The influence of antimicrobial treatments on the cytocompatibility of polyurethane biosensor membranes.

The cytocompatibility of polyurethane membranes was tested following ultraviolet or gamma irradiation as well as treatment with hydrogen peroxide or glutaraldehyde containing solutions. Despite the fact that all of the methods had been recommended for antimicrobial treatment of glucose biosensors, the treatments investigated significantly influenced cytocompatibility characteristics. Cytotoxicity of membrane eluates was not observed following irradiation treatments. This was also the case when the membranes were repeatedly washed following chemical treatment. Cell growth upon the membranes was stimulated to a different extent after gamma and UV irradiation as well as following hydrogen peroxide treatments. Residues of an urea-based hydrogen peroxide inclusion compound caused a restriction in cell growth upon the membranes as was similarly observed with 2 and 4% glutaraldehyde solutions acting over 2 and 4 h, respectively. It is concluded that cytocompatibility in vitro reflecting the host response against a biomaterial in vivo does not only depend upon the material itself but also upon antimicrobial treatments which could have consequences for its bioperformance characteristics.

Biosensors for in vivo glucose measurement: can we cross the experimental stage.

The development of in vivo working glucose sensors needs two decades, so far. The availability of long term functional implantable biosensors for continuous glucose measurings is a basic prerequisite for the individualized optimum insulin treatment of diabetics. Enzymatic electrochemical sensors are described which realize a functional stability over more than 2 years in vitro, however their function in vivo is limited due to certain bioincompatibility expressed by inflammation of the surrounding tissue, exudates, and immun reactions. The paper reflects an overview concerning different sensor covering materials used as more or less suitable diffusion membranes. From experimental studies in animals and human volunteers conclusions are drawn for further developmental steps of biosensors for in vivo use and for the applicability of glucose sensors for transient diagnostic purposes and as a basis for glucose controlled therapeutic measures. The results demonstrate that further progress aimed at long term biostability of implanted biosensors needs to solve technological problems and the serial production of sensors with really comparable qualities as a prerequisite for clinical trials.

The usefulness of a biosensor controlled perfusion cell culture for the investigation of new drugs demonstrated with the marine fungus Kirschsteiniothelia maritima.

The investigation of new biological active substances from limited sources for example from marine organisms needs sensitive and evident test systems. Such a system is the glucose biosensor controlled perfusion cell culture. The glucose consumption of cells is a very sensitive parameter which allows the continuous measurement of external effects of test substances on the cells under in vivo-like conditions. Cytotoxic concentrations of active substances as well as a virus infection lowers the glucose consumption of continuously perfused cells. This effect can be monitored using a glucose biosensor. The influence of drugs and the virus infection can be observed simultaneously in the same system continuously over several days. With two substances isolated from the marine fungus Kirschsteiniothelia maritima investigations for cytotoxic and for antiviral effects are demonstrated.

Sterilization of enzyme glucose sensors: problems and concepts.

A useful method of enzyme glucose sensor sterilization has not only to ensure the needs of sterility assurance but has also to guarantee the functional stability of the sensors. The action of 2 or 3% alkalinized glutaraldehyde solution, as well as gamma irradiation with a dose of 25 kGy caused changes of the in vitro functionality and polymer material irritations, respectively. After a combined treatment by 0.6% hydrogen peroxide solution acting over 4 days with 7 kGy gamma irradiation only a slight loss of sensitivity must be registered. The combination of a specially designed universal homogeneous ultraviolet irradiation over 300 s with a 3 days lasting treatment by an inclusion compound of hydrogen peroxide with tensides in urea (0.15% effective hydrogen peroxide concentration) did not cause any influence on the glucose sensor function in vitro. With all methods tested here, a Bacillus subtilis spore reduction over 8 log(10) cycles from 10(6) to 10(-2) spores per test object on an average could be proved experimentally. In general, if non-thermal methods must be used it seems to be impossible to guarantee a sterility assurance level of 10(-6) as it is demanded by the pharmacopoeias. Consequently, effective concepts to produce sterile glucose biosensors for medical and biological applications should be based not only on final product treatments but should include germ reducing measures in every manufacturing step.

Biosensor-controlled perfusion culture to estimate the viability of cells.

A perfusion cell culture is characterised by the continuous addition of fresh nutrient medium and the withdrawal of an equal volume of used medium, allowing the realisation of cell cultivation conditions that are approximated as closely as possible to the in vivo situation. The combination of a perfusion cell culture with an enzyme glucose biosensor allows the glucose consumption of the cell culture to be monitored continuously. The resulting biosensor-controlled perfusion cell culture is a complex biomonitoring system that is useful for checking the metabolic state of a perfusion cell culture continuously and non-invasively over several days. With this experimental setup, it has been possible to test detrimental external effects on living systems at early stages, in vitro, but under in vivo-like conditions.