Martin Hajnsek

Evaluation of subcutaneous glucose monitoring systems under routine environmental conditions in patients with type 1 diabetes

Continuous and flash glucose monitoring (GM) systems have been established in diabetes care. We compared the sensor performance of 3 commercially available GM systems. A total of 12 patients with type 1 diabetes were included in a single‐centre, open‐label study in which the sensor performance of the Abbott FreeStyle libre (Abbott), Dexcom G4 Platinum (Dexcom) and Medtronic MiniMed 640G (Medtronic) systems over 12 hours was compared during mimicked real‐life conditions (meals, exercise, hypo‐ and hyperglycaemia). Sensor performance was determined by fulfilment of ISO 15197:2013 criteria, calculating mean absolute relative difference (MARD), and was also illustrated using Parkes error grid and Bland–Altman plots. Sensor performance during changes in metabolic variables (lactate, betahydroxybutyrate, glucagon, non‐esterified‐fatty‐acids) was determined by Spearmans rank correlation coefficient testing. The systems fulfilled ISO 15197:2013 criteria by 73.2% (Abbott), 56.1% (Dexcom) and 52.0% (Medtronic). The MARDs ± standard deviation in the entire glycaemic range were 13.2% ± 10.9% (Abbott), 16.8% ± 12.3% (Dexcom) and 21.4% ± 17.6% (Medtronic), respectively. All sensors performed less accurately during hypoglycaemia and best during hyperglycaemia. We did not observe an influence of metabolic variables on sensor performance.

Integrated catheter system for continuous glucose measurement and simultaneous insulin infusion

A new measurement system enables combination of continuous glucose monitoring (CGM) and insulin infusion. A sensor system comprising an optical glucose biosensor and an optical oxygen sensor is integrated into the insulin infusion catheter of an insulin pump. Both sensors rely on near infrared (NIR) phosphorescent porphyrin dyes, wherefore the signals can be read out transcutaneous and non-invasively with a custom-built phase fluorometer measurement module. The spectral properties of the indicator dyes and the optical setup of the measurement module were optimized to enable independent read-out in two channels. Dynamic ranges from 0 mmHg to 160 mmHg oxygen and 0mg/dL to 360 mg/dL glucose (LOD 2mg/dL) are covered by the oxygen and the glucose sensor, respectively. In-vivo measurements in pigs demonstrate good correlation of reference blood glucose levels and glucose values obtained with the presented sensor system. The evaluation of the clinical accuracy of the system with Clarke Error Grid Analysis showed similar results to CGM-devices currently on the market.

First application of a transcutaneous optical single-port glucose monitoring device in patients with type 1 diabetes mellitus

The combination of continuous glucose monitoring (CGM) and continuous subcutaneous insulin infusion can be used to improve the treatment of patients with diabetes. The aim of this study was to advance an existing preclinical single-port system for clinical application by integrating the sensors of a phosphorescence based CGM system into a standard insulin infusion set. The extracorporeal optical phase fluorimeter was miniaturised and is now comparable with commercial CGM systems regarding size, weight and wear comfort. Sensor chemistry was adapted to improve the adhesion of the sensor elements on the insulin infusion set. In-vitro tests showed a linear correlation of R2=0.998 between sensor values and reference glucose values in the range of 0-300mg/dl. Electrical and cytotoxicity tests showed no negative impact on human health. Two single-port devices were tested in each of 12 patients with type 1 diabetes mellitus in a clinical set-up for 12h. Without additional data processing, the overall median absolute relative difference (median ARD) was 22.5%. For some of the used devices the median ARD was even well below 10%. The present results show that individual glucose sensors performance of the single-port system is comparable with commercial CGM systems but further improvements are needed. The new system offers a high extent of safety and usability by combining insulin infusion and continuous glucose measurement in a single-port system which could become a central element in an artificial pancreas for an improved treatment of patients with type 1 diabetes mellitus.

Assessment of skin permeability to topically applied drugs by skin impedance and admittance

OBJECTIVE Pharmacokinetic and pharmacodynamic studies of topically applied drugs are commonly performed by sampling of interstitial fluid with dermal open flow microperfusion and subsequent analysis of the samples. However, the reliability of results from the measured concentration-time profile of the penetrating drug suffers from highly variable skin permeability to topically applied drugs that is mainly caused by inter- and intra-subject variations of the stratum corneum. Thus, statistically significant results can only be achieved by performing high numbers of experiments. To reduce the expenditures needed for such high experiment numbers we aimed to assess the correlation between skin permeability and skin impedance/skin admittance. APPROACH We performed an ex vivo drug penetration study with human skin, based on the hypothesis that inter-subject variations of the respective concentration-time profiles can be correlated with variations of the passive electrical properties of the skin. Therefore, skin impedance and skin admittance were related to the skin permeability to the model drug Clobetasol-17-proprionate. MAIN RESULTS The measured low frequency skin impedance and the skin admittance correlated linearly with the drug concentration-time profiles from dermal sampling. SIGNIFICANCE Skin permeability can be assessed by measuring the passive electrical properties of the skin, which enables correction of skin permeability variations. This allows reduction of experiment numbers in future pharmacokinetic and pharmacodynamic studies with human skin ex vivo and in vivo and leads to diminished study costs.

