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Apical Lesions of Endodontic Origin (ALEO) are initiated by polymicrobial endodontic canal infection. <i>Porphyromonas gingivalis</i> (<i>Pg</i>) and <i>Porphyromonas endodontalis</i> (<i>Pe</i>) lipopolysaccharides (LPS) can induce a pro-inflammatory macrophage response through their recognition by TLR2 and TLR4. However, polarization responses induced by <i>Pg</i> and/or <i>Pe</i> LPS in macrophages are not fully understood. We aimed to characterize the polarization profiles of macrophages differentiated from THP-1 cells following <i>Pg</i> and/or <i>Pe</i> LPS stimulation from reference strain and clinical isolates. A modified LPS purification protocol was implemented and the electrophoretic LPS profiles were characterized. THP-1 human monocytes differentiated to macrophages were stimulated with <i>Pg</i> and <i>Pe</i> LPS. Polarization profiles were characterized through cell surface markers and secreted cytokines levels after 24 h of stimulation. TLR2 and TLR4 cell surfaces and transcriptional levels were determined after 24 or 2 h of LPS stimulation, respectively. LPS from <i>Pg</i> induced a predominant M1 profile in macrophages evidenced by changes in the expression of the surface marker CD64 and pro-inflammatory cytokine profiles, TNF-α, IL-1β, IL-6, and IL-12. <i>Pe</i> LPS was unable to induce a significant response. TLR2 and TLR4 expressions were neither modified by <i>Pg</i> or <i>Pe</i> LPS. <i>Pg</i> LPS, but not <i>Pe</i> LPS, induced a macrophage M1 Profile.