Synthetic ameloblastin peptide stimulates differentiation of human periodontal ligament cells
Synthetic ameloblastin peptide stimulates differentiation of human periodontal ligament cells
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DOI：
10.1016/j.archoralbio.2010.10.012
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发表时间：
2011-04-01
影响因子：
3
通讯作者：
Takata, Takashi
中科院分区：
文献类型：
--
作者：
Kitagawa, Masae;Kitagawa, Shoji;Takata, Takashi
目的：本研究探讨合成的猪成釉素肽的N端区域对人牙周膜细胞（PDLC）增殖和分化的影响。设计：我们使用细胞计数器评估猪成釉素肽对PDLC增殖的影响。为了研究成釉素肽对 PDLC 分化的影响，我们采用 Bessey-Lowry 酶学方法定量分析碱性磷酸酶 (ALP) 活性，采用 Dahl 法定量分析矿化结节形成，并采用 RT-PCR 定量分析矿化相关基因的表达。我们使用抗成釉素抗体来确定 ALP 活性的刺激是否是由该肽引起的。结果：在所有检测浓度下，成釉素肽对细胞增殖的影响与对照相比没有显着差异。然而，该肽以剂量依赖性方式显着刺激 ALP 活性。 ALP 活性被抗成釉细胞蛋白抗体显着抑制，导致 ALP 水平恢复到未治疗条件下的近似水平。当浓度大于 1 ng/ml 时，肽促进 PDLC 矿化结节的形成。并且该肽诱导比对照更高的ALP和骨唾液蛋白(BSP)表达。结论：我们的结果表明成釉素肽上调ALP和BSP水平并且可以增强PDLC的钙化。因此，我们认为N端合成的成釉素肽可促进PDLC的分化活性。 (C) 2010 Elsevier Ltd. 保留所有权利。
Objective: This study investigates the effect of the N-terminal region of a synthetic porcine ameloblastin peptide on the proliferation and differentiation of human periodontal ligament cells (PDLC).Design: We used a cell counter to assess the effect of ameloblastin peptides on the proliferation of PDLC. To investigate the effect of ameloblastin peptides on the differentiation of PDLC, we examined quantitative analysis of alkaline phosphatase (ALP) activity by the Bessey-Lowry enzymological method, mineral nodule formation by Dahl's method, and expression of mineralization-related genes by RT-PCR. We used an anti-ameloblastin antibody to determine whether stimulation of ALP activity was caused by the peptide.Results: At all concentrations examined, the effect of the ameloblastin peptide on cell proliferation was not significantly different compared with the control. However, the peptide significantly stimulated ALP activity in a dose-dependent manner. ALP activity was significantly inhibited by an anti-ameloblastin antibody, which caused ALP levels to revert to their approximate levels in the untreated condition. At concentrations greater than 1 ng/ml, the peptide promoted mineralized nodule formation of PDLC. And the peptide induced higher expressions of ALP and bone sialoprotein (BSP) than the control.Conclusion: Our results show that the ameloblastin peptide upregulate ALP and BSP levels and can enhance calcification of PDLC. Thus, we suggest that the N-terminal synthetic ameloblastin peptide promotes the differentiation activity of PDLC. (C) 2010 Elsevier Ltd. All rights reserved.