Effects of Combined Cyclosporin and Azithromycin Treatment on Human Mononuclear Cells Under Lipopolysaccharide Challenge

Norah Alotaibi ¹ Aminah Alesawy ¹ Marwa Alalshaikh ¹ Faisal E Aljofi ¹ Nada Aldossary ² Nada Alzahrani ³ Omar Omar ⁴ Marwa Madi ^1*

• ¹ Department of Preventive Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Damam, Saudi Arabia
• ² Department of Pathology, College of Medicine, Imam Abdulrahman Bin Faisal University, Damam, Saudi Arabia
• ³ Blood Blood Bank, Laboratory Medicine, King Fahad University Hospital, Al Khobar, Saudi Arabia
• ⁴ Department of Biomedical Dental Science, College of Dentistry, Imam Abdulrahman Bin Faisal University, Damam, Saudi Arabia

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 1000 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 1000 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.The combination of azithromycin with moderate cyclosporin concentrations (200 ng/ml) provides optimal immunomodulatory effects while maintaining cell viability. Higher cyclosporin doses (1000 ng/ml) showed increased cytotoxicity despite enhanced immunomodulation.Clinical Relevance: These findings suggest that optimizing cyclosporin dosage in combination with azithromycin could improve therapeutic outcomes in inflammatory conditions while minimizing adverse effects. The 200 ng/ml cyclosporin concentration appears to offer the best balance between efficacy and safety.