Objective This study was aimed to develop a method combining computed tomography (CT) and fluorescence imaging, allowing identification of microvasculature in anatomical donors and facilitating translational research and education. Methods We investigated homogeneity and radiopacity of 30 different mixtures including radiopaque substances povidone-iodine (Betadine), barium sulfate (BaSO 4 ), and bismuth subsalicylate (Pepto-Bismol) varying in suspension and dilution with agar, latex, or gelatin. Three candidate mixtures were selected for testing the extent of perfusion in renal vasculature to establish methodology. From these candidate mixtures, two were selected for mixture with fluorescein and infusion into cadavers based on their ability to perfuse renal vasculature. The extent to which these two candidate mixtures combined with fluorescein were able to perfuse vasculature in a cadaver head was used to determine which mixture was superior. Results BaSO 4 and bismuth subsalicylate-based mixtures demonstrated superior opacity in vials. In terms of solidifying agents, gelatin-based mixtures demonstrated increased friability and lower melting points compared with the other agents, so only latex and agar-based mixtures were used moving forward past the vial stage. Combinations of BaSO 4 and latex and BaSO 4 and 3% agar were found to perfuse kidneys superiorly to the mixture containing bismuth subsalicylate. Finally, in cadaver heads, the mixture containing BaSO 4 , agar, and fluorescein was found to perfuse the smallest vasculature. Conclusion A final combination of BaSO 4 , 3% agar, and fluorescein proves to be a powerful and novel combination enabling CT imaging, fluorescence imaging, and dissection of vasculature. This paves the way for future translational research and education.

The Division of Anatomical Sciences at the University of Michigan Medical School offers a team‐taught undergraduate anatomy course (Anatomy 403) to 170–200 students each semester. As a supplement to the lectures, the students review the anatomy in the laboratory through demonstration of our extensive plastinated anatomical specimen collection. Understanding of laboratory based anatomy is assessed through cadaveric image based practical exam questions. In the Winter 2017 semester, we created an additional undergraduate course (Anatomy 510) to give students who successfully completed Anatomy 403 an opportunity to teach future students as teaching assistants (TAs). Feedback from the first two semesters of Anatomy 510 suggested that course expectations and outcomes were unclear. We hypothesized that restructuring the teaching course would have a positive impact on TA satisfaction and student engagement in the laboratory. Under the revised structure, TAs create mini lab lesson proposals, develop these proposals, practice the lesson with peers and faculty, and execute the lesson in the Anatomy 403 laboratory. Further, students participated in an orientation at the beginning of the semester to discuss grading and expectations. Under the restructure, students spent more time reviewing with faculty and had increased faculty support for lesson development and setting up the laboratory. Lastly, to ensure a thorough review of the material, online readiness quizzes covering all laboratory structures were developed for the TAs. We are assessing the efficacy of the restructuring of the course through narrative feedback, anonymous survey data from both TAs and students, course evaluations for Anatomy 403 and 510, and 403 student outcomes on the practical exam. Data collection and analysis is ongoing. Preliminary results reveal that Anatomy 510 course expectations were clearer following the restructure of the course. Additionally, Anatomy 403 students felt that the presence of TAs and their mini‐lessons were helpful for their success in the course. These results will contribute to a better understanding of an effective undergraduate anatomy teaching experience.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Cortical injury elicits long-term cytotoxic and cytoprotective mechanisms within the brain and the balance of these pathways can determine the functional outcome for the individual. Cytotoxicity is exacerbated by production of reactive oxygen species, accumulation of iron, and peroxidation of cell membranes and myelin. There are currently no neurorestorative treatments to aid in balancing the cytotoxic and cytoprotective mechanisms following cortical injury. Cell based therapies are an emerging treatment that may function in immunomodulation, reduction of secondary damage, and reorganization of surviving structures. We previously evaluated human umbilical tissue-derived cells (hUTC) in our non-human primate model of cortical injury restricted to the hand area of primary motor cortex. Systemic hUTC treatment resulted in significantly greater recovery of fine motor function compared to vehicle controls. Here we investigate the hypothesis that hUTC treatment reduces oxidative damage and iron accumulation and increases the extent of the microglial response to cortical injury. To test this, brain sections from these monkeys were processed using immunohistochemistry to quantify oxidative damage (4-HNE) and activated microglia (LN3), and Prussian Blue to quantify iron. hUTC treated subjects exhibited significantly reduced oxidative damage in the sublesional white matter and iron accumulation in the perilesional area as well as a significant increase in the extent of activated microglia along white matter pathways. Increased perilesional iron accumulation was associated with greater perilesional oxidative damage and larger reconstructed lesion volume. These findings support the hypothesis that systemic hUTC administered 24 h after cortical damage decreases the cytotoxic response while increasing the extent of microglial activation.

