Kusuyama Joji, Nakamura Toshiaki, Ohnishi Tomokazu, Eiraku Nahoko, Noguchi Kazuyuki, Matsuguchi Tetsuya
Departments of *Oral Biochemistry; and †Periodontology, Graduate School of Medical and Dental Sciences, Kagoshima University.
J Orthop Trauma. 2017 Jul;31(7):S4. doi: 10.1097/01.bot.0000520897.92470.70.
We previously reported that low intensity pulsed ultrasound (LIPUS) promotes marrow stromal cell (MSC) osteogenesis and suppresses the LPS-induced inflammatory response in osteoblasts. Here, we examined the effects of LIPUS on human periodontal ligament-derived stem cells (hPDLSCs) in chronic inflammatory bone disease, such as periodontitis.
hPDLSCs were collected from 3 healthy third molars. hPDLSCs were induced to differentiate by either recombinant BMP2 or BMP9 with or without daily LIPUS treatment (20 min/d). hPDLSCs were also stimulated by Porphyromonas gingivalis-derived LPS (LPS-PG), IL-1beta, and TNF-alpha with or without LIPUS. Matrix mineralization was evaluated by alizarin red S staining. The expression of genes for osteogenic makers and for inflammatory cytokines were analyzed by real time RT-PCR.
LIPUS promoted BMP9-induced osteogenesis of hPDLSCs based on increases in both cell calcification and osteogenic marker expression. In contrast, LIPUS did not affect BMP2-induced osteogenic differentiation. LIPUS-induced Noggin expression was potentially involved in the differential response of the cells. Either LPS-PG, IL-1beta, or TNF-alpha-induced ERK phosphorylation and IL-8, CCL2, and RANKL expression were decreased in LIPUS-treated hPDLSCs. Moreover, the inhibitory effects of LPS-PG and IL-1beta on osteogenesis of hPDLSCs were significantly blocked by LIPUS.
LIPUS is an effective tool to promote osteogenic differentiation under inflammatory conditions.
我们之前报道过，低强度脉冲超声（LIPUS）可促进骨髓基质细胞（MSC）的成骨作用，并抑制脂多糖（LPS）诱导的成骨细胞炎症反应。在此，我们研究了LIPUS对慢性炎症性骨病（如牙周炎）中人类牙周膜来源干细胞（hPDLSCs）的影响。
从3颗健康的第三磨牙中收集hPDLSCs。hPDLSCs在有或无每日LIPUS治疗（20分钟/天）的情况下，用重组骨形态发生蛋白2（BMP2）或骨形态发生蛋白9（BMP9）诱导分化。hPDLSCs也在有或无LIPUS的情况下，受到牙龈卟啉单胞菌来源的LPS（LPS-PG）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的刺激。通过茜素红S染色评估基质矿化情况。通过实时逆转录聚合酶链反应（RT-PCR）分析成骨标志物和炎症细胞因子基因的表达。
基于细胞钙化和成骨标志物表达的增加，LIPUS促进了BMP9诱导的hPDLSCs成骨作用。相比之下，LIPUS不影响BMP2诱导的成骨分化。LIPUS诱导的Noggin表达可能与细胞的不同反应有关。在接受LIPUS治疗的hPDLSCs中，LPS-PG、IL-1β或TNF-α诱导的细胞外信号调节激酶（ERK）磷酸化以及IL-8、趋化因子配体2（CCL2）和核因子κB受体活化因子配体（RANKL）表达均降低。此外，LIPUS显著阻断了LPS-PG和IL-1β对hPDLSCs成骨作用的抑制效果。
LIPUS是在炎症条件下促进成骨分化的有效工具。
