Association of Outdoor Ambient Fine Particulate Matter With Intracellular White Matter Microstructural Properties Among Children

Key Points Question Is annual residential exposure to particulate matter 2.5 μm or less in diameter (PM2.5) associated with neuroimaging diffusion markers of white matter microstructure? Findings This cross-sectional study of 7602 children 9 to 10 years of age found evidence of an association between PM2.5 exposure and hemispheric differences in white matter microstructure. In hemisphere-specific models, adjusted for confounding variables, statistically significant positive associations were observed between PM2.5 and restricted isotropic diffusion, and statistically significant negative associations were observed between PM2.5 and mean diffusivity. Meaning Findings from this study suggest that exposure to PM2.5 may be associated with differences in white matter microarchitecture, supporting a need for further improvements in air quality to protect the developing brain.


Introduction
Ambient airborne particulate matter is composed of suspended particles with an aerodynamic diameter of 2.5 μm or less (PM 2.5 ). 1 Long-term exposure to PM 2.5 is reportedly associated with adverse nervous system effects. 2 Animal studies [3][4][5][6] have indicated that inhaled PM 2.5 leads to neuroinflammation and oxidative stress, which may induce neuronal injury and affect glial support cells. Recent magnetic resonance imaging (MRI) studies have suggested an association of PM 2.5 exposure with brain structure and volume, 7,8 including white matter, [9][10][11][12] which is primarily made up of myelinated axons and glial support cells. 13 The potential impingement on key neurodevelopmental processes by PM 2.5 exposure may cause lifelong health effects. 14 Myelination and improved microstructural organization of white matter pathways continue throughout childhood and into young adulthood, ultimately allowing for improved signal transduction and communication between distal brain regions among cognitive and emotional systems. [15][16][17] Studies have found that air pollution is associated with smaller white matter surface area in children, 11 increases in myo-inositol, a brain metabolite involved in cell membrane and myelination, 18 and reduced fractional anisotropy (FA). 12,19 These studies 11,12,18,19 are primarily based on smaller, localized populations, and results may have limited generalizability. Furthermore, exposure levels in these populations average above the current US Environmental Protection Agency standard of 12 μg/m 3 . 3,9,20 Further research is warranted to examine the potential effects of exposure to levels of PM 2.5 at or below regulatory standards across larger, more geographically diverse populations of children and using more advanced diffusion weighted imaging (DWI) techniques.
By modeling multishell high-angular resolution DWI data with a novel framework called restriction spectrum imaging (RSI), we aimed to characterize associations between ambient PM 2.5 exposure and white matter microarchitecture in children 9 to 10 years of age from the Adolescent Brain Cognitive Development (ABCD) Study. Restriction spectrum imaging adopts a biophysical model that goes beyond conventional diffusion tensor imaging (DTI) techniques to distinguish different types of microstructural tissue compartments, including restricted water bounded by cell membranes (intracellular) and hindered water primarily within the extracellular space, where glial cell bodies and other neural processes increase the tortuosity of diffusion (Figure 1). [22][23][24][25] Restriction spectrum imaging improves sensitivity and specificity in understanding tissue damage 26 and normative changes in microstructural development. 21, 27 Given that particle pollution has been linked with impaired myelination and alteration of glial cells, 28,29 we hypothesized that higher levels of ambient PM 2.5 exposure would be associated with decreased restricted directional intracellular diffusion (rND) (eg, organized myelination) and increased restricted isotropic intracellular diffusion (rN0) (eg, glia and cell bodies). Previous work 21 suggests a moderately positive correspondence between FA and rND and a large correspondence (in opposite directions) between mean diffusivity (MD) and rN0 in age-related white matter changes in early adolescence; thus, we hypothesized that PM 2.5 exposure may be associated with decreased FA and MD. We also examined whether these associations varied by hemisphere and sex, given previous evidence of hemispheric and sex differences in associations between PM 2.5 exposure and health outcomes. 7,8,[30][31][32][33][34]

