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Microplastics in the bloodstream can induce cerebral reserved; exclusive
licensee American
thrombosis by causing cell obstruction and lead to Association for the
Advancement of
neurobehavioral abnormalities Science. No claim to
original U.S.
Government Works.
Haipeng Huang1,2,3, Jiaqi Hou1*, Mingxiao Li1, Fangchao Wei4, Yilie Liao5, Beidou Xi1* Distributed under a
Creative Commons
Human health is being threatened by environmental microplastic (MP) pollution. MPs were detected in the blood- Attribution
stream and multiple tissues of humans, disrupting the regular physiological processes of organs. Nanoscale plas- NonCommercial
tics can breach the blood-­brain barrier, leading to neurotoxic effects. How MPs cause brain functional irregularities License 4.0 (CC BY-­NC).
remains unclear. This work uses high-­depth imaging techniques to investigate the MPs within the brain in vivo. We
show that circulating MPs are phagocytosed and lead these cells to obstruction in the capillaries of the brain cor-
tex. These blockages as thrombus formation cause reduced blood flow and neurological abnormalities in mice.
Our data reveal a mechanism by which MPs disrupt tissue function indirectly through regulation of cell obstruc-
tion and interference with local blood circulation, rather than direct tissue penetration. This revelation offers a lens
through which to comprehend the toxicological implications of MPs that invade the bloodstream.

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INTRODUCTION stroke, or death from any cause in 34 months (18). These potential
Microplastics (MPs) are plastic particles with a diameter of less threats can be life-­threatening compared to the chronic diseases
than 5 mm (1, 2). These particles originate from small plastic pel- caused by MPs. In addition, MPs can cause brain dysfunction. Nano-
lets produced for specific purposes, as well as from the degrada- sized plastic particles can penetrate the blood-­brain barrier (BBB)
tion, weathering, and fragmentation of larger plastic products in and enter brain tissue (19, 20). The interaction between NPs and
the environment (3–5). MPs are ubiquitous worldwide, present in a-­synuclein fibrils can exacerbate the spread of a-­synuclein pathology
various environments ranging from oceans to land and from Ant- in vulnerable brain regions, potentially triggering or worsening con-
arctic ice to human settlements (4). Pollution from MPs is particu- ditions such as Parkinson’s disease and other neurologically related
larly notable in the oceans, where marine organisms such as fish, dementia diseases (21). However, how micron-­sized plastics affect
shellfish, and plankton ingest them, thereby introducing them into brain function remains unclear. Even several studies have proved that
the human food chain (4–6). Substantial amounts of MPs have also treatment with MPs affects behavior and induces phenotypes such as
been found in freshwater systems, including rivers, lakes, and res- anxiety in mice (22, 23). It is supposed that micron-­sized plastics
ervoirs, allowing for contamination of human water sources. In ad- break down into nanosized plastic particles in the body and then en-
dition, MP particles can be transported through the atmosphere ter the brain to exert their effects. It has also been suggested that the
and disseminated into the air, eventually entering the human re- impact of MPs on the brain may be mediated through their effects on
spiratory system (7). Recent studies have indicated that MPs can peripheral tissues, including the modulation of immune inflamma-
directly enter the human bloodstream through the use of plastic tion and glycolipid metabolism (24–27). What is the mode of MPs
medical supplies (8). affecting the brain and the mechanisms that they exert their effects
MPs have been found in human feces and various tissues, includ- remain uncertain.
ing the liver, kidney, placenta, and blood (9–11). Accumulation of The development and innovative application of previously unknown
MPs in organisms can result in tissue dysfunctions and chronic dis- research techniques often open new avenues, providing researchers
eases such as respiratory diseases, immune system disorders, chronic with fresh perspectives and facilitating a deeper understanding of scien-
inflammation, endocrine gland effects leading to hormone imbalance, tific principles. In this study, we applied miniature two-­photon micros-
and metabolic dysfunctions (12–15). In particular, the presence of copy (mTPM) and imaged MPs in the mouse brain in vivo while the
MPs in the bloodstream poses a substantial health challenge. As the animal was awake. With the high-­depth imaging capability, we observed
blood circulates, these MPs may be carried to any organ, especially MPs in the blood vessels of the mouse cerebral cortex. We tracked the
the distal branch vessels. Studies have shown that blood MPs can lead high-­speed movement of MP particles in the blood vessels, revealing a
to acute cardiovascular diseases (16–17). Furthermore, in a study, pa- mechanism by which MPs can induce brain dysfunction and neuro-
tients with carotid artery plaque in which MPs and nanoplastics (NPs) logical impairment.
were detected had a higher risk of a composite of myocardial infarction,

1
RESULTS
State Key Laboratory of Environment Criteria and Risk Assessment, Chinese Re-
search Academy of Environmental Sciences, Beijing, China. 2Institute of Molecular
Blood vessels exhibit strong contrast and weak fluorescence
Medicine, College of Future Technology, Peking University, Beijing, China. 3PKU-­ signals in the imaging of the brain cortex in living mice
Nanjing Institute of Translational Medicine, Nanjing Raygen Health, Nanjing, China.
4
To examine the effects of MP particles in the brain, we chose to first
Department of Pharmacology and Cancer Biology, Duke University, Durham, NC, image brain blood vessels. We hypothesized that, even if MPs are first
USA. 5National University of Singapore, Lower Kent Ridge Road, Singapore, Singapore.
*Corresponding author. Email: xibd@​craes.​org.​cn (B.X.); houjiaqi0325@​163.​com broken down into NPs to pass through the BBB and affect neuron
(J.H.) function, it is needed for NPs to arrive at the local brain vessel and

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try to cross the BBB; thus, we can catch the process by the vessel im- vessels (BV) and blood flow were visible within FOV2 and FOV3
aging. Moreover, an imaging strategy incorporating fluorescent MP (Fig. 1B; refer to movie S1). Selecting an ideal imaging area, fluores-
microspheres can help us to test this hypothesis. cence imaging of deep brain tissue was conducted using two-­photon
To conduct in vivo cerebral vascular imaging in mice, we used an excitation within FOV4 (Fig. 1B).
appropriate imaging system (fig. S1, A and B) (28). Surgical inter- There was no fluorescent signal in the blood, and the vessels
vention was necessary to attach the mouse head assembly onto the and surrounding tissues in the imaging FOV showed high signal
microscope lens (Fig. 1A). Following the surgical procedure, the contrast. Converting the image to three-­dimensional (3D) for sig-
vascular network of the cerebral cortical tissue could be observed nal presentation, we could visualize the vessel channel more clear-
through the imaging window in the head, as depicted in the field of ly (Fig. 1C). The regions of interest (ROIs) were outlined in the
view (FOV) (Fig. 1B) under bright-­field imaging. The expression of image, and the fluorescence intensity was calculated. The fluores-
fluorescent probes in neurons via adeno-­associated virus served as a cence values for the linear ROIs a# and d# exhibited a significant
background signal for vascular imaging, enhancing the contrast decrease in fluorescence intensity at the blood vessels (Fig. 1D). In
of blood vessels. After single-­photon fluorescence excitation, an contrast, the long-­term fluorescence values for ROIs 1# and 4# in-
optimal imaging position was identified within the FOV. Both blood dicated stable fluctuations around a certain value, suggesting that

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Fig. 1. Blood vessels exhibit strong contrast and weak fluorescence signals in brain cortex imaging by using two-­photon microscopy in living mice. (A) Sche-
matic imaging representation by miniaturized two-­photon microscopy to cortical blood vessels (BV). Created using FigDraw.com. (B) Fields of view (FOVs) of the blood
vessels: FOV1#, bright field (scale bar, 1 cm); FOV2#, 488-­nm single-­photon excitation (scale bar, 250 mm); FOV3#, 488-­nm single-­photon excitation (scale bar, 100 mm);
and FOV4#, 920-­nm two-­photon excitation (scale bar, 100 μm, a red arrow indicates the location of blood vessel). (C) Three-­dimensional (3D) mapping of fluorescence
signals within the FOV 4#. (D) Fluorescence signal detections for regions of interest (ROIs) a# and d#. (E) The fluorescence signal is recorded over a long period for fluores-
cence signal detections for ROIs 1# and 4#.

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blood flow did not induce significant or irreversible changes in the that fluorescent MPs that enter the bloodstream are phagocytosed
signal (Fig. 1E and fig. S1, C to E). Together, these experiments sug- by cells, which are labeled with a fluorescent signal. To test this hy-
gest that the imaging system is suitable for targeting fluorescence sig-
pothesis, we directly injected MPs into the bloodstream of mice via
nal detection in blood vessels of the brain in vivo. intravenous injection (Fig. 3C). The MP-­Flash signal was observed
in the cerebral cortical blood vessels within minutes following intra-
MPs were detected in the imaging of cerebral blood venous injection, as compared to gavage administration (Fig. 3D).
vessels in vivo There was no significant difference in the trajectory length of the
Using this imaging system, we attempted to visualize fluorescent MP-­Flash signals observed (Fig. 3E). Unexpectedly, the fluorescence-­
MP particles that may be present in the bloodstream. The typical labeled cells could be detected ~10 min after intravenous injection
blood flow in mice ranges from 2.5 to 18 cm/s, slowing down sig- (Fig. 3F). We compared the timing of MP-­Flash appearance and the
nificantly in capillaries to a few hundred micrometers per second fluorescence-­labeled cells in each experiment, and we found that
(29). Because of the limitations of the imaging FOV, to detect MP the fluorescence cell typically appeared within 6 min of observing
particles moving with the blood, imaging frequencies below 1 Hz the MP-­Flash (Fig. 3F). Moreover, no similar fluorescence cells were
are required. After selecting a clear FOV for vascular imaging, mice observed in the blood under non-­treated conditions. Collectively,
were gavaged with water containing fluorescently labeled MP parti- fluorescent MPs led to the emergence of fluorescence-­labeled cells
cles (Fig. 2, A and B). To increase the likelihood of capturing MPs in the blood, suggesting the MPs that enter the bloodstream are
images, we aimed to maximize the effective imaging time. There- phagocytosed by cells.
fore, imaging was initiated 1 hour after MP treatment (Fig. 2B). Next, we tracked and observed these cells. We named them MP-­
We first observed MPs moving within the bloodstream 2 hours labeled cells (MPL-­Cells). Using ImageJ, we calculated the diame-
and 20 min after treating mice (Fig. 2C; refer to movie S2). The ters of these cells, which were around 21 μm (Fig. 3G and fig. S3B).

