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.
In vivo and in vitro radiotherapy increased dentin enzymatic activity / Cunha S.R.; Maravic T.; Comba A.; Ramos P.A.; Tay F.R.; Pashley D.H.; Fregnani E.R.; Aranha A.C.; Mazzoni A.; Breschi L.. - In: JOURNAL OF DENTISTRY. - ISSN 0300-5712. - ELETTRONICO. - 100:(2020), pp. 103429.1-103429.6. [10.1016/j.jdent.2020.103429]
In vivo and in vitro radiotherapy increased dentin enzymatic activity
Maravic T.;Comba A.;Mazzoni A.;Breschi L.
2020
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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
