Differential expression of matrix metalloproteinase-2 in human coronal and radicular sound and carious dentine

M. Toledano, R. Nieto-Aguilar, R. Osorio, A. Campos, E. Osorio, Franklin Chi Meng Tay, M. Alaminos

Research output: Contribution to journalArticle

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Abstract

Objective: To examine the differential expression of matrix metalloproteinase-2 (MMP-2) in human coronal and radicular sound and carious dentine using combined trichrome staining technique and immunofluorescence approach. Methods: Freshly extracted human premolars were fixed with formaldehyde, demineralised with 10% EDTA (pH 7.4), dehydrated and sectioned for light and immunofluorescence microscopy. Half of the sections were stained with Masson's trichrome and examined with light microscopy to identify regions in the coronal and radicular parts of the teeth that contained sound, caries-affected and caries-infected dentine. The rest of the sections were hybridized with anti-mouse MMP-2 primary antibody and FITC-conjugated secondary antibody. Immunofluorescence of the FITC that was indicative of the distribution of the MMP-2 in coronal and radicular dentine was analysed by fluorescence light microscopy. Results: Trichrome staining revealed a green zone of unaffected sound dentin, red irregular regions of caries-infected dentine and pink regions of caries-affected dentine. Immunofluorescence signals that were indicative of MMP expression were the lowest in sound dentine and most intense in the caries-infected dentine. Caries-affected dentine showed intermediate immunoreactivity. The variations in the intensities of immunofluorescence corresponded well with the distribution of caries-infected and caries-affected dentine in the trichrome-stained sections, for both coronal and radicular dentine. Conclusion: Caries stimulates MMP-2 expression, resulting in the differential expression of this protease in sound, caries-affected and caries-infected dentine. The more intense MMP-2 expression in caries-affected dentine compared with sound dentine may imply more rapid hybrid layer degradation when caries-affected dentine is employed as the substrate for bonded restorations.

Original languageEnglish (US)
Pages (from-to)635-640
Number of pages6
JournalJournal of Dentistry
Volume38
Issue number8
DOIs
StatePublished - Aug 1 2010

Fingerprint

Dental Caries
Dentin
Matrix Metalloproteinase 2
Fluorescent Antibody Technique
Fluorescein-5-isothiocyanate
human MMP2 protein
Fluorescence Microscopy
Light
Staining and Labeling
Antibodies
Bicuspid
Matrix Metalloproteinases
Edetic Acid
Formaldehyde
Microscopy
Tooth
Peptide Hydrolases

Keywords

  • Caries-affected
  • Caries-infected
  • Dentine
  • Immunofluorescence
  • Matrix metalloproteinase
  • MMP-2
  • Trichrome stain

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Differential expression of matrix metalloproteinase-2 in human coronal and radicular sound and carious dentine. / Toledano, M.; Nieto-Aguilar, R.; Osorio, R.; Campos, A.; Osorio, E.; Tay, Franklin Chi Meng; Alaminos, M.

In: Journal of Dentistry, Vol. 38, No. 8, 01.08.2010, p. 635-640.

Research output: Contribution to journalArticle

Toledano, M. ; Nieto-Aguilar, R. ; Osorio, R. ; Campos, A. ; Osorio, E. ; Tay, Franklin Chi Meng ; Alaminos, M. / Differential expression of matrix metalloproteinase-2 in human coronal and radicular sound and carious dentine. In: Journal of Dentistry. 2010 ; Vol. 38, No. 8. pp. 635-640.
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abstract = "Objective: To examine the differential expression of matrix metalloproteinase-2 (MMP-2) in human coronal and radicular sound and carious dentine using combined trichrome staining technique and immunofluorescence approach. Methods: Freshly extracted human premolars were fixed with formaldehyde, demineralised with 10{\%} EDTA (pH 7.4), dehydrated and sectioned for light and immunofluorescence microscopy. Half of the sections were stained with Masson's trichrome and examined with light microscopy to identify regions in the coronal and radicular parts of the teeth that contained sound, caries-affected and caries-infected dentine. The rest of the sections were hybridized with anti-mouse MMP-2 primary antibody and FITC-conjugated secondary antibody. Immunofluorescence of the FITC that was indicative of the distribution of the MMP-2 in coronal and radicular dentine was analysed by fluorescence light microscopy. Results: Trichrome staining revealed a green zone of unaffected sound dentin, red irregular regions of caries-infected dentine and pink regions of caries-affected dentine. Immunofluorescence signals that were indicative of MMP expression were the lowest in sound dentine and most intense in the caries-infected dentine. Caries-affected dentine showed intermediate immunoreactivity. The variations in the intensities of immunofluorescence corresponded well with the distribution of caries-infected and caries-affected dentine in the trichrome-stained sections, for both coronal and radicular dentine. Conclusion: Caries stimulates MMP-2 expression, resulting in the differential expression of this protease in sound, caries-affected and caries-infected dentine. The more intense MMP-2 expression in caries-affected dentine compared with sound dentine may imply more rapid hybrid layer degradation when caries-affected dentine is employed as the substrate for bonded restorations.",
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T1 - Differential expression of matrix metalloproteinase-2 in human coronal and radicular sound and carious dentine

