Investigating neuropathological changes and underlying neurobiological mechanisms in the early stages of primary blast-induced traumatic brain injury: Insights from a rat model.

The utilization of explosives and chemicals has resulted in a rise in blast-induced traumatic brain injury (bTBI) in recent times. However, there is a dearth of diagnostic biomarkers and therapeutic targets for bTBI due to a limited understanding of biological mechanisms, particularly in the early stages. The objective of this study was to examine the early neuropathological characteristics and underlying biological mechanisms of primary bTBI. A total of 83 Sprague Dawley rats were employed, with their heads subjected to a blast shockwave of peak overpressure ranging from 172 to 421 kPa in the GI, GII, and GIII groups within a closed shock tube, while the body was shielded. Neuromotor dysfunctions, morphological changes, and neuropathological alterations were detected through modified neurologic severity scores, brain water content analysis, MRI scans, histological, TUNEL, and caspase-3 immunohistochemical staining. In addition, label-free quantitative (LFQ)-proteomics was utilized to investigate the biological mechanisms associated with the observed neuropathology. Notably, no evident damage was discernible in the GII and GI groups, whereas mild brain injury was observed in the GIII group. Neuropathological features of bTBI were characterized by morphologic changes, including neuronal injury and apoptosis, cerebral edema, and cerebrovascular injury in the shockwave's path. Subsequently, 3153 proteins were identified and quantified in the GIII group, with subsequent enriched neurological responses consistent with pathological findings. Further analysis revealed that signaling pathways such as relaxin signaling, hippo signaling, gap junction, chemokine signaling, and sphingolipid signaling, as well as hub proteins including Prkacb, Adcy5, and various G-protein subunits (Gnai2, Gnai3, Gnao1, Gnb1, Gnb2, Gnb4, and Gnb5), were closely associated with the observed neuropathology. The expression of hub proteins was confirmed via Western blotting. Accordingly, this study proposes signaling pathways and key proteins that exhibit sensitivity to brain injury and are correlated with the early pathologies of bTBI. Furthermore, it highlights the significance of G-protein subunits in bTBI pathophysiology, thereby establishing a theoretical foundation for early diagnosis and treatment strategies for primary bTBI.

Effects of different preservation on the mechanical properties of cortical bone under quasi-static and dynamic compression.

Introduction: Mechanical properties of biological tissue are important for numerical simulations. Preservative treatments are necessary for disinfection and long-term storage when conducting biomechanical experimentation on materials. However, few studies have been focused on the effect of preservation on the mechanical properties of bone in a wide strain rate. The purpose of this study was to evaluate the influence of formalin and dehydration on the intrinsic mechanical properties of cortical bone from quasi-static to dynamic compression. Methods: Cube specimens were prepared from pig femur and divided into three groups (fresh, formalin, and dehydration). All samples underwent static and dynamic compression at a strain rate from 10-3 s-1 to 103 s-1. The ultimate stress, ultimate strain, elastic modulus, and strain-rate sensitivity exponent were calculated. A one-way ANOVA test was performed to determine if the preservation method showed significant differences in mechanical properties under at different strain rates. The morphology of the macroscopic and microscopic structure of bones was observed. Results: The results show that ultimate stress and ultimate strain increased as the strain rate increased, while the elastic modulus decreased. Formalin fixation and dehydration did not affect elastic modulus significantly whereas significantly increased the ultimate strain and ultimate stress. The strain-rate sensitivity exponent was the highest in the fresh group, followed by the formalin group and dehydration group. Different fracture mechanisms were observed on the fractured surface, with fresh and preserved bone tending to fracture along the oblique direction, and dried bone tending to fracture along the axial direction. Discussion: In conclusion, preservation with both formalin and dehydration showed an influence on mechanical properties. The influence of the preservation method on material properties should be fully considered in developing a numerical simulation model, especially for high strain rate simulation.

A neuromuscular human body model for lumbar injury risk analysis in a vibration loading environment.

