基质金属蛋白酶、基质金属蛋白酶组织抑制剂与环孢素 A 诱导的牙龈增生中的炎症：采用人口腔黏膜三维模型的初步体外研究。

Matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases, and inflammation in cyclosporine A-induced gingival enlargement: a pilot in vitro study using a three-dimensional model of the human oral mucosa.

机构信息

Department of Periodontics, The University of Texas Health Science Center, San Antonio, TX, USA.

出版信息

J Periodontol. 2013 May;84(5):634-40. doi: 10.1902/jop.2012.120224. Epub 2012 Aug 30.

DOI:10.1902/jop.2012.120224

PMID:22934840

Abstract

BACKGROUND

It has been suggested that cyclosporine A (CsA) induces gingival enlargement by promoting an increase in the gingival extracellular matrix (ECM). Nonetheless, the variable occurrence of CsA-induced gingival enlargement in patients receiving this medication indicates a multifactorial pathogenesis. Clinical observations suggest that local inflammation is associated with the development and severity of CsA-induced gingival enlargement. Therefore, the purpose of this study is to investigate the effects of CsA and inflammation on the production of ECM homeostatic mediators.

METHODS

The effects of CsA and inflammation (as assessed using interleukin [IL]-1β) on the secretion of mediators involved in ECM homeostasis were determined using fibroblast monolayers and three-dimensional (3D) models of the human oral mucosa. Fibroblast monolayers and 3D cultures were treated with CsA alone or in combination with IL-1β for up to 72 hours, and the secretion of matrix metalloproteinases (MMPs) 1, 2, 3, 8, 9, 10, and 13 and tissue inhibitors of MMPs (TIMPs) 1, 2, and 4 into the culture medium was assessed using enzyme-linked immunoassay-based antibody arrays.

RESULTS

Fibroblast monolayers responded to CsA with no changes in the secretion of ECM mediators. Conversely, 3D cultures responded to CsA treatment with a reduction in MMP-10 secretion. IL-1β alone triggered higher secretory levels of MMPs in both fibroblast monolayers (MMP-3 and MMP-10) and 3D cultures (MMP-9 and MMP-10). Importantly, fibroblast monolayers and 3D cultures treated with a combination of IL-1β and CsA showed a decrease in the MMP-1/TIMP-1 ratio.

CONCLUSIONS

These data support the hypothesis that inflammation may alter the pathogenesis of CsA-induced gingival enlargement by promoting a synergistic decrease in the MMP-1/TIMP-1 ratio.

摘要

背景

有研究表明，环孢素 A（CsA）通过促进细胞外基质（ECM）的增加诱导牙龈肥大。然而，接受这种药物治疗的患者中 CsA 诱导的牙龈肥大的发生率不同，这表明其发病机制具有多因素性。临床观察表明，局部炎症与 CsA 诱导的牙龈肥大的发展和严重程度有关。因此，本研究旨在探讨 CsA 和炎症对 ECM 稳态介质产生的影响。

方法

使用成纤维细胞单层和人口腔黏膜的三维（3D）模型，研究 CsA 和炎症（用白细胞介素[IL]-1β评估）对 ECM 稳态介质分泌的影响。成纤维细胞单层和 3D 培养物用 CsA 单独或与 IL-1β 联合处理，时间长达 72 小时，用基于酶联免疫吸附试验的抗体阵列评估基质金属蛋白酶（MMPs）1、2、3、8、9、10 和 13 以及组织抑制剂 MMPs（TIMPs）1、2 和 4 分泌到培养基中的情况。

结果

CsA 作用于成纤维细胞单层时，ECM 介质的分泌没有变化。相反，3D 培养物对 CsA 处理的反应是 MMP-10 分泌减少。IL-1β 单独作用于成纤维细胞单层（MMP-3 和 MMP-10）和 3D 培养物（MMP-9 和 MMP-10）时，会引发更高的 MMP 分泌水平。重要的是，用 IL-1β 和 CsA 联合处理的成纤维细胞单层和 3D 培养物显示 MMP-1/TIMP-1 比值降低。