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movement of the MP resembled a lightning bolt in a dark vascular We further discovered that the movement of MPL-­Cells in blood
region. The trajectory of the MP appeared shuttle shaped (Fig. 2C). vessels was not simply synchronized with the blood flow. In cer-
In particular, one MP track observed at 3 hours and 4 min resem- tain tests, we noticed that MPL-­Cells would become trapped in
bled a comet with a trailing tail. The imaging time for one frame is specific locations within the blood vessel and remain there for an
208 ms, and the entire MP trajectory was captured within this frame extended period. The movement trajectory of an MPL-­C ell in the
time (Fig. 2D). At the site of MP emergence, there is a significant cortical vasculature is depicted in Fig. 3H (fig. S3C). It took near-
change in fluorescence value, as shown in Fig. 2D. To simplify and ly 10 min for this MPL-­Cell to enter and leave the imaging FOV,
illustrate this phenomenon in the text, we have coined the term however, showing nonuniform motion with only a few seconds
“microplastic flash (MP-­Flash)” to describe the intravascular move- spent in most segments of the trajectory, as indicated in motion
ment of MP particles. In our study with five mice, MP-­Flash was trajectories 1 and 3 (Fig. 3I). Motion trajectory 2 consumed most
observed in four of them (Fig. 2E). All four mice exhibited MP-­ of the time during the entire motion process, despite covering the
flashes 2 hours after water treatment. Among the four mice show- shortest distance. This MPL-­Cell encountered a nearly 90° turn in
ing MP-­Flash, it was detected once in one mouse (Fig. 2E). The motion trajectory 2, which might be the reason leading to its ob-
median time for the appearance of MP-­Flash after drinking was struction. By analyzing similar events, we defined an obstructed
191 min (Fig. 2F). The trajectory lengths of the MP-­Flash were all cell as one that remained trapped at a position in the vessels for
less than 330 μm (Fig. 2G), and the entire trajectory was captured over 2 min. Some MPL-­Cells eventually exited the imaging FOV
in a single imaging frame, lasting less than 208 ms. The median following a period of obstruction due to blood flow (Fig. 3J; refer
length of the MP-­Flash trajectory was 57.5 μm (Fig. 2H and fig. S2A). to movie S4). The trajectories of these cells can be categorized into
There is no linear correlation between the appearance time of these three stages, with some being directly obstructed without progress-
MP-­Flashes and their track lengths (fig. S2B). In addition, the me- ing to stage 1 and later leaving after a period of blockage. Notably,
dian running speed of MP particles during MP-­Flash was calcu- we observed that around 18% of the cells remained blocked in the
lated to be 321 μm/s (Fig. 2I), comparable to the blood flow rate. same position until the end of the imaging session (Fig. 3K and
However, we could not guarantee that the vessels imaged in each fig. S3D). Furthermore, we did not observe MPs attaching to the ves-
mouse were of the same size type, potentially leading to differences sel wall or crossing the vessel wall into the brain tissue.
in MP-­Flash velocity and trajectory length (Fig. 2G). In summary,
through millisecond imaging of deep tissue vessels, we successfully Fluorescent MPL-­Cells were sorted and characterized
detected MP-­Flash in animals and calculated several basic charac- Next, we tried to sort the MPL-­Cells (Fig. 4A). Mouse blood was col-
teristic indices. These results show an effective method for detecting lected from the heart 20 min after injecting the fluorescent MPs. The
MPs moving within the cerebral vasculature of living animals. isolated cells were then analyzed by flow cytometry, and a distinct
population of fluorescein isothiocyanate (FITC)–positive cells was
Circulating MPs are phagocytosed by cells in the blood observed in the injected group compared to that in the untreated
After 3 hours of fluorescent MP treatment, we found that cells la- control group (Fig. 4B and fig. S4, A and B). All five mice that re-
beled with fluorescent signals were observed in the bloodstream ceived injections showed clear FITC-­positive cells (Fig. 4C). Immune
(Fig. 3A and fig. S3A; refer to movie S3). However, in the absence cells are known to have a strong phagocytic ability toward exogenous
of fluorescent MP treatment, these signals were never detected, pathogens and foreign substances (30). Our findings revealed that
even with a longer imaging duration (>3 hours). The fluorescence ~94% of the FITC-­positive cells in the treatment group were positive
intensity of these cells was comparable to that of the MP-­Flash sig- for the cell surface antibody CD45, indicating that they were immune
nal, which was significantly higher than the fluorescence signal cells (Fig. 4D and fig. S4C). To identify the immune cell type, we
from probe-­expressed neuronal cells (Fig. 3B). Thus, we hypothesized further labeled the cells with antibodies F4/80 and Ly6G/C (Fig. 4E

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Fig. 2. MPs were detected in the imaging of cerebral blood vessels in vivo. (A) A suitable FOV for vascular imaging. (B) Schematic representation of the treatment and
imaging design of the mice. Created using FigDraw.com. (C) Captured MP-­Flash images; this FOV is the rectangular FOV framed by the dashed line in (A) (scale bar, 100
μm). (D) The emergence of an MP-­Flash is demonstrated; ROI 1# represents the MP-­Flash, and the peak of wave that exhibits a fluorescent signal is present; ROI 2# repre-
sents the background control, no fluctuation in fluorescence value. (E and F) Comparison of the time of MP-­Flash appearance (n = 10 from mouse 1, n = 8 from mouse 2,
n = 4 from mouse 3, and n = 1 from mouse 4) (E), and the median time of the MP-­Flash occurrence (F). (G and H) Comparison of the length of the MP-­Flash track (G), and
the median length of the MP-­Flash track (H). (I) Calculated MP-­Flash running speed (n = 23 from four mice). Data are represented as means ± SEM.

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Fig. 3. Circulating MPs are phagocytosed by cells in the blood. (A) A representative fluorescent signal labeled cell. (B) Quantitative comparison of fluorescence signals
(n = 10 MP-­Flash, n = 4 fluorescent signal labeled cells, and n = 10 background ROIs) (three independent replicated experiments verified the result). (C) Schematic repre-
sentation of the intravenous injection in mice. Created using FigDraw.com. (D and E) Comparison of MP-­Flash emergence times (n = 10 to 12 from three mice) (D) and
lengths (n = 13 to 20 from three mice) (E) between the two treatments. (F) Comparison of the time of appearance of MP-­Flash and the time of appearance of fluorescence-­
labeled cells after injection (n = 5 experiments from five mice). (G) Diameter statistics for fluorescence-­labeled cells (n = 15 from four mice). (H) A trajectory of a
fluorescence-­labeled cell and the corresponding time. (I) The movement trajectory can be labeled as three motion processes. (J) Statistical analysis of the time taken by
the three processes for equal events (n = 6 events from four mice). (K) Percentage of labeled cells that are consistently obstructed (n = 5 FOVs from five mice). Data are
represented as means ± SEM. ***P < 0.001. n.s., not significant.

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Fig. 4. Fluorescent MPL-­Cells were sorted and characterized. (A) Schematic representation of the MPL-­Cells in cerebral vessels, isolation, and characterization. Created using
FigDraw.com. (B) Representative fluorescence-­activated cell sorting (FACS) analysis plots for fluorescein isothiocyanate (FITC)–positive cells. (C) Quantification of FITC-­positive cells
(n = 5 mice for each group, more than 1 × 105 cells were counted per sample). (D) Percentage of CD45-­positive cells among FITC-­positive cells in MP-­treated mice (n = 4 mice).
(E and F) FACS analysis plots (E) and quantification (F) for the F4/80-­positive and Ly6G/Ly6C-­positive cells (n = 4 mice). (G and H) analysis of the FSC (G) and SSC (H) in the MP-­labeled
neutrophils and control. (I) Illustration of the effects of phagocytosis of plastic microspheres on cell morphology. Data are represented as means ± SEM. **P < 0.01 and ***P < 0.001.

and fig. S4, D and E). The results indicated that the majority of the decrease in side scatter (SSC) (Fig. 4, G and H). These results indicate
immune cells exhibited double positivity for F4/80 and Ly6G/C that the engulfment of MPs affects cell size and surface status, poten-
(~41%) or single positivity for F4/80 (~35%) (Fig. 4F). This suggested tially contributing to MPL-­Cell obstructions (Fig. 4I).
that the cells involved in phagocytosis of MP particles were predomi-
nantly neutrophils (F4/80+ and Ly6G+) and macrophages (F4/80+ Obstructed MPL-­Cells induced long-­time blockages in the
and Ly6G−), supporting our hypothesis as neutrophils and macro- cerebral blood vessels
phages are known to have strong phagocytic capabilities. We com- The imaging data revealed that some MPL-­Cells would exit the im-
pared neutrophils that had phagocytosed MPs with unphagocytosed aging FOV after being obstructed in the cerebral vessels for a period
controls, showing a significant increase in forward scatter (FSC) and a of time. However, there were still MPL-­Cells that remained obstructed

