2nd International Formaldehyde Science Conference

HUMAN BIOMONITORING OF FORMALDEHYDE:

APPLICATION OF THE CBMN ASSAY

Carina Ladeira

Madrid, 19 April 2012

OVERVIEW
Human Biomonitoring Aims of Human Biomonitoring Biomarkers Cytokinesis-Block Micronucleus assay Human Biomonitoring of Formaldehyde

HUMAN BIOMONITORING
Repeated, controlled measurement of chemical or biochemical markers in fluids, tissues or other accessible samples from subjects exposed or exposed in the past or to be exposed to chemical, physical or biological risk factors in workplace and/or the general environment

Manno et al., Toxicology Letters 192, 2010.

AIMS OF HUMAN BIOMONITORING

Individual or collective exposure assessment
Can reveal subtle differences in individual susceptibility to particular agents

Health protection Occupational health risk assessment

HUMAN BIOLOGICAL MEDIA IN BIOMONITORING

Whole blood/serum Urine Adipose tissue (fat) Hair Breast milk Saliva/ sputum Semen

Most common sample collected Common pathway for most chemicals and their metabolites

BIOMARKERS

Alteration in cellular or biochemical components, processes, structure or functions that is measurable in a biological system or sample
National Academy of Sciences

BIOMARKERS - CLASSIFICATION
Exposure
Exogenous chemical or its metabolites

Effect
Biochemical, physiologic or other alteration in a biologic process

Susceptibility
Indicator of an inherent or acquired limitation to responding to a chemical challenge

National Research Council, 1987

Genotoxicity biomarkers are a subtype of biomarkers of effect used to measure specific occupational and environmental exposures or to predict the risk of disease or to monitor the effectiveness of exposure control procedures in subjects to genotoxic chemicals
M N ly m pho cy te s N PB NB U D

B io mar ker o f b reaka ge an d lo ss of ch ro m oso m es

B io m ark er o f D N A m is-rep air, ch ro m oso m e rea rran ge me nt o r telo me re e nd fusio n s

B io mar ker o f g en e am p lific atio n an d /or a lter ed gen e do sag e

Adapted Fenech et al., 2003

Manno et al., 2010

CYTOKINESIS-BLOCK MICRONUCLEUS ASSAY

MICRONUCLEUS
Represent true mutagenic events Increased use in biomonitoring studies Originated from chromosome fragments or lagging whole cromossomes Are a sensitive indicator of damage to chromosomes or the mitotic spindle MN belong to the most sensitive genetic endpoints for the detection of FA induced genotoxicity
Merk & Speit, 1998

Adapted Fenech et al., 2003

FORMALDEHYDE
Classified by IARC as carcinogenic to humans (Group 1), based on sufficient evidence in humans and experimental animals

Relevant workplace exposure to FA also occurs in resins-production industries; anatomy pathology histopathology; and in mortuaries

HUMAN BIOMONITORING OF FORMALDEHYDE

Formaldehyde-resins production and anatomy pathology laboratories Histopathology laboratories

Photo Ionisation Detector

Method NIOSH 2451 Analysis and time-weighted average (TWA8h) Sampling time was 6 to 7 hours FA levels were measured by Gas Chromatography

Method of Photo Ionisation Detection with video recording Detects FA from 1 ppb to 10,000 ppm and performs automatically data log readings from the sensor on a second basis Allows direct association between worker activities and ceiling values

McGlothin, et al., 2005

Source: Ion Science

Environmental Monitoring of FA exposure

Biomonitoring of FA Micronucleus Assay

Culture of peripheral lymphocytes and CytokinesisBlock Micronucleus assay May-Grnwald Giemsa technique Scoring of 1000 lymphocytes from peripheral blood per individual

TWA8h 0.16 ppm Ceiling 1.14 ppm

CONCLUSIONS
Health effects resulting from FA exposure are related more to peaks of high concentrations than to long time exposure at low levels (IARC, 2006; Pyatt et al., 2008) MN in lymphocytes showed a potential genotoxic damage in subjects with long-exposure to FA (Ye et al., 2005) Chromosome damage and effects upon lymphocytes arise because FA escapes from sites of direct contact, such as the mouth, originating nuclear alterations in the lymphocytes of those exposed (Zhang et al., 2009; Orsire et al., 2006; He et al, 1998) Changes in peripheral lymphocytes indicate that the cytogenetic effects triggered by FA can reach tissues faraway from the site of initial contact (Suruda et al., 1993)

CONCLUSIONS
There is a moderate positive correlation between MN frequency and the duration of FA exposure (years of employment) Years of exposure are also relevant for the development of health effects The CBMN assay is a simple, practical, inexpensive and non-invasive screening technique that can be used for clinical prevention and management of workers under occupational carcinogenic risks, namely exposure to a genotoxic agent such as formaldehyde

The association of these cytogenetic effects with FA exposure gives important information to risk assessment process and may also be used to assess health risks for exposed groups

2nd International Formaldehyde Science Conference

HUMAN BIOMONITORING OF FORMALDEHYDE:

APPLICATION OF THE CBMN ASSAY

Carina Ladeira

Madrid, 19 April 2012