In vivo and in vitro radiotherapy increased dentin enzymatic activity

Sandra Ribeiro Cunha; Tatjana Maravic; Allegra Comba; Pedro Augusto Ramos; Franklin R. Tay; David H. Pashley; Eduardo Rodrigues Fregnani; Ana Cecília Aranha; Annalisa Mazzoni; Lorenzo Breschi

doi:10.1016/j.jdent.2020.103429

In vivo and in vitro radiotherapy increased dentin enzymatic activity

Sandra Ribeiro Cunha, Tatjana Maravic, Allegra Comba, Pedro Augusto Ramos, Franklin R. Tay, David H. Pashley, Eduardo Rodrigues Fregnani, Ana Cecília Aranha, Annalisa Mazzoni, Lorenzo Breschi

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

Objectives: The present study investigated the effects of in vitro and in-vivo radiotherapy on endogenous enzymatic activity in dentin using gelatin zymography and in-situ zymography. Methods: Gelatin zymographic assays were performed on protein extracts obtained from dentin powder of sound non-irradiated (NRT), in vitro irradiated (VTRT) and in vivo irradiated (VIRT) human teeth. Their proteolytic activities were quantified using band densitometric evaluation. For in-situ zymography, dentin specimens from NRT, VIRT and VTRT were covered with fluorescein-conjugated gelatin and examined with confocal laser-scanning microscopy. Fluorescence intensity emitted by the hydrolyzed fluorescein-conjugated gelatin was quantified and statistically analyzed. In-situ zymography data were statistically analyzed using Kruskal-Wallis ANOVA and Dunn's multiple comparison procedures (α = 0.05). Results: No difference between in vitro and in vivo radiotherapy treatment was found. Both VTRT and VIRT groups showed increase in MMP-9 expression when compared to NRT group. Significant increases (p < 0.05) in gelatinolytic activity (26 % for VTRT; 55 % for VIRT) were observed when compared to the NRT group. Conclusion: Radiotherapy increase endogenous enzymatic activity in non-restored dentin.

Original language	English (US)
Article number	103429
Journal	Journal of Dentistry
Volume	100
DOIs	https://doi.org/10.1016/j.jdent.2020.103429
State	Published - Sep 2020

Keywords

Endogenous enzymatic activity
Head and neck cancer
Radiotherapy
Zymography

ASJC Scopus subject areas

General Dentistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.jdent.2020.103429

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@article{1abd982688374922900bc99e5dd6df57,

title = "In vivo and in vitro radiotherapy increased dentin enzymatic activity",

abstract = "Objectives: The present study investigated the effects of in vitro and in-vivo radiotherapy on endogenous enzymatic activity in dentin using gelatin zymography and in-situ zymography. Methods: Gelatin zymographic assays were performed on protein extracts obtained from dentin powder of sound non-irradiated (NRT), in vitro irradiated (VTRT) and in vivo irradiated (VIRT) human teeth. Their proteolytic activities were quantified using band densitometric evaluation. For in-situ zymography, dentin specimens from NRT, VIRT and VTRT were covered with fluorescein-conjugated gelatin and examined with confocal laser-scanning microscopy. Fluorescence intensity emitted by the hydrolyzed fluorescein-conjugated gelatin was quantified and statistically analyzed. In-situ zymography data were statistically analyzed using Kruskal-Wallis ANOVA and Dunn's multiple comparison procedures (α = 0.05). Results: No difference between in vitro and in vivo radiotherapy treatment was found. Both VTRT and VIRT groups showed increase in MMP-9 expression when compared to NRT group. Significant increases (p < 0.05) in gelatinolytic activity (26 % for VTRT; 55 % for VIRT) were observed when compared to the NRT group. Conclusion: Radiotherapy increase endogenous enzymatic activity in non-restored dentin.",

keywords = "Endogenous enzymatic activity, Head and neck cancer, Radiotherapy, Zymography",

author = "Cunha, {Sandra Ribeiro} and Tatjana Maravic and Allegra Comba and Ramos, {Pedro Augusto} and Tay, {Franklin R.} and Pashley, {David H.} and Fregnani, {Eduardo Rodrigues} and Aranha, {Ana Cec{\'i}lia} and Annalisa Mazzoni and Lorenzo Breschi",