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in the blood vessels even after an extended imaging duration. We signals and exclusively capture the fluorescence signal of the MPL-­
observed three MPL-­Cells still present within the FOV following Cells. Subsequently, we observed that MPL-­Cells exhibited diverse
nearly 2.5 hours of imaging (Fig. 5A). These three cells exhibited dis- morphological features due to their nonalignment in the same hori-
tinct morphological characteristics, displaying both regular oval and zontal imaging plane. As depicted in Fig. 5B, we captured the imag-
ruffled morphology. To obtain clearer observations of the obstructed ing plane at two different depths, I and II. Altering the imaging depth
cells, we used mice without labeled neurons to minimize background resulted in the transformation of cell I-­a from ellipsoidal to ruffled

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Fig. 5. Obstructed MPL-­Cells induced long-­time blockages in the cerebral blood vessels. (A) Representative MPL-­Cells, time points in which cells appear were marked
on the graph, and time 0 represents the imaging start. (B to D) Imaging of MPL-­Cells sectioned at different depths (B); z-­stack imaging of the MPL-­Cells (C); reconstruction
of MPL-­Cells 3D images, two sections, I and II, of cell correspond to the imaged sections in (B). (D). (E) The density of MPL-­Cells within each two-­photon imaging FOV was
calculated and analyzed at different time points after treatment (n = 10 FOVs from four mice per time point). (F) Showing MPL-­Cell distribution at larger FOV by image
stitching, at 3 hours after MP injection. (G) Long-­time imaging arrangements. Created using FigDraw.com. (H) An MPL-­Cell that has been obstructed in a vessel for 7 days.
(I) The density of MPL-­Cells in the brain cortex at the indicated time points (n = 10 FOVs from four mice per time point). Data are represented as means ± SEM. *P < 0.05.

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II-­a, while cell I-­b changed from ruffled to ellipsoidal II-­b. To gen- blood flow and potentially result in neurological dysfunction (32–34).
erate 3D images of the cells, we conducted z-­stack imaging (Fig. 5C We conducted a 1-­week assay on treated mice (Fig. 5G), and, un-
and fig. S5A). expectedly, 7 days after treatment, the cells were still not cleared
We observed that MPL-­Cells exhibited a vertical morphology, (Fig. 5H), although the density of blocked cells had been significantly
appearing as if they were elongated and inserted into blood vessels reduced (Fig. 5I).
(Fig. 5D; refer to movie S5). Different imaging depths revealed vary-
ing cross sections of the longitudinal depth of the cells, as depicted Cell obstruction is influenced by the size of the
in Fig. 5B (I and II). We speculate that MPL-­Cells may be mostly plastic particles
obstructed at the junction of blood vessels extending from the deep Furthermore, the question arose as to whether the mechanism of
brain toward the cortex with laterally distributed blood vessels with- MPL-­Cells blocking blood vessels is specific to micrometer-­scale
in the cortex, where there would be greater vascular curvature (31). plastics. Does the size of the plastic particles affect the obstruction of
Next, the distribution of MPL-­Cell blockage density in the cortex these labeled cells? Subsequently, mice were treated similarly with
was quantitatively analyzed at different time points after injection 2-­and 0.08-­μm diameter fluorescent plastic particles, we detected
(fig. S5B). The number of blocked cells was significantly higher at 1 hour these fluorescent plastic particles within the blood vessels and, sub-
after treatment compared to half an hour (Fig. 5E). The obstruction sequently, observed cells labeled by them (Fig. 6A). The area of
density remained high at 2 hours after treatment, with no significant detectable fluorescent signals is smaller in cells labeled with 2-­μm
difference to 1 hour. By using a post-­imaging image stitching method, plastic particles compared to those labeled with 5 μm. The morphol-
the distribution of MPL-­Cells across a large-­scale range in the cerebral ogy of the signal in the 2-­μm plastic particle-­labeled cells does not
cortex is presented (Fig. 5F). Upon analysis of the numerous cells, it exhibit a fully spherical shape; instead, it appears as a stronger signal
was observed that these cells predominantly obstructed in the vessels at the core, surrounded by a weaker ring of dispersed signals (Fig.

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that are perpendicular to the cortex, as depicted in Fig. 5B. 6A; refer to movie S6). In contrast, the morphology of the signals in
Moreover, how long will the MPL-­Cells, which are obstructed in cells labeled with 0.08-­μm plastic particles is much less distinct, with
blood vessels, last before being cleared? The brain contains a large no obvious core visible and a weaker signal (Fig. 6A; refer to movie
number of capillaries, and the blockage of these cells may affect S7). Two hours after exposure to plastic particles, we measured the

Fig. 6. Cell obstruction is influenced by the size of the plastic particles. (A) Exposure experiments were conducted on mice using plastic particles of varying sizes, and im-
ages of cells labeled with fluorescent plastics of different sizes were captured during the imaging of blood vessels. (B) The density of plastic-­labeled cells (PL-­Cells) within each
FOV was calculated and analyzed. Data collected at 2 hours after treatment (n = 15 FOVs from four mice for the 5-­μm group and n = 15 FOVs from two mice for the 5-­μm and
0.08-­μm groups). (C) PL-­Cell densities at different time points after exposure were comparatively analyzed (n = 15 FOVs from four mice for the 5-­μm group and n = 15 FOVs from
two mice for the 5-­μm and 0.08-­μm groups). (D) A comparative analysis of PL-­Cell density was performed following exposure to different concentrations of 5 μm MPs (n = 15
FOVs from four mice for group at 50 μg/ml and n = 15 FOVs from two mice for groups at 25 and 5 μg/ml). Data are represented as means ± SEM. *P < 0.05 and ***P < 0.001.

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extent of the cell obstruction in the brain. Unexpectedly, the levels of obstruction. However, upon reducing the final concentration to 5 μg/
PL-­Cell obstruction in the cerebral vasculature of mice exposed ml, a marked decline in cell obstruction levels was observed (Fig. 6D).
to 2-­μm plastic particles were significantly lower. Cells labeled with Notably, cell obstructions within the cerebral vasculature can be ob-
2-­μm particles demonstrated an obstruction level that was over 50% served even in the lowest-­concentration (5 μg/ml) group of MPs.
lower than that of cells labeled with 5-­μm particles (Fig. 6B). Fur-
thermore, the obstruction level in cells labeled with 0.08-­μm plastic MPL-­Cell obstructions as thrombus formation inhibit
particles was even lower, as only one obstructed cell was observed intracerebrovascular blood perfusion
across 15 FOVs in two mice (Fig. 6B). Compared to the 5-­μm group, To investigate the impact of MPL-­Cell obstruction in blood vessels
there was a notable reduction in the obstruction level in the 2-­μm on blood flow, we used laser speckle contrast imaging (LSCI) to
group within 24 hours (Fig. 6C). These findings indicated that not monitor blood flow in the mouse brain. We observed changes in
only did the cells labeled by 2-­μm plastic particles experience lower blood flow at 30 ROIs in the cerebral cortical vessels at various
levels of obstruction, but the obstructed cells were also more readily time points after injection (Fig. 7A and fig. S6). The LSCI results
cleared. Consistent with that, cells labeled by 0.08-­μm plastic parti- indicated that treatment with MPs led to a reduction in blood per-
cles completely vanished within 12 hours (Fig. 6C). These findings fusion levels in the vessels (Fig. 7B), particularly notable at 30 min
suggest that plastic particle size is a crucial factor in regulating after treatment, with significantly lower perfusion levels compared
MPL-­Cell obstruction, when particles are no larger than 5 μm in to 10 min.
diameter, a smaller particle size corresponds to a lower level of Subsequently, we classified the vessels based on their flow levels
obstruction. to further analyze the impact of MPs on vessels with different flow
Exposure concentration is a crucial indicator for evaluating the rates. The ROIs were divided into three groups on the basis of the
biotoxicity of MPs. To enhance comparability with actual human measured blood perfusion levels: perfusion units (PU) greater than

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exposure level (12 μg/ml in the blood) (8), we tested various con- 500, PU greater than 400 but less than 500, and PU less than 400.
centrations using 5-­μm MPs. The obstruction level of MPL-­Cells Among these, the ROIs with PU greater than 500 exhibited no sig-
was not significantly affected when the exposure concentration was nificant changes over the course of measurement (Fig. 7C). Con-
halved (Fig. 6D), suggesting that the original exposure concentra- versely, the other two groups showed a noticeable decrease in blood
tion may have reached a saturation point that leads to MPL-­Cell perfusion levels starting at 10 min after treatment (Fig. 7, D and E),

Fig. 7. MPL-­Cell obstructions as thrombus formation inhibit intracerebrovascular blood perfusion. (A) Representative laser speckle contrast imaging (LSCI) images
at indicated time points after treatment. A black line square identifies an ROI location with perfusion units (PU) > 500, and a white circle identifies an ROI location with
PU < 400. (B) Quantitative statistical analysis of ROIs (n = 30). (C to E) Analysis of ROIs with PU > 500 (n = 8), ROIs with 400 < PU < 500 (n = 9), and ROIs with PU < 400
(n = 13). Three independent replicated experiments verified the results. Data are represented as means ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001.

