In-vitro investigation of compression and catabolic cytokines on human cartilage endplate cells in agarose

Crump, Katherine B.; Bermudez-Lekerika, Paola; Alminawi, Ahmad; Geris, Liesbet; Noailly, Jérôme; Gantenbein, Benjamin (2023). In-vitro investigation of compression and catabolic cytokines on human cartilage endplate cells in agarose. In: Annual Meeting of the Swiss Society of Biomedical Engineering. 13 September.

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Introduction

Intervertebral disc (IVD) degeneration is the main cause of low back cases in young adults [1]. However, the initiating and risk factors are poorly understood as it is a highly multifactorial disease. The cartilage endplate (CEP) covers the top and bottom of the IVD and acts to transmit compressive loads and transport water, nutrients, and waste in and out of the disc. [2] Early CEP degeneration is likely to play a key role in IVD degeneration, but little is known about CEP mechanobiology and its changes in degeneration. [3,4]. Investigating these changes is essential to elucidate how the CEP contributes to IVD pathology. It was hypothesized that CEP cells would behave similarly to articular chondrocytes. Thus, it was predicted that dynamic compression would be sufficient to induce anabolism, while stimulation with pro-inflammatory cytokines would induce catabolism.

Material and Methods

Human CEP cells were expanded until passage 3 or 4, then seeded at a density of 7.5x10 6 cells/ml into 2% agarose carriers (dimensions: 6 mm ⌀ and 3 mm height) and cultured for 5 days for phenotype recovery. Cellagarose carriers were placed in custom-made chambers, stimulated with 10 ng/ml TNF throughout the entirety of the experiment and dynamically compressed under ~7% strain for one hour at 1.5 Hz daily for up to 14 days. Carriers were collected on Days 0, 7, and 14 for downstream analysis of cell viability, metabolism, gene expression, and glycosaminoglycan (GAG) content.

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