Novel Ex Vivo Human Osteochondral Explant Model of Knee and Spine Osteoarthritis Enables Assessment of Inflammatory and Drug Treatment Responses

Jeroen Geurts, Doria Jurić, Miriam Müller, Stefan Schären, Cordula Netzer, Jeroen Geurts, Doria Jurić, Miriam Müller, Stefan Schären, Cordula Netzer

Abstract

Osteoarthritis of the knee and spine is highly prevalent in modern society, yet a disease-modifying pharmacological treatment remains an unmet clinical need. A major challenge for drug development includes selection of appropriate preclinical models that accurately reflect clinical phenotypes of human disease. The aim of this study was to establish an ex vivo explant model of human knee and spine osteoarthritis that enables assessment of osteochondral tissue responses to inflammation and drug treatment. Equal-sized osteochondral fragments from knee and facet joints (both n = 6) were subjected to explant culture for 7 days in the presence of a toll-like receptor 4 (TLR4) agonist and an inhibitor of transforming growth factor-beta (TGF-β) receptor type I signaling. Markers of inflammation, interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), but not bone metabolism (pro-collagen-I) were significantly increased by treatment with TLR4 agonist. Targeting of TGF-β signaling resulted in a strong reduction of pro-collagen-I and significantly decreased IL-6 levels. MCP-1 secretion was increased, revealing a regulatory feedback mechanism between TGF-β and MCP-1 in joint tissues. These findings demonstrate proof-of-concept and feasibility of explant culture of human osteochondral specimens as a preclinical disease model, which might aid in definition and validation of disease-modifying drug targets.

Keywords: bone metabolism; experimental model; inflammation; knee; osteoarthritis; osteochondral; spine.

Conflict of interest statement

The authors declare no conflict of interest. The funding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
Tissue viability after explant culture of osteochondral specimens from (a) knee and (b) facet joint osteoarthritis. Fresh clinical specimens were cut in equal-sized fragments and cultured in osteogenic culture medium for one week. Samples were either left untreated (control) or challenged with 1 μg/mL lipopolysaccharide (inflammation) in the absence and presence of a drug treatment (10 μM TGF-β receptor type I inhibitor).
Figure 2
Figure 2
Assessment of secreted markers of bone metabolism and inflammation under basal and inflammatory conditions. Osteoarthritic specimens were left untreated in osteogenic culture medium (control) or challenged with LPS. Secreted protein levels of (a) pro-Col-I, (b) IL-6 and (c) MCP-1 were determined by ELISA. * p < 0.05, *** p < 0.001 by ratio paired t-test.
Figure 3
Figure 3
Assessment of secreted markers of bone metabolism and inflammation under inflammatory conditions in the presence and absence of TGF-β receptor type I inhibition. Osteoarthritic specimens were challenged with LPS (1 μg/mL) and treated with 10 μM SB-505124. Secreted protein levels of (a) pro-Col-I, (b) IL-6 and (c) MCP-1 were determined by ELISA. * p < 0.05, ** p < 0.05, *** p < 0.001 by ratio paired t-test.

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