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reaching their lowest point at 30 min. These findings suggest that obstruct cerebral blood vessels. By combining multilayer scanning im-
MPL-­Cell obstruction in the vessels affects blood perfusion, particu- aging with 3D reconstruction techniques, we obtained clear images of
larly affecting smaller vessels with lower blood flow. the blocked MPL-­Cells (Fig. 5, B to D). Long-­duration tracking imag-
ing showed the MPL-­Cells present difficulties in clearance for a pro-
Suppressed blood flow in the brain causes longed period of at least 1 week. The obstruction notably hindered
neurobehavioral disorders blood flow in the vessels, resulting in impaired neurological function
Vascular embolism resulted in insufficient blood supply to the brain in the mice. The direct observation of MPL-­Cells provides a mechanis-
tissue, leading to disturbances in neural activity and cognitive impair- tic explanation for MP-­induced neurological dysfunction.
ment in mice (35, 36). We investigated whether obstruction of MPL-­ Nevertheless, the application of this technology remains in its
Cells caused behavioral changes in mice (Fig. 8A). The open-­field early stages, and there are still several deficiencies in data collection
experiment is a widely used method for assessing the exploratory be- and analysis. Visual documentation of MPs in circulating blood was
havior of rodents, particularly in the context of anxiety disorders and obtained (Fig. 2C). Using a millisecond-­scale imaging technique,
other neurological and psychiatric conditions. Mice were assessed in the movement trajectory of the MP-­Flash was captured within 208 ms,
the open field 6 hours after injection (Fig. 8B). The total distance and enabling the calculation of the motion velocity of the MP (Fig. 2I).
speed of movement of MP-­treated mice were significantly lower com- Nevertheless, it is important to acknowledge the constraints as-
pared to those in the control group (Fig. 8, C and D). No significant sociated with this calculation. First, there is uncertainty regarding
difference was observed in the number of entries into the center area whether the motion of the MP-­Flash falls entirely within the imag-
(Fig. 8E). In the Y-­maze test for assessing working memory (Fig. 8F), ing plane, potentially leading to an underestimation of the trajectory
the total distance traveled by MP-­treated mice remained lower than length used for calculation. In addition, despite the utilization of
that in the control group (Fig. 8G), with a significant reduction in high-­speed bidirectional line scan mode for image acquisition, the

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spatial memory (Fig. 8H). These findings also suggest that MPs po- total imaging time for the entire FOV upon completion of the scan
tentially inhibit motor abilities in mice. To investigate this further, a was 208 ms, suggesting that the actual motion time of the MP-­Flash
rotarod test was conducted to evaluate motor coordination. MP-­ may be shorter. These factors may have biased our calculations low.
treated mice exhibited a significant decrease in latency time (Fig. 8I). Moreover, the inherent physical characteristics (diameter, polysty-
Similarly, in the rod-­hanging test, MP-­treated mice showed a signifi- rene material, and spherical) of the MP particles used in our study
cant decrease in hanging time at 1 and 3 days after injection, indicat- present constraints on our imaging techniques and computations,
ing reduced coordination and endurance. However, this difference potentially limiting their generalizability to other varieties of MPs.
was no longer significant on day 7 (Fig. 8J). The effects of MPs on Our findings suggest that the phagocytosis of MPs is mainly by im-
mouse locomotion may influence performance in open field and Y-­ mune cells in the blood. Nevertheless, the underlying reasons for im-
maze tests. These findings indicate that mice display multifaceted ab- mune cells obstructing the vasculature after ingestion of MPs remain
normalities in neurobehavioral regulation, resembling depressive states unclear. Neutrophils express various families of adhesion molecules,
associated with disrupted cerebral blood flow (37, 38). By day 28 after such as integrins and selectins, that interact with their corresponding
MP injection, the rotarod test showed no significant difference (Fig. 8K), ligands on other cells or in the extracellular matrix, leading to the for-
suggesting that the impairment in behavioral abilities caused by mation of transient or stable adhesions (37–39). In addition, the adhe-
MPL-­Cell blockage and altered blood perfusion was restored by 28 days. sion of neutrophil could be induced by the polarization stimulated by
In addition, MPs was found to induce weight loss in mice (Fig. 8L), the pathogens and local signals of tissue damage. Thus, it is hypothe-
potentially due to changes in feeding behavior resulting from al- sized that MPs as xenobiotics cause neutrophils polarization, facilitating
tered locomotor abilities. Last, at 28 days after injection, the recovery their interaction with endothelial cells and resulting in adhesion (40, 41).
of MPL-­Cell obstruction in the cerebral vasculature was observed, In addition, the formation of neutrophil extracellular traps may occur
with a significantly lower density compared to 7 days after injection subsequent to phagocytosis and adhesion (42). As we observed the cel-
(Fig. 8M). These results align with the behavioral experiments con- lular morphology characterized by umbrella-­like folds in the 3D imag-
ducted in mice at 28 days after injection. ing (Fig. 5D). Moreover, it could also explain why the obstructions were
observed in such a fast time; in addition to the role of the vascular envi-
ronment, changes in the adhesive properties of the MPL-­Cells them-
DISCUSSION selves may be key. The ingestion of the larger particles may induce
For an extended period, the application of advanced technology— changes in cell size and physical attributes, potentially leading to swell-
through long-­distance (e.g., astronomical sciences), large-­scale (e.g., ing or increased hardness (43, 44), making it more challenging for them
earth sciences), or microworld (e.g., life sciences) imaging—has served to pass through the vascular stenosis and the turns. Consistent with this,
as a crucial method for scientific researchers to conduct explorations the flow cytometry data showed that the neutrophils with MPs revealed
and summarize the underlying principles governing the various an increase in FSC and a decrease in SSC (Fig. 4, G and H).
phenomena. We used fluorescent MPs and in vivo imaging to detect However, multiple cellular biological processes are involved in
the MPs in the circulatory system of an organism. The MP-­Flash was the MPL-­Cell obstruction, most of which remain unknown. It is
captured in clear trails in their entirety. The establishment of this imag- unclear which cellular signaling pathways are altered following the
ing system enables the visualization of MPs at the in vivo level within phagocytosis of MPs and which cell surface receptors are subse-
the organism while simultaneously providing researchers with a pow- quently affected. Furthermore, the role of these membrane receptors
erful tool for studying the role of MPs in the organism. Using this im- in the process of cell obstruction is not well understood. Whether
aging system, our results demonstrate that immune cells phagocytosed the specificity of the vascular endothelium regulates the obstruction.
the MPs in the bloodstream, termed MPL-­Cells (fig. S7). We captured The extent to which signaling proteins in blood influence this pro-
images of the MPL-­Cells in motion and found that these cells can cess remains to be determined. However, the inhibition of specific

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Fig. 8. Suppressed blood flow in the brain causes neurobehavioral disorders. (A) Schematic representation of the arrangements for behavioral experiments.
Created using FigDraw.com. (B to E) An open-­field experiment was performed on MP-­treated and control mice (n = 10 to 11 mice for each group), representative dia-
grams of mouse locomotor trajectories (B), analytical statistics of the total travel distance (C), movement speeds (D), and the number of trips to the center zone (E). (F to
H) Y-­maze experiment (n = 11 mice for each group), representative diagrams of mouse locomotor trajectories (F), analytical statistics of the total movements (G), and al-
ternation triplet (H). (I) The latency time in the rotarod test of the MP-­treated and control mice (n = 12 mice for each group). (J) Hanging time for MP-­treated mice and
control mice experimented on different days after MP treatment (n = 12 mice for each group). (K) Latency time in the rotarod test at 28 days after MP treatment (n = 12
mice for each group). (L) Body weight change curves of mice affected by MP treatment (n = 11 to 12 mice for each group). (M) Analytical statistics of the obstructed MPL-­
Cell density in the brain at 7 and 28 days after MP treatment (n = 20 FOVs from four mice per time point). Data are represented as means ± SEM. *P < 0.05, **P < 0.01, and
***P < 0.001.

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functional proteins aids in elucidating the cellular biological pro- not completely eliminated. Correspondingly, behavioral deficits in
cesses that may be involved in this phenomenon. For instance, inhib- mice returned to baseline levels after 4 weeks.
iting the functional protein dynamin with the small-­molecule drug It was believed that MPs influence brain function in two primary
Dyngo-­4a may provide insights into how endocytosis affects MPL-­ ways. MPs can indirectly regulate brain function through peripheral
Cell levels. In addition, inhibiting cell adhesion using the small-­ organs, or they might influence the brain by somehow crossing the
molecule drug Milategrast could be used to investigate whether the BBB or interacting directly with it. The MPL-­Cell mechanism can be
obstruction of MPL-­Cells is dependent on the cell adhesion or categorized as a third type of potential regulation in which MPs affects
whether the structural expansion of the cells alone is sufficient to brain function; unlike the previously thought, MPs reach local brain
induce the obstruction phenomena at this scale. Additional further tissues but do not cross the BBB and affect local brain function by
molecular manipulations should be used to explore the underlying regulating blood perfusion. Our study has identified an ingenious
mechanisms. We have existing data indicating that the size of phago- mechanism through which MPs make function to organisms. We
cytosed particles is critical for triggering cellular obstruction (Fig. 6B); have demonstrated a negative effect of this mechanism on neurobehav-
however, it remains unclear whether because particles of varying ioral regulation in the brain; moreover, we also believe that it may also
sizes activate different signaling pathways, thereby influencing cell have detrimental impacts on a broader range of organs. Environment
attachment. In addition, in vitro experiments have demonstrated MPs might be another important risk factor for cardiovascular disease
that the size of phagocytic particles directly affects immune cell (18, 50, 51) and received scant attention so far. Our findings provide a
functionality (43). Extraction and transcriptome sequencing analy- mechanistic explanation for tissue damage induced by MPs, particu-
sis of PL-­Cells induced by plastic particles of differing sizes will help larly regarding their effects on vascular obstruction. This obstruction is
further elucidate this issue. likely to have detrimental consequences for cardiovascular health and
The obstruction of the MPL-­Cells is characterized by dynamic may result in more severe adverse effects, especially in patients with un-

