Morpho-functional effects of different universal dental adhesives on human gingival fibroblasts: an in vitro study

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ORIGINAL ARTICLE

Morpho‑functional effects of different universal dental adhesives on human gingival fibroblasts: an in vitro study Stefano Pagano1 · Guido Lombardo1 · Egidia Costanzi2 · Stefania Balloni2 · Stefano Bruscoli3 · Sara Flamini3 · Maddalena Coniglio1 · Chiara Valenti1 · Stefano Cianetti1 · Lorella Marinucci2  Received: 11 June 2020 / Accepted: 26 October 2020 © The Author(s) 2020

Abstract To analyze the effects of four universal adhesives (Optibond Solo Plus—OB, Universal Bond—UB, Prime&Bond Active— PBA, FuturaBond M + —FB) on human gingival fibroblasts in terms of cytotoxicity, morphology and function. After in vitro exposure for up to 48 h, fibroblast viability was determined by the MTT assay determined, morphology by phase-contrast microscopy and migration by the scratch wound assay. Expression levels of IL1β, IL6, IL8, IL10, TNFα and VEGF genes were assessed by RT-PCR and their protein production by Western blot analysis. Apoptosis and cell cycle were analyzed by flow cytometry. OB and UB induced early morphological changes on fibroblasts (3 h) with extended cell death at 24 h/48 h. Gene expression of collagen type I and fibronectin increased fivefold compared with controls, elastin disappeared and elastase increased threefold, indicating gingival tissue tended to become fibrotic. Only UB and OB increased gene expression of inflammatory markers: IL1β at 3 and 48 h (up to about three times), IL6 and IL8 at 3 h (up to almost four times) which corresponded to the increase of the activated form NF-kB. All adhesives showed an effect on the functionality of fibroblasts with cytotoxic effect time and concentration dependent. Among all the OB and UB adhesives, they showed the greatest cell damage. The in-depth analysis of the effects of universal adhesives and possible functional effects represents an important information for the clinician towards choosing the most suitable adhesive system. Keywords  Dental adhesive · Fibroblast · MTT · Apoptosis · Inflammation · ECM

* Lorella Marinucci [email protected]

Chiara Valenti [email protected]

Stefano Pagano [email protected]

Stefano Cianetti [email protected]

Guido Lombardo [email protected]

1



School of Medicine, Department of Biomedical and Surgical Sciences, Odontostomatological University Centre: Chair Prof. Stefano Cianetti, University of Perugia, S. Andrea Delle Fratte, 06156 Perugia, Italy

2



Department of Experimental Medicine, Section of Biosciences and Medical Embriology, University of Perugia, S. Andrea Delle Fratte, 06156 Perugia, Italy

3



Department of Medicine, Section of Pharmacology, University of Perugia, S. Andrea Delle Fratte, 06156 Perugia, Italy

Egidia Costanzi [email protected] Stefania Balloni [email protected] Stefano Bruscoli [email protected] Sara Flamini [email protected] Maddalena Coniglio [email protected]

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Odontology

Introduction Universal adhesives were developed to solve clinical and practical problems in cons