Paes Leme A F, Bellato C M, Bedi G, Cury A A Del Bel, Koo H, Cury J A
Piracicaba Dental School, UNICAMP, Piracicaba, Brazil.
Caries Res. 2008;42(6):435-43. doi: 10.1159/000159607. Epub 2008 Oct 3.
Previous studies have shown that sucrose promotes changes in the composition of the extracellular matrix (ECM) of plaque-like biofilm (PLB), but its effect on protein expression has not been studied in vivo. Therefore, the protein compositions of ECM of PLB formed with and without sucrose exposure were analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). For this purpose, a crossover study was conducted during two phases of 14 days each, during which a volunteer wore a palatal appliance containing eight enamel blocks for PLB accumulation. In each phase, a 20% sucrose solution or distilled and deionized water (control) were extraorally dripped onto the blocks 8x/day. On the 14th day, the PLB were collected, the ECM proteins were extracted, separated by two-dimensional gel electrophoresis, digested by in-gel trypsin and MALDI-TOF MS analyzed. In the ECM of PLB formed under sucrose exposure, the following changes compared with the control PLB were observed: (1) the presence of upregulated proteins that may be involved in bacterial response to environmental changes induced by sucrose and (2) the absence of calcium-binding proteins that may partly explain the low inorganic concentration found in ECM of PLB formed under sucrose exposure. The findings showing that sucrose affected the ECM protein composition of PLB in vivo provide further insight into the unique cariogenic properties of this dietary carbohydrate.
先前的研究表明,蔗糖可促进菌斑样生物膜(PLB)细胞外基质(ECM)成分的变化,但其对蛋白质表达的影响尚未在体内进行研究。因此,通过二维凝胶电泳和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)分析了在有或无蔗糖暴露情况下形成的PLB的ECM蛋白质组成。为此,进行了一项交叉研究,分为两个阶段,每个阶段14天,在此期间一名志愿者佩戴一个含有八个用于PLB积聚的牙釉质块的腭部矫治器。在每个阶段,每天8次经口外将20%的蔗糖溶液或蒸馏水和去离子水(对照)滴到这些块上。在第14天,收集PLB,提取ECM蛋白质,通过二维凝胶电泳分离,用胶内胰蛋白酶消化并进行MALDI-TOF MS分析。在蔗糖暴露下形成的PLB的ECM中,与对照PLB相比观察到以下变化:(1)存在可能参与细菌对蔗糖诱导的环境变化反应的上调蛋白质,以及(2)不存在钙结合蛋白,这可能部分解释了在蔗糖暴露下形成的PLB的ECM中发现的低无机浓度。这些结果表明蔗糖在体内影响了PLB的ECM蛋白质组成,为深入了解这种膳食碳水化合物独特的致龋特性提供了进一步的依据。