Yi Wang1, Haiyan Wang1, Qingsong Ye2, Jie Ye1, Chunyan Xu3, Leilei Lin1, Hui Deng4, Rongdang Hu5. 1. Orthodontic Department, School of Stomatology, Wenzhou Medical College, China. 2. Department of Orthodontics, James Cook University, Australia. 3. Orthodontic Department, Yiwu Central Hospital, China. 4. Periodontal Department, School of Stomatology, Wenzhou Medical College, China. Electronic address: dh0726@gmail.com. 5. Orthodontic Department, School of Stomatology, Wenzhou Medical College, China. Electronic address: hurongdang@hotmail.com.
Abstract
AIMS: Orthodontic forces are known to aggravate inflammation-induced destruction of the periodontium, but the underlying mechanism has not been elucidated. The present study investigates how inflammation and forces co-regulate periodontium damage. MAIN METHODS: Cultures of MC3T3-E1 osteoblasts were pre-treated with conditioned medium from RAW264.7 macrophages exposed to 100ng/ml Porphyromonas gingivalis (Pg)-LPS. Conditioned medium was analyzed by ELISA for interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Osteoblasts were then subjected to tensile strain (0.5Hz; 1000μ or 3000μ) for 0min, 5min, 15min, 30min, 1h, 3h, and 6h. The cultures were analyzed for mRNA and protein levels of c-fos. Cells were also analyzed for alkaline phosphatase (ALP) activity. KEY FINDINGS: (Pg)-LPS stimulated the secretion of all three cytokines from RAW264.7 cells in a dose- and time-dependent manner. Medium from (Pg)-LPS stimulated cells induced a 10-fold increase in c-fos expression, which decreased to a 4-fold plateau after 3h. In contrast, ALP activity of control osteoblasts decreased during the first 60min, then recovered over the next 4h. Pretreatment with conditioned medium generated the same initial decrease during tensile strain but prevented the recovery. SIGNIFICANCE: Our study showed, for the first time, that the inhibitory effect of inflammation and tensile strain on osteogenicity is associated with the upregulation in c-fos expression. In addition, inflammation may reduce the ability of osteoblasts to restore their osteogenic capacity during sustained tensile stress and contribute to periodontium damage.
AIMS: Orthodontic forces are known to aggravate inflammation-induced destruction of the periodontium, but the underlying mechanism has not been elucidated. The present study investigates how inflammation and forces co-regulate periodontium damage. MAIN METHODS: Cultures of MC3T3-E1 osteoblasts were pre-treated with conditioned medium from RAW264.7 macrophages exposed to 100ng/ml Porphyromonas gingivalis (Pg)-LPS. Conditioned medium was analyzed by ELISA for interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Osteoblasts were then subjected to tensile strain (0.5Hz; 1000μ or 3000μ) for 0min, 5min, 15min, 30min, 1h, 3h, and 6h. The cultures were analyzed for mRNA and protein levels of c-fos. Cells were also analyzed for alkaline phosphatase (ALP) activity. KEY FINDINGS: (Pg)-LPS stimulated the secretion of all three cytokines from RAW264.7 cells in a dose- and time-dependent manner. Medium from (Pg)-LPS stimulated cells induced a 10-fold increase in c-fos expression, which decreased to a 4-fold plateau after 3h. In contrast, ALP activity of control osteoblasts decreased during the first 60min, then recovered over the next 4h. Pretreatment with conditioned medium generated the same initial decrease during tensile strain but prevented the recovery. SIGNIFICANCE: Our study showed, for the first time, that the inhibitory effect of inflammation and tensile strain on osteogenicity is associated with the upregulation in c-fos expression. In addition, inflammation may reduce the ability of osteoblasts to restore their osteogenic capacity during sustained tensile stress and contribute to periodontium damage.
Authors: Andressa Vilas Boas Nogueira; Marjan Nokhbehsaim; Sigrun Eick; Christoph Bourauel; Andreas Jäger; Søren Jepsen; Carlos Rossa; James Deschner; Joni Augusto Cirelli Journal: Mediators Inflamm Date: 2014-05-25 Impact factor: 4.711