Dental materials : official publication of the Academy of Dental Materials · 2026
Trans-dentinal cytotoxicity assessment of adhesive systems on sound and caries-affected dentin using a microphysiological artificial pulp chamber system.
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Álamo Larissa, Stuani Vitor de Toledo, Correa Lígia Espoliar, Braga de Melo Camila Correa da Silva, Soares Igor Paulino Mendes, Anselmi Caroline, de Souza Costa Carlos Alberto, Hebling Josimeri, Soares Diana Gabriela
Аннотация
OBJECTIVE: This study evaluated, in vitro, the trans-dentinal cytotoxicity of a universal adhesive system (Single Bond Universal, SBU) applied onto sound and caries-affected dentin (CAD), using a microphysiological artificial pulp chamber (APC) on-a-chip system.
METHODS: Human dentin discs (0.35 mm) from sound (SD) or microbiologically demineralized teeth (CAD) were used in a co-culture model under simulated pulpal pressure into APC-on-a-chip, with MDPC-23 cells seeded on the pulpal surface and human dental pulp cells (HDPCs) cultured in 3D collagen gels beneath the dentin. SBU was applied on the occlusal surface in self-etch (SE) or etch-and-rinse (ER) mode and LED-cured (continuous mode). After 24 h, cell metabolism (Alamar Blue), viability (Live/Dead), dentin permeability (Lp), morphology (SEM), and HEMA diffusion (UV-vis) were evaluated. Data were analyzed using two-way ANOVA/Tukey or Student's t-tests (p < 0.05).
RESULTS: SBU application reduced MDPC-23 metabolism by 71.2% (SE) and 85.7% (ER) on SD, and 46.8% (SE) and 62.2% (ER) on CAD. HDPCs showed similar trends: decreases of 50.2% and 70.1% on SD, and 38.1% and 61.9% on CAD, respectively. Live/Dead staining confirmed increased cell death, especially in SBU-ER groups. CAD discs had ∼82% lower permeability than SD; ER etching increased permeability in both. SEM showed open tubules in SD and demineralized surfaces in CAD. HEMA diffusion was reduced by 36.5% and 54.7% in CAD for SE and ER, respectively.
SIGNIFICANCE: SBU featured cytotoxicity regardless on dentin condition (SD or CAD), highlighting biological risks of using universal adhesives in deep cavities.
METHODS: Human dentin discs (0.35 mm) from sound (SD) or microbiologically demineralized teeth (CAD) were used in a co-culture model under simulated pulpal pressure into APC-on-a-chip, with MDPC-23 cells seeded on the pulpal surface and human dental pulp cells (HDPCs) cultured in 3D collagen gels beneath the dentin. SBU was applied on the occlusal surface in self-etch (SE) or etch-and-rinse (ER) mode and LED-cured (continuous mode). After 24 h, cell metabolism (Alamar Blue), viability (Live/Dead), dentin permeability (Lp), morphology (SEM), and HEMA diffusion (UV-vis) were evaluated. Data were analyzed using two-way ANOVA/Tukey or Student's t-tests (p < 0.05).
RESULTS: SBU application reduced MDPC-23 metabolism by 71.2% (SE) and 85.7% (ER) on SD, and 46.8% (SE) and 62.2% (ER) on CAD. HDPCs showed similar trends: decreases of 50.2% and 70.1% on SD, and 38.1% and 61.9% on CAD, respectively. Live/Dead staining confirmed increased cell death, especially in SBU-ER groups. CAD discs had ∼82% lower permeability than SD; ER etching increased permeability in both. SEM showed open tubules in SD and demineralized surfaces in CAD. HEMA diffusion was reduced by 36.5% and 54.7% in CAD for SE and ER, respectively.
SIGNIFICANCE: SBU featured cytotoxicity regardless on dentin condition (SD or CAD), highlighting biological risks of using universal adhesives in deep cavities.