Journal of Hazardous Materials 416 (2021) 126124

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Journal of Hazardous Materials

journal homepage: www.elsevier.com/locate/jhazmat

Research Paper

Presence of airborne microplastics in human lung tissue

Luís Fernando Amato-Lourenço a, b, *, Regiani Carvalho-Oliveira a, Gabriel Ribeiro Júnior a,
Luciana dos Santos Galvão c, Rômulo Augusto Ando d, Thais Mauad a, b
a
Department of Pathology, Faculty of Medicine, University of São Paulo, Avenida Dr. Arnaldo, 455, Room 1150, Cerqueira Cesar, 01246903 São Paulo, São Paulo,
Brazil
b
Institute of Advanced Studies (IEA) Global Cities Program, University of São Paulo, Rua da Praça do Relógio, 109, Térreo, Cidade Universitária, 05508-050 São Paulo,
São Paulo, Brazil
c
Chemical Analyses Laboratory, Institute for Technological Research (IPT), Avenida Prof. Almeida Prado, 532 - Butantã, 05508-901 São Paulo, São Paulo, Brazil
d
Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo (IQUSP), Avenida Prof. Lineu Prestes, 748 - Butantã, 05508-000 São Paulo, São
Paulo, Brazil

A R T I C L E I N F O A B S T R A C T

Editor: Dr. R Teresa Plastics are ubiquitously used by societies, but most of the plastic waste is deposited in landfills and in the natural
Airborne microplastics environment. Their degradation into submillimetre fragments, called microplastics, is a growing concern due to
Human lung tissue potential adverse effects on the environment and human health. Microplastics are present in the air and may be
Air pollution
inhaled by humans, but whether they have deleterious effects on the respiratory system remain unknown. In this
Raman: Microspectroscopy
study, we determined the presence of microplastics in human lung tissues obtained at autopsies. Polymeric
Public health
particles (n = 33) and fibres (n = 4) were observed in 13 of 20 tissue samples. All polymeric particles were
smaller than 5.5 µm in size, and fibres ranged from 8.12 to 16.8 µm. The most frequently determined polymers
were polyethylene and polypropylene. Deleterious health outcomes may be related to the heterogeneous char­
acteristics of these contaminants in the respiratory system following inhalation.

1. Introduction their characteristics, including size, format, colour, and polymeric

The high production and consumption of plastics worldwide (Plas­
ticsEurope, 2020), their inadequate disposal and their slow degradation
(Barnes et al., 2009) results in the ubiquitous and long-lasting presence
of microplastics (MPs) in the environment (Kubowicz and Booth, 2017).
MPs are vectors of pollutants (Hartmann et al., 2017) and pathological
microorganisms present in the environment (Foulon et al., 2016) as well
as their own chemical additives, which are added during manufacturing
(Hahladakis et al., 2018). In aquatic habitats, MPs have been shown to
be toxic to several forms of life (Sussarellu et al., 2016; Jeong et al.,
2017; Rochman et al., 2013). To date, there are no in vivo data on po­
tential adverse health effects in humans.
Airborne microplastics have been found in cities, as shown in studies
performed in Shanghai, Paris, and London (Liu et al., 2019; Dris et al.,
2015; Wright et al., 2020). Due to the wide spectrum of MP sizes, it is
highly likely that MPs are inhaled by humans; the gastrointestinal tract
is another possible site of MP entry in organisms.
In this study, we determined the presence of microplastics in human
lung tissue of twenty cases obtained at coroner autopsies and identified
matrix.
2. Experimental
2.1. Participant recruitment and ethics
This study was approved by the Ethical Board of the São Paulo
University Medical School, approval #08304519.4.0000.0065. Written
informed consent was provided by the next of kin. The study was con­
ducted from August 2019 to March 2021.
2.2. Study population
We collected pulmonary tissue samples (parenchymal tissue from the
distal and proximal regions of the left lung) from 20 non-smoking adult
individuals who underwent routine coroner autopsy at São Paulo City
Death Verification Service of São Paulo University for the verification of
cause of death. All individuals lived in São Paulo for more than 10 years.
Via a questionnaire with the next of kin, we obtained information about

* Correspondence to: Faculty of Medicine, University of São Paulo, Dr. Arnaldo Avenue, 455, Room 1150, Cerqueira Cesar, São Paulo, 01246903, Brazil.
E-mail address: luisfamato@usp.br (L.F. Amato-Lourenço).