Stroke results in enduring damage to the brain which is accompanied by innate neurorestorative processes, such as reorganization of surviving circuits. Nevertheless, patients are often left with permanent residual impairments. Cell based therapy is an emerging therapeutic that may function to enhance the innate neurorestorative capacity of the brain. We previously evaluated human umbilical tissue-derived cells (hUTC) in our non-human primate model of cortical injury limited to the hand area of primary motor cortex. Injection of hUTC 24 h after injury resulted in significantly enhanced recovery of fine motor function compared to vehicle treated controls (Moore et al., 2013). These monkeys also received an injection of Bromodeoxyuridine (BrdU) 8 days after cortical injury to label cells undergoing replication. This was followed by 12 weeks of behavioral testing, which culminated 3 h prior to perfusion in a final behavioral testing session using only the impaired hand. In this session, the neuronal activity initiating hand movements leads to the upregulation of the immediate early gene c-Fos in activated cells. Following perfusion-fixation of the brain, sections were processed using immunohistochemistry to label c-Fos activated cells, pre-synaptic vesicle protein synaptophysin, and BrdU labeled neuroprogenitor cells to investigate the hypothesis that hUTC treatment enhanced behavioral recovery by facilitating reorganization of surviving cortical tissues. Quantitative analysis revealed that c-Fos activated cells were significantly increased in the ipsi- and contra-lesional ventral premotor but not the dorsal premotor cortices in the hUTC treated monkeys compared to placebo controls. Furthermore, the increase in c-Fos activated cells in the ipsi- and contra-lesional ventral premotor cortex correlated with a decrease in recovery time and improved grasp topography. Interestingly, there was no difference between treatment groups in the number of synaptophysin positive puncta in either ipsi- or contra-lesional ventral or dorsal premotor cortices. Nor was there a significant difference in the density of BrdU labeled cells in the subgranular zone of the hippocampus or the subventricular zone of the lateral ventricle. These findings support the hypothesis that hUTC treatment enhances the capacity of the brain to reorganize after cortical injury and that bilateral plasticity in ventral premotor cortex is a critical locus for this recovery of function. This reorganization may be accomplished through enhanced activation of pre-existing circuits within ventral premotor, but it could also reflect ventral premotor projections to the brainstem or spinal cord.

Introduction: Stroke is the leading cause of long-term disability in the United States due to impairments that endure after brain injury. While studies in rodent models have evaluated numerous neurorestorative treatments following stroke, none have received FDA approval. We evaluated a therapy using human umbilical tissue-derived cells (hUTC) as a potential neurorestorative treatment in our non-human primate model of cortical injury limited to the hand area of primary motor cortex. Given treatment 24 hours after injury, hUTC treated monkeys showed a significantly greater degree of recovery of fine motor function compared to vehicle treated controls (Moore et al., 2013). To explore the effect of hUTC, histopathological markers of inflammation and oxidative stress were assessed. Hypothesis: Treatment with hUTC will enhance the recruitment of glia to the injury and reduce the cascade of inflammation and oxidative stress. Methods: Using immunohistochemistry, activated microglia (LN3), reactive astrocytes (GFAP), oxidative damage (4HNE), and accumulated hemosiderin (Perls’ Prussian Blue) were quantified in ipsilesional primary motor cortex and underlying white matter. Microglia were counted using unbiased stereology. A Sholl Analysis was performed on traced perilesional astrocytes. The area of oxidative damage and hemosiderin was assessed using densitometry. Results: Compared to vehicle controls, density of activated microglia in the hUTC treated group approached a significant increase in the perilesional gray and white matter (p=0.070; p=0.092). Astrocytes exhibited more complex processes in treated monkeys (p=0.042). Staining for 4HNE was significantly reduced in white matter underlying the lesion in treated monkeys (p=0.033). Lastly, both the area and intensity of Perls’ staining for hemosiderin was significantly reduced in the perilesional area of treated monkeys (p=0.045; p=0.001). Conclusions: Treatment with hUTC resulted in increased activation of microglia and complexity of reactive astrocyte processes as well as reduced post-lesion oxidative damage and hemosiderin deposition. This suggests the hUTC treatment enhanced recovery, in part, by recruitment of glial cells that limited the damage following cortical injury.

