血小板衍生生长因子-BB通过p38信号转导通路介导牙周细胞的迁移。

Ray Angel K, Jones Anne C, Carnes David L, Cochran David L, Mellonig James T, Oates Thomas W

Department of Pathology, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA.

J Periodontol. 2003 Sep;74(9):1320-8. doi: 10.1902/jop.2003.74.9.1320.

BACKGROUND

Intracellular signaling pathways mediate specific responses to growth factors. The manipulation of these pathways ultimately may be used to control the clinical outcomes of periodontal regenerative therapy. The purpose of this study was to examine the role of the p38 signal transduction pathway in the responses of periodontal cells to platelet-derived growth factor-BB (PDGF).

METHODS

Primary cultures of human periodontal ligament cells (PDLs) and gingival fibroblasts (GFs) were used for all experiments. Cell numbers, 3H-thymidine incorporation, and Boyden chamber assays were used to characterize the effects of SB 203580 (SB), a specific inhibitor of the p38 signaling pathway, on cell proliferation and migration. An in vitro wound model also was used to assess the effects of SB. For the in vitro wound assay, triplicate wells were incubated for 1, 3, 5, and 7 days using 0.1% fetal bovine serum (FBS), 10% FBS +/- 10 microM SB, or 20 ng/ml PDGF +/- 10 microM SB. Digital histomorphometric analysis assessed cellular fill within the wound area.

RESULTS

SB specifically inhibited PDGF-induced migration in the Boyden chamber assays without affecting cell proliferation. The wound model data showed similar levels of wound fill for PDLs and GFs in 10% FBS. Relative to 10% FBS, PDLs stimulated with PDGF showed significantly (P < 0.01, analysis of variance) greater wound fill (74%) than GFs (12%). SB inhibited the PDGF-induced wound fill of PDLs and GFs by 64% and 57%, respectively. This inhibition was significant (P < 0.01, ANOVA) only for PDLs. The addition of SB to 10% FBS did not significantly affect the wound fill response of either cell type compared to 10% FBS alone.

CONCLUSIONS

These results demonstrate that periodontal cells possess distinct responses to PDGF that may be altered at the signal transduction level. The manipulation of these responses through the use of inhibitors to specific signaling pathways may enhance our control of periodontal regeneration in the future.

背景

细胞内信号通路介导对生长因子的特定反应。对这些通路的操控最终可能用于控制牙周再生治疗的临床结果。本研究的目的是检测p38信号转导通路在牙周细胞对血小板衍生生长因子-BB（PDGF）反应中的作用。

方法

所有实验均使用人牙周膜细胞（PDL）和牙龈成纤维细胞（GF）的原代培养物。使用细胞数量、3H-胸腺嘧啶核苷掺入法以及Boyden小室试验来表征p38信号通路的特异性抑制剂SB 203580（SB）对细胞增殖和迁移的影响。还使用体外创伤模型来评估SB的作用。对于体外创伤试验，使用0.1%胎牛血清（FBS）、10% FBS ± 10 μM SB或20 ng/ml PDGF ± 10 μM SB将一式三份的孔孵育1、3、5和7天。数字组织形态计量分析评估创伤区域内的细胞填充情况。

结果

在Boyden小室试验中，SB特异性抑制PDGF诱导的迁移，而不影响细胞增殖。创伤模型数据显示，在10% FBS中，PDL和GF的创伤填充水平相似。相对于10% FBS，用PDGF刺激的PDL显示出比GF（12%）显著更高（P < 0.01，方差分析）的创伤填充（74%）。SB分别抑制PDGF诱导的PDL和GF的创伤填充64%和57%。这种抑制仅对PDL具有显著性（P < 0.01，方差分析）。与单独的10% FBS相比，向10% FBS中添加SB对两种细胞类型的创伤填充反应均无显著影响。