Abstract
The use of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC) has recently been investigated for its effectiveness in the prevention of collagen degradation over time and the improvement of resin-dentin bond durability. The objective of the present study was to evaluate the effects of a 0.3 M EDC-containing conditioner on endogenous enzymatic activities within the hybrid layer (HL) created by a self-etch or an etch-and-rinse adhesive after 1 y. The activity within the HL was examined using in situ zymography and confocal laser scanning microscopy after 24 h or 1-y storage in artificial saliva. Dentin specimens were bonded with Clearfil SE Bond (CSE) or XP Bond (XPB). For CSE, the self-etching primer was applied and treated with 0.3 M EDC for 1 min, and then the bonding agent was applied. For XPB, dentin was etched and treated with 0.3 M EDC for 1 min and then bonded with the primer-bonding agent. Control specimens were prepared without EDC treatment. Slices containing the adhesive-dentin interface were covered with fluorescein-conjugated gelatin and observed with a multiphoton confocal microscope. Fluorescence intensity emitted by hydrolyzed fluorescein-conjugated gelatin was quantified, and the amount of gelatinolytic activity was represented by the percentage of green fluorescence emitted within the HL. After 24 h of storage, enzymatic activity was detected by in situ zymography within the HLs of both tested adhesives, with XPB higher than CSE (P < 0.05). Almost no fluorescence signal was detected when specimens were pretreated with EDC compared to controls (P < 0.05). After 1 y of storage, enzymatic activities significantly increased for all groups (excluding XPB control) compared to 24-h storage (P < 0.05), with EDC pretreated specimens exhibiting significantly lower activity than controls (P < 0.05). The present study showed, for the first time, that the use of EDC for both the self-etch and the etch-and-rinse approaches results in the reduction but not complete inhibition of matrix-bound collagenolytic enzyme activities over time in the HL.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 902-908 |
| Number of pages | 7 |
| Journal | Journal of Dental Research |
| Volume | 96 |
| Issue number | 8 |
| DOIs | |
| State | Published - Jul 1 2017 |
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Keywords
- adhesives
- collagen(s)
- dentin
- enzymology
- matrix metalloproteinases
- microscopy
ASJC Scopus subject areas
- Dentistry(all)
Cite this
Substantivity of Carbodiimide Inhibition on Dentinal Enzyme Activity over Time. / Mazzoni, A.; Angeloni, V.; Sartori, N.; Duarte, S.; Maravic, T.; Tjäderhane, L.; Pashley, D. H.; Tay, F. R.; Breschi, L.
In: Journal of Dental Research, Vol. 96, No. 8, 01.07.2017, p. 902-908.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Substantivity of Carbodiimide Inhibition on Dentinal Enzyme Activity over Time
AU - Mazzoni, A.
AU - Angeloni, V.
AU - Sartori, N.
AU - Duarte, S.
AU - Maravic, T.
AU - Tjäderhane, L.
AU - Pashley, D. H.
AU - Tay, F. R.
AU - Breschi, L.
PY - 2017/7/1
Y1 - 2017/7/1
N2 - The use of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC) has recently been investigated for its effectiveness in the prevention of collagen degradation over time and the improvement of resin-dentin bond durability. The objective of the present study was to evaluate the effects of a 0.3 M EDC-containing conditioner on endogenous enzymatic activities within the hybrid layer (HL) created by a self-etch or an etch-and-rinse adhesive after 1 y. The activity within the HL was examined using in situ zymography and confocal laser scanning microscopy after 24 h or 1-y storage in artificial saliva. Dentin specimens were bonded with Clearfil SE Bond (CSE) or XP Bond (XPB). For CSE, the self-etching primer was applied and treated with 0.3 M EDC for 1 min, and then the bonding agent was applied. For XPB, dentin was etched and treated with 0.3 M EDC for 1 min and then bonded with the primer-bonding agent. Control specimens were prepared without EDC treatment. Slices containing the adhesive-dentin interface were covered with fluorescein-conjugated gelatin and observed with a multiphoton confocal microscope. Fluorescence intensity emitted by hydrolyzed fluorescein-conjugated gelatin was quantified, and the amount of gelatinolytic activity was represented by the percentage of green fluorescence emitted within the HL. After 24 h of storage, enzymatic activity was detected by in situ zymography within the HLs of both tested adhesives, with XPB higher than CSE (P < 0.05). Almost no fluorescence signal was detected when specimens were pretreated with EDC compared to controls (P < 0.05). After 1 y of storage, enzymatic activities significantly increased for all groups (excluding XPB control) compared to 24-h storage (P < 0.05), with EDC pretreated specimens exhibiting significantly lower activity than controls (P < 0.05). The present study showed, for the first time, that the use of EDC for both the self-etch and the etch-and-rinse approaches results in the reduction but not complete inhibition of matrix-bound collagenolytic enzyme activities over time in the HL.
AB - The use of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC) has recently been investigated for its effectiveness in the prevention of collagen degradation over time and the improvement of resin-dentin bond durability. The objective of the present study was to evaluate the effects of a 0.3 M EDC-containing conditioner on endogenous enzymatic activities within the hybrid layer (HL) created by a self-etch or an etch-and-rinse adhesive after 1 y. The activity within the HL was examined using in situ zymography and confocal laser scanning microscopy after 24 h or 1-y storage in artificial saliva. Dentin specimens were bonded with Clearfil SE Bond (CSE) or XP Bond (XPB). For CSE, the self-etching primer was applied and treated with 0.3 M EDC for 1 min, and then the bonding agent was applied. For XPB, dentin was etched and treated with 0.3 M EDC for 1 min and then bonded with the primer-bonding agent. Control specimens were prepared without EDC treatment. Slices containing the adhesive-dentin interface were covered with fluorescein-conjugated gelatin and observed with a multiphoton confocal microscope. Fluorescence intensity emitted by hydrolyzed fluorescein-conjugated gelatin was quantified, and the amount of gelatinolytic activity was represented by the percentage of green fluorescence emitted within the HL. After 24 h of storage, enzymatic activity was detected by in situ zymography within the HLs of both tested adhesives, with XPB higher than CSE (P < 0.05). Almost no fluorescence signal was detected when specimens were pretreated with EDC compared to controls (P < 0.05). After 1 y of storage, enzymatic activities significantly increased for all groups (excluding XPB control) compared to 24-h storage (P < 0.05), with EDC pretreated specimens exhibiting significantly lower activity than controls (P < 0.05). The present study showed, for the first time, that the use of EDC for both the self-etch and the etch-and-rinse approaches results in the reduction but not complete inhibition of matrix-bound collagenolytic enzyme activities over time in the HL.
KW - adhesives
KW - collagen(s)
KW - dentin
KW - enzymology
KW - matrix metalloproteinases
KW - microscopy
UR - http://www.scopus.com/inward/record.url?scp=85022187092&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85022187092&partnerID=8YFLogxK
U2 - 10.1177/0022034517708312
DO - 10.1177/0022034517708312
M3 - Article
C2 - 28499097
AN - SCOPUS:85022187092
VL - 96
SP - 902
EP - 908
JO - Journal of Dental Research
JF - Journal of Dental Research
SN - 0022-0345
IS - 8
ER -