In vitro microdialysis recovery and delivery investigation of cytokines as prerequisite for potential biomarker profiling.
Cytokines as immunomodulatory proteins are secreted by immune and tissue cells mediating immune responses, e.g. inflammation. The use of microdialysis as a minimally invasive technique for sampling interstitial fluid might provide the basis for biomarker profiling for diseases and therapy monitoring. The objectives of this investigation were to develop reproducible methods and apply them to define applicability limits to quantify cytokines in microdialysate. In vitro microdialysis recovery and delivery investigations were performed utilising a standardised system exploring analyte adsorption, pH effects, the infuence of cytokine concentration and temperature of the catheter surrounding medium. A Ringer's/human albumin solution was used as microperfusate and catheter surrounding medium; interleukin 6, 8 and 10 (IL-6, IL-8, IL-10) and tumour necrosis factor alpha (TNF-α) served as model cytokines. Microdialysate was sampled (n=3) at flow rates of 0.3-5.0μL/min using 3 linear probes. All samples were measured using a validated flow-cytometry method adapted to microdialysate. Relative recoveries of the individual cytokines decreased exponentially with increasing flow rates and were not influenced by the catheter surrounding medium concentration but recovery of IL-6, IL-10 and TNF-α by the pH value. Relative recovery and relative delivery of IL-8 were of comparable extent and increased with higher temperatures. For the other cytokines, however, negative values occurred for relative delivery probably due to ultrafiltration. Clinical application of microdialysis of cytokines is principally feasible if the many influencing factors are controlled. Since relative delivery determination is only reliable for IL-8, retrodialysis or similar calibration methods must be avoided. As future perspective, in vivo μD feasibility should next be demonstrated.
Department of Clinical Pharmacy and Biochemistry, Freie Universitaet Berlin, Berlin, Germany.
Claudia
C
Kirbs
Department of Clinical Pharmacy and Biochemistry, Freie Universitaet Berlin, Berlin, Germany. Electronic address: charlotte.kloft@fu-berlin.de.
Charlotte
C
Kloft
D000327:Adsorption; D015415:Biomarkers / Q000032:analysis; D016207:Cytokines / Q000032:analysis*; D004867:Equipment Design; D005240:Feasibility Studies; D005434:Flow Cytometry; D006863:Hydrogen-Ion Concentration; D016753:Interleukin-10 / Q000032:analysis; D015850:Interleukin-6 / Q000032:analysis; D016209:Interleukin-8 / Q000032:analysis; D017551:Microdialysis / Q000295:instrumentation; D015203:Reproducibility of Results; D013696:Temperature; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D015415:Biomarkers; D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10
Cytokine; Interleukin 10; Interleukin 6; Interleukin 8; Microdialysis; Tumour necrosis factor alpha