Use of crosslinkers to inactivate dentin MMPs

R. Seseogullari-Dirihan, F. Apollonio, A. Mazzoni, L. Tjaderhane, David Henry Pashley, L. Breschi, A. Tezvergil-Mutluay

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)423-432
Number of pages10
JournalDental Materials
Volume32
Issue number3
DOIs
StatePublished - Mar 1 2016

Fingerprint

Dentin
Matrix Metalloproteinases
Gelatin
Assays
Riboflavin
Grape Seed Extract
Matrix Metalloproteinase 8
Proteins
Curcumin
Confocal microscopy
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Glutaral
Fluorescein
Rhus
Powders
Hydrolysis
Peptide Hydrolases
Collagen
Metalloproteases

Keywords

  • Collagen crosslinker
  • Cycteine cathepsins
  • Dentin
  • Gluteraldehyde
  • Matrix metalloproteinase

ASJC Scopus subject areas

  • Materials Science(all)
  • Dentistry(all)
  • Mechanics of Materials

Cite this

Seseogullari-Dirihan, R., Apollonio, F., Mazzoni, A., Tjaderhane, L., Pashley, D. H., Breschi, L., & Tezvergil-Mutluay, A. (2016). Use of crosslinkers to inactivate dentin MMPs. Dental Materials, 32(3), 423-432. https://doi.org/10.1016/j.dental.2015.12.012

Use of crosslinkers to inactivate dentin MMPs. / Seseogullari-Dirihan, R.; Apollonio, F.; Mazzoni, A.; Tjaderhane, L.; Pashley, David Henry; Breschi, L.; Tezvergil-Mutluay, A.

In: Dental Materials, Vol. 32, No. 3, 01.03.2016, p. 423-432.

Research output: Contribution to journalArticle

Seseogullari-Dirihan, R, Apollonio, F, Mazzoni, A, Tjaderhane, L, Pashley, DH, Breschi, L & Tezvergil-Mutluay, A 2016, 'Use of crosslinkers to inactivate dentin MMPs', Dental Materials, vol. 32, no. 3, pp. 423-432. https://doi.org/10.1016/j.dental.2015.12.012
Seseogullari-Dirihan R, Apollonio F, Mazzoni A, Tjaderhane L, Pashley DH, Breschi L et al. Use of crosslinkers to inactivate dentin MMPs. Dental Materials. 2016 Mar 1;32(3):423-432. https://doi.org/10.1016/j.dental.2015.12.012
Seseogullari-Dirihan, R. ; Apollonio, F. ; Mazzoni, A. ; Tjaderhane, L. ; Pashley, David Henry ; Breschi, L. ; Tezvergil-Mutluay, A. / Use of crosslinkers to inactivate dentin MMPs. In: Dental Materials. 2016 ; Vol. 32, No. 3. pp. 423-432.
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abstract = "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.",
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AU - Tjaderhane, L.

AU - Pashley, David Henry

AU - Breschi, L.

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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.

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