Primary human mesothelial cell culture in the evaluation of the inflammatory response to different sclerosing agents used for pleurodesis

原代人间皮细胞培养在评估胸膜固定术所用不同硬化剂的炎症反应中的应用

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作者:Michal Mierzejewski, Magdalena Paplinska-Goryca, Piotr Korczynski, Rafal Krenke

Abstract

The mechanisms of chemical pleurodesis are still not fully explained. We aimed to evaluate the feasibility of using primary biopsy-derived human mesothelial cells to establish an in vitro culture and to assess the response of pleural mesothelial cells to different sclerosing agents. Talc, povidone-iodine, doxycycline, and TGF-β were used at different doses to stimulate pleural mesothelial cells. After 6 and 24 h, mRNA expression of interleukin (IL)-1β, IL-6, IL-8, TGF-β, MCP-1, IL-17A, and MMP9 was measured in cultured cells, and the protein level of IL-1β, IL-6, and IL-8 was measured in the culture supernatant. The most pronounced response was observed after talc exposure. It was expressed as an increase in IL-1β concentration in culture supernatant after 24 h of higher talc dose stimulation compared to 6 h of stimulation (17.14 pg/ml [11.96-33.32 pg/ml] vs. 1.84 pg/ml [1.81-1.90 pg/ml], p = 0.02). We showed that culture pleural mesothelial cells isolated from pleura biopsy specimens is feasible. Inflammatory responses of mesothelial cells to different sclerosants were highly variable with no consistent pattern of mesothelium reaction neither in terms of different sclerosing agents nor in the time of the most significant reaction. We demonstrated that pro-inflammatory mesothelial response includes an increase in IL-1β mRNA expression and protein production. This may suggest the role of IL-1β in the formation and maintenance of the inflammatory response during pleurodesis.

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