Brain Sham Research Articles

Targeting of nanoparticles to the cerebral vasculature after traumatic brain injury.

Traumatic brain injury has faced numerous challenges in drug development, primarily due to the difficulty of effectively delivering drugs to the brain. However, there is a potential solution in targeted drug delivery methods involving antibody-drug conjugates or nanocarriers conjugated with targeting antibodies. Following a TBI, the blood-brain barrier (BBB) becomes permeable, which can last for years and allow the leakage of harmful plasma proteins. Consequently, an appealing approach for TBI treatment involves using drug delivery systems that utilize targeting antibodies and nanocarriers to help restore BBB integrity. In our investigation of this strategy, we examined the efficacy of free antibodies and nanocarriers targeting a specific endothelial surface marker called vascular cell adhesion molecule-1 (VCAM-1), which is known to be upregulated during inflammation. In a mouse model of TBI utilizing central fluid percussion injury, free VCAM-1 antibody did not demonstrate superior targeting when comparing sham vs. TBI brain. However, the administration of VCAM-1-targeted nanocarriers (liposomes) exhibited a 10-fold higher targeting specificity in TBI brain than in sham control. Flow cytometry and confocal microscopy analysis confirmed that VCAM-1 liposomes were primarily taken up by brain endothelial cells post-TBI. Consequently, VCAM-1 liposomes represent a promising platform for the targeted delivery of therapeutics to the brain following traumatic brain injury.

Acute gastrointestinal permeability after traumatic brain injury in mice precedes a bloom in Akkermansia muciniphila supported by intestinal hypoxia

Traumatic brain injury (TBI) increases gastrointestinal morbidity and associated mortality. Clinical and preclinical studies implicate gut dysbiosis as a consequence of TBI and an amplifier of brain damage. However, little is known about the association of gut dysbiosis with structural and functional changes of the gastrointestinal tract after an isolated TBI. To assess gastrointestinal dysfunction, mice received a controlled cortical impact or sham brain injury and intestinal permeability was assessed at 4 h, 8 h, 1 d, and 3 d after injury by oral administration of 4 kDa FITC Dextran prior to euthanasia. Quantification of serum fluorescence revealed an acute, short-lived increase in permeability 4 h after TBI. Despite transient intestinal dysfunction, no overt morphological changes were evident in the ileum or colon across timepoints from 4 h to 4 wks post-injury. To elucidate the timeline of microbiome changes after TBI, 16 s gene sequencing was performed on DNA extracted from fecal samples collected prior to and over the first month after TBI. Differential abundance analysis revealed that the phylum Verrucomicrobiota was increased at 1, 2, and 3 d after TBI. The Verrucomicrobiota species was identified by qPCR as Akkermansia muciniphila, an obligate anaerobe that resides in the intestinal mucus bilayer and produces short chain fatty acids (e.g. butyrate) utilized by intestinal epithelial cells. We postulated that TBI promotes intestinal changes favorable for the bloom of A. muciniphila. Consistent with this premise, the relative area of mucus-producing goblet cells in the medial colon was significantly increased at 1 d after injury, while colon hypoxia was significantly increased at 3 d. Our findings reveal acute gastrointestinal functional changes coupled with an increase of beneficial bacteria suggesting a potential compensatory response to systemic stress after TBI.

Neuroprotective Effects of a Serotonin Receptor Peptide Following Sham vs. Mild Traumatic Brain Injury in the Zucker Rat.

