Inubushi T, Tanaka E, Rego E B, Kitagawa M, Kawazoe A, Ohta A, Okada H, Koolstra J H, Miyauchi M, Takata T, Tanne K
Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan.
J Periodontol. 2008 Oct;79(10):1984-90. doi: 10.1902/jop.2008.080081.
The purpose of this study was to investigate the effects of low-intensity pulsed ultrasound (LIPUS) stimulation on the proliferation and differentiation of cementoblast lineage cells.
An immortalized human periodontal ligament cell line (HPL) showing immature cementoblastic differentiation was used. Cultured HPL cells were subjected to LIPUS exposure (frequency = 1 MHz; pulsed 1:4; intensity = 30 mW/cm(2)) or sham exposure for 15 minutes per day. Expression levels of alkaline phosphatase (ALP), type I collagen (Col-I), runt-related gene 2 (Runx2), bone sialoprotein (BSP), osteocalcin (OCN), and osteopontin (OPN) mRNA were analyzed with real-time polymerase chain reaction analysis. Furthermore, ALP activity, collagen synthesis, and protein level of Runx2 were examined after 6 days of LIPUS exposure.
mRNA and protein levels of ALP, Col-I, and Runx2 were significantly increased by LIPUS exposure compared to controls, whereas BSP, OCN, and OPN mRNA expression could not be detected in HPL cells, irrespective of LIPUS exposure.
LIPUS enhanced ALP activity, collagen synthesis, and Runx2 expression of HPL cells, which provides important insight into the promotion of early cementoblastic differentiation of immature cementoblasts.
本研究旨在探讨低强度脉冲超声(LIPUS)刺激对成牙骨质细胞系细胞增殖和分化的影响。
使用显示未成熟成牙骨质细胞分化的永生化人牙周膜细胞系(HPL)。培养的HPL细胞每天接受LIPUS照射(频率=1MHz;脉冲1:4;强度=30mW/cm²)或假照射15分钟。用实时聚合酶链反应分析碱性磷酸酶(ALP)、I型胶原(Col-I)、 runt相关基因2(Runx2)、骨唾液蛋白(BSP)、骨钙素(OCN)和骨桥蛋白(OPN)mRNA的表达水平。此外,在LIPUS照射6天后检测ALP活性、胶原合成和Runx2的蛋白水平。
与对照组相比,LIPUS照射显著提高了ALP、Col-I和Runx2的mRNA和蛋白水平,而无论是否进行LIPUS照射,在HPL细胞中均未检测到BSP、OCN和OPN mRNA的表达。
LIPUS增强了HPL细胞的ALP活性、胶原合成和Runx2表达,这为促进未成熟成牙骨质细胞的早期成牙骨质细胞分化提供了重要的见解。