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The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method
Mineralized and sound dentin matrices contain inactive preforms of proteolytic enzymes that may be activated during the demineralization cycle. In this study, we tested the hypothesis that protease inhibitors (PI) preserve demineralized collagen fibrils and other constituents of the dentin matrix an...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5705123/ https://www.ncbi.nlm.nih.gov/pubmed/29182665 http://dx.doi.org/10.1371/journal.pone.0188277 |
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author | Nurrohman, Hamid Carneiro, Karina M. M. Hellgeth, John Saeki, Kuniko Marshall, Sally J. Marshall, Grayson W. Habelitz, Stefan |
author_facet | Nurrohman, Hamid Carneiro, Karina M. M. Hellgeth, John Saeki, Kuniko Marshall, Sally J. Marshall, Grayson W. Habelitz, Stefan |
author_sort | Nurrohman, Hamid |
collection | PubMed |
description | Mineralized and sound dentin matrices contain inactive preforms of proteolytic enzymes that may be activated during the demineralization cycle. In this study, we tested the hypothesis that protease inhibitors (PI) preserve demineralized collagen fibrils and other constituents of the dentin matrix and thereby affect the potential for remineralization. Artificial carious lesions with lesion depths of 140 μm were created with acetate buffer (pH = 5.0, 66 hours), and remineralized using a polymer-induced-liquid-precursor (PILP) process (pH = 7.4, 14 days) containing poly(aspartic acid) (pAsp) as the process-directing agent. De- and remineralizing procedures were performed in the presence or absence of PI. Ultrastructure and mechanical recovery of demineralized dentin following PILP remineralization were examined and measured in water with atomic force microscopy (AFM) and nanoindentation. Nanomechanical properties of hydrated artificial lesions had a low elastic modulus (E(R) <0.4 GPa) extending about 100 μm into the lesion, followed by a sloped region of about 140 μm depth where values reached those of normal dentin (18.0–20.0 GPa). Mapping of mineral content by both micro-FTIR and micro x-ray computed tomography correlated well with modulus profiles obtained by nanoindentation. Tissue demineralized in the presence of PI exhibited higher elastic moduli (average 2.8 GPa) across the lesion and comprised a narrow zone in the outer lesion with strongly increased modulus (up to 8 GPa; p < 0.05), which might be related to the preservation of non-collagenous proteins that appear to induce calcium phosphate mineral formation even under demineralizing physical-chemical conditions. However, mechanical aspects of remineralization through the elastic modulus change, and the micromorphological aspects with SEM and TEM observation were almost identical with PILP treatments being conducted in the presence or absence of PI. Thus, the application of the protease inhibitors (PI) seemed to be less effective in promoting the remineralization of demineralized dentin. |
format | Online Article Text |
id | pubmed-5705123 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-57051232017-12-08 The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method Nurrohman, Hamid Carneiro, Karina M. M. Hellgeth, John Saeki, Kuniko Marshall, Sally J. Marshall, Grayson W. Habelitz, Stefan PLoS One Research Article Mineralized and sound dentin matrices contain inactive preforms of proteolytic enzymes that may be activated during the demineralization cycle. In this study, we tested the hypothesis that protease inhibitors (PI) preserve demineralized collagen fibrils and other constituents of the dentin matrix and thereby affect the potential for remineralization. Artificial carious lesions with lesion depths of 140 μm were created with acetate buffer (pH = 5.0, 66 hours), and remineralized using a polymer-induced-liquid-precursor (PILP) process (pH = 7.4, 14 days) containing poly(aspartic acid) (pAsp) as the process-directing agent. De- and remineralizing procedures were performed in the presence or absence of PI. Ultrastructure and mechanical recovery of demineralized dentin following PILP remineralization were examined and measured in water with atomic force microscopy (AFM) and nanoindentation. Nanomechanical properties of hydrated artificial lesions had a low elastic modulus (E(R) <0.4 GPa) extending about 100 μm into the lesion, followed by a sloped region of about 140 μm depth where values reached those of normal dentin (18.0–20.0 GPa). Mapping of mineral content by both micro-FTIR and micro x-ray computed tomography correlated well with modulus profiles obtained by nanoindentation. Tissue demineralized in the presence of PI exhibited higher elastic moduli (average 2.8 GPa) across the lesion and comprised a narrow zone in the outer lesion with strongly increased modulus (up to 8 GPa; p < 0.05), which might be related to the preservation of non-collagenous proteins that appear to induce calcium phosphate mineral formation even under demineralizing physical-chemical conditions. However, mechanical aspects of remineralization through the elastic modulus change, and the micromorphological aspects with SEM and TEM observation were almost identical with PILP treatments being conducted in the presence or absence of PI. Thus, the application of the protease inhibitors (PI) seemed to be less effective in promoting the remineralization of demineralized dentin. Public Library of Science 2017-11-28 /pmc/articles/PMC5705123/ /pubmed/29182665 http://dx.doi.org/10.1371/journal.pone.0188277 Text en © 2017 Nurrohman et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Nurrohman, Hamid Carneiro, Karina M. M. Hellgeth, John Saeki, Kuniko Marshall, Sally J. Marshall, Grayson W. Habelitz, Stefan The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title | The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title_full | The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title_fullStr | The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title_full_unstemmed | The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title_short | The role of protease inhibitors on the remineralization of demineralized dentin using the PILP method |
title_sort | role of protease inhibitors on the remineralization of demineralized dentin using the pilp method |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5705123/ https://www.ncbi.nlm.nih.gov/pubmed/29182665 http://dx.doi.org/10.1371/journal.pone.0188277 |
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