Morikawa Taiki, Matsuzaka Kenichi, Nakajima Kei, Yasumura Toshihiko, Sueishi Kenji, Inoue Takashi
Department of Orthodontics, Tokyo Dental College, 2-9-18, Misaki-cho, Chiyoda-ku, Tokyo, 101-0061, Japan.
Department of Clinical Pathophysiology, Tokyo Dental College, 2-9-18, Misaki-cho, Chiyoda-ku, 101-0061, Tokyo, Japan.
Arch Oral Biol. 2016 Oct;70:158-164. doi: 10.1016/j.archoralbio.2016.06.021. Epub 2016 Jun 23.
This study investigated the expression of receptor activator of nuclear factor-κB ligand (RANKL) in periodontal ligament (PDL) cells co-cultured with dental pulp (DP) cells following mechanical stress in vitro. Furthermore, the expression of prostaglandin (PG) E and substance P (SP) by the PDL cells and by the DP cells were also examined.
PDL and DP cells were obtained from 10 rats. The experimental group consisted of PDL cells subjected to centrifugal force as mechanical stress and co-cultured with DP cells. The 3 control groups of PDL cells were: 1) PDL cells without mechanical stress, 2) PDL cells treated with mechanical stress and 3) PDL cells co-cultured with DP cells. The 2 control groups of DP cells were: 1) DP cells without mechanical stress and 2) DP cells co-cultured with PDL cells. In each group, both cells were examined at day 1 and day 3, and mRNA levels of RANKL by PDL cells were analyzed using Real time quantitative Reverse Transcription (RT)-PCR. Furthermore, RANKL expression was observed using Immunofluorescence staining. PGE and SP expression levels by PDL cells and DP cells were characterized by ELISA analysis.
The expression of RANKL by PDL cells under mechanical stress increased by co-culture with DP cells. PGE and SP expressions were increased in the group of PDL cells subjected to mechanical stress and co-cultured with DP cells.
DP cells may facilitate the expression of RANKL in PDL cells under mechanical stress via PGE and SP.
本研究调查体外机械应力作用后,与牙髓(DP)细胞共培养的牙周膜(PDL)细胞中核因子κB受体活化因子配体(RANKL)的表达。此外,还检测了PDL细胞和DP细胞中前列腺素(PG)E和P物质(SP)的表达。
从10只大鼠获取PDL细胞和DP细胞。实验组为受到离心力作为机械应力作用并与DP细胞共培养的PDL细胞。PDL细胞的3个对照组分别为:1)无机械应力的PDL细胞;2)经机械应力处理的PDL细胞;3)与DP细胞共培养的PDL细胞。DP细胞的2个对照组分别为:1)无机械应力的DP细胞;2)与PDL细胞共培养的DP细胞。每组在第1天和第3天检测两种细胞,使用实时定量逆转录(RT)-PCR分析PDL细胞中RANKL的mRNA水平。此外,采用免疫荧光染色观察RANKL表达。通过酶联免疫吸附测定(ELISA)分析PDL细胞和DP细胞中PGE和SP的表达水平。
机械应力作用下,PDL细胞与DP细胞共培养后RANKL表达增加。在受到机械应力并与DP细胞共培养的PDL细胞组中,PGE和SP表达增加。
DP细胞可能通过PGE和SP促进机械应力作用下PDL细胞中RANKL的表达。