Sampling insulin in different tissue compartments using microdialysis: methodological aspects

Abstract

Sampling the concentration of insulin in human skin using microdialysis is challenging because of low intracutaneous concentrations and low recovery, presumably due to adsorption of insulin to the microdialysis system. In this study, we aimed to (1) measure how the concentration of insulin varies in three different tissue compartments (intracutaneous, subcutaneous and intravenous) and (2) to study how much insulin is adsorbed to the microdialysis catheter membranes and tubing during a typical microdialysis experiment, both in vivo and in vitro. We hypothesized that (1) the concentration of insulin decreases from the intravenous compartment to the intracutaneous and subcutaneous tissue, and that (2) adsorption of insulin to the microdialysis membrane and tubing impairs the recovery of insulin from the tissue. In this experimental study, microdialysis catheters were inserted intracutaneously, subcutaneously and intravenously in 11 healthy subjects. Systemic endogenous hyperinsulinemia was induced by intake of an oral glucose load. Insulin concentration was measured in the dialysate and in the extracted samples from the catheter membrane and tubings. In vitro microdialysis was performed to investigate the temporal resolution of the adsorption. After an oral glucose load insulin concentration increased intravenously, but not in the intracutaneous or subcutaneous compartments, while glucose, lactate and pyruvate concentrations increased in all compartments. The adsorption of insulin to the microdialysis membrane in vivo was highest in the intravenous compartment (p = 0.01), compared to the intracutaneous and subcutaneous compartments. In vitro, the adsorption to the microdialysis membrane was highest one hour after sampling, then the concentration gradually decreased after three and five hours of sampling. The concentration of insulin in peripheral tissues is low, probably due to decreasing tissue vascularity. Adsorption of insulin to the microdialysis membrane is modest but time-dependent. This finding highlights the importance of a stabilization time for the microdialysis system before sampling tissue analytes.

Investigation of proteins important for microcirculation using in vivo microdialysis after glucose provocation: a proteomic study

“…Microdialysis catheters (100 kDa CMA 71 and 20 kDa CMA 67, 10 mm membrane, M dialysis AB, Stockholm, Sweden) were inserted intracutaneous in the volar skin of the non-dominant forearm, subcutaneous in the periumbilical adipose tissue and intravenous in a peripheral vein in each of the eight subjects. The procedure for insertion of microdialysis catheters has previously been described in detail 8 , 9 , 14 . Before insertion, all catheters were connected to microinjection pumps (CMA 107, CMA AB, Uppsala, Sweden) and perfused for 90 min to ensure adequate function and reduce the risk of obstructing bubbles in the microdialysis tubing.…”

“…This is partially to increase access to glucose, through a process primarily believed to be mediated by endothelial insulin dependent vasodilation. Measuring the concentration of insulin in the human skin using microdialysis has however appeared to be challenging, because the concentration of insulin in peripheral tissues (intracutaneous and subcutaneous) is low, probably related to decreasing tissue vascularity and blood flow 14 . Altogether, it is known that glucose provocation has microcirculatory effects, partly dependent on the vascular effects of insulin, but the effects on changes in local protein expression are lacking.…”