BACKGROUND AND OBJECTIVE: Long-term intensive exposure to whole-body vibration substantially increases the risk of low back pain and degenerative diseases in special occupational groups, like motor vehicle drivers, military vehicle occupants, aircraft pilots, etc. This study aims to establish and validate a neuromuscular human body model focusing on improvement of the detailed description of anatomic structures and neural reflex control, for lumbar injury analysis in vibration loading environments. METHODS: A whole-body musculoskeletal in Opensim codes was first improved by including a detailed anatomic description of spinal ligaments, non-linear intervertebral disc, and lumbar facet joints, and coupling a proprioceptive feedback closed-loop control strategy with GTOs and muscle spindles modeling in Python codes. Then, the established neuromuscular model was multi-levelly validated from sub-segments to the whole model, from regular movements to dynamic responses to vibration loadings. Finally, the neuromuscular model was combined with a dynamic model of an armored vehicle to analyze occupant lumbar injury risk in vibration loadings due to different road conditions and traveling velocities. RESULT: Based on a series of biomechanical indexes, including lumbar joint rotation angles, the lumbar intervertebral pressures, the displacement of the lumbar segments, and the lumbar muscle activities, the validation results show that the present neuromuscular model is available and feasible in predicting lumbar biomechanical responses in normal daily movement and vibration loading environments. Furthermore, the combined analysis with the armored vehicle model predicted similar lumbar injury risk to the experimental or epidemiologic studies. The preliminary analysis results also showed that road types and travelling velocities have substantial combined effects on lumbar muscle activities, and indicated that intervertebral joint pressure and muscle activity indexes can need to be jointly considered for lumbar injury risk evaluation. CONCLUSION: In conclusion, the established neuromuscular model is an effective tool to evaluate vibration loading effects on injury risk of the human body and assist vehicle design vibration comfort by directly concerning the human body injury itself.

[Study on injury parameters of severe blast injury in mice at plain and plateau based on equivalent trauma].

OBJECTIVE: To explore the establishment of the interconvertible injury parameters of same severe blast injury in mice at plain and plateau. METHODS: A total of 157 C57BL/6 male mice were randomly divided into plain control group (8 mice), plain injury group (77 mice), plateau control group (8 mice) and plateau injury group (64 mice) according to random number table method. The mice in plateau control group and plateau blast injury group had been placed in animal experimental low-pressure oxygen chamber to simulate 4 000 meters plateau environment for 5 days in advance. Then the mice in plain blast injury group and plateau blast injury group were put into biological shock tube, respectively. Different pressures of the driving section were selected to establish the severe blast injury models in mice at plain and 4 000 meters plateau to reach approximately 70% mortality within 72 hours. The equivalent traumatic condition at 24 hours after blast injury in different groups was verified by the series of experiments including gross autopsy, lung wet/dry weight ratio (W/D), hematoxylin-eosin (HE) staining and histological scoring. RESULTS: The mice mortality were basically consistent between the plain injury group (65%) and plateau injury group (75%) when 5.4 MPa and 4.0 MPa of the driving section pressures were chosen, respectively. Compared with the corresponding control groups, the lungs showed massive hemorrhage (patchy and diffuse) with significant pulmonary edema in both plain 5.4 MPa-injured group and the plateau 4.0 MPa-injured group at 24 hours after blast injury. Compared with the plateau control group, the pulmonary W/D ratio were significantly increased in the plateau injury group (5.579±0.646 vs. 4.476±0.076, P < 0.05), while the difference between plateau injury group and the plain control group was not statistically significant (5.303±1.020 vs. 4.015±0.144, P > 0.05). Also, compared with the corresponding control groups, the analysis of lung histopathological sections showed that there were several pathological changes including large alveolar rupture and fusion, thickened alveolar walls, and a small amount of inflammatory cell infiltration in the alveolar lumen in the groups of plain 5.4 MPa and plateau 4.0 MPa. In addition, the histopathological scores of lung in the groups of plain 5.4 MPa and plateau 4.0 MPa were significantly higher than that in corresponding control group (8.67±0.82 vs. 1.67±0.52, 9.00±1.10 vs. 2.17±0.41, both P < 0.05), however, there was no statistical difference for the above score between plain blast injury group and plateau blast injury group. CONCLUSIONS: The pressures of driving section 5.4 MPa and 4.0 MPa are injury parameters to establish equivalent severe blast injury in mice at plain and plateau, respectively, which can be converted to each other. This study provides support for the application and evaluation of prevention and treatment technology for severe blast injury in special environment.

Research on skeletal muscle impact injury using a new rat model from a bioimpact machine.

