An Evaluation of Different 3D Cultivation Models on Expression Profiles of Human Periodontal Ligament Fibroblasts with Compressive Strain

• Published in: International journal of molecular sciences 2022-02, Vol.23 (4), p.2029
• Main Authors: Schröder, Agnes, Schöniger, Ricarda, Oeldemann, Juliane, Spanier, Gerrit, Proff, Peter, Jantsch, Jonathan, Kirschneck, Christian, Ullrich, Niklas
• Format: Article
• Language: English
• Subjects: 3D cell culture; Angiogenesis; Biomechanics; Bone remodeling; Cardiovascular disease; Cell culture; Cells, Cultured; Collagen Type I - genetics; Collagen Type I - metabolism; Compressive properties; Cyclooxygenase 2 - genetics; Cyclooxygenase 2 - metabolism; Extracellular matrix; Fibroblasts; Fibroblasts - cytology; Fibroblasts - metabolism; Gels; Gene expression; Gene regulation; Genes; Humans; Hydrogels; Inflammation; Interleukin 6; Interleukin-6 - genetics; Interleukin-6 - metabolism; Investigations; Ligaments; Ligands; Mechanical stimuli; Orthodontics; Osteoprotegerin; Osteoprotegerin - genetics; Osteoprotegerin - metabolism; PDLF; Periodontal ligament; Periodontal Ligament - cytology; Physiology; Pressure; Pressure effects; Primary Cell Culture - methods; RANK Ligand - genetics; RANK Ligand - metabolism; Scaffolds; Spheroids; Tissue Engineering - methods; Tissue Scaffolds - chemistry; TRANCE protein; Two dimensional models; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23042029

The effects of compressive strain during orthodontic treatment on gene expression profiles of periodontal ligament fibroblasts (PDLFs) have mostly been studied in 2D cell culture. However, cells behave differently in many aspects in 3D culture. Therefore, the effect of pressure application on PDLFs in different 3D structures was investigated. PDLFs were either conventionally seeded or embedded into different 3D structures (spheroids, Mebiol gel, 3D scaffolds) and exposed to compressive force or incubated without pressure. For one 3D scaffold (POR), we also tested the effect of different compressive forces and application times. Expression of an angiogenic gene ( ), a gene involved in extracellular matrix synthesis ( ), inflammatory genes ( , ), and genes involved in bone remodelling ( , ) were investigated by RT-qPCR. Depending on the used 3D cell culture model, we detected different effects of compressive strain on expression profiles of PDLFs. was downregulated in all investigated 3D culture models. Angiogenetic and proinflammatory genes were regulated differentially between models. In 3D scaffolds, regulation of bone-remodelling genes upon compressive force was contrary to that observed in 3D gels. 3D cell culture models provide better approximations to in vivo physiology, compared with conventional 2D models. However, it is crucial which 3D structures are used, as these showed diverse effects on the expression profiles of PDLFs during mechanical strain.