Orthodontic Section, Faculty of Dentistry, Department of Preventive Dentistry, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand.
Faculty of Dentistry, Department of Oral Surgery, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand.
Cell Biol Int. 2018 Sep;42(10):1349-1357. doi: 10.1002/cbin.11019. Epub 2018 Jul 3.
The facial and long bones have distinct developmental origins, structures, and cellular compositions. This study aimed to compare the in vitro responses of human mandible and long bone osteoblasts to low-magnitude, high-frequency (LMHF) mechanical vibration in terms of expression of mediators of bone remodeling. Osteoblast-like cell cultures were prepared from iliac crest and mandibular bone specimens from three individuals and cultured in osteogenic induction media. Induction of mature osteoblastic phenotypes was confirmed by analysis of DNA content, alkaline phosphatase activity and gene expression every 3 days for 27 days. Based on gene expression, mature osteoblasts formed by day 15 of induction culture. After 15 days of culture in induction media, mature osteoblasts were subjected to vibration (0, 30, or 60 Hz) for 30 min every 24 h. After 48 h, RANKL, OPG, IL-1β, IL-6 and TGF-β gene, and protein expression were determined by real-time PCR analysis of total cellular mRNA and ELISAs of the cell supernatants. Both iliac and mandible osteoblasts responded to LMHF vibration: IL-1β and RANKL mRNA were downregulated and IL-6 mRNA was upregulated. However, TGF- β mRNA was unaltered and OPG mRNA was upregulated in iliac osteoblasts, whereas both TGF-β and OPG mRNA were downregulated in mandible osteoblasts. As a result, LMHF reduced the RANKL/OPG mRNA ratio in iliac osteoblasts but did not alter the RANKL/OPG mRNA ratio in mandible osteoblasts. This study suggests mature iliac osteoblasts exhibit a more potent anti-resorptive response to vibration, while this tendency was not obviously apparent in mature mandible osteoblasts.
面部和长骨具有明显不同的发育起源、结构和细胞组成。本研究旨在比较人下颌骨和长骨成骨细胞对低幅度、高频率(LMHF)机械振动的体外反应,从骨重塑介质的表达方面进行比较。从 3 个人的髂嵴和下颌骨标本中制备成骨细胞样细胞培养物,并在成骨诱导培养基中培养。通过分析 DNA 含量、碱性磷酸酶活性和基因表达,每 3 天对培养物进行一次分析,持续 27 天,以确认成熟成骨细胞表型的诱导。诱导培养 15 天后形成成熟的成骨细胞。在诱导培养基中培养 15 天后,将成熟的成骨细胞用 0、30 或 60Hz 的振动(每 24 小时 30 分钟)处理。48 小时后,通过实时 PCR 分析总细胞 mRNA 和细胞上清液的 ELISA 测定 RANKL、OPG、IL-1β、IL-6 和 TGF-β基因和蛋白表达。髂骨和下颌骨成骨细胞均对 LMHF 振动有反应:IL-1β 和 RANKL mRNA 下调,IL-6 mRNA 上调。然而,TGF-βmRNA 不变,OPG mRNA 在髂骨成骨细胞中上调,而 TGF-β和 OPG mRNA 在下颌骨成骨细胞中均下调。结果,LMHF 降低了髂骨成骨细胞中的 RANKL/OPG mRNA 比值,但没有改变下颌骨成骨细胞中的 RANKL/OPG mRNA 比值。本研究表明成熟的髂骨成骨细胞对振动表现出更强的抗吸收反应,而这种趋势在下颌骨成骨细胞中并不明显。