Study Population
Data were obtained from baseline assessments (September 1, 2016, to October 15, 2018) of the ABCD Study (2020 National Institute of Mental Health Data Archive 3.0 data release), a cohort study of participants 9 to 10 years of age in the US. [35][36][37] The ABCD Study implemented identical protocols for recruitment and neuroimaging of all participants at 21 study sites across the US. 35,[38][39][40][41][42][43] The primary inclusion criteria were age and English proficiency; exclusion criteria included severe sensory, intellectual, medical, or neurologic issues that would affect valid data collection (eMethods in the Supplement). Data analysis was performed from September 15, 2020, to June 30, 2021. Study sites obtained approval from their local institutional review boards, and centralized institutional review board approval was obtained from the University of California, San Diego. All parents or caregivers provided written informed consent; each child provided written assent. All data were deidentified before use. This study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline. 44 We further excluded participants with nonvalid addresses, low-quality or missing MRI, and incidental abnormal MRI findings (eMethods in the Supplement). Within-family nonindependence Illustration of the biological components of white matter in an imaging voxel and schematic representation of the 2 diffusion tensor imaging (DTI) modeling approaches used in this study: DTI and restricted spectrum imaging (RSI). Diffusion tensor imaging measures extracellular water diffusion across a voxel. Primary DTI outcomes include fractional anisotropy (FA) and mean diffusivity (MD). Although the magnitude and direction of these outcomes allow for inferences to be made regarding axonal structure and integrity, DTI only allows for quantification of a single principal direction of diffusion and does not allow for characterization of the relative contribution of neurite orientations within a single voxel. In contrast, RSI is a biophysical model that allows for estimates of compartmentalized hindered and restricted water diffusion. Primary RSI outcomes include total hindered diffusion (hD) as well as restricted isotropic intracellular diffusion (rN0) and restricted directional intracellular diffusion (rND), which together provide greater insight into the biological properties of the microstructure of white matter tissue. The hindered compartment could encompass diffusion within intracellular spaces that allow for diffusion greater than the diffusion length scale (typically approximately 10 μm for human DTI). Given that rN0 and rND are normalized with respect to the hD compartments, changes in restricted compartments are relative to the other compartments. Previous studies using both common DTI and novel RSI metrics have shown similarities in directionality of rND and FA but opposite associations between rN0 and MD metrics in white matter during childhood. 21 Created with BioRender.com. was managed by randomly including 1 sibling per family (eFigure 1 in the Supplement). The final analytic sample included 7602 participants (Table; eFigure 1 in the Supplement).

Estimation of PM 2.5 Exposure
The methods used to estimate residential PM 2.5 exposure have been previously described. 7 Daily estimates of hybrid spatiotemporal PM 2.5 models were used to aggregate daily PM 2.5 exposure

MRI Acquisition and Processing
The MRI data collection was harmonized across the 21 sites using 3 T scanners (Siemens Prisma, General Electric 750, Philips). 39,45 The diffusion-weighted acquisition was conducted as previously described. 45 After preprocessing of DWIs (eMethods in the Supplement), RSI was used to fit fiber orientation density functions to model rND, rN0, and total hindered diffusion (hD) (eg, primarily extracellular space around neurites). 45 The DTI outcomes included FA and MD. Major white matter tracts were labeled with AtlasTrack using prior probabilities and orientation of long-range projection fibers. 46 Probability estimates for each white matter tract were used to calculate weighted means of the RSI and DTI measures for all white matter fibers as well as key association, commissural, and projection fiber tracts, 45

Sensitivity Analysis
We conducted sensitivity analyses adjusting models for population density and proximity to major roadways. We also evaluated the addition of random slopes by ABCD Study site to investigate geographic variability in the associations between PM 2.5 exposure and DWI outcomes. Finally, we tested a possible interaction between PM 2.5 exposure and assigned sex at birth.

Statistical Analysis
We used hierarchical mixed-effects models with random intercepts by study site. Natural cubic splines for PM 2.5 were fit with 2 knots at 7.05 and 8.31 μg/m 3 derived from tertiles of exposure. We used an interaction term of PM 2.5 × hemisphere and then fit hemisphere-stratified models (eMethods in the Supplement). All models were adjusted for covariates selected based on a directed acyclic graph, including sex, child's age, parent-declared race and ethnicity, highest educational level of any household member, household income, parental employment status, a mean score of a 3-item assessment of parent perspectives of neighborhood safety, imaging device manufacturer, handedness, and motion artifact indexed by framewise displacement (eFigure 3 and eTable 3 in the Supplement). Analyses were performed using R, version 4.0.2 (R Foundation for Statistical Computing). An α = .05 was chosen, a priori, before any models were fit or analyzed as a threshold for significance. All reported P values are 1-sided.

