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

Abstract

Introduction Periprosthetic 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. 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 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.

Dental and Orthopaedic Implant Loosening: Overlap in Gene Expression Regulation

“…Detailed histomorphological characteristics were not performed in the current study due to a limited number of cells per donor. Hartmann et al found conglomerates of PPFs in frozen sections of transwell cultures forming a polylayer structure (34). This coincides with findings of Sabater et al (27), showing that cultivating fibroblasts in transwell systems lead to an increased number of cells and higher cell mass compared to cultures on standard well bottoms, despite the smaller surface of the transwell membranes.…”

“…Morphological characterization of PPFs in transwell cultures had been performed by our group (34). Detailed histomorphological characteristics were not performed in the current study due to a limited number of cells per donor.…”

[Retracted] Exosome: Function and Application in Inflammatory Bone Diseases

“…PGE2 is a paracrine factor released from mesenchymal stem cells with powerful immunomodulatory functions and upregulates IL-10 secreted by macrophages and stimulates M2 macrophage polarization to accelerate the recovery of damaged tissues [ 44 ]. However, PGE2 also promotes RANKL expression in periprosthetic fibroblasts [ 45 ].…”

“…Several studies demonstrated that many MMPs might lead to prosthetic loosening and osteolysis through pathological extracellular matrix degradation and periprosthetic connective tissue/bone remodeling [ 89 – 92 ]. Periprosthetic fibroblast-like cells express MMP-13 in response to induction of wear particles, and the increase in matrix metalloproteinases destroys the periprosthetic tissue [ 45 ]. Blocking TNF- α and IL-1 β receptors does not attenuate the damaging effects of exosomes on the extracellular matrix, suggesting that the mechanism of increased matrix metalloproteinase production caused by exosomes is independent of the TNF- α -induced inflammatory response.…”