Diffuse Axonal Injury Research Articles

Background Diffuse axonal injury (DAI), a severe subtype of traumatic brain injury (TBI), lacks reliable early diagnostic biomarkers, contributing to poor clinical outcomes. Systemic metabolic pathway dysregulation in DAI remains poorly characterized, limiting targeted therapeutic strategies. Objectives Identify DAI-specific metabolic network disruptions and evaluate their diagnostic and prognostic utility. Methods In this prospective cohort study, serum metabolomics profiling, pathway enrichment analysis, and machine learning were integrated with clinical assessments in 64 adults with acute TBI (30 DAI, 34 non-DAI). Untargeted metabolomics via UPLC-LTQ-Orbitrap MS identified differential metabolites, which were mapped to biological pathways using MetaboAnalyst 5.0. Diagnostic and prognostic performance of pathway-based models was assessed using ROC analysis. Results DAI patients exhibited distinct metabolic perturbations, with significant dysregulation in mitochondrial fatty acid oxidation (FAO) and phospholipid metabolism. Key discriminative metabolites included carnitine C8:1 (VIP = 3.26) and lysophosphatidylcholine 22:3 sn-2, which correlated with Marshall CT scores ( ρ = 0.62, p < 0.001) and pupillary reflex loss. A multi-parameter model integrating FAO and phospholipid degradation markers achieved superior diagnostic accuracy (AUC = 0.927, 95% CI: 0.86–0.98) compared to clinical models (AUC = 0.744). Pathway disruptions further predicted 3-month functional outcomes (GOSE AUC = 0.912). Conclusion DAI involves systemic metabolic network dysfunction centered on mitochondrial energetics and lipid metabolism. Pathway-centric biomarkers enhance diagnostic precision and prognostication, offering a novel framework for biomarker-driven management of TBI. These findings highlight mitochondrial FAO and phospholipid homeostasis as potential therapeutic targets, addressing a critical gap in DAI care.

Isolated oculomotor nerve palsy (ONP) following mild traumatic brain injury (TBI) is rare and often presents diagnostic challenges. Typically associated with diffuse axonal injury and poor prognosis, ONP lacks comprehensive radiological documentation when no skull base fractures along the course of the 3rd Nerve or brainstem injuries are evident. This study explores the diagnostic utility of contrast-enhanced magnetic resonance imaging (CEMRI) in identifying ONP cases in mild TBI patients. A retrospective analysis was conducted on six patients diagnosed with isolated ONP after mild TBI, with no evident skull base fractures along the course of the 3rd Nerve or brainstem findings. All patients underwent CE-MRI to identify structural or vascular anomalies along the course of the third cranial nerve. Clinical presentations, imaging findings, and outcomes were meticulously documented and reviewed by a neuroradiologist. CE-MRI findings revealed consistent abnormalities in all cases. Thickening, blooming, and post-contrast enhancement of the cisternal portion of the third cranial nerve were observed, with two cases demonstrating extension into orbital segments. Despite the absence of fractures or direct injuries, partial recovery was noted in most cases, facilitated by targeted steroid therapy in some instances. These imaging patterns suggest indirect mechanisms such as traction, vascular compromise, or intraneural hemorrhage as potential causes of ONP. High-resolution CE-MRI proves instrumental in diagnosing isolated ONP in mild TBI patients, even without conventional radiological indicators. Early imaging and intervention may improve recovery outcomes. This study underscores the significance of including CE-MRI in evaluation protocols for ONP. It highlights the importance of further research to unravel the underlying pathophysiology and optimize therapeutic approaches for these patients.

Concussion-related symptoms, such as impaired balance, slower processing speed, attention deficits, memory dysfunction, and irritability, are thought to result from diffuse axonal injury (DAI), characterized by selective damage to white matter axons. Axons subjected to this mechanical stretch injury exhibit diverse pathological changes, including disruption of axonal transport, neurofilament compaction and degradation, myelin sheath disruption, and loss of sodium channels required for action potential generation and propagation. These distinct forms of axonal pathology may evolve differentially over time and preferentially localize to specific white matter tracts. In this study, we employed the clinically relevant ferret model of concussion using the closed head impact model of engineered rotational acceleration (CHIMERA). 55 male ferrets were randomly allocated to sham or injury groups and then to either 24 h, 72 h, or 14d survival time points. We confirmed that axonal transport disruption and neurofilament pathology represent independent processes, with minimal colocalization but a shared peak of around 72 h following injury. Furthermore, we observed a persistent loss of ankyrin-G, a critical anchoring protein for sodium channels at the node of Ranvier, up to 14d postinjury, suggesting that the resultant impairment in axonal transmission may underlie many concussion symptoms. Indeed, injured ferrets displayed significant deficits in balance, working memory, spatial memory, and recognition memory. These findings demonstrate that the CHIMERA model in ferrets recapitulates key axonal pathologies and their associated clinical manifestations following concussion. This model offers a valuable platform for investigating the temporal evolution of axonal injury and developing targeted therapeutic interventions to mitigate concussion-related deficits.

