Enamel matrix derivative inhibits TNF-alpha-induced apoptosis in osteoblastic MC3T3-E1 cells.

Enamel matrix derivative inhibits TNF-alpha-induced apoptosis in osteoblastic MC3T3-E1 cells.
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发表时间：
2005
期刊：
Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics
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通讯作者：
Jianing He;Yiming King;Jin Jiang;K. Safavi;L. Spångberg;Qiang Zhu
Jianing He;Yiming King;Jin Jiang;K. Safavi;L. Spångberg;Qiang Zhu
中科院分区：
其他
文献类型：
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作者：
Jianing He;Yiming King;Jin Jiang;K. Safavi;L. Spångberg;Qiang Zhu

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目的探讨釉基质衍生物（elaminalmatrixderivatives，EMD）对肿瘤坏死因子-α（TNF-α）诱导的成骨细胞MC 3 T3-E1凋亡的影响。研究设计MC 3 T3-E1细胞以5000/cm 2的初始密度在含有10%胎牛血清（FBS）的Dulbecco改良的Eagle培养基（DMEM）中培养，并使其贴壁24小时。然后将培养基更换为含0.5%FBS的DMEM。16小时后，用单独的EMD（100 μ g/mL）、单独的肿瘤坏死因子α（TNF-α）（20 ng/mL）、单独的转化生长因子β 1（TGF-β 1）（10 ng/mL）、TNF-α加TGF-β 1或TNF-α加EMD处理细胞。用DMEM和0.5%FBS培养的细胞作为对照。孵育24小时后，通过末端脱氧核苷酸转移酶缺口末端标记（TUNEL）试验评估细胞凋亡，并通过细胞死亡酶联免疫吸附试验（ELISA）定量。结果TUNEL法和ELISA法均显示TNF-α诱导MC 3 T3-E1细胞凋亡。TNF-α使细胞死亡增加约2倍，这被EMD和TGF-β 1减弱。结论EMD与TGF-β 1一样，对炎症诱导的成骨细胞凋亡具有保护作用。
OBJECTIVE The purpose of this study was to examine the effect of enamel matrix derivative (EMD) on TNF-alpha-induced apoptosis in osteoblastic MC3T3-E1 cells. STUDY DESIGN MC3T3-E1 cells were cultured at an initial density of 5000/cm 2 in Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS) and allowed to adhere for 24 hours. Medium was then changed into DMEM with 0.5% FBS. After 16 hours, cells were treated with EMD (100 microg/mL) alone, tumor necrosis factor alpha (TNF-alpha) (20 ng/mL) alone, transforming growth factor beta 1 (TGF-beta1) (10 ng/mL) alone, TNF-alpha plus TGF-beta1, or TNF-alpha plus EMD. Cells cultured with DMEM and 0.5% FBS served as control. Following 24-hour incubation, apoptosis was assessed by terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) assay, and quantified by cell death enzyme-linked immunosorbent assay (ELISA). RESULTS Both TUNEL assay and cell death ELISA show that TNF-alpha induces apoptosis in MC3T3-E1 cells. TNF-alpha increases cell death by approximately 2-fold, which is attenuated by both EMD and TGF-beta1. CONCLUSION Like TGF-beta1, EMD protects osteoblasts from inflammation-induced apoptosis.