Al-Sharabi Niyaz, Mustafa Manal, Ueda Minoru, Xue Ying, Mustafa Kamal, Fristad Inge
Department of Clinical Dentistry, University of Bergen, Bergen, Norway.
Oral Health Centre of Expertise in Western Norway, Bergen, Norway.
Dent Traumatol. 2017 Feb;33(1):19-26. doi: 10.1111/edt.12277. Epub 2016 May 4.
To evaluate the effect of MSC-conditioned medium (CM) on the secretion of pro- and anti-inflammatory cytokines from dental pulp cells (hDPC) in vitro, and on the gene expression in vivo after replantation of rat molars.
hDPC were cultured in CM for 24 h, and the concentration of interleukin IL-10, IL-4, IL-6, and IL-8, regulated on activation, normal T Cell expressed and secreted (RANTES), and prostaglandin E (PGE ) in the media were measured by multiplex assay and ELISA, respectively. Expression of cyclooxygenase-2 (COX-2) was also examined by Western blot analysis after 24 h. Left and right maxillary first rat molars (n = 20) were elevated for 2 min and then replanted with or without application of CM into the tooth sockets. Levels of IL-1β, IL-10, IL-4, IL-6, and IL-8, and tumor necrosis factor-alpha (TNF-α) mRNA were evaluated by real-time qRT-PCR 3 and 14 days following tooth replantation.
The production of IL-8, IL-10, and IL-6, RANTES and PGE by cells cultured in CM was significantly higher than production by cells cultured in standard medium (DMEM). At day 3 following replantation in vivo, the levels of IL-1β and IL-6, and TNF-α mRNA were significantly lower in the CM-treated replanted teeth compared with control teeth. Further, at day 3, the IL-6/IL-10 ratio was significantly lower in the CM-treated replanted teeth compared with control. At day 14 following replantation, no differences in the mRNA ratios were detected between the pulp tissues of replanted and control teeth.
These findings indicated that CM promotes secretion of pro- and anti-inflammatory cytokines from hDPCin vitro and attenuates the initial inflammatory response in the rat dental pulp in vivo following tooth replantation.
评估间充质干细胞条件培养基（CM）对体外牙髓细胞（hDPC）促炎和抗炎细胞因子分泌的影响，以及对大鼠磨牙再植后体内基因表达的影响。
将hDPC在CM中培养24小时，分别通过多重检测和酶联免疫吸附测定法测量培养基中白细胞介素IL-10、IL-4、IL-6、IL-8、正常T细胞激活后表达和分泌的调节趋化因子（RANTES）以及前列腺素E（PGE）的浓度。24小时后还通过蛋白质免疫印迹分析检测环氧合酶-2（COX-2）的表达。将20只大鼠的左右上颌第一磨牙抬起2分钟，然后在牙槽窝中植入或不植入CM进行再植。在牙齿再植后3天和14天，通过实时定量逆转录聚合酶链反应评估IL-1β、IL-10、IL-4、IL-6、IL-8以及肿瘤坏死因子-α（TNF-α）mRNA的水平。
在CM中培养的细胞产生的IL-8、IL-10、IL-6、RANTES和PGE明显高于在标准培养基（DMEM）中培养的细胞。在体内再植后第3天，与对照牙相比，CM处理的再植牙中IL-1β和IL-6以及TNF-α mRNA的水平明显较低。此外，在第3天，与对照相比，CM处理的再植牙中IL-6/IL-10的比值明显较低。在再植后第14天，再植牙和对照牙的牙髓组织之间未检测到mRNA比值的差异。
这些发现表明，CM在体外促进hDPC分泌促炎和抗炎细胞因子，并在大鼠牙齿再植后体内减弱牙髓的初始炎症反应。