Evaluation of functional brain damage using resting-state functional magnetic resonance imaging in patients with diffuse axonal injury admitted to the ICU

Diffuse axonal injury (DAI) arises as a severe consequence of traumatic brain injury (TBI), characterized by axonal misalignment, stretching, or shearing. These events trigger pathophysiological cascades, including axonal depolarization, metabolic perturbations, cellular swelling, cytotoxic edema, and apoptosis. While DAI typically involves multifocal brain damage, it predominantly affects the central third of the brain, with the corpus callosum and brainstem being especially prone to injury. Clinically, DAI presents a heterogeneous spectrum of manifestations, from acute loss of consciousness to cognitive deficits and persistent coma. Predicting outcomes remains challenging, and no standardized treatment protocol has been established. The authors report a case of a 23-year-old male victim of a high-speed traffic accident, who progressed to a comatose state and was diagnosed with grade III DAI. Subsequently, the patient underwent successful extubation and showed favorable recovery progress.

Disclosure: C. Gandhi: None. S. Sankanagoudar: None. S. Tiwari: None. M. Garg: None. D.K. Jha: None. R. Shukla: None.Background: Traumatic Brain Injury (TBI) survivors experience neurocognitive deficits, metabolic disturbances, and reduced quality of life due to undiagnosed hypopituitarism, yet, pituitary function assessment is not a part of routine protocol. This study aims to study the prevalence, type, and trend of pituitary dysfunction while identifying specific predictors (trauma severity, CT severity, copeptin, anti-GFAP antibody) and to observe changes in body composition at baseline and 6-months in male subjects with TBI due to Road Traffic Accidents. Methodology: This 6-month prospective study included 70 patients. TBI severity (GCS score), CT severity (Marshall and Rotterdam score), Pituitary hormone assessment including copeptin, and BMD-DXA was done within 5 days post-trauma. At the 6-month follow-up, 44 patients were reassessed, pituitary hormones including a Glucagon Stimulation test, anti-GFAP antibody and repeat BMD-DXA was done. Results: In the acute phase, 83% had pituitary dysfunction, most common being low testosterone in 81% while cortisol was least affected in 2.9% only. Diffuse axonal injury correlated positively with prolactin (r=0.27, p=0.02). Basal cistern obliteration was identified as an independent predictor of pituitary dysfunction (p=0.03). On follow-up, 75% had pituitary dysfunction with isolated deficits (59%) more common than multiple axes involvement (16%). GHD was observed in 57%, although none had IGF-1 levels below -2 SDS. Peak GH showed no correlation with baseline or follow-up IGF-1. Baseline copeptin did not predict long-term pituitary dysfunction. Anti-GFAP antibodies at 6-month were positive in 70% but showed no association with GHD or hypopituitarism. TBI severity, Rotterdam score (p<0.01), Marshall score (p<0.01) and basal cistern obliteration (p<0.01) were significantly associated with the number of axes in acute and long-term phase. Basal skull fractures (p=0.03) were identified as independent predictor of GHD, while cerebral oedema (p=0.06) and contusions (p=0.08) demonstrated possible association. Elevated baseline LH [18.71 vs 2.28, p=0.04] was observed in patients who succumbed to TBI. Body-composition analysis revealed increase in appendicular lean mass/height2 with no significant changes in body fat, lean mass or visceral adipose tissue on follow-up. Conclusion: Trend was towards recovery of gonadotroph and thyrotroph axis with new-onset deficits in somatotroph and corticotroph axes. Radiological parameters emerged as strong predictors, while copeptin and anti-GFAP antibodies did not predict pituitary dysfunction. Severe trauma was associated with multiple axes involvement. IGF-1 in adult GHD were found to be unreliable, emphasizing the need for dynamic testing. Acute phase LH may serve as a potential biomarker for mortality. An increase in appendicular lean mass indicates some recovery in physical strength over time.Presentation: Saturday, July 12, 2025

Unraveling the link between brain injury and enhanced artistic skills.