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changes. In certain instances, MPL-­Cells could not remain stable derlying conditions similar to myocardial infarction. Notably, a recent
after obstruction and exhibit slow movement within the vessel. Fol- study on humans by Marfella et al. (18) supports this correlation.
lowing a period of interaction with the vessel wall, they may detach However, it is premature to directly apply this mechanism to
and continue to move with the blood flow (Fig. 3I). On the other human research systems. Humans and mice have different immune
hand, an MPL-­Cell that has become stably obstructed can impede systems, coagulation systems, and cardiovascular and cerebrovas-
the movement of newly arriving MPL-­Cells, causing them to co- cular circulatory systems (52–55). The circulating blood volume in
alesce and increase the area of the obstruction (movie S4). However, humans is ~1200 times greater than that of mice, and, notably, sig-
most of the newly arriving MPL-­Cells disengage and continue mov- nificantly different vascular diameters would greatly reduce the de-
ing after a certain duration. Notably, an MPL-­Cell that has become gree of MPL-­Cell obstruction in humans. The internal diameter of
stably obstructed will not be affected by the presence of a new MPL-­ the coronary arteries in the human heart is about 4 mm, whereas,
Cell nor will it be displaced. This stability likely arises from a rela- in mice, it measures less than 100 μm (56, 57). Consequently, there
tively stronger bond formed between the MPL-­Cell and the vessel is uncertainty regarding whether MPs will induce or influence the
wall. Similarly, a developing thrombus recruits unstimulated plate- obstruction in human blood vessels. However, the diameter of hu-
lets. Thrombus formation is a dynamic process in which some plate- man capillary microvessels at their narrowest point measures only
lets adhere to the developing thrombus, while others separate from 8 to 10 μm, which closely resembles the 8-­to 9-­μm diameter of the
it, influenced by shear, flow, and turbulence (45, 46). However, the terminal branches of venous vessels in mice (58–60). Moreover, we
progression from MPL-­Cell to thrombus may represent a distinct can deduce that the presence of MPs in the bloodstream may more
pathway compared to the classical thrombus formation triggered by likely induce obstruction in areas where there are sedimentations
vascular injury, which initiates platelet activation through subendo- in the lining of blood vessels. These preexisting conditions could
thelial collagen (47). The changes induced by MP in the MPL-­Cell hinder the transit of MPL-­C ells through narrowed vascular re-
itself, leading to obstruction, should be the primary causative event gions, thereby increasing the likelihood of obstruction. This may
and first to occur. Afterward, the immune factors released by MPL-­ significantly elevate the risk for specific populations, particularly
Cells may collaborate with tissue factors, triggering a secondary those with a history of thrombotic conditions, such as cerebrovas-
pathway that initiates platelet activation and, subsequently, activates cular infarction and myocardial infarction. In these patients, MPL-­
a proteolytic cascade that generates thrombin. Visualizing platelets Cells are more likely to cause obstructions at the narrowed sites of
during the obstruction of MPL-­Cells through fluorescent labeling blood vessels. Obesity is increasingly as a prevalent human disease,
will help address this question. and it can contribute to the accumulation of cholesterol and lipids
The obstruction of MPL-­Cells rapidly decreased blood perfusion in the walls of blood vessels (61, 62). Therefore, it is essential to
levels in the brain. We did not observe longer-­term changes, and the investigate the potential obstructive effects of MPL-­C ells in hu-
data at 50 min already showed no significant differences from the mans. Moreover, humans are exposed to a variety of MP sizes, and
data at 30 min (Fig. 7B). In addition, subgroup analyses of different the combinatorial effects of different MP sizes on the likelihood of
flow vessels suggest that the blockage effect is more pronounced inducing infarctions remain to be estimated. The use of larger mam-
in narrower vessels with lower perfusion rates (Fig. 7D), supporting mals or animal models that more closely resemble the human cir-
the idea that the inhibition of blood perfusion is caused by cellular culatory system, such as nonhuman primates, is thus crucial for
obstruction. The specific structure of the vascular network in the studying this process.
cortex means that obstruction of key nodes can directly affect perfu- In this study, we investigated the effects of MPs exposure over a
sion levels in a specific area of tributaries (48, 49). This explains the maximum duration of 28 days. The long-­term consequences of MPL-­
results that the bottleneck period of inhibition occurs rapidly. The Cell obstruction and the cumulative effects of multiple repeated MPs
blocked cells, which were significantly reduced after 4 weeks, were exposures remain unknown. Collectively, studies indicate that the

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human circulatory system is at significant risk due to direct exposure anesthetized with 1.5% isoflurane in air at a flow rate of 0.4 liters/min
to MPs (63, 64). If medical injection devices are not rapidly and thor- and kept on a 37°C heating pad during surgery. The specific cerebral
oughly improved, then the direct infiltration of MPs into the human cortical region for imaging was identified using stereotactic coordi-
bloodstream may become a persistent and potentially recurrent issue. nates (motor cortex area, from bregma: anteroposterior, 0.5 mm; me-
The potential long-­term effects of MPs on neurological disorders such diolateral, 1.2 mm). The skull over the ROI was thinned using a
as depression and cardiovascular health are concerning. This study of- high-­speed micro-­drill under a dissection microscope, with inter-
fers a theoretical foundation and a focused direction for understand- mittent drilling and application of normal saline to prevent overheat-
ing the potential health risks associated with MPs in this context. ing and damage to the underlying cerebral tissue. After removing the
Increased investment in this area of research is urgent and essential to external layer of compact bone and most of the spongy bone layer,
fully comprehend the health risks posed by MPs in human blood. the area was further thinned until a smooth, expanded area was
achieved, ensuring that the center was thin enough for high-­quality
imaging. A drop of saline was applied, and a 3-­mm-­diameter glass
METHODS coverslip, sterilized in 70% ethanol, was placed on the window. Den-
Mouse husbandry and experimentation tal cement was then applied around the glass coverslip. All subjects
All experiments involving animals conformed to the rules of the As- were housed individually in separate cages following the surgery. The
sociation for Assessment and Accreditation of Laboratory Animal Care mice were used in the experiment after a postoperative recovery pe-
and the Guide for the Care and Use of Laboratory Animals published by riod of 2 to 4 weeks.
the US National Institutes of Health (NIH Publication eighth edition, AAV9-­hSyn-­DIO-­mitoEGFP (100 nl; BrainVTA, no. PT-­2619)
update 2011). All procedures were approved by the Animal Care Com- and AAV9-­CaMKII-­Cre (100 nl; BrainVTA, no. PT-­0220) were in-
mittee of Charles River (PA23053002-­0) and PKU-­Nanjing Institute of jected into the motor cortex at coordinates—anteroposterior, 0.5 mm;

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Translational Medicine (IACUC-­2021-­023). All mice were housed in a mediolateral, 1.2 mm; dorsoventral, −0.3 mm—at a rate of 50 nl/min.
temperature-­controlled (20° to 22°C) and specific pathogen–free ani- A 3-­mm-­diameter glass coverslip was implanted and secured on the
mal facility, three to five per cage, and maintained on a 12-­hour light/ motor cortex. The mice were used in the experiment 2 to 4 weeks after
dark cycle, with water and food available ad libitum. postoperative recovery.
Eight-­week-­old male wild-­type (WT) C57BL/6J mice were pro-
cured from Vital River Laboratory. We used a 5-­μm-­diameter fluo- Imaging and recording
rescently labeled polystyrene plastic microsphere, MPs, a commonly After undergoing surgery, mice fitted with a transparent cranial window
used material in the field (65–69) (Baseline, catalog no.7-­3-­0500). were selected for further experiments. The mice were securely head
On the basis of previous research, mice were assigned randomly to fixed on the imaging stage, and single-­photon imaging was initially con-
untreated or gavaged with 100 μl of MP water mixture at a dose of ducted to determine the appropriate vascular FOV. Subsequently,
2 mg/ml. Experimenters were informed about the grouping of mice. mTPM was used. A suitable FOV containing a clear image of blood ves-
MPs can enter the human bloodstream through medical supplies sels located 20 to 50 μm beneath the pial surface was identified. The
(8, 70–72), about 12 μg of MPs per milliliter of blood have been de- mTPM and FOV were then stabilized, and changes following the ad-
tected in human blood. The need to simulate the human condition in ministration of MP treatment were observed within the same imaging
the concentration settings of the experiment was taken into account. FOV. The fast high-­resolution mTPM large FOV model featured a head-
We would like to bring mouse blood MPs to this level by injection. piece weight of 2.45 g, a lens numerical aperture of 0.5, and an FOV of
An adult mouse of 30 g has a blood volume of about 2 ml. We in- 520 μm by 440 μm. Images were captured to visualize cortical blood
jected 100 μl of MPs at a concentration of 1 mg/ml intravenously into vessels surveillance through either a time-­lapse xy imaging stack at a
the mouse, and the diluted final concentration after entering the frame rate of 4.8 frames per second or multiplane imaging stacks con-
bloodstream should be blood of about 50 μg/ml. In the experiments sisting of 40 planes at 2.5-­μm intervals, with 4.8 frames per second. Each
involving varying concentrations of MPs exposure, the injection vol- mTPM model was equipped with a water-­immersion miniature objec-
ume remained constant, while the MP concentrations administered tive and a 920-­nm femtosecond fiber laser. Fluorescence imaging excita-
were diluted to 0.5 and 0.1 mg/ml. In the exposure experiments with tion was achieved with 150-­fs laser pulses (80 MHz) at 920 nm for the
different sizes of plastic particles, 2-­μm-­diameter (Baseline, catalog fluorescently labeled MPs, with a power of ~25 mW after passing
no. 7-­3-­0200) and 80-­nm-­diameter (Baseline, catalog no. 7-­3-­0008) through the objective.
fluorescently labeled polystyrene plastics were used.
Male WT mice, 8 weeks old, were procured from Vital River Labo- Blood sample processing and white blood cell isolation
ratory. After a 2-­week acclimatization period to the rearing environ- Upon receipt, ~700 μl of fresh blood samples were received in EDTA
ment, the mice were used for behavioral detection experiments. To tubes, within 2 hours of collection at room temperature (RT) mixed
minimize the impact of injection manipulation on mice, WT mice with 6.3 ml of 10× red blood cell (RBC) lysis buffer (Beyotime, cata-
designated for behavioral assays were not permitted to undergo behav- log no. C3702). RBC lysis of whole blood was performed to isolate
ioral testing until at least 6 hours after the completion of intravenous leukocytes. Samples were lysed at RT for 10 min, continuously mix-
injection. Generally, all injections in both groups of mice were admin- ing on a tube rotator. Cells were then centrifuged at 300g for 5 min
istered within a controlled timeframe of 1 hour. Control mice received and washed with Sorter Buffer (PBS and 1 mM EDTA). The cells
an equivalent volume of phosphate-­buffered saline (PBS) solution. were washed twice with PBS and then used for the next step.