https://doi.org/10.1016/j.jhazmat.2021.126124
Received 9 April 2021; Received in revised form 11 May 2021; Accepted 12 May 2021
Available online 24 May 2021
0304-3894/© 2021 Elsevier B.V. All rights reserved.
L.F. Amato-Lourenço et al.
previous occupations and underlying diseases. We reviewed autopsy
reports and lung histological findings. Lung tissue collection was per­
formed between August and October 2019.
2.3. Quality control and evaluation of sample processing
A plastic-free approach was applied to guarantee the quality of the
results, allowing us to evaluate the sources of variability and error and
ensure the reliability of the collected data. All procedures, from lung
tissue collection to Raman spectroscopy analysis, followed the protocols
recommended by Rochman et al. (2019). In brief, all materials were
washed thoroughly using Milli-Q® water three times. All glass materials
were cleaned with a 1:1 nitric acid solution. All materials and samples
were covered with aluminium foil (before and after processing) to avoid
airborne sample contamination. Cotton laboratory coats (100%) and
clean nitrile gloves were used during all procedures. The samples were
processed in a clean laminar flow cabinet. A procedural blank was used
for every ten samples during the whole procedure. Additionally, lung
tissue samples from 2 stillborns (weighting 5.4 g and 3.9 g) were pro­
cessed and analysed along with the other samples to demonstrate
whether airborne microplastic contamination had occurred during au­
topsy.t.
2.3.1. Sample digestion and filtering
The samples were individually frozen (− 20 ◦ C) in glass vials covered
with aluminum foil and glass lid until the digestion procedure. Then, the
tissues were digested for 12 h at 60 ◦ C using the enzymatic mixture
Corolase® 7089 (20 UHb/mL for lung samples) (Catarino et al., 2017).
For the filtration procedure, it was necessary to use a density sepa­
ration procedure (1.5 g cm− 3) using ZnCl2 P.A. Imhof et al., (2012) to
minimize clogging of the silver filters. After the addition of ZnCl2, the
solution was stirred for 15 min and held for 45 min. Then, the super­
natant was filtered using silver membrane filters (25 mm in diameter
and 0.45 µm pore size - Millipore®). Silver membrane filters were
chosen due to their efficiency in the identification of polymeric materials
using Raman spectrometry (Wright et al., 2019). The membrane filters
were oven-dried (covered) at 60 ◦ C for 24 h.
2.4. Raman spectroscopy
Filter membranes were first manually inspected under light micro­
scopy using a ×10 objective and morphologically characterized under a
×100 objective. Raman spectra were obtained with a Witec Alpha 300
R® coupled to an optical microscope with an excitation value of λ0 =
633 nm obtained by using a solid-state laser. The use of a Zeiss lens
(x100) with a numerical aperture (NA = 0.90) and 1 mm of focal dis­
tance, with the λ0 = 633 nm excitation wavelength guarantees a spatial
resolution of 0.5 µm.
The obtained spectra were processed using KnowItAll 2020 software
(John Wiley and Sons, Inc.® New Jersey, USA). The results were
compared using two spectral libraries developed for research on
microplastics (SLoPP and SLoPP-E Raman Spectral Libraries) (Munno
et al., 2020).
2.5. Microphotograph analysis
The particle size measurements were calculated from the micro­
photographs obtained during Raman spectroscopy using Image-Pro Plus
4.5.0.29 software (Media Cybernetics, Silver Spring, USA).
2.6. Statistical analysis
Descriptive analyses were performed using SPSS 17.0 software (SPSS
Inc., Chicago, IL, USA).
2
Journal of Hazardous Materials 416 (2021) 126124
3. Results
The median age of the 20 decedents was 78.5 years, ranging from 48
to 94 years; there were 7 males and 13 females. Demographic infor­
mation is presented in Table 1.
All samples were collected from non-smokers, 35% of whom were ex-
smokers but had not smoked in more than 15 years. One individual had a
history of chronic lung disease of unknown duration.
The selected decedents had a mean municipality residence time of
52.8 (±3.98) years and a mean residence address time of 34.15 (±4.70)
years.
Lung tissue was collected in proximal and distal regions of the left
superior lobe. The mean weight of the collected lung tissue was 3.28 g
(2.04–4.45 g). Individual data are shown in Table 1.
Lung histology results were reviewed, and the main findings are
shown in Table 1.
In total, 31 synthetic polymer particles and fibres were observed in
thirteen of the twenty autopsied decedents, of which 87.5% were par­
ticles (all fragments) and 12.5% were fibres (length to width ratio > 3).
The mean particle size was 3.92 (±0.67) µm, ranging from 1.60 to 5.56
µm, while the mean fibre length was 11.23 (±1.96) µm, ranging from
8.12 to 16.80 µm. We also identified 5 natural polymer particles with
mean value of 3.44 (±1.35) µm ranging from 1.98 to 5.42 µm.
No particles/fibres were observed in the two stillborn lung samples
that have been used to demonstrate whether airborne microplastic
contamination had occurred during autopsy and analytical procedures.
Only one polymeric particle (polypropylene) was detected in the pro­
cedural blanks. This particle had a size of 50.25 µm and was grey in
colour.
Polypropylene was the most frequent polymer (35.1%), followed by
polyethylene (24.3%); cotton (16.2%); polyvinyl chloride and cellulose
acetate (5.4%); and polyamide, polyethylene co-polypropylene, poly­
styrene, polystyrene-co-polyvinyl chloride, and polyurethane (2.7%).
When compared to the reference spectral library of plastic particles, the
detected MP particles/fibres had signs of weathering. The Raman
spectra of weathered microplastics are distinctly different from the
spectra of fresh standard samples; many peaks in the Raman spectra of
weathered microplastics are weakened and even invisible (Dong et al.,
2020). For this reason, some databases containing the Raman spectra of
weathered MPs are being elaborated (Munno et al., 2020; Dong et al.,
2020). In this study we have used the database of Munno et al. (2020).
Microphotographs and the Raman spectra of the main types of
microplastics found in the lungs are presented in Fig. 1. Table 2 reports
the morphology, colour, and chemical characterization of the particles/
fibres. Figure data on all the analysed decedents are presented in the
Supplementary Information.
4. Discussion
This is the first study to identify and characterize the presence of
microplastics in human lung tissue using Raman spectroscopy. We found
polymeric particles with diameters less than 5 µm and, at a much lower
frequency, fibres (4 units) in more than 50% of the analysed lungs with
signs of weathering. Our data confirm that the respiratory system is an
exposure route, and the lungs are a site of accumulation of MPs in
humans.
The characterization of MP particles indicated that polypropylene
and polyethylene were the most common particles found in lung tissue.
Both polymers are the most frequently used types of plastics in
manufacturing (PlasticsEurope, 2020); they are present in food pack­
aging, automotive parts, sweets and snack packaging, and bank notes,
among others. Further, cotton particles were found in 16.2% of the
samples. Natural fibres are often dyed and coated in chemicals such as
flame retardants, which may reduce their biodegradability, when they
are used in clothing production (Chen and Jakes, 2001). It is possible
that the type of polymers found in the lungs are associated to indoor
L.F. Amato-Lourenço et al. Journal of Hazardous Materials 416 (2021) 126124

