in vitro inhalation

In vitro Inhalation

The inhalation route is the most common way in which the body is exposed to airborne materials (e.g., airborne contaminants, diesel exhaust, inhaled therapeutics) either unintentionally or deliberately.

The latest achievements in human cell culture exposure and test systems has led to the development of new in vitro assay systems that are more predictive and appropriate for the screening of the toxicity of a wide variety of compounds (chemicals, nanomaterials, pharmaceuticals, feed additives, tobacco products). These new in vitro toxicology systems have been developed using animal or human based cellular systems in agreement with the 21st Century Toxicology recommendations.

The use of in vitro testing is the first primary method of choice for toxicity screening of inhaled materials since it provides a number of advantages compared to in vivo inhalation studies; they are cost and time efficient and involve no ethical issues. In addition, these models offer great flexibility as they are available for different regions of the respiratory tract (i.e., nasal, trachea, bronchial and pulmonary regions). Although these in vitro lung models have a limitation, namely that they do not fully reflect the complexity of the whole respiratory system, they do provide broad knowledge related to inhalation toxicology, since they represent the human cellular structure and tissue architecture of the respiratory system.

 

In vitro human inhalation model

 

Our in vivo inhalation toxicity team has extensive experience in in vivo toxicology inhalation studies using state-of-the-art technology (link web inhalation). Therefore, we are able to transfer our expertise in the generation of a homogeneous, controlled and stable test atmosphere from in vivo to in vitro exposure systems.

Vivotecnia is working on in vitro inhalation toxicity services using lung cell lines and 3D airway epithelium models:

  • Proximal lung tissue models: EpiAirwayTM (MatTek Corporation2); MucilAirTM (Epithelix3)
  • Distal lung tissue models: EpiAlveolarTM (MatTek Corporation2); SmallAirTM (Epithelix3)

These 3D airway epithelium models maintain the human tissue structure and cellular morphology. The air-liquid-interface (ALI) exposure represents a more realistic exposure scenario for inhaled materials into the lung compared to submerged exposures.

Therefore, the combination of 3D airway epithelium models and ALI exposure has the following advantages:

  • It offers new possibilities to test biological responses of inhaled materials under physiologically relevant conditions.

It not only provides an accurate assessment of acute respiratory toxicity, but also predicts the in vivo responses of respiratory toxins in humans.

 

In vitro Inhalation Services

Vivotecnia provides a wide range of endpoints to be assessed in this in vitro human inhalation model:

  • Mechanism-based assessment:
    • Cell viability (e.g., trypan blue dye exclusion)
    • Cell metabolism (e.g., MTT)
    • Membrane leakage such a loss of lactate deshidrogenase (LDH) assays
    • Oxidative stress
    • Inflammation by measurement of pro-inflammatory mediators
    • Cellular stress responses
    • Tissue damage and matrix degradation
    • Genotoxicity assessment (e.g., DNA damage by COMET assay)
    • Gene expression/miRNAs
  • Functional analysis:
    • Integrity of the airway epithelium tight junctions by TEER (transepithelial electrical resistance)
    • Morphology
    • Cilia beating frequency
    • Mucociliary clearance

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