Effect of lactic acid and proteolytic enzymes on the release of organic matrix components from human root dentin.

S. Z. Dung, Richard Gregory, Y. Li, G. K. Stookey

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Abstract

The mechanisms of organic matrix breakdown in the root caries process are not well understood. Therefore, the combined and separate effects of lactic acid and proteolytic enzymes on the degradation of human dentin collagen, glycoproteins, proteoglycans and phosphoproteins were investigated in the present study. Dentin powder was pretreated with lactic acid (pH 4.0), distilled and deionized (dd) water (pH 7.0) and EDTA/guanidine HCl (pH 7.4) for 24 h. Pellets of acid- or dd water-pretreated dentin powder were washed, dried, and then treated with trypsin, bacterial or mammalian tissue collagenase, or control buffer for 3 h. The released dentin proteins were analyzed by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting to identify degraded type I collagen, proteoglycans, glycoproteins and phosphoproteins. All water and acid pretreatment and enzyme treatment groups demonstrated two collagen fragment bands with molecular weights at approximately 79 kD. Further studies showed that the 79 kD proteins from acid-pretreated dentin collagen were degraded by tissue collagenase, suggesting that endogenous collagenase may be involved in the degradation of root dentin collagen. Dentin proteoglycans were detectable in all the treatment groups by protein slot blotting. Relatively few distinct glycoproteins and proteoglycans, and no phosphoproteins were detected by immunoblotting. Results from this study suggest that both acids and proteolytic enzymes from either host or microbial origin are important in the degradation of human dentin matrix and the mechanisms involved in the release of various noncollagenous proteins may be different.

Original languageEnglish
Pages (from-to)483-489
Number of pages7
JournalCaries Research
Volume29
Issue number6
StatePublished - 1995

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Dentin
Lactic Acid
Peptide Hydrolases
Proteoglycans
Phosphoproteins
Collagenases
Collagen
Glycoproteins
Acids
Immunoblotting
Powders
Water
Proteins
Root Caries
Guanidine
Collagen Type I
Edetic Acid
Sodium Dodecyl Sulfate
Trypsin
Polyacrylamide Gel Electrophoresis

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Effect of lactic acid and proteolytic enzymes on the release of organic matrix components from human root dentin. / Dung, S. Z.; Gregory, Richard; Li, Y.; Stookey, G. K.

In: Caries Research, Vol. 29, No. 6, 1995, p. 483-489.

Research output: Contribution to journalArticle

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