Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures

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European Journal of Medical Research Open Access

RESEARCH

Fibroblast‑like cells change gene expression of bone remodelling markers in transwell cultures Eliza S. Hartmann1†, Sabine Schluessel1†, Miriam I. Köhler1, Felicitas Beck1, Julia I. Redeker1, Burkhard Summer2, Veronika Schönitzer3, Andreas Fottner1 and Susanne Mayer‑Wagner1*

Abstract  Introduction:  Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplas‑ ties. Various studies have examined PPF behavior in monolayer culture systems. However, the periprosthetic tissue is a three-dimensional (3D) mesh, which allows the cells to interact in a multidirectional way. The expression of bone remodeling markers of fibroblast-like cells in a multilayer environment changes significantly versus monolayer cultures without the addition of particles or cytokine stimulation. Gene expression of bone remodeling markers was therefore compared in fibroblast-like cells from different origins and dermal fibroblasts under transwell culture condi‑ tions versus monolayer cultures. Methods:  PPFs from periprosthetic tissues (n = 12), osteoarthritic (OA) synovial fibroblast-like cells (SFs) (n = 6), and dermal fibroblasts (DFs) were cultured in monolayer (density 5.5 × 103/cm2) or multilayer cultures (density 8.5 × ­105/ cm2) for 10 or 21 days. Cultures were examined via histology, TRAP staining, immunohistochemistry (anti-S100a4), and quantitative real-time PCR. Results:  Fibroblast-like cells (PPFs/SFs) and dermal fibroblasts significantly increased the expression of RANKL and significantly decreased the expression of ALP, COL1A1, and OPG in multilayer cultures. PPFs and SFs in multilayer cultures further showed a higher expression of cathepsin K, MMP-13, and TNF-α. In multilayer PPF cultures, the mRNA level of TRAP was also found to be significantly increased. Conclusion:  The multilayer cultures are able to induce significant expression changes in fibroblast-like cells depend‑ ing on the nature of cellular origin without the addition of any further stimulus. This system might be a useful tool to get more in vivo like results regarding fibroblast-like cell cultures. Keywords:  Implant material, Fibroblast-like cells, Transwell culture, Bone remodeling, Aseptic loosening

*Correspondence: [email protected]‑muenchen.de † Eliza S. Hartmann and Sabine Schluessel have contributed equally to this work 1 Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU Munich, Marchioninistr. 15, 81377 Munich, Germany Full list of author information is available at the end of the article

Background Various materials are used in implant applications. However, the fail of implants always leads to the common process of periprosthetic osteolysis, which is a multifactorial process [1–3]. Osteoclastogenesis is induced due to an inflammatory response to miscellaneous implant particles. A pseudomembrane forms, which consists of various cellular compounds predominantly of fibroblasts, osteoclasts, giant cells, inflammator