Table 1
Demographic information of the 20 decedents. *Histological analysis not performed.
Case Demographic data Underlying diseases Cause of death Histological findings Pulmonary Died at
tissue weight hospital?
Age at Gender Occupation
(g) (yes/no)
death

1 55 M Bricklayer Diabetes, systemic arterial Pulmonary oedema * 3.04 No

assistant hypertension, hypertensive
cardiomyopathy, alcoholism
2 89 M Salesman Systemic arterial hypertension, Pulmonary Pulmonary 2.06 No
malignant prostate cancer thromboembolism thromboembolism, senile
emphysema, pneumonia
3 79 M Construction Stroke, prostate cancer Massive pulmonary * 3.56 Yes
supervisor thromboembolism
4 74 M Accountant Diabetes, systemic arterial Aspiration pneumonia Bacterial pneumonia 4.23 No
hypertension, stroke
5 86 F Housewife Ruptured brain aneurysm Cerebral haemorrhage Senile emphysema 2.10 Yes
6 59 M Serigrapher Old myocardial infarction Acute myocardial Pulmonary oedema 2.64 No
infarction
7 79 M Bricklayer Systemic arterial hypertension, Pulmonary Pulmonary 3.50 Yes
assistant stroke thromboembolism thromboembolism,
bronchopneumonia
8 78 F Housewife Systemic arterial hypertension Bronchopneumonia Bronchopneumonia 2.23 No
9 75 F Quality Heart disease, chronic lung Hydropericardium Emphysema, chronic 2.83 No
inspector disease, cardiac failure bronchitis
10 79 F Housekeeper Systemic arterial hypertension, Pulmonary oedema Pulmonary oedema 3.89 No
atherosclerotic hypertensive
heart disease, stroke
11 48 M Security guard Diabetes, systemic arterial Pulmonary oedema Chronic passive congestion 5.42 No
hypertension, heart disease, of the lung
12 94 F Housewife Dementia Bilateral * 2.57 Yes
bronchopneumonia
13 74 F Librarian Systemic arterial hypertension, Hypovolemic shock Normal histology 3.86 No
ruptured abdominal aortic
aneurysm
14 90 F Teacher Systemic arterial hypertension, Pulmonary Pulmonary oedema 4.42 Yes
heart disease thromboembolism
15 67 F Farmer Systemic arterial hypertension, Abdominal Pulmonary oedema 3.50 Yes
heart disease, stroke, haemorrhage
alcoholism,
16 60 F Housekeeper Diabetes, systemic arterial Pulmonary oedema Pulmonary oedema 2.04 No
hypertension, acute myeloid
leukaemia, stroke
17 79 F Masseuse Diabetes, systemic arterial Subarachnoid Normal histology 2.94 No
hypertension, cerebral haemorrhage
aneurysm
18 88 F Housekeeper Diabetes, stroke, systemic Cerebral ischemic Bronchopneumonia 3.53 No
arterial hypertension, infarction
generalized atherosclerosis
19 89 F Cooker Systemic arterial hypertension, Pulmonary Pulmonary 4.45 No
stroke thromboembolism thromboembolism
20 78 F Housewife Systemic arterial hypertension, Acute peritonitis Pulmonary oedema 2.71 No
stroke, focal necrosis of the
small intestine

