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

Eliza S Hartmann,Veronika Schönitzer,Sabine Schluessel,Susanne Mayer-Wagner,Julia I Redeker,Felicitas Beck,Miriam I Köhler,Burkhard Summer,Andreas Fottner

doi:10.1186/s40001-020-00453-y

Abstract

IntroductionPeriprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplasties. 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 conditions versus monolayer cultures.MethodsPPFs 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.ResultsFibroblast-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.ConclusionThe multilayer cultures are able to induce significant expression changes in fibroblast-like cells depending 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.

Highlights

Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplas‐ ties
We showed that periprosthetic fibroblast-like cells (PPFs) co-cultured with PBMCs in transwell cultures resulted in a considerably more osteoclastogenic gene expression profile, being closer to in vivo findings of periosteolytic tissues [17]
The gene expression of Receptor activator of nuclear factor-κB ligand (RANKL)

Summary

Introduction

Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplas‐ ties. 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 compared in fibroblast-like cells from different origins and dermal fibroblasts under transwell culture condi‐ tions versus monolayer cultures. A pseudomembrane forms, which consists of various cellular compounds predominantly of fibroblasts, osteoclasts, giant cells, inflammatory cells, and macrophages [4]. The periprosthetic tissue is lined by a synovium-like interface membrane, which consists of synovial-like lining cells. The synoviallike lining cells form the surface of the synovium-like interface, which consists mainly of periprosthetic fibroblast-like cells (PPFs). The PPFs have been shown to play an active role in periprosthetic osteolysis [7]

Methods

Results

Conclusion