Low-Intensity Pulsed Ultrasound Induces Osteogenic Differentiation of Human Periodontal Ligament Cells Through Activation of Bone Morphogenetic Protein-Smad Signaling

被引:28
|
作者
Yang, Zun [1 ,3 ]
Ren, Leixi [4 ]
Deng, Feng [1 ,3 ]
Wang, Zhibiao [2 ]
Song, Jinlin [1 ,3 ]
机构
[1] Chongqing Med Univ, Affiliated Hosp Stomatol, Chongqing 401147, Peoples R China
[2] Chongqing Med Univ, Coll Biomed Engn, Chongqing 401147, Peoples R China
[3] Chongqing Key Lab Oral Dis & Biomed Sci, Chongqing, Peoples R China
[4] Chongqing Med Univ, Childrens Hosp, Dept Stomatol, Chongqing 401147, Peoples R China
基金
中国国家自然科学基金;
关键词
basic science; bone morphogenetic protein-Smad; human periodontal ligament cells; low-intensity pulsed ultrasound; osteogenic differentiation; signal transduction; ALKALINE-PHOSPHATASE ACTIVITY; STEM-CELLS; IN-VITRO; OSTEOBLAST DIFFERENTIATION; ENDOTHELIAL-CELLS; GROWTH-FACTORS; TGF-BETA; EXPRESSION; REGENERATION; FIBROBLASTS;
D O I
10.7863/ultra.33.5.865
中图分类号
O42 [声学];
学科分类号
070206 ; 082403 ;
摘要
Objectives-Low-intensity pulsed ultrasound (US) can accelerate fracture healing and osteogenic differentiation. The aim of this study was to investigate the osteogenic effect of low-intensity pulsed US on human periodontal ligament cells and to determine whether bone morphogenetic protein (BMP)-Smad signaling was involved. Methods-Human periodontal ligament cells were exposed to low-intensity pulsed US at a frequency of 1.5 MHz and intensity of 90 mW/cm(2) for 20 min/d. Osteogenic differentiation was determined by assaying alkaline phosphatase (ALP) and calcium deposition. Expression of BMP-2, BMP-6, and BMP-9 was detected by real-time polymerase chain reaction analysis. Phosphorylated Smad was detected by western blotting; Smad in the cells was labeled by an immunofluorescent antibody and observed by laser-scanning confocal microscopy. Results-The optical density of ALP stimulated by US at 1.5 MHz and 90 mW/cm2 for 20 min/d was significantly higher than in other groups (P < .01); therefore, this dosage was considered optimal for promoting osteogenic differentiation. After 13 days of US exposure, ALP increased gradually after 5 days, peaked at 11 days, and decreased at 13 days, with a significant difference compared with the control group (P < .05). Osteocalcin production increased from 9 to 13 days and peaked at 15 days, with a significant difference compared with the control group (P < .05). BMP-2 and BMP-6 increased dynamically after exposure for 13 days. BMP-2 increased 6.07-fold at 3 days, 6.39-fold at 11 days, and 5.97-fold at 13 days. BMP-6 expression increased 6.82-fold at 1 day and 51.5-fold at 3 days and decreased thereafter. BMP-9 was not expressed. Phospho-Smad1/5/8 expression was significantly increased after exposure (P < .05) and transferred from the cytoplasm into the nuclei. Conclusions-Low-intensity pulsed US effectively induced osteogenic differentiation of human periodontal ligament cells, and the BMP-Smad signaling pathway was involved in the mechanism.
引用
收藏
页码:865 / 873
页数:9
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