), Rho GDP-dissociation inhibitor 2 (Rho-GDI2, Spot 11) and L-lactate dehydrogenase B chain (L-LDH, Spot 15).More importantly, we revealed that SOD2 was required to maintain monocyte differentiation into functional OCs and may become a potential target for regulating the efficiency of OTM in the future.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Military Stomatology, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, People's Republic of China.

ABSTRACT
Bone homeostasis is maintained by the balance of osteoblasts (OBs) and osteoclasts (OCs). Increased activity of OCs not only contributes to pathological bone resorption, such as osteoporosis and periodontitis, but also is responsible for physiological conditions like orthodontic tooth movement (OTM). However, the detailed mechanism by which orthodontic force promotes the formation of OCs is still poorly understood. In this study, we confirmed that static force promoted the differentiation of human cord monocytes (HMNCs) into OCs depending on loading time and magnitude. Protein expression profiles among HMNCs, HMNCs subjected to static force and mature OCs were established via 2-DE and MALDI-TOF-MS analyses. Total respective protein spot numbers of 549 ± 13, 612 ± 19 and 634 ± 16 were detected in each of the gels by image analysis. The five proteins identified were plasminogen activator inhibitor 2 (PAI-2, Spot 1), peroxiredoxin-6 (PRD-6, Spot 3), manganese superoxide dismutase (SOD2, Spot 6), Rho GDP-dissociation inhibitor 2 (Rho-GDI2, Spot 11) and L-lactate dehydrogenase B chain (L-LDH, Spot 15). More importantly, we revealed that SOD2 was required to maintain monocyte differentiation into functional OCs and may become a potential target for regulating the efficiency of OTM in the future.

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Pick up effective loading time and magnitude of static force.(A), TRAP mRNA and protein expression for six force levels with five different force duration times evaluated via real-time PCR and western blotting. (B), (C), HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium renew. OC formation was analysed by TRAP staining (B) after loading for 3 and 5 days, and bone resorption pits were examined via Toluidine-blue staining (C) under the same loading conditions for 9 days. Cont, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3; SF, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium was renewed. magnification × 100 and × 200. Data represent mean ± SD.*P < 0.05, n = 6.
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f2: Pick up effective loading time and magnitude of static force.(A), TRAP mRNA and protein expression for six force levels with five different force duration times evaluated via real-time PCR and western blotting. (B), (C), HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium renew. OC formation was analysed by TRAP staining (B) after loading for 3 and 5 days, and bone resorption pits were examined via Toluidine-blue staining (C) under the same loading conditions for 9 days. Cont, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3; SF, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium was renewed. magnification × 100 and × 200. Data represent mean ± SD.*P < 0.05, n = 6.

Mentions: To determine the appropriate loading time and magnitude of static force application on HMNCs differentiation, six different levels of magnitude and five different duration times were tested with regard to the expression level of TRAP mRNA and protein. Real-time PCR assay and western blotting analysis showed that static force promoted TRAP expression depending on the loading time and magnitude, especially under 150 kpa loading for 1.5 h (Fig. 2a). We next tested the effect of static force on differentiation of HMNCs. HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded under 150 kpa static force for 1.5 h every 3 days after the medium was renewed. The results showed that TRAP-positive cells increased significantly with loading static force at day 3 compared with the non-loading group, and many multinuclear OCs were formed in the loading group at day 5 (Fig. 2b and Supplementary Fig. 2). More importantly, bone-resorbing pits were observed in the loading group, while almost no pits could be seen in the non-loading group (Fig. 2c). These results suggested that the 150 kpa static force loading for 1.5 h significantly promotes differentiation of HMNCs into functional-OCs.


Manganese superoxide dismutase is required to maintain osteoclast differentiation and function under static force.