exposures, since most of the people in cities stay more hours indoor than
outdoor (Al Horr et al., 2016). Further, the number of fibres is usually
higher in indoor than outdoor environments (Dris et al.,). Accordingly,
(Dris et al., 2017) found in indoor samples in Paris predominance of
polypropylene and no polyester derived MPs, which is similar to our
findings.
Although fibres are the most frequently found MP type in air samples
(Dris et al., 2015; Wright et al., 2020), our study identified mainly
polymeric particles in lung tissue. Wright et al. (2020) described, in
addition to MP fibres, the presence of many smaller particles in London
air samples that were not further characterized due to the lower
analytical threshold of the Fourier transform infrared (FTIR) instrument
employed in that study (~20 µm). Deposition of exogenous particulates
in the lungs depends on size, aerodynamic diameter, charge, density,
and flow rate (Carvalho et al., 2011). The particles found in this study,
which ranged from 1.60 to 5.58 µm in size, indeed have the capability to
reach the bronchial-alveolar regions by mechanisms of inertial impac­
tion and sedimentation. For fibres, this capability is regulated predom­
inantly by the actual diameter, length and density (Donaldson et al.,
1993). Only fibres with a physical diameter smaller than 3 µm, even if
they are very long, can reach the alveolar region (Donaldson et al.,
1993), which may explain the lower number of fibres found in this
study. The characteristics of the fibres found in our samples in relation to
width (<3 µm), length and biopersistence/resistance suggest pathogenic
potential (Warheit et al., 2001). Our results are in contrast with data
presented by Pauly et al. (1998). Studying lung cancer samples, the
authors detected higher amounts of fibres identified as plastics by light
microscopy than we did. Currently, the identification of polymeric fibres
by only visual methods is prone to criticism. Further, those authors did
not use a totally plastic-free protocol. We cannot exclude, however, that
there is indeed a higher number of plastic fibres in patients with lung
cancer than in individuals with non-neoplastic disease.
Ragusa et al. (2021) identified MP particles in human placental tis­
sue, with sizes ranging from ~5–10 µm. The authors suggested that MP
particles reached the placenta via inhalation or gastrointestinal trans­
location. Pulmonary translocation to systemic circulation has been
demonstrated for fine and ultrafine particles (Elder and Oberdörster,
2006; Peters et al., 2006). Intranasal instillation of 1.1-µm polystyrene

