We carry out the toxicological assessment of emissions of our products through established, standardized and validated tests that are in-line with those used for pharmaceutical and food products.
Reduction in harmful or potentially harmful constituents (HPHCs) itself does not necessarily mean reduced toxicity to human beings. A biological approach is essential to estimate the level of toxicity. We evaluate toxicity by in silico (using computer simulations), in vitro (using bacteria or cultured cells) and in vivo (using living organisms) assays as representative methodologies. Generally, we evaluate toxicity by in silico and in vitro methods, and we use in vivo assays only where required by regulators or there is no possible way to gather the necessary information using in silico/in vitro models or previously published data.
In-vitro toxicological assessment
In-vitro toxicological assessment is the scientific analysis to evaluate the effects of chemical substances on bacteria or cultured cells. “In -vitro” means “within the glass” when translated from Latin. This indicates that the various conditions are artificially controlled in what was historically a glass test tube. In-vitro toxicological assessment is generally used to detect the potential toxicity of chemicals and to confirm the absence of certain toxic properties in the early stages of development of products such as pharmaceuticals, cosmetics, agricultural substances and food additives.
We carry out in-vitro toxicological testing to help understand the risks associated with the use of our products, thereby guaranteeing that our products undergo adequate scientific assessments. In recent years we have also investigated the possibility to obtain regulatory certifications of potential risk reduction and exposure reduction claims in countries and regions where legislation requires such certifications – striving to address our consumers’ need for more information.
Potential hazards associated with health outcomes such as cancer may be investigated through standardized and validated tests that are in-line with the safety assessments used for pharmaceutical and food products. These tests investigate functional change or damage in bacteria and cultured cells. We primarily conduct Ames, NRU, and MN tests (in vitro core battery tests)
In-vivo toxicological assessment
In-vivo toxicological assessment is the scientific analysis to evaluate the effects of chemical substances on entire living organisms, such as laboratory animals. “In-vivo” means “within the living” in Latin, indicating that the various conditions are not artificially controlled in contrast to in-vitro. The effects on humans cannot be fully evaluated by in-vitro assessments. Therefore, the in-vivo assessments will be conducted when it is necessary or mandated by regulation.
In certain situations when animal disease models are needed to assess the risk of disease, in-vivo studies using laboratory animals are conducted for RRP.
An example of in-vivo studies is the microscopic examination of animal tissues (histopathological analysis).
Image of alveolar cells observed under a microscope
Ensuring ethical integrity
In general, we don’t test our products using animals and we use alternative test methods and strategies whenever possible. In cases where it cannot be avoided, e.g. when governments require the use of animal testing for specific investigations related to the assessment of RRP, we contract independent laboratories that are accredited and internationally recognized for their high standards of research and animal welfare
Our research activities are carried out in an ethical manner and comply with all relevant laws, regulations, and industry standards.
We are also using novel methodologies, such as “organ on a chip technologies” and “3D tissue culture models”, to further limit the need for animal testing.
3D culture model of human bronchial tissue and direct aerosol exposure system
We are developing novel in-vitro test methods for aerosols generated from tobacco products. For example, a three-dimensional culture model of human bronchial tissue is adopted to investigate the effects of the aerosols on the respiratory system of tobacco product users. The exposure of this in-vitro tissue was performed using the direct aerosol exposure system which delivers smoking machine generated aerosols directly to the apical surface of the in-vitro tissue. The combination of a three-dimensional culture model and direct aerosol exposure system enables an approximation reproduction of the situation in the respiratory epithelium of the product users. This test method is anticipated to be used more frequently as an alternative to animal inhalation testing in the future.
Direct aerosol exposure system
Image of 3D model of human bronchial tissues
Regulatory Toxicology and Pharmacology, 2018
Regulatory Toxicology and Pharmacology, 2018
Approach using state-of-the-art technologies
To ensure product safety, we are also proactively working on the development of new evaluation methods by introducing most recent research methods..
Organ-on-a-chip technology
Interest in the organ-on-a-chip technology, a new system culturing human cells on a microfluidic device, has grown in recent years as a cutting-edge tool for pre-clinical studies. Organ-on-a-chip enables us to mimic the local physiological environment in humans; therefore, it can provide more informative insights for mechanism research and have the potential to reduce the use of animals.
Image of Nuclei and Plasma membrane on the chip
Exposure simulation model
The computer simulation model can estimate the deposition amount of components in vapor/smoke, at each part of the respiratory tract under the actual usage condition of our products. This model enables the biological assays to reflect the actual consumer vaping/smoking situation.
High content analyses
High Content Analysis (HCA) is a cell-based assay that combines microscopy and image analysis. Using various fluorescent probes, HCA can capture morphological changes of individual cells and various changes inside cells as images, and perform quantitative analysis of the changes to be analyzed. Using this technology, we have been elucidating the multiple key molecular responses in DNA damage, oxidative stress, or protein damage, where the consumption of a tobacco product could lead to cellular impacts such as genotoxicity or cytotoxicity. Through this research, we are aiming to construct an evaluation system that considers whether the effects confirmed at the cellular level are possible mechanisms in the human body.
This is a microscopic image of cells and specific particles stained with a fluorescent dye. Substances that are not found in the cells in the left photo are present in the cells in the right photo. Using the microscopic images, HCA can provide a quantitative analysis of the cellular responses caused by exposure to tobacco products.
Omics analyses
"Omics analysis" means a method of comprehensively investigating various molecules, such as DNA, RNA, proteins, and metabolites, which compose the living body, based on genomic information by combining analytical technology with computational science.
Omics analysis makes it possible to analyze the biological effects of tobacco products at the molecular level, allowing a deeper understanding of such effects. In addition, it would make it possible to understand how the biological effects of RRP differ from cigarettes in detail at the molecular level.
Computer analysis and an example of analysis results