Introduction: Skeletal muscle impact injury occurs frequently during sports, falls, and road traffic accidents. From the reported studies on skeletal muscle injury, it is difficult to determine the injury parameters. Therefore, we developed a new model of gastrocnemius impact injury in rats with a bioimpact machine, with which the experimental operation could be conducted in feasibility from the recorded parameters. Through this novel model, we study the skeletal muscle impact injury mechanisms by combining temporal and spatial variation. Methods: The gastrocnemius of anesthetized rats was injured by a small pneumatic-driven bioimpact machine; the moving speed and impact force were determined, and the whole impact process was captured by a high-speed camera. We observed the general condition of rats and measured the changes in injured calf circumference, evaluating calf injuries using MRI, gait analysis system, and pathology at different times after the injury. Results: The gastrocnemius was injured at an impact speed of 6.63 m/s ± 0.25 m/s and a peak force of 1,556.80 N ± 110.79 N. The gait analysis system showed that the footprint area of the RH limb decreased significantly on the first day and then increased. The calf circumference of the injured limb increased rapidly on the first day post-injury and then decreased in the next few days. MRI showed edema of subcutaneous and gastrocnemius on the first day, and the area of edema decreased over the following days. HE staining showed edema of cells, extensive hyperemia of blood vessels, and infiltration of inflammatory cells on the first day. Cell edema was alleviated day by day, but inflammatory cell infiltration was the most on the third day. TEM showed that the sarcoplasmic reticulum was dilated on the first day, the mitochondrial vacuolation was obvious on the second day, and the glycogen deposition was prominent on the fifth day. Conclusion: In our experiment, we developed a new and effective experimental animal model that was feasible to operate; the injured area of the gastrocnemius began to show "map-like" changes in the light microscope on the third day. Meanwhile, the gastrocnemius showed a trend of "edema-mitochondrial vacuolation-inflammatory cell aggregation" after impact injury.

Experimental Study on Intracranial Pressure and Biomechanical Response in Rats under the Blast Wave.

Explosion overpressure propagates extracranially and causes craniocerebral injury after being transmitted into the brain. Studies on the extent of skull to reduce impact overpressure are still lacking. Therefore, it is necessary to study the relationship between intracranial pressure (ICP) and external field pressure and the situation of craniocerebral injury under the blast wave. Pressure sensor of ϕ 1.2 mm was disposed 3 mm posterior to the bregma of rat skull, and type I biological shock tube (BST-I) was used as the source of injury while a side-on air pressure sensor was installed at the horizontal position of the ICP sensor. Eleven groups of blast experiments with peak air overpressure ranging from 167 kPa to 482 kPa were performed to obtain the variation law of ICP and injury of rats. Data measured by sensors show that the peak pressure formed in the rat brain are lower than the external air overpressure; the differential pressure between the inside and outside of the brain is 27-231 kPa. When side-on air overpressure is ≤363 kPa, ICP is ≤132 kPa, and the hemorrhage area of the rat's brain is <15%, the injury is minor. When side-on air overpressure is 363 kPa-401 kPa, ICP range is from 132 kPa to 248 kPa, hemorrhage area is about 15%-20%, and the injury increases. When side-on air overpressure is 401 kPa-435 kPa, ICP range from 248 kPa to 348 kPa, the hemorrhage area is about 20%-24%, and the injury is serious. When side-on air overpressure ≥482 kPa, the peak ICP surged to 455 kPa and the peak negative ICP reached -84 kPa, the hemorrhage area exceeded 26%. When the external blast wave is weak, skull can absorb the blast wave better, reducing the pressure by 81.4%, when the external shockwave is strong, skull only reduces the pressure by 5.6%, but both can play certain protective role. The fitting curve of air overpressure and ICP can be used to predict the changes of ICP under different external blast overpressure. Analysis of cranial injury showed that the area of cranial hemorrhage with extremely severe injury increased by 107.9% compared with mild injury, increased by 53.3% compared with moderate injury, and increased by 21.6% compared with severe injury. This work may provide references for the dynamic response of biological cranial and brain injury mechanism under the effect of blast wave.

Effects of altitude changes on mild-to-moderate closed-head injury in rats following acute high-altitude exposure.

Mild-to-moderate closed-head injury (mmCHI) is an acute disease induced by high-altitudes. It is general practice to transfer patients to lower altitudes for treatment, but the pathophysiological changes at different altitudes following mmCHI remain unknown. The present study simulated acute high-altitude exposure (6,000 m above sea level) in rats to establish a model of mmCHI and recorded their vital signs. The rats were then randomly assigned into different altitude exposure groups (6,000, 4,500 and 3,000 m) and neurological severity score (NSS), body weight (BW), brain magnetic resonance imaging (MRI), brain water content (BWC) and the ratio of BW/BWC at 6, 12 and 24 h following mmCHI, and the glial fibrillary acidic protein levels were analysed in all groups. The results revealed that within the first 24 h following acute high-altitude exposure, mmCHI induced dehydration, brain oedema and neuronal damage. Brain injury in rats was significantly reversed following descent to 4,500 m compared with the results from 6,000 or 3,000 m. The results indicated that subjects should be transported as early as possible. Furthermore, avoiding large-span descent altitude was beneficial to reduce neurological impairment. The examination of brain-specific biomarkers and MRI may further be useful in determining the prognosis of high-altitude mmCHI. These results may provide guidance for rescuing high altitude injuries.