Sensitivity Analysis
The addition of 1 or both of our sensitivity covariates (distance to major roadways and population density) did not improve model fit or change associations. We did not see any meaningful variability in the random slopes across ABCD Study sites. In addition, no associations with sex were found for any outcomes.

Discussion
This cross-sectional analysis used data from a diverse cohort of 7602 children 9 to 10 years of age located at 21 geographically diverse locations across the US. Our objective was to characterize associations between annual ambient PM 2.5 exposure and white matter microarchitecture. We found evidence of an interaction with hemisphere for nearly every white matter tract analyzed. In hemisphere-stratified models, higher PM 2.5 exposure was associated with increased rN0 in 2 tracts in the left hemisphere only and bilaterally in 2 tracts. Higher exposure was associated with decreases in MD in 3 tracts in the left hemisphere only, bilaterally in 4 tracts, and in the corpus callosum.
Significant hemisphere-specific associations were not observed between PM 2.5 and rND, hD, or FA.
These findings suggest that higher PM 2.5 exposure is linked to increases in cellular barriers in white matter (reflected by decreases in MD), including increases in the isotropic compartment (reflected by increases in rN0), which may indicate changes to the cellular composition of key white matter tracts. To our knowledge, this is the first study to investigate how PM 2.5 is associated with RSI-measured restricted and hindered water diffusion. The robustness of these findings is supported by strict quality control criteria for MRI inclusion, selection of covariates to control confounding, and sensitivity analyses (eMethods in the Supplement). Although the observed associations are small, it is plausible that repeated daily exposure to ambient PM 2.5 across adolescence may have important implications for long-term neurophysiologic health outcomes of today's youth. 48 Our findings suggest that PM 2.5 exposure may be associated with changes in intracellular microarchitecture of frontoparietal and limbic white matter circuitry, important for attention (SLF), emotional processing (UNC, CGH, and SLF), and memory (FX and SLF). [49][50][51][52] Particle pollution may pass from the lungs into the bloodstream to infiltrate the blood brain barrier or create systematic secondary effects through inflammation. 4 Microglia, the resident immune cells of the central nervous  system, respond to pollutants and cause inflammatory activation. 28 Activated microglia in animal and cell models exhibit increases in somal size compared with resting microglia. 53 The rN0 reflects diffusion bound within cell membranes in spherical structures less than approximately 10 μm or within multiple cylindrical structures oriented such that diffusion is occurring equally in all directions.

