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Extraction and Separation of Collagen on SDS PAGE to Determine Effect of Whitening Strips on Teeth
Author(s) -
MaricheBanos David,
Rotsides Photis,
Keenan Kelly
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.530.27
Subject(s) - tooth whitening , chemistry , dentin , chromatography , saliva , coomassie brilliant blue , enamel paint , demineralization , polyacrylamide gel electrophoresis , extraction (chemistry) , acetic acid , sodium metabisulfite , dentistry , pepsin , staining , biochemistry , food science , medicine , enzyme , organic chemistry , pathology
In the United States, billions of dollars are spent each year of tooth whitening products including treatments in dentist's offices as well as whitening strips which are sold over the counter. Both of these products use hydrogen peroxide as the active ingredient which has long been known to damage proteins. The outermost part of teeth is the enamel which contains several soluble proteins and the dentin underneath contains nearly 90 to 95 % collagen. The goal of this project was to extract collagen from both untreated teeth as well as those treated with whitening strips and examine the collagen on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). Teeth were placed into artificial saliva and either treated with whitening strips for one or three rounds according to manufacturer's instructions or were untreated. Following a treatment, teeth were placed in artificial saliva for 30 minutes. Following treatments, teeth were dialyzed against 10% EDTA, pH 9.0 for two weeks with several changes of EDTA solution. The teeth were then dialyzed against water for two weeks with several changes. The tooth remnants were placed in liquid nitrogen, hammered into shards and ground into a powder using a blender. The powder was suspended in 0.5 M acetic acid and is the collagen fraction (COL); it was treated with pepsin for four hours, neutralized with TRIS, centrifuged and the supernatant was the pepsin soluble collagen fraction (PSC). Gels of varying concentrations of acrylamide were prepared and COL and PSC fractions were loaded. Two modified procedures for the Coomassie Blue staining protocol were used. In one, the washing solution did not contain methanol and this caused the COL proteins to turn a pink color. In addition, the gels were stained with the Oriole fluorescent stain. In order to determine the type of collagen present, standards made from collagen I and III were also used. Results will be presented that shows the level of COL in treated and untreated teeth. Support or Funding Information Funding was provided by the Research and Professional Development Grant from Stockton University. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .