The self-renewal dental pulp stem cell microtissues challenged by a toxic dental monomer
| Autor: | Drago Skrtic, Gili Kaufman, Naima Makena Kiburi |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Adult
0301 basic medicine extracellular matrix Primary Cell Culture Immunocytochemistry Biophysics self-renewal Composite Resins Biochemistry Molecular Bases of Health & Disease Cell Line Polyethylene Glycols Extracellular matrix Young Adult 03 medical and health sciences 0302 clinical medicine Polymethacrylic Acids stomatognathic system SOX2 Dental pulp stem cells medicine Dentin Humans mineralization Molecular Scaffolds & Matrices Cell Self Renewal Molecular Biology Research Articles Dental Pulp Basement membrane Chemistry Stem Cells SOXB1 Transcription Factors Cell Biology Dentinogenesis Therapeutics & Molecular Medicine dental pulp stem cells Cell biology Phenotype 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis TEGDMA cytotoxicity Pulp (tooth) Stem cell Signal Transduction Transcription Factors |
| Zdroj: | Bioscience Reports |
| ISSN: | 1573-4935 0144-8463 |
| DOI: | 10.1042/bsr20200210 |
| Popis: | Dental pulp stem cells (DPSCs) regenerate injured/diseased pulp tissue and deposit tertiary dentin. DPSCs stress response can be activated by exposing cells to the monomer triethyleneglycol dimethacrylate (TEGDMA) and inducing the DNA-damage inducible transcript 4 (DDIT4) protein expression. The goal of the present study was to determine the impact of TEGDMA on the ability of DPSCs to maintain their self-renewal capabilities, develop and preserve their 3D structures and deposit the mineral. Human primary and immortalized DPSCs were cultured in extracellular matrix/basement membrane (ECM/BM) to support stemness and to create multicellular interacting layers (microtissues). The microtissues were exposed to the toxic concentrations of TEGDMA (0.5 and 1.5 mmol/l). The DPSCs spatial architecture was assessed by confocal microscopy. Mineral deposition was detected by alizarin red staining and visualized by stereoscopy. Cellular self-renewal transcription factor SOX2 was determined by immunocytochemistry. The microtissue thicknesses/vertical growth, surface area of the mineralizing microtissues, the percentage of area covered by the deposited mineral, and the fluorescence intensity of the immunostained cells were quantified ImageJ. DDIT4 expression was determined by a single molecule RNA-FISH technique and the cell phenotype was determined morphologically. DDIT4 expression was correlated with the cytotoxic phenotype. TEGDMA affected the structures of developing and mature microtissues. It inhibited the deposition of the mineral in the matrix while not affecting the SOX2 expression. Our data demonstrate that DPSCs retained their self-renewal capacity although their other functions were impeded. Since the DPSCs pool remained preserved, properties effected by the irritant should be restored by a proper rescue therapy. |
| Databáze: | OpenAIRE |
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