Multimodal mass spectrometry unravels knee osteoarthritis pathways in a human infrapatellar fat pad ex vivo model

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Osteoarthritis (OA) is the most prevalent rheumatic disease in elderly people. However, joint injuries such as cartilage defects (CD) and metabolic disorders can accelerate OA onset in young adults. Several joint tissues are involved in OA progression, including the infrapatellar fat pad (IPFP). In this work we studied the IFPF’s spatial molecular composition by mass spectrometry-based approaches to unveil inflammatory-driven mechanisms in early and late OA. Human IFPF biopsies from CD and OA patients were exposed to IL1β and TNFα treatment to mimic the joint inflammatory environment. The secretome and proteome were analyzed by using a Q-Exactive orbitrap in data independent (DIA) acquisition mode. MALDI-MSI was employed to assess the lipid response (Rapiflex, resolution 30 m). Results indicated that both patient groups responded to inflammation stimulus via increase of PTGS2 and oxidative stress marker (SOD2). FABP5 protein was significantly upregulated in CD secretome implying a potential lipid dysregulation. Aligned with these results, MALDI-MSI showed a high abundance of phosphatidylinositol species whereas OA explants displayed increased lysophosphatidylcholine levels. Both groups presented reduced cardiolipin levels, emphasizing the relevance of mitochondrial homeostasis in OA. Together, these findings underscore unique lipid and protein profiles in early OA upon inflammation highlighting the importance of multi-omics for identifying pathogenic pathways and biomarkers.

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