TY - JOUR
T1 - Use of crosslinkers to inactivate dentin MMPs
AU - Seseogullari-Dirihan, R.
AU - Apollonio, F.
AU - Mazzoni, A.
AU - Tjaderhane, L.
AU - Pashley, D.
AU - Breschi, L.
AU - Tezvergil-Mutluay, A.
N1 - Funding Information:
This study was supported by grant #8126472 from the Academy of Finland to AT-M (PI), EVO funding of Turku University Hospital to AT-M (PI) and by FINDOS Mobility Grant awarded to Dr. Seseogullari Dirihan as part of her PhD thesis. The authors wish to thank Mr. Aurelio Valmori for photographical assistance. The authors do not have a financial interest in products, equipment, and companies cited in the manuscript.
Publisher Copyright:
© 2015 Academy of Dental Materials.
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Objectives This study evaluated the endogenous matrix metalloproteinase (MMP) activity of demineralized dentin matrix following 1 or 5 min pretreatment by various collagen crosslinkers. Generic MMP activity assay, total protein analysis, in situ zymography, gelatin zymography and multiplex bead technology were used to evaluate matrix-bound MMP activity. Methods Six different crosslinkers; glutaraldehyde, riboflavin/UVA, riboflavin-5-monophospate/UVA, sumac berry extract, grape seed extract, and curcumin were used. Demineralized dentin beams were pretreated with respective crosslinkers for 1 or 5 min. Demineralized dentin beams with no crosslinker pretreatment served as control. The reduction in the total activity of dentin matrices were measured using generic MMP activity assay. Dentin slabs were used for in situ zymography and evaluated by using hydrolysis of self-quenched fluorescein-conjugated gelatin under confocal microscopy. Dentin beam extracts were used for total protein assay and multiplex analysis and powder extracts were used for gelatin zymography. Results MMP activity in crosslinker pretreated samples decreased significantly between 21% and 70%, whereas untreated control samples' activity increased up to 84%. Zymograms confirmed a decrease in the gelatinolytic activity and in the amount of extractable total protein content. Multiplex analysis of extracts of crosslinker-treated dentin showed a reduction in the MMP-8, MMP-2 and MMP-9 release. Significance The result of this work suggests that the effect of the crosslinkers is source-dependent. The use of crosslinkers for as little as 1 min on demineralized dentin can inactivate the endogenous protease activity of dentin matrices.
AB - Objectives This study evaluated the endogenous matrix metalloproteinase (MMP) activity of demineralized dentin matrix following 1 or 5 min pretreatment by various collagen crosslinkers. Generic MMP activity assay, total protein analysis, in situ zymography, gelatin zymography and multiplex bead technology were used to evaluate matrix-bound MMP activity. Methods Six different crosslinkers; glutaraldehyde, riboflavin/UVA, riboflavin-5-monophospate/UVA, sumac berry extract, grape seed extract, and curcumin were used. Demineralized dentin beams were pretreated with respective crosslinkers for 1 or 5 min. Demineralized dentin beams with no crosslinker pretreatment served as control. The reduction in the total activity of dentin matrices were measured using generic MMP activity assay. Dentin slabs were used for in situ zymography and evaluated by using hydrolysis of self-quenched fluorescein-conjugated gelatin under confocal microscopy. Dentin beam extracts were used for total protein assay and multiplex analysis and powder extracts were used for gelatin zymography. Results MMP activity in crosslinker pretreated samples decreased significantly between 21% and 70%, whereas untreated control samples' activity increased up to 84%. Zymograms confirmed a decrease in the gelatinolytic activity and in the amount of extractable total protein content. Multiplex analysis of extracts of crosslinker-treated dentin showed a reduction in the MMP-8, MMP-2 and MMP-9 release. Significance The result of this work suggests that the effect of the crosslinkers is source-dependent. The use of crosslinkers for as little as 1 min on demineralized dentin can inactivate the endogenous protease activity of dentin matrices.
KW - Collagen crosslinker
KW - Cycteine cathepsins
KW - Dentin
KW - Gluteraldehyde
KW - Matrix metalloproteinase
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U2 - 10.1016/j.dental.2015.12.012
DO - 10.1016/j.dental.2015.12.012
M3 - Article
C2 - 26764177
AN - SCOPUS:84958768558
SN - 0109-5641
VL - 32
SP - 423
EP - 432
JO - Dental Materials
JF - Dental Materials
IS - 3
ER -