Skin impedance measurements support ex-vivo penetration studies for topical applied drugs

Open flow microperfusion can be used to assess the amount of a topically applied drug directly in the skin. Large intra- and interindividual differences in skin proper- ties can lead to biased results. In this study we propose a method based on skin impedance measurements to assess the properties of the skin barrier and thus compensate for vary- ing permeability characteristics.

Monitoring tissue oxygen heterogeneities and their influence on optical glucose measurements in an animal model

The purpose of this study was to characterize the heterogeneity of oxygen partial pressure in different adipose tissue zones and to assess the possibility of compensating these heterogeneities during optical glucose measurements. In this proof of concept study, the heterogeneity of oxygen partial pressure was determined in the adipose tissue of a pig by using 48 oxygen sensors in 3 zones of the abdominal region at two different blood oxygen levels. Sensor oxygen values correlated well with reference blood oxygen values and we identified heterogeneities in oxygen partial pressure among the defined zones of the abdominal region. Significant differences in the mean oxygen partial pressure were found when comparing the three abdominal zones but no significant differences were found when comparing two sensors located in close proximity (on one cannula). The low heterogeneity on one cannula allows the compensation of physiological oxygen variations for optical glucose measurements by using an additional oxygen sensor in close proximity to the glucose sensor. In addition, this setup can be used to continuously monitor tissue oxygenation e.g. in patients with adipose tissue dysfunction or serve limb ischemia.

Optical biosensor system with integrated microfluidic sample preparation and TIRF based detection

There is a steadily growing demand for miniaturized bioanalytical devices allowing for on-site or point-of-care detection of biomolecules or pathogens in applications like diagnostics, food testing, or environmental monitoring. These, so called labs-on-a-chip or micro-total analysis systems (μ-TAS) should ideally enable convenient sample-in – result-out type operation. Therefore, the entire process from sample preparation, metering, reagent incubation, etc. to detection should be performed on a single disposable device (on-chip). In the early days such devices were mainly fabricated using glass or silicon substrates and adapting established fabrication technologies from the electronics and semiconductor industry. More recently, the development focuses on the use of thermoplastic polymers as they allow for low-cost high volume fabrication of disposables. One of the most promising materials for the development of plastic based lab-on-achip systems are cyclic olefin polymers and copolymers (COP/COC) due to their excellent optical properties (high transparency and low autofluorescence) and ease of processing. We present a bioanalytical system for whole blood samples comprising a disposable plastic chip based on TIRF (total internal reflection fluorescence) optical detection. The chips were fabricated by compression moulding of COP and microfluidic channels were structured by hot embossing. These microfluidic structures integrate several sample pretreatment steps. These are the separation of erythrocytes, metering of sample volume using passive valves, and reagent incubation for competitive bioassays. The surface of the following optical detection zone is functionalized with specific capture probes in an array format. The plastic chips comprise dedicated structures for simple and effective coupling of excitation light from low-cost laser diodes. This enables TIRF excitation of fluorescently labeled probes selectively bound to detection spots at the microchannel surface. The fluorescence of these detection arrays is imaged using a simple set-up based on a digital consumer camera. Image processing for spot detection and intensity calculation is accomplished using customized software. Using this combined TIRF excitation and imaging based detection approach allowes for effective suppression of background fluorescence from the sample, multiplexed detection in an array format, as well as internal calibration and background correction.

Sensor for early detection of wound infection.

We have developed a system to diagnose wound infection in a very early stage. Based on an electrochemical hydrogen peroxide sensor, the system determines the enzyme activity of myeloperoxidase, which is a biomarker for wound infection. The sensor system was evaluated in vitro and with samples of actual wound fluid. We show that the infection status of wounds can be reliably detected.

icath - Integrated Catheter for Simultaneous Glucose Measurement and Insulin Infusion.

The integration of a glucose sensor into the insulin infusion catheter of an insulin pump allows for simultaneous insulin delivery and continuous glucose monitoring at the same spot in subcutaneous tissue. Invivo experiments in pigs have been conducted with the presented single-port system. It could be shown that the glucose concentration measured subcutaneously in the interstitial liquid follows the glucose profile in blood and that insulin infusion at the site of measurement doesn’t influence the glucose concentration.

Diagnostic Lab-on-a-Chip System Based on Fluorescence Imaging and Integrating Sample Preparation.

In this contribution we describe a microfluidic chip combining plasma separation, sample metering, dissolution/incubation with reagents stored on-chip and optical detection. The system allows defining the incubation time and works under constant externally applied pressure using only passive valves for actuation. This allowed the realization of a bioanalytical device for whole blood samples comprising a disposable plastic chip using TIRF (total internal reflection fluorescence) based optical detection of biochemical binding events.