Traumatic brain injury and neurodegenerative diseases: the role of axonal injury and amyloid-β.

Electric pruning shears have become increasingly popular in Türkiye, particularly in regions where viticulture and olive cultivation are common. Although these tools offer ergonomic advantages, they also pose a significant risk of hand injuries. While injuries from other agricultural machinery have been documented in the literature, electric pruning shear injuries have not been systematically studied. Clinical observations of increased emergency admissions during the pruning season prompted this study. We retrospectively reviewed 28 patients admitted to our emergency department between November 2024 and March 2025 due to electric pruning shear injuries. Demographic data, injury mechanisms, affected anatomical zones, and associated tissue damage were recorded. Injuries were classified according to the volar hand injury zones and Ishikawa's classification. The time of injury, use of protective equipment, and patient training status were also evaluated. All patients sustained single-digit injuries distal to the metacarpophalangeal joint. The average age was 53.6 years, and 92.9% were male. Injuries predominantly affected the non-dominant hand (82.1%), with the index finger being the most commonly injured (39.1%). Twenty-seven patients were not wearing protective gloves, and none had received formal training in device use. Neurovascular injury was present in 20 cases, and distal circulation was absent at admission in 13 patients. Smoking was reported by 78.5% of patients. All injuries occurred during daylight hours, with a peak incidence around 2 p.m. Three patients were under 18, and five were over 70 years old, highlighting both the accessibility and physical risks associated with the device. Electric pruning shear injuries have emerged as a significant occupational hazard in agricultural regions. Ease of access, lack of training, and low awareness of protective measures contribute to their increasing frequency. Public health strategies such as safety regulations, mandatory training programs, and awareness campaigns are urgently needed. Further prospective studies are needed to evaluate functional outcomes and broader seasonal injury patterns.

BackgroudTraumatic brain injury (TBI) ranks among the leading causes of death worldwide. However, the association between hemoglobin variation (ΔHb) and hospital mortality in TBI patients at high altitude remains uninvestigated.MethodThis retrospective cohort study was conducted from January 2020 to March 2025 in the Tibetan Plateau region, enrolling 191 patients who resided at an average altitude of 3,000 m. ΔHb (peak Hb-nadir Hb) during the hospitalization, related covariates and hospital mortality were collected. Backward stepwise multivariable logistic regression was used to select key variables. The non-linear relationship between ΔHb and mortality was investigated using the multivariable fractional polynomial (MFP) method. The threshold effects of ΔHb were explored through two-piecewise logistic regression models.ResultsLogistic regression showed that ΔHb was independently and significantly associated with hospital mortality (OR = 1.08, 95% CI: 1.02–1.15, P = 0.005) after adjustment for nadir Hb, diffuse axonal injury and GCS (Glasgow coma scale) score. A cubic non-linear relationship between ΔHb and hospital mortality was revealed (P for non-linearity = 0.010), with an inflection point at 19.8 g/L. Additionally, an interaction effect between ΔHb and GCS score was found (P = 0.035).ConclusionsIn the clinical management of high-altitude TBI patients, our findings suggest that those with a ΔHb > 20 g/L, and a substantially elevated ΔHb and a low GCS score have an increased risk of mortality. A study investigating interventional strategies aimed at reducing ΔHb in TBI patients is warranted.

The central nervous system is characterized by its limited regenerative potential, yet striking examples of functional recovery after injury in animal models and humans highlight its capacity for repair. Little is known about repair of pathways/circuits after traumatic brain injury (TBI), which results in disruption of connectivity. Here we utilize a mouse model of diffuse traumatic axonal injury (Impact-acceleration TBI) in order to explore, for the first time, the evolution of structural and functional changes in the terminal fields of the injured visual system. Retinal ganglion cell (RGC) axons and synapses were genetically labeled via AAV transduction, while anterograde and transsynaptic tracers were used to mark terminals and postsynaptic cells. Functional connectivity and visual integrity were assessed by monitoring c-Fos expression following light stimulation and pattern-reversal visual evoked potentials (pVEPs). Our findings demonstrate that, although TAI results in approximately a 50% loss of RGC axons and terminals, surviving RGCs undergo collateral sprouting, a form of compensatory branching of surviving axons, that restores terminal density to pre-injury levels. Transsynaptic tracing and c-Fos mapping confirmed the reestablishment of connectivity, which was also associated with significant improvements in visual function as measured by pVEPs. Interestingly, the recovery process exhibited sexual dimorphism, with female mice showing delayed or incomplete repair. Moreover, collateral sprouting proceeded normally inSarm1knockout mice, evidence of some independence from Wallerian degeneration. Our findings show that collateral sprouting may be an important mechanism of circuit repair in TAI and may represent a promising target for therapeutic interventions.