Novel-graded traumatic brain injury model in rats induced by closed head impacts.

Traumatic brain injury (TBI) is a leading cause of death and disability worldwide. Due to the heterogeneity of human TBI, none of the available animal models can reproduce the entire spectrum of TBI. This study was designed to develop a novel-graded TBI rat model which is induced by closed head impacts (CHI) with reproducible brain damage and neurological dysfunction. A total of 75 male Sprague-Dawley rats (200 ± 20 g) were randomly equally divided into five groups: the Sham, 0.5, 0.6, 0.7 and 0.8 MPa groups. A custom-made, air-driven injury apparatus was used to induce CHIs (from 0.5 to 0.8 MPa). The kinematic parameters during the procedure were recorded by a force sensor and a high-speed camera. Mortality rate, duration of unconsciousness (latency period of righting reflex), modified neurological severity score (mNSS) and whole brain water content (BWC) were examined. Pathological changes were evaluated by hematoxylin-eosin (HE) stain and immunohistochemical stain for amyloid precursor protein (APP). The impact force and speed were 785.3 ± 14.12 N and 5.71 m/s in the 0.5 MPa group, 837.72 ± 10.41 N and 6.06 m/s in the 0.6 MPa group, 857.65 ± 11.11 N and 6.25 m/s in the 0.7 MPa group, and 955.6 ± 16.35 N and 6.67 m/s in the 0.8 MPa group. The periods of loss of righting reflex in 0.6-0.8 MPa groups were significantly higher than that in the Sham group. The mNSS score and BWC of the 0.8 MPa group remained higher 24 h after injury than other groups. Brain damage was indicated by increased APP expression in TBI rats. In conclusion, the newly developed CHI rat model was a highly controlled and reproducible graded TBI model, and provided a useful tool to investigate the underlying mechanism and therapeutic effects of TBI with various injury severities.

Biomechanical Responses and Injury Characteristics of Knee Joints under Longitudinal Impacts of Different Velocities.

BACKGROUND AND OBJECTIVE: Knee joint collision injuries occur frequently in military and civilian scenarios, but there are few studies assessing longitudinal impacts on knee joints. In this study, the mechanical responses and damage characteristics of knee longitudinal collisions were investigated by finite element analysis and human knee impact tests. MATERIALS AND METHODS: Based on a biocollision test plateau, longitudinal impact experiments were performed on 4 human knee joints (2 in the left knee and 2 in the right knee) to measure the impact force and stress response of the bone. And then a finite element model of knee joint was established from the Chinese Visible Human (CVH), with which longitudinal impacts to the knee joint were simulated, in which the stress response was determined. The injury response of the knee joint-sustained longitudinal impacts was analyzed from both the experimental model and finite element analysis. RESULTS: The impact experiments and finite element simulation found that low-speed impact mainly led to medial injuries and high-speed impact led to both medial and lateral injuries. In the knee joint impact experiment, the peak flexion angles were 13.8° ± 1.2, 30.2° ± 5.1, and 92.9° ± 5.45 and the angular velocities were 344.2 ± 30.8 rad/s, 1510.8 ± 252.5 rad/s, and 9290 ± 545 rad/s at impact velocities 2.5 km/h, 5 km/h, and 8 km/h, respectively. When the impact velocity was 8 km/h, 1 knee had a femoral condylar fracture and 3 knees had medial tibial plateau fractures or collapse fractures. The finite element simulation of knee joints found that medial cortical bone stress appeared earlier than the lateral peak and that the medial bone stress concentration was more obvious when the knee was longitudinally impacted. CONCLUSION: Both the experiment and FE model confirmed that the biomechanical characteristics of the injured femur and medial tibia are likely to be damaged in a longitudinal impact, which is of great significance for the prevention and treatment of longitudinal impact injuries of the knee joint.

Experimental Study of Thoracoabdominal Injuries Suffered from Caudocephalad Impacts Using Pigs.