While cognitive decline is observed in the normal aging monkey, neurons are not lost with age. Instead, frontal white matter is lost as myelin degenerates and both correlate with age-related cognitive decline. As age-related myelin damage increases, there should be an increase in clearance of damaged myelin by microglial phagocytosis. In this study, brains of behaviorally tested rhesus monkeys were assessed using unbiased stereology to quantify the density of activated microglia (LN3 antibody positive) and phagocytic microglia (galectin-3 (Gal-3) antibody positive) in three white matter regions: the corpus callosum, cingulum bundle (CGB), and frontal white matter (FWM). LN3 cell density was significantly increased in the CGB, whereas Gal-3 cell density was significantly increased in all regions. Increases in Gal-3 cell density in the FWM were associated with cognitive impairment. In the FWM of old animals, Gal-3-positive microglia were classified by morphological subtype as ramified, hypertrophic, or amoeboid. The densities of hypertrophic and amoeboid microglia significantly correlated with cognitive impairment. Finally, microglia were double-labeled with LN3 and Gal-3 showing that 91% of Gal-3 cells were also LN3 positive, thus expressing an “activated” phenotype. Furthermore, 15% of all double-labeled cells formed phagocytic cups. Overall, these results suggest that microglia become activated in white matter with age where the majority express a phagocytic phenotype. We hypothesize that age-related phagocytic activation of microglia is a response to accumulating myelin pathology. The association of Gal-3 in the FWM with cognitive impairment may reflect regional differences in damage or dysfunction of normal clearance mechanisms.

The objective of our study is to determine the utility of 3D printed models to understand the layers of the abdominal wall and peritoneum. The clinical importance of the abdominal wall structures and their continuity with other regions of the body is an important topic to medical gross anatomy students. Through dissection, these layers can be appreciated by an experienced anatomist, but the subtlety is often lost by new learners of anatomy. To supplement anatomical dissection, which is irreplaceable, we created a 3D printed model of a cross section of the abdomen with an exaggerated version of each layer. This model represents layers using rigid polylactic acid (PLA) 3D printed filament, Play‐doh, and flexible thermoplastic polyurethane (TPU) 3D printed filament. The model was presented to 50 medical gross anatomy students at three optional sessions (16–18 students per session). In each session, the fascial planes of anterolateral abdominal wall were first reviewed. Next, students were placed into groups of 3–4 to assemble a three‐piece model of the abdomen. The large group reviewed each part of the model and what it is meant to represent together. Then, students worked in small groups to color and label a 2D artistic representation of the model to validate the 3D concept into a 2D document that may be used to study beyond the session. Once the abdomen was thoroughly reviewed using the 3D printed model as well as 2D artistic representations of each of the model structures, students were given an assessment in their small groups where they were asked to identify the layers of the thoracic wall and pleura/pericardium on 2D artistic representations of the thorax. This assessment probed the ability of the students to apply a concept of fascial layers from the abdomen to the thorax. On average, the group performance on the assessment was 93%. Common misconceptions were identified and clarified in the large group following a review of the answers. Finally, the students received a survey 3–4 days after their session. This survey, a modified Likert scale, explored the effectiveness of the instructor, the model, and the session. Of 50 students, 30% of attendees responded to the survey. All students agreed that the instructor explained the material clearly, encouraged students to participate, and answered questions clearly. A large proportion (87%) of students agreed that the model and the session helped them better understand the material of the course and overall topic. Finally, 67% of students agreed that they were likely to recommend the session to other students. This study provides a framework for a larger study to examine the effectiveness of these 3D models to understand the fascial and peritoneal layers of the abdomen and thorax. Overall, students responded positively to the sessions and found that the most effective part of the session was the 3D model, which allows for a tactile medium to observe and comprehend the fascial layers and their continuity with other structures. We conclude that this preliminary data supports the effective use of 3D printed models as a supplement to anatomical dissection in understanding fascial layers of the abdomen.