Elevated intracranial pressure (ICP) is a complication of severe traumatic brain injury (TBI) that carries a risk of secondary brain injury. This study investigated the association between ICP burden and brain injury patterns on MRI in children with severe TBI. Secondary analysis of the Approaches and Decisions in Acute Pediatric TBI (ADAPT) study, which included children with severe TBI (Glasgow Coma Scale score < 9) who received a clinical MRI within 30 days of injury. We excluded patients who had ICP monitoring less than 24 hours, were missing ICP data for greater than 40% of monitoring time, or who underwent craniectomy. None. ICP burden was defined as the trapezoidal area under the curve of hourly ICP greater than 20 mm Hg. ICP was standardized to total monitoring time, and patients were categorized to four levels of ICP burden. MRI was evaluated for number of diffuse axonal injury (DAI) microhemorrhages, intracerebral hemorrhage (ICH) volume, contusion volume, and number of regions with ischemia. Fisher exact or chi-square tests were used to test the independence between ICP burden and MRI injury amount. Of the 220 patients, 156 (71%) had DAI, 31 (14%) had ICH, 161 (73%) had contusions, and 70 (32%) had ischemia on MRI. Most patients (180, 82%) experienced episodes of ICP greater than 20 mm Hg. Contusion volume (p = 0.02) and number of regions with ischemia (p = 0.007) were associated with ICP burden, but we failed to identify such an association for DAI or ICH. Severe (but not mild or moderate) ICP burden was associated with presence of ischemia (odds ratio, 4.64 [95% CI, 1.30-19.5]; p = 0.02). Elevated ICP was prevalent in the ADAPT cohort. Ischemia and contusion were associated with the burden of ICP. Further research is needed to determine temporal relationships between elevated ICP and ischemia.

IntroductionTraumatic Brain Injury (TBI) often leads to lasting cognitive and functional deficits, with Traumatic Axonal Injury (TAI) being a significant prognostic factor. This study investigated white matter microstructural changes in moderate-to-severe TBI, focusing on the presence and number of cerebral microbleeds (MBs) using diffusion tensor imaging (DTI).Materials and methods51 participants were recruited and categorized into three groups: 17 controls, 17 TBI patients with MBs (MBP), and 17 TBI patients without MBs (MBN). Age matching was applied to minimize confounding effects. MRI scans were acquired using a 3 T Siemens MAGNETOM Prisma scanner, and DTI data were preprocessed using FSL software. Whole white matter and corpus callosum masks were reconstructed using FreeSurfer, while tractography-based methods were implemented with FSL. Fractional anisotropy (FA) and mean diffusivity (MD) were extracted and compared across groups. Group-level voxel-wise statistical analysis was conducted using Tract-Based Spatial Statistics (TBSS), and generalized linear models (GLiMs) were applied to assess the effects of age, sex and MB number on DTI parameters.ResultsSignificant decrease in FA (p = 0,008 − 0,042) and increases in MD (p = 0,004 − 0,016) were observed in the WM masks when comparing the MBP group with the controls. In the TBSS analysis FA (p = 0,008) and MD (p = 0,005) showed significant differences between the MBP-CON comparison, while FA (p = 0,012) and MD (p = 0,043) were significantly different between the MBP and MBN groups. Moreover, a significant FA decrease was observed in the corpus callosum when comparing the MBP and MBN groups (p = 0,007). Additionally, an increasing number of microbleeds was significantly associated with altered DTI metrics in across all white matter masks.ConclusionOur findings highlight MBs as potential markers of more extensive white matter injury in moderate-to-severe TBI. The increase in MBs suggests even greater white matter damage, indicating a progression of microstructural alterations. On a global scale, tractography enhances the sensitivity in detecting structural alterations compared to traditional segmentation techniques. Examination of central white matter areas holds significant importance in uncovering the relevance of MBs.

