@article{ummarino_vitro_2023,
	title = {An \textit{in vitro} model for osteoarthritis using long‐cultured inflammatory human macrophages repeatedly stimulated with {TLR} agonists},
	issn = {0014-2980, 1521-4141},
	url = {https://onlinelibrary.wiley.com/doi/10.1002/eji.202350507},
	doi = {10.1002/eji.202350507},
	abstract = {Abstract
            
              Osteoarthritis ({OA}) is characterized by an abundance of inflammatory M1‐like macrophages damaging local tissues. The search for new potential drugs for {OA} suffers from the lack of appropriate methods of long‐lasting inflammation. Here we developed and characterized an
              in vitro
              protocol of long‐lasting culture of primary human monocyte‐derived macrophages differentiated with a combination of M‐{CSF}+{GM}‐{CSF} that optimally supported long‐cultured macrophages ({LC}‐Mϕs) for up to 15 days, unlike their single use. Macrophages repeatedly stimulated for 15 days with the {TLR}2 ligand Pam3CSK4 ({LCS}‐Mϕs), showed sustained levels over time of {IL}‐6, {CCL}2, and {CXCL}8, inflammatory mediators that were also detected in the synovial fluids of {OA} patients. Furthermore, macrophages isolated from the synovia of two {OA} patients showed an expression profile of inflammation‐related genes similar to that of {LCS}‐Mϕs, validating our protocol as a model of chronically activated inflammatory macrophages. Next, to confirm that these {LCS}‐Mϕs could be modulated by anti‐inflammatory compounds, we employed dexamethasone and/or celecoxib, two drugs widely used in {OA} treatment, that significantly inhibited the production of inflammatory mediators. This easy‐to‐use
              in vitro
              protocol of long‐lasting inflammation with primary human macrophages could be useful for the screening of new compounds to improve the therapy of inflammatory disorders.},
	pages = {2350507},
	journaltitle = {European Journal of Immunology},
	shortjournal = {Eur J Immunol},
	author = {Ummarino, Aldo and Pensado‐López, Alba and Migliore, Roberta and Alcaide‐Ruggiero, Lourdes and Calà, Nicholas and Caputo, Michele and Gambaro, Francesco M. and Anfray, Clément and Ronzoni, Flavio L. and Kon, Elizaveta and Allavena, Paola and Torres Andón, Fernando},
	urldate = {2023-10-10},
	date = {2023-10-06},
	langid = {english},
}