Accelerated cognitive decline frequently complicates traumatic brain injury. Obesity and type 2 diabetes mellitus drive peripheral inflammation which may accelerate traumatic brain injury-associated neurodegeneration. The Zucker rat harbors G-protein coupled receptor agonist IgG autoantibodies and in vitro neurotoxicity caused by these autoantibodies was prevented by a novel synthetic fragment of the serotonin 2A receptor. The aim of the present study was to test whether genetic obesity manifested in Zucker diabetic fatty rat is associated with greater spatial memory impairment before and after mild traumatic brain injury compared to Zucker lean rats. Furthermore, we investigated whether these neurodegenerative complications can be lessened by administration of a novel putative neuroprotective peptide comprised of a fragment of the second extracellular loop of the serotonin 2A receptor. Age-matched lean and fatty diabetic Zucker rats were tested in the Morris water maze (spatial memory) prior to receiving a sham-injury or lateral fluid percussion (LFP) mild traumatic brain injury. Behavioral testing was repeated at 1-week, 1-month, and 3-month intervals following injury. A synthetic peptide consisting of a portion of the 5-hydroxytryptamine (serotonin) 2A receptor (2 mg/kg) (vehicle, or an inactive scrambled version of the peptide (2 mg/kg)) was administered via intraperitoneal route every other day for 7 days after sham or LFP injury to lean rats or 7 days before and after sham or LFP injury to fatty rats. Mild traumatic brain injury impaired recall of spatial memory in fatty and lean rats. Zucker fatty rats subjected to sham-injury or mild TBI experienced a significantly greater longitudinal decline in recall of spatial memory compared to lean Zucker rats. A synthetic peptide fragment of the 5-hydroxytryptamine 2A receptor significantly enhanced acquisition of spatial learning and it appeared to strengthen recall of spatial learning (one-week) after sham injury in Zucker rats. These data suggest that the Zucker diabetic fatty rat is a suitable animal model to investigate the role of metabolic factor(s) in accelerated cognitive decline. A novel synthetic peptide comprised of a fragment of the second extracellular loop of the human serotonin 2A receptor appeared to have neuroprotective effects on both acquisition and recall of spatial memory in subsets of Zucker rats, with relatively greater benefit in sham-injured, lean Zucker rats.

High-Definition Transcranial Direct Current Stimulation in the Right Ventrolateral Prefrontal Cortex Lengthens Sustained Attention in Virtual Reality.

Due to the current limitations of three-dimensional (3D) simulation graphics technology, mind wandering commonly occurs in virtual reality tasks, which has impeded it being applied more extensively. The right ventrolateral prefrontal cortex (rVLPFC) plays a vital role in executing continuous two-dimensional (2D) mental paradigms, and transcranial direct current stimulation (tDCS) over this cortical region has been shown to successfully modulate sustained 2D attention. Accordingly, we further explored the effects of electrical activation of the rVLPFC on 3D attentional tasks using anodal high-definition (HD)-tDCS. A 3D Go/No-go (GNG) task was developed to compare the after effects of real and sham brain stimulation. Specifically, GNG tasks were periodically interrupted to assess the subjective perception of attentional level, behavioral reactions were tracked and decomposed into an underlying decision cognition process, and electroencephalography data were recorded to calculate event-related potentials (ERPs) in rVLPFC. The p-values statistically indicated that HD-tDCS improved the subjective mentality, led to more cautious decisions, and enhanced neuronal discharging in rVLPFC. Additionally, the neurophysiological P300 ERP component and stimulation being active or sham could effectively predict several objective outcomes. These findings indicate that the comprehensive approach including brain stimulation, 3D mental paradigm, and cross-examined performance could significantly lengthen and robustly compare sustained 3D attention.

N6-methyladenosine RNA is modified in the rat hippocampus following traumatic brain injury with hypothermia treatment.

Recent studies have suggested a role for N6-methyladenosine (m6A) modification in neurological diseases. Hypothermia, a commonly used treatment for traumatic brain injury, plays a neuroprotective role by altering m6A modifications. In this study, methylated RNA immunoprecipitation sequencing (MeRIP-Seq) was applied to conduct a genome-wide analysis of RNA m6A methylation in the rat hippocampus of Sham and traumatic brain injury (TBI) groups. In addition, we identified the expression of mRNA in the rat hippocampus after TBI with hypothermia treatment. Compared with the Sham group, the sequencing results of the TBI group showed that 951 different m6A peaks and 1226 differentially expressed mRNAs were found. We performed cross-linking analysis of the data of the two groups. The result showed that 92 hyper-methylated genes were upregulated, 13 hyper-methylated genes were downregulated, 25 hypo-methylated genes were upregulated, and 10 hypo-methylated genes were downregulated. Moreover, a total of 758 differential peaks were identified between TBI and hypothermia treatment groups. Among these differential peaks, 173 peaks were altered by TBI and reversed by hypothermia treatment, including Plat, Pdcd5, Rnd3, Sirt1, Plaur, Runx1, Ccr1, Marveld1, Lmnb2, and Chd7. We found that hypothermia treatment transformed some aspects of the TBI-induced m6A methylation landscape of the rat hippocampus.

