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.
Cunha S.R., Maravic T., Comba A., Ramos P.A., Tay F.R., Pashley D.H., et al. (2020). In vivo and in vitro radiotherapy increased dentin enzymatic activity. JOURNAL OF DENTISTRY, 100, 1-6 [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.
