Importance of material evaluation prior to the construction of devices for in vitro techniques.

The development and validation of new in vitro methods in the field of toxicology have gained more importance in recent years due to stricter guidelines for animal testing, especially in the European Union. Consequently, advances in the construction of technical devices for the exposure of cell or tissue cultures to test substances are necessary. Here, to obtain reliable results, it is important to exclusively use materials that do not interfere with the cell viability. Thus, similar to the biomaterials testing of medical devices which is regulated in the Directive 93/42/EEC, the biocompatibility of the materials has to be verified prior to the construction of such devices. We present here a novel approach for biomaterials testing which allows the quantitative and qualitative assessment of cytotoxicity of material samples. Stainless steel and silicone are often used for laboratory equipment, due to their high chemical, thermal and mechanical resistance. However, our results highlight that some types of silicone may have adverse effects on cultured cells. Moreover, special methods for the surface treatment of metals may also be a critical factor for in vitro devices. Therefore, the testing of all materials coming in contact with cell cultures is highly recommended.

The CULTEX RFS: a comprehensive technical approach for the in vitro exposure of airway epithelial cells to the particulate matter at the air-liquid interface.

The EU Regulation on Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) demands the implementation of alternative methods for analyzing the hazardous effects of chemicals including particulate formulations. In the field of inhalation toxicology, a variety of in vitro models have been developed for such studies. To simulate the in vivo situation, an adequate exposure device is necessary for the direct exposure of cultivated lung cells at the air-liquid interface (ALI). The CULTEX RFS fulfills these requirements and has been optimized for the exposure of cells to atomized suspensions, gases, and volatile compounds as well as micro- and nanosized particles. This study provides information on the construction and functional aspects of the exposure device. By using the Computational Fluid Dynamics (CFD) analysis, the technical design was optimized to realize a stable, reproducible, and homogeneous deposition of particles. The efficiency of the exposure procedure is demonstrated by exposing A549 cells dose dependently to lactose monohydrate, copper(II) sulfate, copper(II) oxide, and micro- and nanoparticles. All copper compounds induced cytotoxic effects, most pronounced for soluble copper(II) sulfate. Micro- and nanosized copper(II) oxide also showed a dose-dependent decrease in the cell viability, whereby the nanosized particles decreased the metabolic activity of the cells more severely.

Analytical in vitro approach for studying cyto- and genotoxic effects of particulate airborne material.

In the field of inhalation toxicology, progress in the development of in vitro methods and efficient exposure strategies now offers the implementation of cellular-based systems. These can be used to analyze the hazardous potency of airborne substances like gases, particles, and complex mixtures (combustion products). In addition, the regulatory authorities require the integration of such approaches to reduce or replace animal experiments. Although the animal experiment currently still has to provide the last proof of the toxicological potency and classification of a certain compound, in vitro testing is gaining more and more importance in toxicological considerations. This paper gives a brief characterization of the CULTEX® Radial Flow System exposure device, which allows the exposure of cultivated cells as well as bacteria under reproducible and stable conditions for studying cellular and genotoxic effects after the exposure at the air-liquid or air-agar interface, respectively. A commercial bronchial epithelial cell line (16HBE14o-) as well as Salmonella typhimurium tester strains were exposed to smoke of different research and commercial available cigarettes. A dose-dependent reduction of cell viability was found in the case of 16HBE14o- cells; S. typhimurium responded with a dose-dependent induction of revertants. The promising results recommend the integration of cellular studies in the field of inhalation toxicology and their regulatory acceptance by advancing appropriate validation studies.