note = "Funding Information: We would like to thank FAPESP, Funda{\c c}{\~a}o de Amparo {\`a} Pesquisa do Estado de S{\~a}o Paulo for the PhD Scholarship to the first author [grant number 2015/20297-3 and 2017/01777-0 ]. The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article. Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2020",

month = sep,

doi = "10.1016/j.jdent.2020.103429",

language = "English (US)",

volume = "100",

journal = "Journal of Dentistry",

issn = "0300-5712",

publisher = "Elsevier BV",

}

TY - JOUR

T1 - In vivo and in vitro radiotherapy increased dentin enzymatic activity

AU - Cunha, Sandra Ribeiro

AU - Maravic, Tatjana

AU - Comba, Allegra

AU - Ramos, Pedro Augusto

AU - Tay, Franklin R.

AU - Pashley, David H.

AU - Fregnani, Eduardo Rodrigues

AU - Aranha, Ana Cecília

AU - Mazzoni, Annalisa

AU - Breschi, Lorenzo

N1 - Funding Information: We would like to thank FAPESP, Fundação de Amparo à Pesquisa do Estado de São Paulo for the PhD Scholarship to the first author [grant number 2015/20297-3 and 2017/01777-0 ]. The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article. Publisher Copyright: © 2020 Elsevier Ltd Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/9

Y1 - 2020/9

N2 - Objectives: The present study investigated the effects of in vitro and in-vivo radiotherapy on endogenous enzymatic activity in dentin using gelatin zymography and in-situ zymography. Methods: Gelatin zymographic assays were performed on protein extracts obtained from dentin powder of sound non-irradiated (NRT), in vitro irradiated (VTRT) and in vivo irradiated (VIRT) human teeth. Their proteolytic activities were quantified using band densitometric evaluation. For in-situ zymography, dentin specimens from NRT, VIRT and VTRT were covered with fluorescein-conjugated gelatin and examined with confocal laser-scanning microscopy. Fluorescence intensity emitted by the hydrolyzed fluorescein-conjugated gelatin was quantified and statistically analyzed. In-situ zymography data were statistically analyzed using Kruskal-Wallis ANOVA and Dunn's multiple comparison procedures (α = 0.05). Results: No difference between in vitro and in vivo radiotherapy treatment was found. Both VTRT and VIRT groups showed increase in MMP-9 expression when compared to NRT group. Significant increases (p < 0.05) in gelatinolytic activity (26 % for VTRT; 55 % for VIRT) were observed when compared to the NRT group. Conclusion: Radiotherapy increase endogenous enzymatic activity in non-restored dentin.

AB - Objectives: The present study investigated the effects of in vitro and in-vivo radiotherapy on endogenous enzymatic activity in dentin using gelatin zymography and in-situ zymography. Methods: Gelatin zymographic assays were performed on protein extracts obtained from dentin powder of sound non-irradiated (NRT), in vitro irradiated (VTRT) and in vivo irradiated (VIRT) human teeth. Their proteolytic activities were quantified using band densitometric evaluation. For in-situ zymography, dentin specimens from NRT, VIRT and VTRT were covered with fluorescein-conjugated gelatin and examined with confocal laser-scanning microscopy. Fluorescence intensity emitted by the hydrolyzed fluorescein-conjugated gelatin was quantified and statistically analyzed. In-situ zymography data were statistically analyzed using Kruskal-Wallis ANOVA and Dunn's multiple comparison procedures (α = 0.05). Results: No difference between in vitro and in vivo radiotherapy treatment was found. Both VTRT and VIRT groups showed increase in MMP-9 expression when compared to NRT group. Significant increases (p < 0.05) in gelatinolytic activity (26 % for VTRT; 55 % for VIRT) were observed when compared to the NRT group. Conclusion: Radiotherapy increase endogenous enzymatic activity in non-restored dentin.

KW - Endogenous enzymatic activity

KW - Head and neck cancer

KW - Radiotherapy

KW - Zymography

UR - http://www.scopus.com/inward/record.url?scp=85087969470&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85087969470&partnerID=8YFLogxK

U2 - 10.1016/j.jdent.2020.103429

DO - 10.1016/j.jdent.2020.103429

M3 - Article

C2 - 32673637

AN - SCOPUS:85087969470

SN - 0300-5712

VL - 100

JO - Journal of Dentistry

JF - Journal of Dentistry

M1 - 103429

ER -

In vivo and in vitro radiotherapy increased dentin enzymatic activity

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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