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Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics

Abstract : Objectives: Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of three-dimensional (3D) culture of osteoblasts to study mechanobiology. Materials and methods: The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription. Results: The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable. Limitations: The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days. Conclusion: The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis.
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Submitted on : Tuesday, May 12, 2020 - 4:16:07 PM
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Damien Brezulier, Pascal Pellen-Mussi, Sylvie Tricot-Doleux, Agnès Novella, Olivier Sorel, et al.. Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics. European Journal of Orthodontics, Oxford University Press (OUP), 2020, ⟨10.1093/ejo/cjaa017⟩. ⟨hal-02533165⟩

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