Inhalation Toxicology
The challenge
Inhalation is a major exposure route for humans, and evaluating inhalation toxicity potential is vital when establishing the safety of new chemicals and products. Typically, inhalation toxicity assessments are conducted in rodents. However, due to physiological differences between humans and rodents, the results of these in vivo studies may not accurately reflect human responses. Increasingly, regulators are calling for alternatives to animal studies, as part of a worldwide push that will both reduce costs and provide more human-relevant results.
At ImmuONE, we specialise in the developing and advancing in vitro assessments. We specialise in lung cell assays that provide detailed information regarding the responses of epithelial and immune cells to inhaled substances.
Our Solutions
Macrophage toxicology
ImmuONE specialises in assessing alveolar macrophage responses. These tissue-resident immune cells are the first line of defence against invading respiratory pathogens, and it is important to know how the substances that we inhale affect them. If a particular substance provokes a strong macrophage reaction, we can reasonably assume that that this substance could cause harm to human lung tissue. At ImmuONE, we can analyse the impact of substances on the lung using either in vitro rat macrophages or our human alveolar macrophage-like models ImmuPHAGE™ and ImmuLUNG™.
Our high content image analysis (HCIA) assays offer comprehensive insight into the effects of inhaled substances on lung macrophages and their morphology. Characteristics that we assess include cell size, extent of vacuolation, cell shape and mitochondrial activity.
Epithelial toxicology
ImmuONE’s HCIA assays can also be used to evaluate how compounds affect alveolar epithelial cells.
Some inhaled substances may damage the alveolar epithelial barrier, potentially leading to pneumonia, acute lung injury and acute respiratory distress syndrome. Using our model ImmuLUNG™, which combines alveolar epithelial and alveolar macrophage-like cells, we can evaluate the effects of compounds on epithelial barrier integrity.
Our standard tests include:
- Trans-epithelial electrical resistance (TEER) measures the tightness of the junctions between epithelial cells, which reflects the integrity of the epithelial barrier.
- Paracellular marker permeability: Paracellular marker permeability assays involve exposing a layer of epithelial cells to a marker molecule in solution, then measuring the movement of the marker across the cell layer. These assays reveal how easily substances can pass through the spaces between epithelial cells.
- Lactate dehydrogenase (LDH): The LDH enzyme is released into the cell culture medium upon damage to the plasma membrane. Using the LDH assay, we can determine the level of plasma membrane damage in a cell population.
- Mitochondrial activity assays (e.g. PrestoBlue): We can use the PrestoBlue reagent to indirectly assess cell viability and cytotoxicity.
- Enzyme-linked immunosorbent assay (ELISA): ELISA determines the concentration of soluble cytokine and chemokine proteins.
- Histological and immunohistochemistry analysis: Using histological analysis, we can visualise cellular organisation, mucus and the structure of the pseudostratified columnar epithelium.
Genetic toxicology (Join the waiting list)
ImmuONE will soon offer genetic toxicology services using industry standard techniques like TG 487, the in vitro mammalian cell micronucleus test. To be the first to find out more, join the waiting list below:
Other services we offer
Mitochondrial activity assays
We can indirectly assess cell viability and cytotoxicity by looking at the mitochondrial activity of alveolar macrophages. Cell cultures are exposed to a reagent (e.g. Presto Blue, alamarBlue and MTT, MTS, WST, XTT). The mitochondria of the macrophages reduce the non-fluorescent blue dye within the reagent to a fluorescent pink dye, and the fluorescent signal is measured.
We can indirectly assess cell viability and cytotoxicity by looking at the mitochondrial activity of alveolar macrophages. Cell cultures are exposed to a reagent (e.g. Presto Blue, alamarBlue and MTT, MTS, WST, XTT). The mitochondria of the macrophages reduce the non-fluorescent blue dye within the reagent to a fluorescent pink dye, and the fluorescent signal is measured.
ATP assays
ATP assays can be used to measure cell viability by measuring the amount of ATP released by cells. Several detection methods can be used, such as colorimetric, fluorescent and bioluminescent.
ATP assays can be used to measure cell viability by measuring the amount of ATP released by cells. Several detection methods can be used, such as colorimetric, fluorescent and bioluminescent.
Flow cytometry viability assessment
Flow cytometry allows for a fast and reliable viability assessment via the use of fluorescent dyes.
Flow cytometry allows for a fast and reliable viability assessment via the use of fluorescent dyes.
Apoptosis assessment
An apoptosis assay detects and quantifies the events related to cell death, and there are a range of assays that ImmuONE can run to this effect.
An apoptosis assay detects and quantifies the events related to cell death, and there are a range of assays that ImmuONE can run to this effect.
Cell cycle analysis
Assays that analyse the cell cycle generally involve flow cytometry to measure cellular DNA content, and during this process a fluorescent dye that binds to DNA is released.
Assays that analyse the cell cycle generally involve flow cytometry to measure cellular DNA content, and during this process a fluorescent dye that binds to DNA is released.
Autophagy
Autophagy is the body’s way of repairing and recycling old and damaged cell parts. ImmuONE can help carry out autophagy assessments to identify under which conditions autophagy occurs and whether it can be modulated.
Autophagy is the body’s way of repairing and recycling old and damaged cell parts. ImmuONE can help carry out autophagy assessments to identify under which conditions autophagy occurs and whether it can be modulated.
Oxidative stress
Oxidative stress is a phenomenon caused by an imbalance between production and accumulation of oxygen reactive species (ROS) in cells and tissues and the ability of a biological system to detoxify these reactive products. Oxidative stress can be measured directly by quantifying the number of reactive toxic oxygen species found, or indirectly by measuring levels of DNA/RNA damage, lipid peroxidation and protein oxidation/nitration.
Oxidative stress is a phenomenon caused by an imbalance between production and accumulation of oxygen reactive species (ROS) in cells and tissues and the ability of a biological system to detoxify these reactive products. Oxidative stress can be measured directly by quantifying the number of reactive toxic oxygen species found, or indirectly by measuring levels of DNA/RNA damage, lipid peroxidation and protein oxidation/nitration.