Impact of Periprosthetic Fibroblast-Like Cells on Osteoclastogenesis in Co-Culture with Peripheral Blood Mononuclear Cells Varies Depending on Culture System.
Miriam I KoehlerEliza S HartmannSabine SchluesselFelicitas BeckJulia I RedekerBaerbel SchmittMarina UngerMartijn van GriensvenBurkhard SummerAndreas FottnerSusanne Mayer-WagnerPublished in: International journal of molecular sciences (2019)
Co-culture studies investigating the role of periprosthetic fibroblasts (PPFs) in inflammatory osteoclastogenesis reveal contrary results, partly showing an osteoprotective function of fibroblasts and high OPG expression in monolayer. These data disagree with molecular analyses of original periosteolytic tissues. In order to find a more reliable model, PPFs were co-cultivated with peripheral blood mononuclear cells (PBMCs) in a transwell system and compared to conventional monolayer cultures. The gene expression of key regulators of osteoclastogenesis (macrophage colony-stimulating factor (MCSF), receptor activator of NF-κB ligand (RANK-L), osteoprotegerin (OPG), and tumor necrosis factor alpha (TNFα)) as well as the ability of bone resorption were analyzed. In monolayer co-cultures, PPFs executed an osteoprotective function with high OPG-expression, low RANK-L/OPG ratios, and a resulting inhibition of osteolysis even in the presence of MCSF and RANK-L. For transwell co-cultures, profound changes in gene expression, with a more than hundredfold decrease of OPG and a significant upregulation of TNFα were observed. In conclusion, we were able to show that a change of culture conditions towards a transwell system resulted in a considerably more osteoclastogenic gene expression profile, being closer to findings in original periosteolytic tissues. This study therefore presents an interesting approach for a more reliable in vitro model to examine the role of fibroblasts in periprosthetic osteoclastogenesis in the future.
Keyphrases
- gene expression
- bone loss
- poor prognosis
- lps induced
- rheumatoid arthritis
- nuclear factor
- dna methylation
- total hip
- total hip arthroplasty
- extracellular matrix
- genome wide
- signaling pathway
- binding protein
- oxidative stress
- inflammatory response
- long non coding rna
- bone mineral density
- machine learning
- electronic health record
- big data
- single molecule
- autism spectrum disorder
- toll like receptor
- case control
- single cell
- intellectual disability
- data analysis
- artificial intelligence
- wound healing