To know the caudocephalad impact- (CCI-) induced injuries more clearly, 21 adult minipigs, randomly divided into three groups: control group (n = 3), group I (n = 9), and group II (n = 9), were used to perform the CCI experiments on a modified deceleration sled. Configured impact velocity was 0 m/s in the control group, 8 m/s in group I, and 11 m/s in group II. The kinematics and mechanical responses of the subjects were recorded and investigated. The functional change examination and the autopsies were carried out, with which the injuries were evaluated from the Abbreviated Injury Scale (AIS) and the Injury Severity Score (ISS). The subjects in group I and group II experienced the caudocephalad loading at the peak pelvic accelerations of 108.92 ± 58.87 g and 139.13 g ± 78.54 g, with the peak abdomen pressures, 41.24 ± 16.89 kPa and 63.61 ± 65.83 kPa, respectively. The injuries of the spleen, lung, heart, and spine were detected frequently among the tested subjects. The maximal AIS (MAIS) of chest injuries was 4 in group I and 5 in group II, while both the MAIS of abdomen injuries in group I and group II were 5. The ISS in group II was 52.71 ± 6.13, significantly higher than in group I, 26.67 ± 5.02 (p < 0.05). The thoracoabdomen CCI injuries and the mechanical response addressed presently may be useful to conduct both the prevention studies against military or civilian injuries.

Preliminary study on alterations of altitude road traffic in China from 2006 to 2013.

INTRODUCTION: Road traffic can play an important role in strengthening regional economic activities, especially at high altitude, and it is necessary to know important traffic-related information. Although previous studies reported on road traffic in China, there has been little research on high-altitude road traffic to date. METHOD: The annual official census of road traffic safety from 2006 to 2013 was used to obtain data on the general population, registered drivers, registered vehicles, newly built roads, road traffic accidents (RTAs), mortality rate per 100 000 populations and per 10 000 vehicles in high-altitude provinces, including Tibet, Qinghai, Xinjiang, Gansu, Yunnan, Sichuan, and Chongqing. These provincial data were reviewed retrospectively, with the national data as the reference. Statistical analysis (i.e., t test) was used to compare the estimated average annual change rate of population, number of registered drivers, registered vehicles, and newly built roads in high-altitude provinces with the national rates. RESULTS: Compared with the national data, there are significantly higher annual rates of population growth in Tibet and Xinjiang, registered drivers in Gansu, registered vehicles in Gansu, Sichuan, and Chongqing, and newly built roads in Tibet and Qinghai. Among the investigated provinces, Tibet, Qinghai, and Yunnan had a higher proportion of the roads with the high class. RTAs and RTA-induced casualties in the high-altitude provinces indicated a decreasing trend. The mortality rate per 10 000 vehicles and per 100 000 populations showed a decreasing trend, while the RTA-related mortality rate in Tibet, Qinghai, Xinjiang and Gansu remained high. CONCLUSIONS: Major changes for road traffic in high-altitude provinces have occurred over the past decade; however, the RTA-related mortality rate in high-altitude provinces has remained high. This study furthers understanding about road traffic safety in China; further studies on road traffic safety at high altitude should be performed.

Mechanical Characterization of Immature Porcine Brainstem in Tension at Dynamic Strain Rates.

BACKGROUND: Many brain injury cases involve pediatric road traffic accidents, and among these, brainstem injury causes disastrous outcomes. A thorough understanding of the tensile characterization of immature brainstem tissue is crucial in modeling traumatic brain injury sustained by children, but limited experimental data in tension is available for the immature brain tissue at dynamic strain rates. MATERIAL AND METHODS: We harvested brainstem tissue from immature pigs (about 4 weeks old, and at a developmental stage similar to that of human toddlers) as a byproduct from a local slaughter house and very carefully prepared the samples. Tensile tests were performed on specimens at dynamic strain rates of 2/s, 20/s, and 100/s using a biological material instrument. The constitutive models, Fung, Ogden, Gent, and exponential function, for immature brainstem tissue material property were developed for the recorded experimental data using OriginPro 8.0 software. The t test was performed for infinitesimal shear modules. RESULTS: The curves of stress-versus-stretch ratio were convex in shape, and inflection points were found in all the test groups at the strain of about 2.5%. The average Lagrange stress of the immature brainstem specimen at the 30% strain at the strain rates of 2, 20, and 100/s was 273±114, 515±107, and 1121±197 Pa, respectively. The adjusted R-Square (R2) of Fung, Ogden, Gent, and exponential model was 0.820≤R2≤0.933, 0.774≤R2≤0.940, 0.650≤R2≤0.922, and 0.852≤R2≤0.981, respectively. The infinitesimal shear modulus of the strain energy functions showed a significant association with the strain rate (p<0.01). CONCLUSIONS: The immature brainstem is a rate-dependent material in dynamic tensile tests, and the tissue becomes stiffer with increased strain rate. The reported results may be useful in the study of brain injuries in children who sustain injuries in road traffic accidents. Further research in more detail should be performed in the future.