Application of non-invasive brain stimulation with the use of repetitive transcranial magnetic stimulation (rTMS) or transcranial direct current stimulation (tDCS) in the treatment of dysphagia following an ischemic stroke – analysis of research reports

Stroke is one of the significant problems and causes of death, in particular in highly developed countries. It is also the most common cause of dysphagia. This study is devoted to the analysis of publications from the last decade concerning research on the use of non-invasive brain stimulation (TMS, tDCS) in the treatment of dysphagia following an ischemic stroke. The following databases were searched for publications: PUBMED, Polish Scientific Journals Database, EBSCO, ScienceDirect. Out of 358 articles found, only two met all the inclusion conditions. The studies discussed in this article included patients who had their first unilateral ischemic stroke, followed by dysphagia. Findings from pilot studies on the effectiveness of the use of tDCS in the treatment of dysphagia following an ischemic stroke were analysed. Fourteen patients aged 50-92 were randomly assigned to two groups. The study group was treated with anodic stimulation, and the control group with sham brain stimulation. The results showed that patients in the study group obtained a statistically significant result, indicating an improvement in the swallowing function as measured by the Dysphagia Score and Severity Scale. The above data may indicate the effectiveness of the use of tDCS in the treatment of dysphagia. Fifty patients were qualified for the research on the effectiveness of rTMS in the treatment of dysphagia. Three groups were created: the first group treated with high frequency rTMS, where patients received rTMS stimulation – 3Hz; the second group with low frequency rTMS – 1Hz; and the control group. The effectiveness of the therapy used was assessed on the fifth day and after 1, 2 and 3 months. After 5 days the groups where active rTMS stimulation was used showed greater improvement in the swallowing function compared to the sham stimulation group. Improved results in the Standardized Swallowing Assessment were also recorded after 3 months in the 1Hz and 3Hz groups, but this did not apply to the control group. In three groups, the results of the water swallow test and the degree of dysphagia improved after 3 months. This analysis shows that non-invasive brain stimulation using tDCS and rTMS in the treatment of dysphagia is associated with improved swallowing function. However, the small number of studies conducted in this area does not allow for extrapolation of their results.

Zolpidem Profoundly Augments Spared Tonic GABAAR Signaling in Dentate Granule Cells Ipsilateral to Controlled Cortical Impact Brain Injury in Mice.

Type A GABA receptors (GABAARs) are pentameric combinations of protein subunits that give rise to tonic (ITonicGABA) and phasic (i.e., synaptic; ISynapticGABA) forms of inhibitory GABAAR signaling in the central nervous system. Remodeling and regulation of GABAAR protein subunits are implicated in a wide variety of healthy and injury-dependent states, including epilepsy. The present study undertook a detailed analysis of GABAAR signaling using whole-cell patch clamp recordings from mouse dentate granule cells (DGCs) in coronal slices containing dorsal hippocampus at 1–2 or 8–13 weeks after a focal, controlled cortical impact (CCI) or sham brain injury. Zolpidem, a benzodiazepine-like positive modulator of GABAARs, was used to test for changes in GABAAR signaling of DGCs due to its selectivity for α1 subunit-containing GABAARs. Electric charge transfer and statistical percent change were analyzed in order to directly compare tonic and phasic GABAAR signaling and to account for zolpidem’s ability to modify multiple parameters of GABAAR kinetics. We observed that baseline ITonicGABA is preserved at both time-points tested in DGCs ipsilateral to injury (Ipsi-DGCs) compared to DGCs contralateral to injury (Contra-DGCs) or after sham injury (Sham-DGCs). Interestingly, application of zolpidem resulted in modulation of ITonicGABA across groups, with Ipsi-DGCs exhibiting the greatest responsiveness to zolpidem. We also report that the combination of CCI and acute application of zolpidem profoundly augments the proportion of GABAAR charge transfer mediated by tonic vs. synaptic currents at both time-points tested, whereas gene expression of GABAAR α1, α2, α3, and γ2 subunits is unchanged at 8–13 weeks post-injury. Overall, this work highlights the shift toward elevated influence of tonic inhibition in Ipsi-DGCs, the impact of zolpidem on all components of inhibitory control of DGCs, and the sustained nature of these changes in inhibitory tone after CCI injury.

