Dental Pulp Fibroblasts Response after Stimulation with HEMA and Adhesive System

Abstract

This study evaluated in vitro cell viability and metabolism, nitric oxide release and production of chemokines by cultured human dental pulp fibroblasts (DPF) under contact with HEMA and Single Bond. Cultures of DPF were established by means of an explant technique. Once plated, cells were kept under contact with increasing concentrations of HEMA (10, 100 and 1000 nM) or Single Bond (SB) [10-fold serially diluted in culture medium (10 -4 , 10 -3 and 10 -2 v/v)] and also with polymerized SB components. Cytotoxicity was assessed by Trypan Blue exclusion method and MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Nitric oxide release on cell supernatant was detected by Griess Method whereas chemokines (CXCL12 and CXCL8) were detected by ELISA. RT-qPCR was employed for chemokines gene expression analysis. Cytotoxic tests showed significant differences for SB 10 -2 . None of the tested materials significantly altered NO levels. Protein levels of CXCL12 were significantly decreased only by HEMA. On the other hand, while CXCL12 mRNA remained unaltered, gene expression of CXCL8 had significant decrease with all materials, except for polymerized SB. In conclusion, Single Bond and HEMA at various concentrations, decreased expression and production of molecules involved in inflammatory processes and, therefore, the use of adhesive systems such as pulp capping materials must be viewed with caution due to its large cytotoxic effect when in close contact with the pulp.

Role of Lipopolysaccharide, Derived from Various Bacterial Species, in Pulpitis—A Systematic Review

“…P. intermedia (ATC 25611) LPS enhances the expression of IL-8 mRNA and its release in human dental pulp fibroblasts with a peak at 12 h [ 92 ].…”

Dental Pulp Fibroblast: A Star Cell

“…Pulp cells, especially human [200][201][202] and animal 203,204 fibroblasts, are the models of choice for biocompatibility testing of dental materials, the cytotoxic effects of which directly affect the dental pulp 205 . Furthermore, DPFs are highly sensitive to toxic substances and are therefore ideal to elucidate the possible adverse effects of restorative [206][207][208] , endodontic [209][210][211] , and novel therapeutic materials [212][213][214][215] . It must be borne in mind that for cell cultures to be considered an acceptable model, it is necessary to demonstrate that the response of cells to the tested materials can be reproduced, that pulp cell cultures can be easily established, and that cell lines can be standardized 205 .…”

Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials

“…Under patient informed consent, one male and two female subjects (18 -25 years) donated three third molars, one of each patient. Dental pulp was removed under aseptic conditions and tissues were minced for fibroblasts cultures using an explant technique (1,19,20).…”

“…The effect of restorative materials together with remnant dentine thickness displays central attention considering the pulp vitality after restorative procedures. Therefore, biological effects of restorative materials on dental pulp cells should be considered, especially due to the large number of adhesive products and techniques becoming regularly available for the dental practice (1,2). The protection of the dentin-pulp complex consists in the application of specific materials between the restorative material and the dentin to avoid additional aggression to the pulp tissue.…”

“…However, unpolymerized HEMA is released by RMGIC in the early 24 h after polymerization and this substance compromises RMGIC biocompatibility (17). Several studies demonstrate that RMGIC are cytotoxic to different cell types and tissues due to the release of HEMA (5,6,17,18) and it causes serious damage to pulp cells (1), that in turn can lead to immune responses in different degrees of severity resulting in pulp inflammation (7,8).…”