Univ Rennes, CHU Rennes, Pole Odontologie, Rennes, France.
ISCR, CNRS-UMR 6226, Rennes, France.
Eur J Orthod. 2020 Sep 11;42(4):387-395. doi: 10.1093/ejo/cjaa017.
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.
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.
The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable.
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.
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.
力学生物学现象是正畸治疗和生物材料植入过程中细胞和组织反应的一个主要因素。更好地理解这些现象将提高我们治疗的效果。本工作的目的是验证一种三维（3D）成骨细胞培养模型，以研究力学生物学。
hFOB 1.19 细胞系传统地在平面上或称为球体的聚集体中培养。它们被嵌入 0.8%低熔点琼脂糖 VII 中，并放置在聚对苯二甲酸乙二醇酯 Transwell 插入物中。通过可调重量施加 1 和 4 g/cm2 的压缩力。通过测量直径、监测葡萄糖水平和进行 Hoechst/碘化丙啶染色来评估增殖。通过酶联免疫吸附试验（ELISA）集中研究促炎介质白细胞介素（IL）-6 和 IL-8 以及骨重塑因子骨保护素，评估可溶性因子的合成。通过定量逆转录聚合酶链反应（qRT-PCR）评估骨标志物的转录。
3D 模型显示出良好的细胞活力，并允许进行 IL 给药。此外，可以分析三种基因表达谱。
该模型允许分析常规标志物；为了更好地理解成骨细胞力学生物学，需要更大的探索。然而，它只能进行 3 天的分析。
对 3D 成骨细胞培养施加恒定压缩力所获得的结果验证了该模型系统，可用于探索生物分子释放和分析基因转录。特别是，它强调了成骨标志物表达的紊乱。