CGRP monoclonal antibody prevents the loss of diffuse noxious inhibitory controls (DNIC) in a mouse model of post-traumatic headache.

Determine the role of calcitonin-gene related peptide in promoting post-traumatic headache and dysregulation of central pain modulation induced by mild traumatic brain injury in mice. Mild traumatic brain injury was induced in lightly anesthetized male C57BL/6J mice by a weight drop onto a closed and unfixed skull, which allowed free head rotation after the impact. We first determined possible alterations in the diffuse noxious inhibitory controls, a measure of net descending pain inhibition called conditioned pain modulation in humans at day 2 following mild traumatic brain injury. Diffuse noxious inhibitory control was assessed as the latency to a thermally induced tail-flick that served as the test stimulus in the presence of right forepaw capsaicin injection that provided the conditioning stimulus. Post-traumatic headache-like behaviors were assessed by the development of cutaneous allodynia in the periorbital and hindpaw regions after mild traumatic brain injury. We then determined if intraperitoneal fremanezumab, an anti-calcitonin-gene related peptide monoclonal antibody or vehicle administered 2 h after sham or mild traumatic brain injury induction could alter cutaneous allodynia or diffuse noxious inhibitory control responses on day 2 post mild traumatic brain injury. In naïve and sham mice, capsaicin injection into the forepaw elevated the latency to tail-flick, reflecting the antinociceptive diffuse noxious inhibitory control response. Periorbital and hindpaw cutaneous allodynia, as well as a loss of diffuse noxious inhibitory control, was observed in mice 2 days after mild traumatic brain injury. Systemic treatment with fremanezumab blocked mild traumatic brain injury-induced cutaneous allodynia and prevented the loss of diffuse noxious inhibitory controls in mice subjected to a mild traumatic brain injury. Sequestration of calcitonin-gene related peptide in the initial stages following mild traumatic brain injury blocked the acute allodynia that may reflect mild traumatic brain injury-related post-traumatic headache and, additionally, prevented the loss of net descending inhibition within central pain modulation pathways. As loss of conditioned pain modulation has been linked to multiple persistent pain conditions, dysregulation of descending modulatory pathways may contribute to the persistence of post-traumatic headache. Additionally, evaluation of the conditioned pain modulation/diffuse noxious inhibitory controls response may serve as a biomarker of vulnerability for chronic/persistent pain. These findings suggest that early anti-calcitonin-gene related peptide intervention has the potential to be effective both for the treatment of mild traumatic brain injury-induced post-traumatic headache, as well as inhibiting mechanisms that may promote post-traumatic headache persistence.

Cognitive effects and acceptability of non-invasive brain stimulation on Alzheimer’s disease and mild cognitive impairment: a component network meta-analysis

ObjectivesTo compare cognitive effects and acceptability of repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) in patients with Alzheimer’s disease (AD) or mild cognitive impairment (MCI), and...

Design of a Novel Paired Associative Nerve Stimulation System and Treatment Strategy for Incomplete Spinal Cord Injury: A Preliminary Study.

Paired associative nerve stimulation (PANS) was proposed as a potential nerve rehabilitation treatment strategy. However, few relevant documents are available regarding the strategy, and only a few clinical studies have involved healthy people. To determine the feasibility of the neurorehabilitation treatment and to estimate the effect of PANS on nerve plasticity for individuals with incomplete spinal cord injury (iSCI), a design combining repetitive transcranial magnetic stimulation (rTMS) with trans-spinal electrical stimulation was developed for treating individuals with iSCI in this pilot case study. First, a novel PANS system with multiple stimulation modes was designed and verified with resistors and a metal coil as load. Then, the system was applied to three individuals with iSCI, and five types of paired associative stimulation was performed to confirm the feasibility of the system and determine the most effective treatment strategy. The preliminary result showed that 20-Hz rTMS combined with cathodal trans-spinal direct current stimulation (tsDCS) had the greatest effect on corticospinal excitability. Next, stimulations of 20-Hz rTMS (brain) and sham (spine) as well as sham (brain) and cathode tsDCS (spine) were administered to individuals with iSCI, and the results revealed that paired associative stimulation of brain and spine was more effective than only 20-Hz rTMS brain stimulation or cathodal tsDCS stimulation for corticospinal plasticity.