AU - Toledano, M.

AU - Nieto-Aguilar, R.

AU - Osorio, R.

AU - Campos, A.

AU - Osorio, E.

AU - Tay, Franklin Chi Meng

AU - Alaminos, M.

PY - 2010/8/1

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N2 - Objective: To examine the differential expression of matrix metalloproteinase-2 (MMP-2) in human coronal and radicular sound and carious dentine using combined trichrome staining technique and immunofluorescence approach. Methods: Freshly extracted human premolars were fixed with formaldehyde, demineralised with 10% EDTA (pH 7.4), dehydrated and sectioned for light and immunofluorescence microscopy. Half of the sections were stained with Masson's trichrome and examined with light microscopy to identify regions in the coronal and radicular parts of the teeth that contained sound, caries-affected and caries-infected dentine. The rest of the sections were hybridized with anti-mouse MMP-2 primary antibody and FITC-conjugated secondary antibody. Immunofluorescence of the FITC that was indicative of the distribution of the MMP-2 in coronal and radicular dentine was analysed by fluorescence light microscopy. Results: Trichrome staining revealed a green zone of unaffected sound dentin, red irregular regions of caries-infected dentine and pink regions of caries-affected dentine. Immunofluorescence signals that were indicative of MMP expression were the lowest in sound dentine and most intense in the caries-infected dentine. Caries-affected dentine showed intermediate immunoreactivity. The variations in the intensities of immunofluorescence corresponded well with the distribution of caries-infected and caries-affected dentine in the trichrome-stained sections, for both coronal and radicular dentine. Conclusion: Caries stimulates MMP-2 expression, resulting in the differential expression of this protease in sound, caries-affected and caries-infected dentine. The more intense MMP-2 expression in caries-affected dentine compared with sound dentine may imply more rapid hybrid layer degradation when caries-affected dentine is employed as the substrate for bonded restorations.

AB - Objective: To examine the differential expression of matrix metalloproteinase-2 (MMP-2) in human coronal and radicular sound and carious dentine using combined trichrome staining technique and immunofluorescence approach. Methods: Freshly extracted human premolars were fixed with formaldehyde, demineralised with 10% EDTA (pH 7.4), dehydrated and sectioned for light and immunofluorescence microscopy. Half of the sections were stained with Masson's trichrome and examined with light microscopy to identify regions in the coronal and radicular parts of the teeth that contained sound, caries-affected and caries-infected dentine. The rest of the sections were hybridized with anti-mouse MMP-2 primary antibody and FITC-conjugated secondary antibody. Immunofluorescence of the FITC that was indicative of the distribution of the MMP-2 in coronal and radicular dentine was analysed by fluorescence light microscopy. Results: Trichrome staining revealed a green zone of unaffected sound dentin, red irregular regions of caries-infected dentine and pink regions of caries-affected dentine. Immunofluorescence signals that were indicative of MMP expression were the lowest in sound dentine and most intense in the caries-infected dentine. Caries-affected dentine showed intermediate immunoreactivity. The variations in the intensities of immunofluorescence corresponded well with the distribution of caries-infected and caries-affected dentine in the trichrome-stained sections, for both coronal and radicular dentine. Conclusion: Caries stimulates MMP-2 expression, resulting in the differential expression of this protease in sound, caries-affected and caries-infected dentine. The more intense MMP-2 expression in caries-affected dentine compared with sound dentine may imply more rapid hybrid layer degradation when caries-affected dentine is employed as the substrate for bonded restorations.

KW - Caries-affected

KW - Caries-infected

KW - Dentine

KW - Immunofluorescence

KW - Matrix metalloproteinase

KW - MMP-2

KW - Trichrome stain

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