Andrukhov Oleh, Gemperli Anja C, Tang Yan, Howald Nadia, Dard Michel, Falkensammer Frank, Moritz Andreas, Rausch-Fan Xiaohui
Competence Centre of Periodontal Research, Bernhard Gottlieb School of Dentistry, Medical University of Vienna, Austria; Division of Conservative Dentistry, Periodontology and Prophylaxis, Bernhard Gottlieb School of Dentistry, Medical University of Vienna, Vienna, Austria.
Institut Straumann AG, Basel, Switzerland.
Dent Mater. 2015 Jul;31(7):822-32. doi: 10.1016/j.dental.2015.04.004. Epub 2015 May 13.
Enamel matrix derivative (EMD) is an effective biomaterial for periodontal tissue regeneration and might stimulate angiogenesis. In order to clarify mechanisms underlying its biological activity, we separated two EMD fractions with different molecular weight protein components and investigated their effects on human umbilical vein endothelial cells (HUVECs) in vitro.
Fraction Low-Molecular Weight (LMW) included proteins with a molecular weight (M.W.)<8kDa. Fraction LMW-depleted included proteins with M.W.>8kDa and lower than approximately 55kDa. The effect of EMD fractions on proliferation/viability, apoptosis, migration and expression of angiopoetin-2 (ang-2), von Willebrand factor (vWF), E-selectin, intracellular adhesion molecules 1 (ICAM-1), vascular endothelial growth factor (VEGF) receptors Flt-1 and KDR was investigated.
The proliferation/viability of HUVECs was inhibited by both LMW and LMW-depleted at concentrations 100μg/ml, whereas EMD slightly increased cell proliferation/viability. The expression of all investigated proteins was up-regulated by EMD. However, differences in the effect of EMD fractions on the protein expression were observed. The effect of LMW-depleted on the expression of ICAM-1 and E-selectin was markedly higher compared to LMW. In contrast, the expression of vWF and VEGF receptors Flt-1 and KDR was primarily affected LMW than by LMW depleted. The expression of ang-2 was not influenced by LMW and LMW-depleted. HUVECs migration was stimulated more strongly by LMW than by EMD and LMW-depleted.
Our in vitro study shows that the proteins composing EMD have different and specific biological activities and consequently have the ability to cover different aspects of EMD's biological and clinical effects.
釉基质衍生物（EMD）是一种用于牙周组织再生的有效生物材料，可能会刺激血管生成。为了阐明其生物活性的潜在机制，我们分离了两种具有不同分子量蛋白质成分的EMD组分，并在体外研究了它们对人脐静脉内皮细胞（HUVECs）的影响。
低分子量（LMW）组分包含分子量（M.W.）<8kDa的蛋白质。去除LMW的组分包含分子量>8kDa且低于约55kDa的蛋白质。研究了EMD组分对增殖/活力、凋亡、迁移以及血管生成素-2（ang-2）、血管性血友病因子（vWF）、E-选择素、细胞间黏附分子1（ICAM-1）、血管内皮生长因子（VEGF）受体Flt-1和KDR表达的影响。
在浓度为100μg/ml时，LMW和去除LMW的组分均抑制了HUVECs的增殖/活力，而EMD则轻微增加了细胞增殖/活力。EMD上调了所有研究蛋白的表达。然而，观察到EMD组分对蛋白表达的影响存在差异。与LMW相比，去除LMW的组分对ICAM-1和E-选择素表达的影响明显更高。相反，vWF以及VEGF受体Flt-1和KDR的表达主要受LMW影响，而不是去除LMW的组分。LMW和去除LMW的组分均未影响ang-2的表达。LMW比EMD和去除LMW的组分更强烈地刺激了HUVECs的迁移。
我们的体外研究表明，构成EMD的蛋白质具有不同的特定生物活性，因此有能力涵盖EMD生物和临床效应的不同方面。
