Frontiers in oral health · 2025
Effects of combined cyclosporin and azithromycin treatment on human mononuclear cells under lipopolysaccharide challenge.
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Alotaibi Norah, Alesawy Aminah, Alalshaikh Marwa, Aljofi Faisal E, Aldossary Nada, Alzahrani Nada, Omar Omar, Madi Marwa
Аннотация
OBJECTIVE: To evaluate the combined effects of azithromycin and varying concentrations of cyclosporin on peripheral blood mononuclear cells (PBMCs) under lipopolysaccharide (LPS) stimulation.
MATERIALS AND METHODS: PBMCs were isolated from four healthy donors and treated with cyclosporin at concentrations of (50, 200, and 1,000 ng/ml) either alone or in combination with azithromycin (0.4 µg/ml), with and without 100 ng ml LPS derived from Porphyromonas gingivalis. Total cell count, cell viability, and lactate dehydrogenase (LDH) activity were assessed at day 1 and 3. While the inflammatory mediators, including IL-6, IL-1β, IL-18, and IgA levels were assessed by ELISA at day 3. Statistical analysis included two-way ANOVA to analyze the effects of the drugs and the presence of LPS (the two independent variables), followed by Tukey's HSD post-hoc test. Multiple linear regression models evaluating treatment effects, LPS exposure, and time points, with assessment of two-way interactions. Models were adjusted for relevant covariates and verified for statistical assumptions, with significance set at p < 0.05.
RESULTS: Lower cyclosporin concentrations (50 and 200 ng/ml) combined with azithromycin maintained higher cell counts and showed reduced cytotoxicity compared to 1,000 ng/ml under LPS exposure. The 200 ng/ml cyclosporin-azithromycin combination demonstrated optimal results, reducing IL-6 and IL-1β levels while maintaining cell viability. Higher concentrations elevated IgA levels, particularly with LPS stimulation, suggesting enhanced immune response modulation.
CONCLUSION: The combination of azithromycin with moderate cyclosporin concentrations (200 ng/ml) provides optimal immunomodulatory effects while maintaining cell viability. Higher cyclosporin doses (1,000 ng/ml) showed increased cytotoxicity despite enhanced immunomodulation.
MATERIALS AND METHODS: PBMCs were isolated from four healthy donors and treated with cyclosporin at concentrations of (50, 200, and 1,000 ng/ml) either alone or in combination with azithromycin (0.4 µg/ml), with and without 100 ng ml LPS derived from Porphyromonas gingivalis. Total cell count, cell viability, and lactate dehydrogenase (LDH) activity were assessed at day 1 and 3. While the inflammatory mediators, including IL-6, IL-1β, IL-18, and IgA levels were assessed by ELISA at day 3. Statistical analysis included two-way ANOVA to analyze the effects of the drugs and the presence of LPS (the two independent variables), followed by Tukey's HSD post-hoc test. Multiple linear regression models evaluating treatment effects, LPS exposure, and time points, with assessment of two-way interactions. Models were adjusted for relevant covariates and verified for statistical assumptions, with significance set at p < 0.05.
RESULTS: Lower cyclosporin concentrations (50 and 200 ng/ml) combined with azithromycin maintained higher cell counts and showed reduced cytotoxicity compared to 1,000 ng/ml under LPS exposure. The 200 ng/ml cyclosporin-azithromycin combination demonstrated optimal results, reducing IL-6 and IL-1β levels while maintaining cell viability. Higher concentrations elevated IgA levels, particularly with LPS stimulation, suggesting enhanced immune response modulation.
CONCLUSION: The combination of azithromycin with moderate cyclosporin concentrations (200 ng/ml) provides optimal immunomodulatory effects while maintaining cell viability. Higher cyclosporin doses (1,000 ng/ml) showed increased cytotoxicity despite enhanced immunomodulation.