3
L.F. Amato-Lourenço et al.
Fig. 1. Microphotographs and Raman spectra of the main types of microplastics found in lung tissues. Bar scale – 10 µm.
microspheres in mice resulted in time-dependent translocation to the
spleen (Eyles et al., 2001). It is therefore likely, due to the mechanisms
of particle deposition in the lungs, that the larger particles found in the
placenta (~10 µm) originated from gastrointestinal translocation.
What are the biological impacts of MP particles in the lungs? From a
toxicological point of view, all particles smaller than 10 µm in diameter
have the potential for biological activity in susceptible individuals
(Heyder, 2004). Occupational exposure to nylon flock induced inter­
stitial lung disease in exposed workers (Kern et al., 1998); however, the
exposure condition was quite different from those associated with
chronic exposure to weathered microplastics. MP particles could induce
local pro-inflammatory effects driven by macrophages or epithelial cells,
be transported to the pleura or to the systemic circulation system. Such
situations will depend on particle/fibre size, burden, density, charge,
and type in addition to individual factor; these situations are especially
affected by environmental changes in microplastics, physicochemical
transformations and the coatings (eco-coronas) (Ramsperger et al.,
2020) plastics acquire during weathering. Further, the presence of fibres
larger than 8–10 µm of a biopersistent/bioresistant material such as
plastic (Pauly et al., 1998) can induce macrophagic activation and the
release of many biologically active inflammatory mediators (Novais
Barbosa and Pereira Vasconcelos, 2020). (Goodman et al., 2021)
cultured human lung epithelial cells with polystyrene spheres, which
caused the inhibition of cell proliferation and major changes in cell
morphology. To date, there is no major in vitro or in vivo information
about the toxicity of environmentally weathered MPs particle or fibres
in human lungs, which is important because environmental exposure
seems to promote the cellular internalization of microplastics (Ram­
sperger et al., 2020).
It has been estimated that an average male performing light activity
would potentially inhale up to 272 MP particles/day (Vianello et al.,
2019). We found approximately 0.56 MP particles/g of lung tissue.
Considering that the mean weight of a set of normal adult lungs is
4
Journal of Hazardous Materials 416 (2021) 126124
approximately 840 g (Molina and DiMaio, 2012), there were 470 par­
ticles in both lungs. This number suggests that a large proportion of
inhaled MPs is retained in the upper airway.
We could not identify any link between occupation and number of
MPs in the lungs. In fact, the patient that had more MPs in the lungs had
diagnosed dementia and probably no occupational activities recently.
Thirteen patients died outside of the hospital, but we could not identify
any differences in number of MPs between hospitalized and non-
hospitalized patients. It must be stressed however, that the number of
studied cases was too small to identify trustworthy associations or
differences.
This study has certain limitations. Although inhalation was the most
likely exposure route, we cannot exclude that some MP particles may
have reached the lungs via systemic translocation since MP particles
have previously been detected in placental tissue. We adopted a plastic-
free protocol throughout the experiments. Because of the presence of MP
fibres and particles in the air, we used processual blanks to detect air
contamination. We found a single 50.25 µm polypropylene particle in
one of the blanks, but no particles of this size were found in the lungs.
This confirmed the validity of our results since an inhaled 50.25-µm
particle would not infiltrate the lung parenchyma; it would be retained
in the upper airway, such as the nose. Further, not finding MPs in the
stillborn lung tissue samples confirmed the soundness of our plastic-free
protocol. Due to the maximum spatial resolution (0.5 µm) of Raman
spectroscopy setup, we were not able to detect MP nanoparticles. These
particles could also be present in the lungs and have a high possibility of
causing biological damage by internalization and translocation (Thorley
et al., 2014).
In summary, we characterized the environmental MPs present in
human lungs for the first time. Weathered particles derived from the
most widely consumed plastics, such as polypropylene and poly­
ethylene, predominated and were found more frequently than fibres.
Extensive research will be needed to elucidate the potential adverse
L.F. Amato-Lourenço et al. Journal of Hazardous Materials 416 (2021) 126124

Table 2 Appendix A. Supporting information

Morphology and chemical characterization of the identified particles/fibres.
Decedent Particle/ Size Polymer matrix Colour Supplementary data associated with this article can be found in the
fibre (µm) online version at doi:10.1016/j.jhazmat.2021.126124.
3 Fibre 16.80 Polypropylene White/grey
3 Particle 2.82 Cotton Brown References
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4 Particle 4.20 Polyethylene Transparent Impact of indoor environmental quality on occupant well-being and comfort: a
4 Particle 2.84 Cotton White review of the literature. Int. J. Sustain. Built Environ. 5 (1), 1–11. https://doi.org/
6 Fibre 8.93 Cellulose acetate Grey 10.1016/j.ijsbe.2016.03.006.
6 Particle 4.30 Polyvinyl chloride Black Barnes, D.K.A., Galgani, F., Thompson, R.C., Barlaz, M., 2009. Accumulation and
8 Fibre 11.08 Polypropylene Blue fragmentation of plastic debris in global environments. Philos. Trans. R. Soc. Lond. B
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This study was financially supported by the São Paulo Research 675, 462–471. https://doi.org/10.1016/j.scitotenv.2019.04.110 (Jul 20, Epub 2019
Foundation (FAPESP- project number: 19/03397-5). We would like to Apr 9).
thank the São Paulo City Death Verification Service (SVOC) staff and the
Verbal Autopsy Team for the technical support.

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