Guo T, Zhang L, Konermann A, Zhou H, Jin F, Liu W - Sci Rep (2015)

Pick up effective loading time and magnitude of static force.(A), TRAP mRNA and protein expression for six force levels with five different force duration times evaluated via real-time PCR and western blotting. (B), (C), HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium renew. OC formation was analysed by TRAP staining (B) after loading for 3 and 5 days, and bone resorption pits were examined via Toluidine-blue staining (C) under the same loading conditions for 9 days. Cont, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3; SF, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium was renewed. magnification × 100 and × 200. Data represent mean ± SD.*P < 0.05, n = 6.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4306132&req=5

f2: Pick up effective loading time and magnitude of static force.(A), TRAP mRNA and protein expression for six force levels with five different force duration times evaluated via real-time PCR and western blotting. (B), (C), HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium renew. OC formation was analysed by TRAP staining (B) after loading for 3 and 5 days, and bone resorption pits were examined via Toluidine-blue staining (C) under the same loading conditions for 9 days. Cont, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3; SF, HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded with 150 kpa static force for 1.5 h every 3 days after the medium was renewed. magnification × 100 and × 200. Data represent mean ± SD.*P < 0.05, n = 6.
Mentions: To determine the appropriate loading time and magnitude of static force application on HMNCs differentiation, six different levels of magnitude and five different duration times were tested with regard to the expression level of TRAP mRNA and protein. Real-time PCR assay and western blotting analysis showed that static force promoted TRAP expression depending on the loading time and magnitude, especially under 150 kpa loading for 1.5 h (Fig. 2a). We next tested the effect of static force on differentiation of HMNCs. HMNCs were cultured in induction medium with 1α, 25-(OH)2D3 and simultaneously loaded under 150 kpa static force for 1.5 h every 3 days after the medium was renewed. The results showed that TRAP-positive cells increased significantly with loading static force at day 3 compared with the non-loading group, and many multinuclear OCs were formed in the loading group at day 5 (Fig. 2b and Supplementary Fig. 2). More importantly, bone-resorbing pits were observed in the loading group, while almost no pits could be seen in the non-loading group (Fig. 2c). These results suggested that the 150 kpa static force loading for 1.5 h significantly promotes differentiation of HMNCs into functional-OCs.

Bottom Line: Total respective protein spot numbers of 549 ± 13, 612 ± 19 and 634 ± 16 were detected in each of the gels by image analysis.The five proteins identified were plasminogen activator inhibitor 2 (PAI-2, Spot 1), peroxiredoxin-6 (PRD-6, Spot 3), manganese superoxide dismutase (SOD2, Spot 6), Rho GDP-dissociation inhibitor 2 (Rho-GDI2, Spot 11) and L-lactate dehydrogenase B chain (L-LDH, Spot 15).More importantly, we revealed that SOD2 was required to maintain monocyte differentiation into functional OCs and may become a potential target for regulating the efficiency of OTM in the future.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Military Stomatology, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, People's Republic of China.

ABSTRACT
Bone homeostasis is maintained by the balance of osteoblasts (OBs) and osteoclasts (OCs). Increased activity of OCs not only contributes to pathological bone resorption, such as osteoporosis and periodontitis, but also is responsible for physiological conditions like orthodontic tooth movement (OTM). However, the detailed mechanism by which orthodontic force promotes the formation of OCs is still poorly understood. In this study, we confirmed that static force promoted the differentiation of human cord monocytes (HMNCs) into OCs depending on loading time and magnitude. Protein expression profiles among HMNCs, HMNCs subjected to static force and mature OCs were established via 2-DE and MALDI-TOF-MS analyses. Total respective protein spot numbers of 549 ± 13, 612 ± 19 and 634 ± 16 were detected in each of the gels by image analysis. The five proteins identified were plasminogen activator inhibitor 2 (PAI-2, Spot 1), peroxiredoxin-6 (PRD-6, Spot 3), manganese superoxide dismutase (SOD2, Spot 6), Rho GDP-dissociation inhibitor 2 (Rho-GDI2, Spot 11) and L-lactate dehydrogenase B chain (L-LDH, Spot 15). More importantly, we revealed that SOD2 was required to maintain monocyte differentiation into functional OCs and may become a potential target for regulating the efficiency of OTM in the future.

Show MeSH
Related in: MedlinePlus