JAMA Network Open | Environmental Health
Therefore, our findings of increased rN0 may reflect differences in the number or size of glial cells (eg, oligodendrocytes, oligodendrocyte precursor cells, astrocytes, and/or microglia) in white matter tracts (Figure 4). 13 Because increases in the restricted signal fraction along white matter tracts have been associated with normative development in the ABCD Study cohort, it is unclear to what extent these microstructural associations with PM 2.5 reflect an acceleration of developmental processes vs an inflammatory response. 21 A clinical study 54 examining rN0 in patients with Parkinson disease found a 9.09% increase in rN0 in the bilateral thalamus of patients compared with healthy controls, pointing to the possibility that rN0 changes may reflect mechanisms underlying symptom origins.
Further experimental animal studies using both cellular and neuroimaging techniques are warranted to better understand the implications of white matter microstructural changes.
Previous research 21 has found that rND and FA tend to show similar patterns, whereas rN0 and MD are inversely associated in white matter tracts during early adolescence. Given these known patterns, the negative associations between PM 2.5 and MD observed here suggest an increased barrier to water diffusion, which is congruent with potential increases in the number or size of support cells. Given that MD quantifies the magnitude of diffusion, whereas FA depends on the overall directionality of diffusion within a voxel, 55 our findings suggest that PM 2.5 may be increasing the number of cellular boundaries but not changing cellular processes that contribute to unidirectional water diffusion, such as axonal organization and/or myelination. Moreover, given the known associations between FA and rND metrics, a lack of association between PM 2.5 and these 2 markers corroborates this conclusion.
Despite congruent findings between DTI and RSI outcomes, the notable negative associations between PM 2.5 and MD in the current study contrast with a previous positive association noted in a cohort of children 9 to 10 years of age from Rotterdam, the Netherlands. 9 PM 2.5 composition varies Hypothesized neurobiological underpinning of particulate matter 2.5 μm or less in diameter (PM 2.5 )associated increase in restricted isotropic intracellular diffusion (rN0). Given that rN0 represents diffusion within cells, it is hypothesized that increases of rN0 in white matter at 9 to 10 years may reflect an increase in the size or number of support and glial cells in response to exposure to PM 2.5 . An overall increase or change in cell numbers within a given white matter region could also contribute to overall decreases in mean diffusivity as measured by diffusion tensor imaging. hD indicates total hindered diffusion; rND, restricted directional intracellular diffusion; RSI, restriction spectrum imaging. by geographic location, and previous evidence 2,56 suggests that certain PM 2.5 components are differentially detrimental to health, which may explain these contrasting findings. Participants from the previous study 9 were also exposed to much higher overall levels of PM 2.5 exposure (mean, 16.5 μg/m 3 ), which may also contribute to this discrepancy because higher levels of exposure may lead to more severe cellular or myelin disruption (reflected by increases in MD). Finally, the current study and the study by Lubczyńska et al 9 used different diffusion-MRI acquisition parameters; the previous study 9 used a single shell DTI sequence, whereas the current study used a multishell high-angular resolution DWI sequence with various b values, allowing for increased sensitivity and specificity. 21,57 Although the exact mechanisms underlying central nervous system asymmetries remain unknown, 58 structural and functional differences have been noted between the 2 cerebral hemispheres at the macroscopic, microstructural, and molecular levels. 59 A previous study 58 of asymmetry in neurologic disorders suggests that typical asymmetries develop between the hemispheres, which may ultimately result in greater hemispheric differences in vulnerability to brain pathologic conditions. A previous study 58 suggests that brain asymmetries occur via differences in functional genetic pathways of microtubule regulation, neurogenesis, and axonogenesis, which are involved in neuronal development and organization and the manifestation of hemispheric differences in gene expression. Thus, these asymmetries in brain structure and function may contribute to the hemisphere-specific patterns observed in this study.

Limitations
This study has limitations. Because of limits in the ABCD Study air pollution data available in the 3.0 release, participants experienced varying time lags between their air pollution exposure estimation (2016) and MRI at the baseline study visit (2016-2018); an assumption was therefore made that the spatial distribution of air pollution estimates remained stable during this 2-year period. This assumption is supported by previous research that indicates that the spatial distribution of estimates of annual mean air pollution concentrations (using the current estimation methods) remained relatively stable in the US between 2008 and 2016. 60 Future data releases from the ABCD Study are expected to contain full lifetime histories of air pollution exposure, which will allow for more temporal precision in cross-sectional analyses and longitudinal investigations.
Ambient outdoor PM 2.5 exposure at a primary residence does not provide a full picture of a child's yearly air pollution exposure. Data on indoor air pollution, school air pollution, and time at the residence, although not currently available, would further clarify how PM 2.5 exposure is associated with white matter connectivity. Similarly, despite efforts to account for confounding in our analyses, it is possible that unmeasured confounders and residual confounding have introduced biases in the associations reported here.
In addition, this study was limited to PM 2.5 exposure, yet other types of air pollution, including nitric dioxide, may affect the morphological features and development of children's brains and their mental health. 9,19,61 In previous studies 9,19 with multipollutant analyses, single pollutant associations tended to become weakened by the addition of 1 of more pollutants; thus, the possibility exists that the PM 2.5 associations reported here are somewhat biased and inflated by the inclusion of only a single air pollutant in our models. When data become available for the ABCD Study population, future analyses are planned to elucidate potential associations among ambient nitric dioxide , ozone , PM 2.5 components, and DWI outcomes.

Conclusions
To our knowledge, this was the first multisite US study to find associations between annual PM 2.5 exposure and white matter microarchitecture. Most of the study population experienced PM 2.5 exposure at or below 12 μg/m 3 , which is within US Environmental Protection Agency standards.
These findings have important public health implications, given the ubiquity of PM 2.5 exposure and its potential effects on white matter connectivity in children across the US.