Abstract 361: The Augmentation of Oxygen-sensitive Generation of Oxygen Radicals in Mitochondria and the Attenuation of Hyperoxia-induced Oxidative Stress After Cardiac Arrest

Introduction: Hyperoxia can induce oxidative stress resulting in organ injuries after cardiac arrest (CA). Mitochondrial reactive oxygen species (ROS) are key to understanding its mechanism, however the augmentation of oxygen-sensitive ROS generation caused by CA ( Fig 1 ) has not been well described. Methods: Rats were assigned into: 1) sham with normoxia, 30% O2; 2) sham with hyperoxia, 100% O2; 3) CA with normoxia; and 4) CA with hyperoxia (n=15 for each group). CA was induced by 10-minute asphyxia and CPR was delivered. Neurological deficit score (NDS), histological lung injury score (LIS), survival time were obtained from the CA groups. Carbonyl protein as an oxidative stress indicator was measured at 2 hrs after resuscitation. H2O2 generation from isolated mitochondria were measured ex vivo at a normoxic and hyperoxic condition set up with nitrogen and air saturated medium. Results: Between the CA groups, the normoxia group had a higher 48hr-survival rate (77%), lower NDS (359±140), and lower LIS (4.3±2.9) compared to those (28%, 452±85, 14±2, respectively) of the hyperoxia group (p<0.01, p<0.05, and p<0.01, respectively). In the brain, lung, and kidney, CA augmented the hyperoxia-induced increase in carbonyl protein (absolute change: sham brain, 0.18 nmol/mg protein vs. CA brain, 0.61 nmol/mg protein; sham lung, 0.29 nmol/mg protein vs. CA lung, 0.72 nmol/mg protein; sham kidney, 0.04 nmol/mg protein vs. CA kidney, 0.55 nmol/mg protein, respectively). In the brain and kidney isolated mitochondria, CA augmented the hyperoxia-induced increase in H2O2 (absolute change: sham brain, 18 pmol/mg protein vs. CA brain, 60 pmol/mg protein; sham kidney, 16 pmol/mg protein vs. CA kidney, 45 pmol/mg protein, respectively). Conclusions: The normoxic therapy may attenuate organ damage and improve survival of CA. The reduction of augmented mitochondrial ROS generation, which is oxygen-sensitive, would be an important mechanism and therapeutic target of post-CA care.

Longitudinal Characterization of Blood-Brain Barrier Permeability after Experimental Traumatic Brain Injury by In Vivo 2-Photon Microscopy.

Vasogenic brain edema (VBE) formation remains an important factor determining the fate of patients with traumatic brain injury (TBI). The spatial and temporal development of VBE, however, remains poorly understood because of the lack of sufficiently sensitive measurement techniques. To close this knowledge gap, we directly visualized the full time course of vascular leakage after TBI by in vivo 2-photon microscopy (2-PM). Male C57BL/6 mice (n = 6/group, 6-8 weeks old) were assigned randomly to sham operation or brain trauma by controlled cortical impact. A cranial window was prepared, and tetramethylrhodamine-dextran (TMRM, MW 40,000 Da) was injected intravenously to visualize blood plasma 4 h, 24 h, 48 h, 72 h, or seven days after surgery or trauma. Three regions with increasing distance to the primary contusion were investigated up to a depth of 300 μm by 2-PM. No TMRM extravasation was detected in sham-operated mice, while already 4 h after TBI vascular leakage was significantly increased (p < 0.05 vs. sham) and reached its maximum at 48 h after injury. Vascular leakage was most pronounced in the vicinity of the contusion. The rate of extravasation showed a biphasic pattern, peaking 4 h and 48-72 h after trauma. Taken together, longitudinal quantification of vascular leakage after TBI in vivo demonstrates that VBE formation after TBI develops in a biphasic manner suggestive of acute and delayed mechanisms. Further studies using the currently developed dynamic in vivo imaging modalities are needed to investigate these mechanisms and potential therapeutic strategies in more detail.

Pharmacological, Psychological, and Noninvasive Brain Stimulation Interventions for Treating Depression After Stroke.

Pharmacological, Psychological, and Noninvasive Brain Stimulation Interventions for Treating Depression After Stroke.

