A time-resolved fibrosis model – in vitro assessment of antifibrotic implant coatings

Abstract

Wound healing after implant surgery represents a spontaneous repair process that, when disturbed, can lead to a pathological situation known as fibrosis, characterized by excessive proliferation, increased extracellular matrix deposition, and scarring of the tissue. Tissue-specific cell culture models simulating physiological or pathophysiological cell responses are valuable research tools in the field of fibrosis, among others. In such in vitro systems, endpoint measurements are typically used to assess cell viability and/or proliferation. However, this approach is usually time consuming, costly and often leads to the destruction of the sample. Additionally, these conventional in vitro systems provide only a single snapshot of the respective sample at the end of the experiment. Therefore, we aimed to evolve the fibrotic disease in vitro model established by Stahnke et al. To this end, we chose a time-resolved, non-invasive approach based on cell impedance measurements and bright-field microscopy that allows continuous monitoring of the cells. Such measurements can be directly conducted using the xCELLigence RTCA eSight system. This system was used to characterize the cellular responses of the fibrosarcoma cell line HT-1080 and primary fibroblasts isolated from the Tenon’s capsule of human donors to TGF-β1, a key cytokine involved in the pathogenesis of fibrosis. The data shows that compared to Tenon’s fibroblasts the response of HT-1080 cells to TGF- β1 is less pronounced and only detectable within a narrow time frame. Thus, HT-1080 do not appear to be a suitable cell culture model for the assessment of antifibrotic implant coatings.

Keywords

• tgf-β1
• proliferation
• cell impedance
• fibrosis