Surgery Flow cytometry

All surgical procedures were performed under sterile conditions, and Cells were stained with specific surface or intracellular antibodies
all reagents administered to the animals were sterile. Mice were (shown in the following) in staining buffer (1.0% bovine serum

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albumin). Cells were washed three times by PBS. The antibodies 2 min. Each mouse session comprised three trials, and the hanging
used were anti-­CD45 (BioLegend, 47-­0451-­80), anti-­F4/80 (Life time was averaged. To ensure consistency in testing, experiments
Technologies, 11-­4801-­81), and anti-­Ly6G/Ly6C (BD Biosciences, typically commenced at 19:00 and concluded by 22:00 on each ex-
561103). Cells were analyzed on an LSRFortessa (BD) flow cytom- perimental day.
eter, and data were analyzed using FlowJo X software.
Quantification and statistical analysis
Laser speckle contrast imaging Imaging data were processed with Fiji (v1.54). All data are shown as the
Mice were imaged under isoflurane anesthesia (3% induction, 1 to means ± SEM. All boxes showed the median of the data. n denotes the
1.5% maintenance, in oxygen) by head fixed, and the skull was ex- number of biological replicates in each experiment, and it is provided
posed. Camera focus and exposure time were adjusted to achieve in the corresponding figure legends. All statistical analyses were con-
optimal imaging conditions. After locking the imaging FOV, mice ducted using GraphPad Prism (v8.0.2). All two-­group comparisons
were injected intravenously with MPs. Tests were performed at were calculated using Student’s t test (two-­tailed, paired, or unpaired).
different time points after injection. For each imaging, time-­length The results were considered statistically significant when P < 0.05.
1-­min blood perfusion levels were continuously recorded. The mean
perfusion level within 1 min for each ROI was calculated.
Supplementary Materials
Open field The PDF file includes:
Figs. S1 to S7
The dimensions of the open field for the mice are 50 cm in length,
Legends for movies S1 to S7
50 cm in width, and 40 cm in height. The laboratory environment is
maintained in a quiet state with controlled lighting at about 8 lux. Other Supplementary Material for this manuscript includes the following:

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Before the experiment, the mice were acclimatized to the laboratory Movies S1 to S7
environment for at least 30 min to minimize stress response to the
unfamiliar surroundings. At the start of the experiment, the mice REFERENCES AND NOTES
were gently placed in a corner of the open field, with each mouse 1. A. Banerjee, W. L. Shelver, Micro-­and nanoplastic induced cellular toxicity in mammals:
A review. Sci. Total Environ. 755, 142518 (2021).
starting from the same initial position. The behavioral activities of 2. W. Huang, B. Song, J. Liang, Q. Niu, G. Zeng, M. Shen, J. Deng, Y. Luo, X. Wen, Y. Zhang,
the mice were recorded via filming for a duration of 7 min. The se- Microplastics and associated contaminants in the aquatic environment: A review on their
quence in which the mice from the two experimental groups under- ecotoxicological effects, trophic transfer, and potential impacts to human health.
went the experiment was randomized. After the completion of the J. Hazard. Mater. 405, 124187 (2021).
3. K. L. Law, R. Narayan, Reducing environmental plastic pollution by designing polymer
experiment for each mouse, any residual urine in the experimental
materials for managed end-­of-­life. Nat. Rev. Mater. 7, 104–116 (2022).
area was cleaned using alcohol. Following the cleanup, the alcohol 4. J. R. Jambeck, R. Geyer, C. Wilcox, T. R. Siegler, M. Perryman, A. Andrady, R. Narayan,
odor was allowed to dissipate before proceeding with the next mouse. K. L. Law, Plastic waste inputs from land into the ocean. Science 347, 768–771 (2015).
5. L. C. M. Lebreton, J. van der Zwet, J.-­W. Damsteeg, B. Slat, A. Andrady, J. Reisser, River
Y-­maze plastic emissions to the world’s oceans. Nat. Commun. 8, 15611 (2017).
6. A. A. Mamun, T. A. E. Prasetya, I. R. Dewi, M. Ahmad, Microplastics in human food
The Y-­maze consisted of a Y-­shaped compartment (21 cm by 7 cm chains: Food becoming a threat to health safety. Sci. Total Environ. 858 (Pt. 1),
by 15.5 cm), it is three arms of equal length. The mice were acclima- 159834 (2023).
tized to the laboratory environment for 3 days before the experi- 7. L. F. Amato-­Lourenço, R. Carvalho-­Oliveira, G. R. Júnior, L. Dos Santos Galvão, R. A. Ando,
ment to familiarize themselves with the room and reduce stress. The T. Mauad, Presence of airborne microplastics in human lung tissue. J. Hazard. Mater.
416, 126124 (2021).
arms of the maze were labeled A, B, and C. Mice were gently placed
8. H. A. Leslie, M. J. M. van Velzen, S. H. Brandsma, A. D. Vethaak, J. J. Garcia-­Vallejo,
in the fixed arm facing the center of the maze. They were given M. H. Lamoree, Discovery and quantification of plastic particle pollution in human blood.
8 min to explore the maze freely. The maze was thoroughly cleaned Environ. Int. 163, 107199 (2022).
with alcohol between experimental mice. 9. P. Wu, S. Lin, G. Cao, J. Wu, H. Jin, C. Wang, M. H. Wong, Z. Yang, Z. Cai, Absorption,
distribution, metabolism, excretion and toxicity of microplastics in the human body and
health implications. J. Hazard. Mater. 437, 129361 (2022).
Rotarod test 10. Y. Yang, E. Xie, Z. Du, Z. Peng, Z. Han, L. Li, R. Zhao, Y. Qin, M. Xue, F. Li, K. Hua, X. Yang,
The rotarod test was used to evaluate the mice’s ability to maintain Detection of various microplastics in patients undergoing cardiac surgery.
balance on a rotating rod. As the rotational speed increased, it be- Environ. Sci. Technol. 57, 10911–10918 (2023).
came more challenging for the mice to stay balanced. Before the test, 11. S. Massardo, D. Verzola, S. Alberti, C. Caboni, M. Santostefano, E. E. Verrina, A. Angeletti,
F. Lugani, G. M. Ghiggeri, M. Bruschi, G. Candiano, N. Rumeo, M. Gentile, P. Cravedi,
the mice underwent a 2-­day training period, with three sessions per
S. La Maestra, G. Zaza, G. Stallone, P. Esposito, F. Viazzi, N. Mancianti, E. La Porta, C. Artini,
day, where they were trained to walk on a rotating rod at a speed of MicroRaman spectroscopy detects the presence of microplastics in human urine and
5 rpm/min for 3 min each time. During the actual test, the mice kidney tissue. Environ. Int. 184, 108444 (2024).
were placed on the rotarod, and the rotational speed was gradually 12. H. Huang, J. Hou, Y. Liao, F. Wei, B. Xing, Polyethylene microplastics impede the innate
increased from 5 rpm/min to 30 rpm/min over a period of 4 min. immune response by disrupting the extracellular matrix and signaling transduction.
iScience 26, 107390 (2023).
The latency time, indicating when the mice fell due to their inability 13. H. Huang, F. Wei, S. Qiu, B. Xing, J. Hou, Polystyrene microplastics trigger adiposity in mice
to maintain balance, was recorded during the acceleration. by remodeling gut microbiota and boosting fatty acid synthesis. Sci. Total Environ.
890, 164297 (2023).
Rod-­hanging test 14. H. T. Pinheiro, C. M. Donald, R. G. Santos, R. Ali, A. Bobat, B. J. Cresswell, R. Francini-­Filho,
Mice were allowed to grasp a horizontally maintained 2-­mm-­diameter R. Freitas, G. F. Galbraith, P. Musembi, T. A. Phelps, J. P. Quimbayo, T. E. A. L. Quiros,
B. Shepherd, P. V. Stefanoudis, S. Talma, J. B. Teixeira, L. C. Woodall, L. A. Rocha, Plastic
wooden rod with their four paws, positioned about 50 cm above a thick pollution on the world’s coral reefs. Nature 619, 311–316 (2023).
layer of soft bedding. The time until the mice fell from the rod was 15. A. Das, The emerging role of microplastics in systemic toxicity: Involvement of reactive
recorded. Following each fall, the mice were permitted to recover for oxygen species (ROS). Sci. Total Environ. 895, 165076 (2023).