Pharmacological, psychological, and non-invasive brain stimulation interventions for treating depression after stroke.

Pharmacological, psychological, and non-invasive brain stimulation interventions for treating depression after stroke.

Empathy to emotional voices and the use of real-time fMRI to enhance activation of the anterior insula

The right anterior insula (AI), known to have a key role in the processing and understanding of social emotions, is activated during tasks that involve the act of empathising. Neurofeedback provides individuals with a visualisation of their own brain activity, enabling them to regulate and modify this activity. Following previous research investigating the ability of individuals to up-regulate right AI activity levels through neurofeedback, we investigated whether this could be similarly accomplished during an empathy task involving auditory stimuli of human positive and negative emotional expressions. Twenty participants, ten with feedback from right anterior insula and ten with feedback from a sham brain region, participated in two sessions that included sixteen neurofeedback runs and four transfer runs. Results showed that for the second session participants in the right AI neurofeedback group demonstrated better ability to up-regulate their right AI compared to the control group who received sham feedback. Examination of the relationship between individual participants’ empathic traits and their ability to up-regulate right AI activity showed that participants low on empathic traits produced a greater increase in activation of right AI by the end of training. Moreover, the response to positively valenced audio stimuli was greater than for negatively valenced stimuli. These results have implications for therapeutic training of empathy in populations with limited empathic response.

P23 Investigating synaptopathy following traumatic brain injury in a preclinical model

ObjectivesThe impact of Traumatic Brain Injury (TBI) on the synapse is poorly understood. We examined the impact of TBI on the distribution of two postsynaptic density proteins (PSD95 and SAP102) at single-synapse resolution.Subjects44 male transgenic knock-in mice, aged 8–16 weeks, expressing fluorescently labelled PSD95 and SAP102.MethodsMice were randomised to a mild lateral fluid percussion injury (LFPI) or sham treatment and brain sections were examined at 7 and 28 days. Using high resolution confocal microscopy and machine learning algorithms, the synaptic puncta density, size and intensity were mapped across 222 brain regions. Microglia and presynaptic changes were indexed using Iba1 and SV2A immunostaining. Pearson correlation and t-tests were used (significance p&lt;0.05).ResultsWe found a significant reduction in synaptic puncta density at 28 days post-injury in brain regions distal to the injury site including the hippocampus. PSD95 and SAP102 density changes had a strong positive correlation at 28 days (r=0.8; p&lt;0.0001). We also observed evidence of synapse recovery in the ipsilateral cortex between 7 and 28 days. Puncta density had a positive correlation with SV2A (r=0.7; p&lt;0.01) and a negative correlation with (r=−0.6; p&lt;0.001) Iba1 count.ConclusionsFocal LFPI induced progressive region-specific loss of synapses for which microglia may play a role. Our study highlights the value of brain-wide synaptome mapping technology and suggests a capacity for synaptic recovery which could be a therapeutic target.

The Effects of Chicken Embryo Brain Extract on Sciatic Nerve Regeneration of Male Rat; An Experimental Study

Objective:To determine the effects of chicken embryo brain extract (BE) on transects sciatic nerve in male rats. Methods:Thirty adult male Sprague-Dawley rats weighing 200 to 250 g, were randomized into three groups treated with (1) sham surgery, (2) normal saline (NS), and (3) brain extract (BE). The BE was taken from incubating chick embryos at day 8. The sciatic nerve was exposed and sharply transected at the mid thigh level. Immediate epineurial repair was then performed. The BE treated animals were given 400 µl/kg of the chick embryo BE intraperitoneal, once daily, for 2 weeks. All animals were evaluated by sciatic functional index (SFI), electrophysiology, histology, and immunohistochemistry at days 28, 90 after surgery. Results:The mean SFI difference between BE and NS groups at days 28, 60 and 90 after surgery was statistically significant (p=0.086). The mean number of myelinated fibers in the BE group was significantly greater than that of the NS group on days 28 and 90 after surgery (p=0.034). At days 28 and 90 after surgery, the mean nerve conduction velocity (NCV) in the BE group was significantly faster than that of the NS group (p=0.041). Conclusion: These results indicate for the first time that chick embryo brain extract can enhance peripheral nerve regeneration in rat.