Moderate and severe traumatic brain injury: abbreviated injury scale scoring and coding of traumatic axonal injury from early MRI.

Abusive head trauma (AHT) is a leading cause of death in infants and toddlers. The objective of this study was to conduct an age-stratified comparison between children with AHT and accidental Traumatic brain injury TBI (aTBI) in the Approaches and Decisions in Acute Pediatric Traumatic Brain Injury (ADAPT) data. Children with severe TBI and an intracranial pressure monitor placed at a study site in the United States were enrolled from February 1, 2014, to September 31, 2017, and subjects <5 years of age and admitted to a US site were selected for analysis. Subjects were stratified by mechanism ('definite' or 'probable' concern for AHT classified as AHT; 'possible' or 'no' concern for AHT as aTBI) and age (<1 year, 1-2.9-years-, 3-4.9-years). Clinical data including epidemiological, clinical events, and imaging that occurred before monitor placement were compared. Of the 313 subjects (n = 111 AHT), apnea, seizures, and bilateral fixed pupils were more frequently observed in AHT (35.1% vs. 21.8%, p = 0.01; 43.2% vs. 20.8, p < 0.001; 31.5% vs. 15.8%, p = 0.008). Subdural hemorrhages, midline shift, and ischemia were more frequently observed in AHT (96.4% vs. 73.1%, p < 0.001; 54.1% vs. 35.0%, p = 0.001; 40.9% vs. 12.2%, p < 0.001) while contusion, subarachnoid hemorrhage and diffuse axonal injury were less frequently observed (20.2% vs. 49.7%; 38.5% vs. 58.4%; 3.7% vs. 20.8%, all p < 0.001). Among the patients <1 year-old, there was no difference in apnea and seizures between AHT and aTBI (40.6% vs. 34.3%, p = 0.53; 44.9% vs. 40.0%, p = 0.63) while ischemia was more commonly observed in AHT (47.1% vs. 20.0%, p < 0.001). AHT subjects exhibited unique clinical characteristics and radiological findings compared to aTBI, even after this age-stratified comparison. Further study is needed on the effects of both guidelines-based and novel therapies for this vulnerable and unique patient population.

Diffuse axonal injury (DAI) is a leading cause of traumatic brain injury (TBI) morbidity and has well-studied molecular pathobiology. Historically, white matter DAI studies indicated unmyelinated axons are more susceptible to injury than myelinated axons, with myelin posited to protect axons from diffuse TBI shear/tensile forces through unresolved mechanisms. Similarly, preclinical studies have also identified gray matter DAI localized to the perisomatic domain (i.e., the unmyelinated axon initial segment [AIS] and first one-to-two nodes of Ranvier). With these concepts in mind, we hypothesized unmyelinated segments are selectively vulnerable to TBI-mediated shear/tensile forces and serve as initiating sites for DAI pathobiology. Using murine midline fluid percussion injury, neocortical layer V pyramidal cell perisomatic domains at the gray-white matter interface were spatiotemporally examined for initiating pathology using antibodies to cytoskeletal proteins to demarcate unmyelinated segments and amyloid precursor protein (i.e., the gold-standard DAI marker) to identify injury. In cells expressing yellow fluorescent protein to enhance injury visualization, axonal swellings were observed simultaneously within perisomatic unmyelinated segments (e.g., AIS; nodes) as well as immediately adjacent myelinated segments, indicating concomitant reactive axonal changes. These data suggest non-selective axonal susceptibility and that myelin may not protect against diffuse injury forces. While expanding DAI topography to the gray-white matter junction, these findings also have implications for action potential initiation, axonal protein trafficking, and cortical circuit connectivity. Furthermore, studies are needed to determine if DAI pathological mechanisms are shared between white and gray matter axons, which have common and differentiating cytoarchitectural components.

FSS in CTE triggers neuronal apoptosis through Piezo1-induced Ca2+ homeostasis disruption.