Huang et al., Sci. Adv. 11, eadr8243 (2025) 22 January 2025 14 of 16

S c i e n c e A d v a n c e s | R e s e ar c h A r t i c l e

16. N. G. Posnack, Plastics and cardiovascular disease. Nat. Rev. Cardiol. 18, 69–70 (2021). 41. H. C. A. Drexler, M. Vockel, C. Polaschegg, M. Frye, K. Peters, D. Vestweber, Vascular
17. X. Zhu, C. Wang, X. Duan, B. Liang, E. G. Xu, Z. Huang, Micro-­and nanoplastics: A new endothelial receptor tyrosine phosphatase: Identification of novel substrates related to
cardiovascular risk factor? Environ. Int. 171, 107662 (2023). junctions and a ternary complex with EPHB4 and TIE2. Mol. Cell. Proteomics
18. R. Marfella, F. Prattichizzo, C. Sardu, G. Fulgenzi, L. Graciotti, T. Spadoni, N. D’Onofrio, 18, 2058–2077 (2019).
L. Scisciola, R. L. Grotta, C. Frigé, V. Pellegrini, M. Municinò, M. Siniscalchi, F. Spinetti, 42. C. C. Winterbourn, A. J. Kettle, M. B. Hampton, Reactive oxygen species and neutrophil
G. Vigliotti, C. Vecchione, A. Carrizzo, G. Accarino, A. Squillante, G. Spaziano, D. Mirra, function. Annu. Rev. Biochem. 85, 765–792 (2016).
R. Esposito, S. Altieri, G. Falco, A. Fenti, S. Galoppo, S. Canzano, F. C. Sasso, G. Matacchione, 43. P. Sharma, A. Vijaykumar, J. V. Raghavan, S. R. Rananaware, A. Alakesh, J. Bodele,
F. Olivieri, F. Ferraraccio, I. Panarese, P. Paolisso, E. Barbato, C. Lubritto, M. L. Balestrieri, J. U. Rehman, S. Shukla, V. Wagde, S. Nadig, S. Chakrabarti, S. S. Visweswariah, D. Nandi,
C. Mauro, A. E. Caballero, S. Rajagopalan, A. Ceriello, B. D’Agostino, P. Iovino, G. Paolisso, B. Gopal, S. Jhunjhunwala, Particle uptake driven phagocytosis in macrophages and
Microplastics and nanoplastics in atheromas and cardiovascular events. N. Engl. J. Med. neutrophils enhances bacterial clearance. J. Control. Release 343, 131–141 (2022).
390, 900–910 (2024). 44. J. Schumann, It is all about fluidity: Fatty acids and macrophage phagocytosis.
19. C.-­S. Yang, C.-­H. Chang, P.-­J. Tsai, W.-­Y. Chen, F.-­G. Tseng, L.-­W. Lo, Nanoparticle-­based in Eur. J. Pharmacol. 785, 18–23 (2016).
vivo investigation on blood-­brain barrier permeability following ischemia and 45. B. Furie, B. C. Furie, Mechanisms of thrombus formation. N. Engl. J. Med. 359, 938–949 (2008).
reperfusion. Anal. Chem. 76, 4465–4471 (2004). 46. C. Dubois, L. Panicot-­Dubois, J. F. Gainor, B. C. Furie, B. Furie, Thrombin-­initiated platelet
20. X. Liu, Y. Zhao, J. Dou, Q. Hou, J. Cheng, X. Jiang, Bioeffects of inhaled nanoplastics on activation in vivo is vWF independent during thrombus formation in a laser injury model.
neurons and alteration of animal behaviors through deposition in the brain. Nano Lett. J. Clin. Invest. 117, 953–960 (2007).
22, 1091–1099 (2022). 47. P. Mangin, C. L. Yap, C. Nonne, S. A. Sturgeon, I. Goncalves, Y. Yuan, S. M. Schoenwaelder,
21. Z. Liu, A. Sokratian, A. M. Duda, E. Xu, C. Stanhope, A. Fu, S. Strader, H. Li, Y. Yuan, C. E. Wright, F. Lanza, S. P. Jackson, Thrombin overcomes the thrombosis defect
B. G. Bobay, J. Sipe, K. Bai, I. Lundgaard, N. Liu, B. Hernandez, C. B. Rickman, S. E. Miller, associated with platelet GPVI/FcRγ deficiency. Blood 107, 4346–4353 (2006).
A. B. West, Anionic nanoplastic contaminants promote Parkinson’s disease-­associated 48. T. Wälchli, J. Bisschop, P. Carmeliet, G. Zadeh, P. P. Monnier, K. De Bock, I. Radovanovic,
α-­synuclein aggregation. Sci. Adv. 9, eadi8716 (2023). Shaping the brain vasculature in development and disease in the single-­cell era.
22. X. Chen, L. Xu, Q. Chen, S. Su, J. Zhuang, D. Qiao, Polystyrene micro-­and nanoparticles Nat. Rev. Neurosci. 24, 271–298 (2023).
exposure induced anxiety-­like behaviors, gut microbiota dysbiosis and metabolism 49. T. Wälchli, A. Wacker, K. Frei, L. Regli, M. E. Schwab, S. P. Hoerstrup, H. Gerhardt,
disorder in adult mice. Ecotoxicol. Environ. Saf. 259, 115000 (2023). B. Engelhardt, Wiring the vascular network with neural cues: A CNS perspective. Neuron

