Assessing the effect of triethyleneglycol dimethacrylate on tissue repair in 3D organotypic cultures

Abstract

Leachables from dental restoratives induce toxicity in gingival and pulp tissues and affect tissue regeneration/healing. Appropriate testing of these materials requires a platform that mimics the in vivo environment and allows the architectural self-assembly of cells into tissue constructs. In this study, we employ a new 3D model to assess the impact of triethyleneglycol dimethacrylate (TEGDMA) on early organization and advanced recruitment/accumulation of immortalized mouse gingival fibroblasts (GFs) and dental papilla mesenchymal cells (DPMCs) in extracellular matrix. We hypothesize that TEGDMA (1) interferes with the developmental architecture of GFs and DPMCs, and (2) inhibits the deposition of mineral. To test these hypotheses, GFs and DPMCs were incubated with the soluble TEGDMA at concentrations (0-2.5) mmol/L. Diameter and thickness of the constructs were determined by microscopic analysis. Cell differentiation was assessed by immunocytochemistry and the secreted mineral detected by alizarin-red staining. TEGDMA interfered with the development of GFs and/or DPMCs microtissues in a dose-dependent manner by inhibiting growth of inter-spherical cell layers and decreasing spheroid size (four to six times). At low/moderate TEGDMA levels, GFs organoids retained their structures while reducing thickness up to 21%. In contrast, at low TEGDMA doses, architecture of DPMC organoids was altered and thickness decreased almost twofold. Overall, developmental ability of TEGDMA-exposed GFs and DPMCs depended on TEGDMA level. GFs constructs were more resistant to structural modifications. The employed 3D platform was proven as an efficient tool for quantifying the effects of leachables on tissue repair capacities of gingiva and dental pulp.

Bioenergetic Impairment of Triethylene Glycol Dimethacrylate- (TEGDMA-) Treated Dental Pulp Stem Cells (DPSCs) and Isolated Brain Mitochondria are Amended by Redox Compound Methylene Blue

“…It was reported that TEGDMA adversely affects cellular metabolism, signaling, differentiation, and dentinogenesis, while it also interferes with wound healing and tissue repair [28,40,[68][69][70][71]; it induces oxidative stress [46][47][48], apoptosis and/or necrosis [42,43,49,72]. Due to its amphiphilic character, TEGDMA can penetrate the membranes of human gingival fibroblast and human oral epithelium of the mouth, resulting in an inflammatory response [73].…”

“…Due to its amphiphilic character, TEGDMA can penetrate the membranes of human gingival fibroblast and human oral epithelium of the mouth, resulting in an inflammatory response [73]. Furthermore, TEGDMA triggers time-and concentration-dependent toxicity in mouse gingival fibroblasts and dental papilla mesenchymal cells in three-dimensional cultures [71].…”

The self-renewal dental pulp stem cell microtissues challenged by a toxic dental monomer

“…Commonly used as a diluent monomer in resin-based dental composites, the unpolymerized TEGDMA can easily leach out from the restorations, interact with the dental pulp tissue and cells. TEGDMA penetrates into the cytosol through the membrane of mammalian cells with the capability to negatively influence cell growth and development [10][11][12][13] and increases the level of ROS in the tissue [10,14].…”

N-Acetyl Cysteine Modulates the Inflammatory and Oxidative Stress Responses of Rescued Growth-Arrested Dental Pulp Microtissues Exposed to TEGDMA in ECM

“…At concentrations 1.5 to 3.0 mmol/L, TEGDMA affects pulp cell growth by increasing the number of cells arrested in G1 and G2 cycles [ 18 , 19 ] and interferes with the development of their progenitor dental papilla microtissues in a dose-dependent manner. The latter is due to the prevention of cell interaction and the creation of the spherical and inter-spherical interactions in the extracellular matrix microenvironment [ 20 ].…”