Postpartum increases in cerebral edema and inflammation in response to placental ischemia during pregnancy

Reduced placental blood flow results in placental ischemia, an initiating event in the pathophysiology of preeclampsia, a hypertensive pregnancy disorder. While studies show increased mortality risk from Alzheimer’s disease, stroke, and cerebrovascular complications in women with a history of preeclampsia, the underlying mechanisms are unknown. During pregnancy, placental ischemia, induced by reducing uterine perfusion pressure (RUPP), leads to cerebral edema and increased blood-brain barrier (BBB) permeability; however whether these complications persist after delivery is not known. Therefore, we tested the hypothesis that placental ischemia contributes to postpartum cerebral edema and neuroinflammation. On gestational day 14, time-pregnant Sprague Dawley rats underwent Sham (n = 10) or RUPP (n = 9) surgery and brain tissue collected 2 months post-delivery. Water content increased in posterior cortex but not hippocampus, striatum, or anterior cerebrum following RUPP. Using a rat cytokine multi-plex kit, posterior cortical IL-17, IL-1α, IL-1β, Leptin, and MIP2 increased while hippocampal IL-4, IL-12(p70) and RANTES increased and IL-18 decreased following RUPP. Western blot analysis showed no changes in astrocyte marker, Glial Fibrillary Acidic Protein (GFAP); however, the microglia marker, ionized calcium binding adaptor molecule (Iba1) tended to increase in hippocampus of RUPP-exposed rats. Immunofluorescence staining revealed reduced number of posterior cortical microglia but increased activated (Type 4) microglia in RUPP. Astrocyte number increased in both regions but area covered by astrocytes increased only in posterior cortex following RUPP. BBB-associated proteins, Claudin-1, Aquaporin-4, and zonular occludens-1 expression were unaltered; however, posterior cortical occludin decreased. These results suggest that 2 months postpartum, neuroinflammation, along with decreased occludin expression, may partly explain posterior cortical edema in rats with history of placental ischemia.

Expression of NGF and its functional implication in the lung injury induced by cerebral ischemia

BackgroundThe purpose of this study is to investigate the expression of nerve growth factor (NGF) in the lung injury induced by cerebral ischemia.MethodsAdult healthy female Sprague‐Dawley rats were randomly divided into two groups: sham group and brain ischemia (BI) induced lung injury group. Lung injury was induced by BI and neurological severity score (NSS) was evaluated. Triphenyltetrazolium chloride (TTC) staining, brain edema, ratio of wet/dry of lung tissue, hematoxylin‐eosin (HE) staining, as well as pulmonary edema were measured, respectively. The expression of mRNA and protein of NGF in lung tissues was detected by qPCR and western blot at 14 days post injury respectively. The immunohistochemical method was used to localize the expression of NGF in the lung tissues.ResultsLung injury from BI was successfully induced. The localization of NGF was detected mainly in the lung epithelial tissue following BI, also in vascular endothelial tissues. Compared with the sham group, the expression level of mRNA and protein for NGF was increased significantly after BI.ConclusionThe increase of NGF in the lung tissue after BI showed that NGF has been involved in the repair of injured lung, and the underlying mechanism of BI induced lung injury.

Transplantation of feeder‐free human induced pluripotent stem cell–derived cortical neuron progenitors in adult male Wistar rats with focal brain ischemia

The use of human induced pluripotent stem cells (hiPSCs) eliminates the ethical issues associated with fetal or embryonic materials, thus allowing progress in cell therapy research for ischemic stroke. Strict regulation of cell therapy development requires the xeno-free condition to eliminate clinical complications. Maintenance of hiPSCs with feeder-free condition presents a higher degree of spontaneous differentiation in comparison with conventional cultures. Therefore, feeder-free derivation might be not ideal for developing transplantable hiPSC derivatives. We developed the feeder-free condition for differentiation of cortical neurons from hiPSCs. Then, we evaluated the cells' characteristics upon transplantation into the sham and focal brain ischemia on adult male Wistar rats. Grafts in lesioned brains demonstrated polarized reactivity toward the ischemic border, indicated by directional preferences in axonal outgrowth and cellular migration, with no influence on graft survival. Following the transplantation, forelimb asymmetry was better restored compared with controls. Herein, we provide evidence to support the use of the xeno-free condition for the development of cell therapy for ischemic stroke.