Downloaded from https://www.science.org on February 19, 2025

23. H. S. Shin, S. H. Lee, H. J. Moon, Y. H. So, H. R. Lee, E.-­H. Lee, E.-­M. Jung, Exposure to 87, 271–296 (2015).
polystyrene particles causes anxiety-­, depression-­like behavior and abnormal social 50. S. Rajagopalan, P. J. Landrigan, Pollution and the heart. N. Engl. J. Med. 385, 1881–1892 (2021).
behavior in mice. J. Hazard. Mater. 454, 131465 (2023). 51. A. D. Vethaak, J. Legler, Microplastics and human health. Science 371, 672–674 (2021).
24. M. Prüst, J. Meijer, R. H. S. Westerink, The plastic brain: Neurotoxicity of micro-­and 52. J. Zschaler, D. Schlorke, J. Arnhold, Differences in innate immune response between man
nanoplastics. Part. Fibre Toxicol. 17, 24 (2020). and mouse. Crit. Rev. Immunol. 34, 433–454 (2014).
25. I. Varó, A. Perini, A. Torreblanca, Y. Garcia, E. Bergami, M. L. Vannuccini, I. Corsi, 53. R. D. Hodge, T. E. Bakken, J. A. Miller, K. A. Smith, E. R. Barkan, L. T. Graybuck, J. L. Close,
Time-­dependent effects of polystyrene nanoparticles in brine shrimp Artemia B. Long, N. Johansen, O. Penn, Z. Yao, J. Eggermont, T. Höllt, B. P. Levi, S. I. Shehata,
franciscana at physiological, biochemical and molecular levels. Sci. Total Environ. B. Aevermann, A. Beller, D. Bertagnolli, K. Brouner, T. Casper, C. Cobbs, R. Dalley, N. Dee,
675, 570–580 (2019). S.-­L. Ding, R. G. Ellenbogen, O. Fong, E. Garren, J. Goldy, R. P. Gwinn, D. Hirschstein,
26. J. Ding, S. Zhang, R. M. Razanajatovo, H. Zou, W. Zhu, Accumulation, tissue distribution, C. D. Keene, M. Keshk, A. L. Ko, K. Lathia, A. Mahfouz, Z. Maltzer, M. M. Graw, T. N. Nguyen,
and biochemical effects of polystyrene microplastics in the freshwater fish red tilapia J. Nyhus, J. G. Ojemann, A. Oldre, S. Parry, S. Reynolds, C. Rimorin, N. V. Shapovalova,
(Oreochromis niloticus). Environ. Pollut. 238, 1–9 (2018). S. Somasundaram, A. Szafer, E. R. Thomsen, M. Tieu, G. Quon, R. H. Scheuermann, R. Yuste,
27. S. Zhang, J. Ding, R. M. Razanajatovo, H. Jiang, H. Zou, W. Zhu, Interactive effects of S. M. Sunkin, B. Lelieveldt, D. Feng, L. Ng, A. Bernard, M. Hawrylycz, J. W. Phillips, B. Tasic,
polystyrene microplastics and roxithromycin on bioaccumulation and biochemical status H. Zeng, A. R. Jones, C. Koch, E. S. Lein, Conserved cell types with divergent features in
in the freshwater fish red tilapia (Oreochromis niloticus). Sci. Total Environ. 648, 1431–1439 human versus mouse cortex. Nature 573, 61–68 (2019).
(2019). 54. A. Wessels, D. Sedmera, Developmental anatomy of the heart: A tale of mice and man.
28. W. Zong, R. Wu, S. Chen, J. Wu, H. Wang, Z. Zhao, G. Chen, R. Tu, D. Wu, Y. Hu, Y. Xu, Y. Wang, Physiol. Genomics 15, 165–176 (2003).
Z. Duan, H. Wu, Y. Zhang, J. Zhang, A. Wang, L. Chen, H. Cheng, Miniature two-­photon 55. B. S. Buetow, M. A. Laflamme, in Cardiovascular, Comparative Anatomy and Histology
microscopy for enlarged field-­of-­view, multi-­plane and long-­term brain imaging. (Academic Press, ed. 2, 2018), chap. 10, pp. 163–189.
Nat. Methods 18, 46–49 (2021). 56. J. T. Dodge Jr., B. G. Brown, E. L. Bolson, H. T. Dodge, Lumen diameter of normal human
29. E. Parzy, S. Miraux, J.-­M. Franconi, E. Thiaudière, In vivo quantification of blood velocity in coronary arteries. Influence of age, sex, anatomic variation, and left ventricular
mouse carotid and pulmonary arteries by ECG-­triggered 3D time-­resolved magnetic hypertrophy or dilation. Circulation 86, 232–246 (1992).
resonance angiography. NMR Biomed. 22, 532–537 (2009). 57. S. Sawall, J. Beckendorf, C. Amato, J. Maier, J. Backs, G. V. Velde, M. Kachelrieß,
30. S. Gordon, Phagocytosis: An immunobiologic process. Immunity 44, 463–475 (2016). J. Kuntz, Coronary micro-­computed tomography angiography in mice. Sci. Rep.
31. V. Coelho-­Santos, A. Y. Shih, Postnatal development of cerebrovascular structure and the 10, 16866 (2020).
neurogliovascular unit. Wiley Interdiscip. Rev. Dev. Biol. 9, e363 (2020). 58. B. Müller, S. Lang, M. Dominietto, M. Rudin, G. Schulz, H. Deyhle, M. Germann, F. Pfeiffer,
32. K. Kisler, A. R. Nelson, A. Montagne, B. V. Zlokovic, Cerebral blood flow regulation and C. David, T. Weitkamp, High-­resolution tomographic imaging of microvessels. Proc. SPIE
neurovascular dysfunction in Alzheimer disease. Nat. Rev. Neurosci. 18, 419–434 (2017). 7078, Developments in X-­Ray Tomography VI, 70780B (2008).
33. D. Attwell, A. M. Buchan, S. Charpak, M. Lauritzen, B. A. Macvicar, E. A. Newman, Glial and 59. H. Gong, D. Xu, J. Yuan, X. Li, C. Guo, J. Peng, Y. Li, L. A. Schwarz, A. Li, B. Hu, B. Xiong,
neuronal control of brain blood flow. Nature 468, 232–243 (2010). Q. Sun, Y. Zhang, J. Liu, Q. Zhong, T. Xu, S. Zeng, Q. Luo, High-­throughput dual-­colour
34. A. J. S. Webb, D. J. Werring, New insights into cerebrovascular pathophysiology and precision imaging for brain-­wide connectome with cytoarchitectonic landmarks at the
hypertension. Stroke 53, 1054–1064 (2022). cellular level. Nat. Commun. 7, 12142 (2016).
35. V. Rajeev, Y. L. Chai, L. Poh, S. Selvaraji, D. Y. Fann, D.-­G. Jo, T. M. De Silva, G. R. Drummond, 60. B. Xiong, A. Li, Y. Lou, S. Chen, B. Long, J. Peng, Z. Yang, T. Xu, X. Yang, X. Li, T. Jiang, Q. Luo,
C. G. Sobey, T. V. Arumugam, C. P. Chen, M. K. P. Lai, Chronic cerebral hypoperfusion: A H. Gong, Precise cerebral vascular atlas in stereotaxic coordinates of whole mouse brain.
critical feature in unravelling the etiology of vascular cognitive impairment. Front. Neuroanat. 11, 128 (2017).
Acta Neuropathol. Commun. 11, 93 (2023). 61. K. Malekmohammad, E. E. Bezsonov, M. Rafieian-­Kopaei, Role of lipid accumulation and
36. G. Bieri, A. B. Schroer, S. A. Villeda, Blood-­to-­brain communication in aging and inflammation in atherosclerosis: Focus on molecular and cellular mechanisms.
rejuvenation. Nat. Neurosci. 26, 379–393 (2023). Front. Cardiovasc. Med. 8, 707529 (2021).
37. M. Michael, M. Parsons, New perspectives on integrin-­dependent adhesions. Curr. Opin. 62. C. Hotoleanu, Association between obesity and venous thromboembolism.
Cell Biol. 63, 31–37 (2020). Med. Pharm. Rep. 93, 162–168 (2020).
38. G. L. Burn, A. Foti, G. Marsman, D. F. Patel, A. Zychlinsky, The meutrophil. Immunity 63. L. Zhu, M. Ma, X. Sun, Z. Wu, Y. Yu, Y. Kang, Z. Liu, Q. Xu, L. An, Microplastics entry into the
54, 1377–1391 (2021). blood by infusion therapy: Few but a direct pathway. Environ. Sci. Technol. Lett. 11, 67–72
39. W. M. Nauseef, N. Borregaard, Neutrophils at work. Nat. Immunol. 15, 602–611 (2014). (2024).
40. A. Margraf, G. Germena, H. C. A. Drexler, J. Rossaint, N. Ludwig, B. Prystaj, S. Mersmann, 64. P. Li, Q. Li, Y. Lai, S. Yang, S. Yu, R. Liu, G. Jiang, J. Liu, Direct entry of micro(nano)plastics
K. Thomas, H. Block, W. Gottschlich, C. Liu, P. W. Krenn, H. Haller, B. Heitplatz, into human blood circulatory system by intravenous infusion. iScience 26, 108454 (2023).
M. M. Z. Brickwedde, M. Moser, D. Vestweber, A. Zarbock, The integrin-­linked kinase is 65. W. Gu, S. Liu, L. Chen, Y. Liu, C. Gu, H.-­Q. Ren, B. Wu, Single-­cell RNA sequencing reveals
required for chemokine-­triggered high-­affinity conformation of the neutrophil size-­dependent effects of polystyrene microplastics on immune and secretory cell
β2-­integrin LFA-­1. Blood 136, 2200–2205 (2020). populations from zebrafish intestines. Environ. Sci. Technol. 54, 3417–3427 (2020).

Huang et al., Sci. Adv. 11, eadr8243 (2025) 22 January 2025 15 of 16

S c i e n c e A d v a n c e s | R e s e ar c h A r t i c l e

66. T. Luo, C. Wang, Z. Pan, C. Jin, Z. Fu, Y. Jin, Maternal polystyrene microplastic exposure Acknowledgments: We thank H.Cheng and the PKU-­Nanjing Institute of Translational
during gestation and lactation altered metabolic homeostasis in the dams and their F1 Medicine for support of fluorescence microscopy image collection, analysis, sharing reagents,
and F2 offspring. Environ. Sci. Technol. 53, 10978–10992 (2019). and providing access to instruments; and the technicians in the Charles River Laboratory
67. Y. Jin, L. Lu, W. Tu, T. Luo, Z. Fu, Impacts of polystyrene microplastic on the gut barrier, Animal Research Center for assistance with animal experiments. We thank all of the volunteers
microbiota and metabolism of mice. Sci. Total Environ. 649, 308–317 (2019). and nurses who participated in this study. Funding: The study was funded by the
68. L. Lu, Z. Wan, T. Luo, Z. Fu, Y. Jin, Polystyrene microplastics induce gut microbiota Fundamental Research Funds for the Central Public-­interest Scientific Institution
dysbiosis and hepatic lipid metabolism disorder in mice. Sci. Total Environ. (2022YSKY-­34) (J.H. and H.H.) and the National Natural Science Foundation of China
631-­632, 449–458 (2018). (51908524) (B.X.). Author contributions: Conceptualization: H.H., J.H., and B.X. Methodology:
69. H. Huang, J. Hou, C. Yu, F. Wei, B. Xi, Microplastics exacerbate tissue damage and promote H.H. Software: H.H. and F.W. Validation: H.H., J.H., B.X., F.W., and Y.L. Formal analysis: J.H., B.X.,
carcinogenesis following liver infection in mice. Ecotoxicol. Environ. Saf. 286, 117217 and Y.L. Investigation: H.H., J.H., M.L., and Y.L. Resources: H.H., J.H., and M.L. Data Curation: H.H.,
(2024). B.X., M.L., and Y.L. Writing—original draft: H.H., J.H., and B.X. Writing—review and editing: H.H.,
70. S. Liu, Y. Yang, Z. Du, C. Wang, L. Li, M. Zhang, S. Ni, Z. Yue, K. Yang, H. Gao, Y. Zeng, Y. Qin, J.H., M.L., and F.W. Visualization: H.H., J.H., B.X., and F.W. Supervision: J.H. and B.X. Project
J. Li, C. Yin, M. Zhang, Percutaneous coronary intervention leads to microplastics entering administration: J.H. and B.X. Funding acquisition: H.H., J.H., and B.X. Competing interests: The
the blood: Interventional devices are a major source. J. Hazard. Mater. 476, 135054 authors declare that they have no competing interests. Data and materials availability: All
(2024). data needed to evaluate the conclusions in the paper are present in the paper and/or the
71. S. V. L. Leonard, C. R. Liddle, C. A. Atherall, E. Chapman, M. Watkins, S. D. J. Calaminus, Supplementary Materials.
J. M. Rotchell, Microplastics in human blood: Polymer types, concentrations and
characterisation using μFTIR. Environ. Int. 188, 108751 (2024). Submitted 18 July 2024
72. D.-­W. Lee, J. Jung, S.-­A. Park, Y. Lee, J. Kim, C. Han, H.-­C. Kim, J. H. Lee, Y.-­C. Hong, Accepted 16 December 2024
Microplastic particles in human blood and their association with coagulation markers. Published 22 January 2025
Sci. Rep. 14, 30419 (2024). 10.1126/sciadv.adr8243

Downloaded from https://www.science.org on February 19, 2025

Huang et al., Sci. Adv. 11, eadr8243 (2025) 22 January 2025 16 of 16