Glucose concentration in human subcutaneous adipose tissue: comparison between forearm and abdomen.

There is still controversy about the relation between the glucose concentration in the subcutaneous (sc.) adipose tissue and the blood plasma. Depending on the technique applied, the glucose concentration in sc. tissue varies between 50% and 100% of the plasma glucose concentration. In the present study the sc. glucose concentration of forearm and abdomen in seven healthy volunteers was compared with plasma glucose by applying the microdialysis technique with very low flow rates. A microdialysis probe implanted into the subcutaneous tissue of abdomen and forearm was perfused with a flow rate of 1 microl/4 min. The dialysate was sampled in three 2-h-fractions in the fasting state and in one 2-h-fraction during a hyperglycemic clamp (216.9+/-3.4 mg/dl) (mean +/- SEM). The mean recoveries of the plasma glucose were 91.1+/-4.1% in the forearm and 82.7+/-18.0% in the abdomen. The recoveries in the sc. tissue of abdomen and arm were not significantly different. However, the arm showed significantly (p < 0.014) less interindividual variance (range 73.2- 103.2%) than the abdomen (range 50.6-117.1%) and appears to be the preferable implantation site. The recovery remained constant during the investigation.

Amperometric biosensor for in vivo glucose sensing based on glucose oxidase immobilized in a redox hydrogel.

A potentially implantable glucose sensor, based on glucose oxidase immobilized in a redox hydrogel, is considered. The redox hydrogel consisted of glucose oxidase immobilized in a cross-linkable poly(vinylpyridine) complex of [Os(bis-bipyridine)2Cl]+1/+2 that communicates electrically with the flavin adenine dinucleotide (FADH2) redox centres of the glucose oxidase. The implantable electrode consisted of a Teflon insulated platinum wire (0.25 mm diameter) which was coated at the tip with a cross-linked redox polymer/glucose oxidase film and covered with a thin layer of polycarbonate. In a three-electrode system at +400 mV (Ag/AgCl) the response to increasing glucose concentrations in isotonic phosphate buffer and human plasma was approximately 0.2-0.3 nA/mM, linear in the range between 0 and 15 mM glucose. No oxygen dependence was observed. To determine the in vivo performance, the electrode was implanted into the subcutaneous tissue of a dog. The sensor currents after an oral glucose load paralleled the plasma glucose measurements, with a time lag of 10 min. Three-day implantations in cultured cells showed that the electrode did not affect the growth and differentiation of cell monolayers.

The function of a hydrogen peroxide-detecting electroenzymatic glucose electrode is markedly impaired in human sub-cutaneous tissue and plasma.

Electroenzymatic glucose sensors implanted into sub-cutaneous (s.c.) tissue of human subjects and experimental animals exhibit lower sensitivities to glucose than in buffer solutions before implantation. The mechanism of the decrease of sensitivity is not known. Sensors used in this study were fabricated from platinum wires (diameter 0.125 mm) with covalently bound glucose oxidase at the tip of the wire. After coating the tip with polyurethane, wires were placed into 27 gauge steel needles. Sensors were operated potentiostatically at 700 mV against Ag/AgCl pseudo-reference electrodes. These sensors were implanted s.c. in 6 diabetic patients for 7 h. In 4 patients, sensors were responsive to successive increases of plasma glucose levels. Mean sensitivity to glucose in s.c. tissue was 29% of in vitro sensitivity. In 2 patients there was a sudden decrease of sensor currents, unrelated to glucose, shortly after implantation. Sensors were inhibited in human plasma to a similar extent. When sensors were exposed to native plasma and to plasma ultrafiltrate (mol. wt. < 10 kDa) for 10 h, identical decreases of signals were found. Exposure to dialysed plasma (mol. wt. > 12 kDa) caused much less decrease of sensor signals. Losses of sensor sensitivities to glucose in s.c. tissue and in plasma were totally reversible upon re-exposure of sensors to buffer solutions. We conclude that sensor inactivation in plasma and possibly in s.c. tissue is caused by low molecular weight substances not retained by the polyurethane membrane.