Traumatic brain injury (TBI) is a complex and often-devastating condition. This disease involves damage to cerebral structures: meninges (dura, arachnoid, pia), cerebral cortex, white matter tracts, and deeper structures (basal ganglia, brainstem), along with mechanisms including contusions, hematomas (epidural/subdural), diffuse axonal injury from shear forces, secondary edema compromising blood-brain barrier, and ischemia/hemorrhage caused by vascular disruption. The pathophysiological process of TBI above varies significantly among individuals. However, prevalent TBI treatments still focus on symptomatic management, such as surgical intervention represented by craniotomy, medical management represented by osmotic agents for cerebral edema, supportive care represented by oxygen therapy, and adjuvant therapies represented by hypothermia. Worse still, traditional therapies often yield unfavorable outcomes and indulge the potential onset of long-term neurodegenerative diseases (NDDs). On the other side, Glymphatic System (GS), discovered as a clearance system in the brain, has made tremendous progress over the past decade. Dysfunction of the GS has been implicated in various central nervous system (CNS) diseases including TBI. The discovery of the GS offers new perspectives for the pathophysiological process of TBI, particularly unveiling the truth of the development of diphasic brain edema following TBI. Impressively, with the GS maturing, unprecedented therapeutic strategies ensue. For instance, the GS might explain sleep deprivation after TBI strikes in part and strongly validate the prospect of sleep therapy, then provide insights into the enigma of sleep. Also, nor-adrenergic inhibition facilitates CSF-ISF exchange and glymphatic outflow, significantly attenuating brain edema. AQP4, the guardian and regulator of brain capacity at the end-foot of astrocyte, which can modulate its array and amounts aligning with nor-adrenergic signal, is indispensable in this process. Moreover, neurons have gained prominence in the brain's clearance system. Exploring the relationship between the GS and TBI will likely to blaze the new trail for advancing our understanding of TBI.

IntroductionTauopathies, including Alzheimer’s disease (AD), feature abnormal accumulations of hyperphosphorylated Tau protein; however, their biomarker potential in traumatic brain injury (TBI) is not well-defined. This study investigated whether cerebrospinal fluid (CSF) phosphorylated Tau at threonine-217 (pTau-217) could serve as an early biomarker for severe TBI (sTBI).MethodsCSF samples from 26 sTBI patients, collected between 6 and 240 h post-injury, and 19 healthy controls were analyzed using an optimized direct enzyme-linked immunosorbent assay (ELISA; sensitivity <4.7 pg/mL) for pTau-217 detection, complemented by Western blot validation. Temporal analysis, ROC curves, and trajectory clustering were used for interpretation.ResultsCSF pTau-217 levels were significantly elevated in sTBI patients at 6, 12, 18, 24, and 48 h post-injury compared to controls (p < 0.05–p < 0.001), peaking around 18 h (~65 ng/mL) before declining to near-control levels by 120 h. ROC analyses showed AUC of 0.78 (6–12 h) and 0.83 (24–48 h). Clustering identified a subgroup with sustained high pTau-217, associated with diffuse axonal injury and worse 6-month outcomes. A significant inverse correlation was observed between CSF pTau-217 at 24–48 h and GOSE (ρ = –0.67, p < 0.01).DiscussionThese findings indicate that CSF pTau-217 is a sensitive and early biomarker of acute tau pathology in sTBI. Its diagnostic performance and association with axonal injury and outcome support its utility, though longitudinal validation in larger cohorts is required to confirm clinical relevance.

Diffuse axonal brain injury (DAI) is a common, debilitating consequence of traumatic brain injury, yet its detection and severity grading remain challenging in clinical and experimental settings. This study evaluated the sensitivity of diffusion tensor imaging (DTI), histology, and neurological severity scoring (NSS) in assessing injury severity in a rat model of isolated DAI. A rotational injury model induced mild, moderate, or severe DAI in male and female rats. Neurological deficits were assessed 48 h after injury via NSS. Magnetic resonance imaging, including DTI metrics, such as fractional anisotropy (FA), relative anisotropy (RA), axial diffusivity (AD), mean diffusivity (MD), and radial diffusivity (RD), was performed prior to tissue collection. Histological analysis used beta amyloid precursor protein immunohistochemistry. Sensitivity and variability of each method were compared across brain regions and the whole brain. Histology was the most sensitive method, requiring very small groups to detect differences. Anisotropy-based MRI metrics, especially whole-brain FA and RA, showed strong correlations with histology and NSS and demonstrated high sensitivity with low variability. NSS identified injury but required larger group sizes. Diffusivity-based MRI metrics, particularly RD, were less sensitive and more variable. Whole-brain FA and RA were the most sensitive MRI measures of DAI severity and were comparable to histology in moderate and severe groups. These findings support combining NSS and anisotropy-based DTI for non-terminal DAI assessment in preclinical studies.