Comparison of three techniques for the collection of in vivo acquired enamel pellicle
Date
1996
DOI
Authors
Ross, Kenneth E.
Version
OA Version
Citation
Abstract
The in vivo acquired enamel pellicle forms within two hours through selective adsorption of salivary proteins on the surface of a tooth, and is composed of saliva, gingival crevicular fluid, and microbial and cellular products. The acquired enamel pellicle serves many functions: acting as a protective barrier, regulating the remineralization and demineralization processes, and modulating bacterial adherence.
Characterizing the components of the acquired enamel pellicle and its function has been hindered by the relatively small amount of material collected for analysis. This study was undertaken to evaluate different collection methods in an attempt to establish the most accurate way to remove pellicle from the tooth surface after two hours. Three methods of collection were utilized: a scaler to remove material from enamel, pumice to remove material from enamel, and hydroxyapatite disks to adsorb proteins in situ. Samples were analyzed by amino acid analysis, reversed phase high performance liquid chromatography, and protein quantification.
The amount of protein obtained using the scaler varied among differing donors. The scaler method gave similar results for the two donors studied. This was indicated through the similar peaks on the reversed phase chromatograms, although relative amounts of the peaks differed among donors. The pumice method also gave similar results (i. e. similar peaks) for the two individual donors, while relative amounts of the peaks differed. Very little protein was adsorbed with the hydroxyapatite disk collection method and there was variability of elution patterns between donors.
The amino acid analysis in all three methods revealed high levels of glutamic acid/glutamine and glycine. Varying degrees of proline, leucine, valine, serine, alanine, and tyrosine were discovered. Only the hydroxyapatite disks showed sulfur containing amino acids. The amino acid compositions of the pellicles differed from those of whole saliva, parotid secretion, and submandibular secretion.
The chromatography patterns of pellicles from the three methods differed. The largest peak from the glass-wool was found at 16.6 minutes, with most peaks found between 7.7 and 16.6 min. For the pumice, most of the peaks fell between 7.7 and 18.4 min., with the largest peak at 18.4 min.
This study suggests that the pumice and glass-wool methods appear to be the most reliable options at this time in retrieving the proteins previously reported within the two hour pellicle samples. The pumice and glass-wool collection methods also afford the largest samples to date with which to work. Differences between the pellicles collected using the scaler, pumice, and hydroxyapatite disks may reflect different adsorption properties of pellicle proteins to glass and pumice, and differences in protein solubilities.
Description
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Thesis (M.Sc.D.)--Boston University, Henry M. Goldman School of Dental Medicine, 1996 (Periodontology and Oral Biology).
Includes bibliographical references: (leaves 46-59).
Thesis (M.Sc.D.)--Boston University, Henry M. Goldman School of Dental Medicine, 1996 (Periodontology and Oral Biology).
Includes bibliographical references: (leaves 46-59).
License
This work is protected by copyright. Downloading is restricted to the BU community. If you are the author of this work and would like to make it publicly available, please contact open-help@bu.edu.