Degradation of insoluble bovine collagen and human dentine collagen pretreated in vitro with lactic acid, pH 4.0 and 5.5.
The purpose of the study was to test the hypothesis that both insoluble pure type I collagen from bovine Achilles tendon and dentine collagen in root dentine powder from human teeth required acid pretreatment for subsequent degradation by trypsin, a non-specific protease. Pure type I collagen or dentine powder was treated with lactic acid, at pH 4 or 5.5, or distilled, deionized water (pH 7) as a negative control. After incubation at 37 degrees C for 24 h, extracts of pure type I collagen solutions were analysed for soluble collagen with the hydroxyproline assay. Extracts of dentine powder solution were analysed for Ca2+, total protein, final pH, and hydroxyproline. Residual, undegraded pellets were washed and then treated with trypsin or collagenase. After 24 h of incubation, the soluble fractions from the enzyme-treated pure type I collagen and dentine powder solutions were analysed for hydroxyproline. Results showed that almost no pure type I collagen was degraded during acid pretreatment. Trypsin degraded significantly more pure type I collagen in the pH 4-treated group than in the other groups. Collagenase degraded about 70% of the pure type I collagen irrespective of acid pretreatment. While acid pretreatment at pH 4 did not degrade dentine collagen, data from Ca2+ analyses and collagen breakdown by trypsin suggested that pretreatment at pH 4 demineralized and denatured dentine collagen so that the collagen could be subsequently degraded by enzymes. After pretreatment at pH 4, about 27 and 57% of the dentine collagen was degraded by trypsin and collagenase, respectively, in contrast to minimal degradation of non-acid-treated dentine collagen by the same enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)
Dung
S Z
SZ
Department of Oral Biology, School of Dentistry, Indiana University, Indianapolis 46202, USA.
Li
Y
Y
Dunipace
A J
AJ
Stookey
G K
GK
D000125:Achilles Tendon / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000032:analysis; D002417:Cattle; D003094:Collagen / Q000032:analysis / Q000737:chemistry*; D017364:Collagenases / Q000737:chemistry; D003804:Dentin / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006909:Hydroxyproline / Q000032:analysis; D007773:Lactates / Q000737:chemistry*; D019344:Lactic Acid; D010750:Phosphoproteins / Q000032:analysis; D011506:Proteins / Q000032:analysis; D012995:Solubility; D014357:Trypsin / Q000737:chemistry
D007773:Lactates; D010750:Phosphoproteins; D011506:Proteins; C017630:phosphophoryn; D019344:Lactic Acid; D003094:Collagen; D014357:Trypsin; D017364:Collagenases; D006909:Hydroxyproline; D002118:Calcium