Meloxicam Effect on Leukocyte Migration under Shear Stress: A New Perfused Triple Chamber (Hofbauer Chamber) Assay Simulating an in Vitro Vascular Wall

Abstract

Background

Leukocyte migration plays a tremendous role during inflammation. Different drugs are able to influence migration of leukocytes. Several authors have attempted to establish methods for investigation of leukocyte migration. Migration of leukocytes under conditions simulating natural blood flow and shear stress is poorly understood. Moreover, leukocyte migration in the simultaneous presence of endothelial cells and smooth muscle cells (SMC) has not yet been examined thoroughly. The aim of the current study was to create a new three-dimensional assay investigating migration during simulated perfusion. The assay was used for investigating the effect of meloxicam on leukocyte migration under shear stress.

Methods

Different concentrations of meloxicam (0.09, 0.9, and 9.0 ng/mL) were used in a newly developed triple migration chamber system for investigation of leukocyte migration (n=7). This new system allows inclusion of SMC- layers. Assay precision tests were done using intra-assay (within-day variability) and interassay (day-to-day variability) controls.

Results

Intra-assay and interassay controls showed reliable results (coefficient of variation: intra-assay 8.53%; interassay 11.39%). Meloxicam in a clinically relevant concentration of 0.9 ng/mL was able to reduce the migration of leukocytes under shear stress (71 ± 14% SD; P<0.05), whereas higher and lower concentrations showed a dose dependent effect.

Conclusion

This new perfusion system allows investigation of drugs, such as meloxicam, on leukocyte migration under shear stress. Moreover, this assay may be used for studies on other cell-cell interactions, as well as on drug influences.