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Effects of Sucrose on the Extracellular Matrix of Plaque-Like Biofilm Formed in vivo, Studied by Proteomic AnalysisPaes Leme A.F.a · Bellato C.M.b · Bedi G.c · Del Bel Cury A.A.a · Koo H.c · Cury J.A.a
aPiracicaba Dental School, UNICAMP, and bCENA/University of São Paulo, Piracicaba, Brazil; cUniversity of Rochester Medical Center, Rochester, N.Y., USA Corresponding Author
Prof. Jaime A. Cury
Piracicaba Dental School, PO Box 52
13414-903 Piracicaba, SP (Brazil)
Tel./Fax +55 19 2106 5302
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 8×/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.
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