Vessels of the umbilical cord: an anatomo-microscopic study in normal and pathological newborns

Intrauterine growth restriction (IUGR) due to placental insufficiency is associated with blood-flow redistribution. Placental insufficiency in late-onset IUGR often goes undetected by umbilical artery Doppler scan. Despite a broad general body of literature referencing placentas of IUGR pregnancies, there are no report on the structural characteristics of the vessels of the umbilical cord in IUGR. Thus, the aim of the present study was to compare the microscopic anatomy of the umbilical arteries and veins in normal, IUGR and small for gestational age (SGA) newborns. Twenty six umbilical cords were taken from 10 IUGR, 5 SGA, and 11 controls newborns. The histological and morphological examination was performed with EE, Azan–Mallory, Sirius Red stains and morphometric evaluation was performed through a computer image analysis approach. In the controls, the umbilical artery shows a muscular tunica, organized by two layers, an outer one with circularly arranged cells, and an inner one, with irregularly arranged cells. In the IUGR longitudinal muscular fibers are observable. In IUGR the percentage of the muscular fibers of the umbilical artery was greater with respect of SGA and controls. In IUGR and SGA the percentage of the muscular fibers was minor with respect to controls. In the umbilical artery in IUGR and SGA the elastic fibers and collagen I was major and collagen III was minor with respect to SGA. In the umbilical vein the collagen III was major in IUGR and SGA with respect to controls. These data agree with those of intrauterine life, in which a major thickness of the abdominal aortic wall was observed in fetuses with abnormalities of Doppler flussimetry. The rearrangement of umbilical artery may affect the mechanical properties of these vessels and disturb fetal blood circulation

Brainstem lesions in permanent vegetative state

Vegetative state has mainly been associated with widespread damage of cortex, white matter and/or thalamus, whereas brainstem findings are usually absent or include focal, slight and non-haemorrhagic lesions. Functional integrity of the brainstem is considered to be necessary for preservation of sleep/wake cycle (arousal and wakefulness), particularly midline structures of the upper pons and midbrain. Awareness, instead, mainly relies on the integrity of the cerebral cortex, white matter, basal ganglia and thalamus. We present here the neuroanatomical study of a case of post-traumatic permanent vegetative state which is characterized by a quite complex damage also at the level of the brainstem. After a car accident a young woman was referred to Neurosurgical Unit in coma and paraplegia due to head injury and C2 fracture. Spontaneous sleep/wake cycle recovered in the following months. Magnetic resonance, performed one month after the trauma, showed injuries of midbrain, thalamus and left frontal lobe. The patient died more than 15 years after the trauma. Macroscopic examination showed generalized hypotrophy of cerebral hemispheres, cerebellum and brainstem. Partial degeneration of the basal ganglia and severe volumetric reduction of the hemispheric white matter were observed. Wide and bilateral thalamic necrosis and degeneration was found. Transverse sections of the brainstem showed degeneration of the descending and ascending systems of motor and sensitive fibres, and neuronal loss in the inferior olivary complex, pontine nuclei, red nucleus and substantia nigra. In the midbrain, areas of necrosis with reactive astrogliosis extended from the mesencephalic aqueduct towards the interpeduncular fossa, affecting the periaqueductal grey matter, and the dorsal and median raphe nuclei. On the basis of the neuroanatomic description of the damaged areas, we discuss the role of the midbrain reticular activating system and possible plasticity responses in the mechanisms of arousal/wakefulness

Forensic Clinical Anatomy of Spine in Child Abuse

Forensic Clinical Anatomy of Child Abuse includes studies of Functional and/or Biomechanical Anatomy which are performed on cadavers to verify compatibility of lesions with accidental dynamics. Moreover, some kinds of damages following Child Abuse are strictly anatomical in nature and require morphological/morphometric methods of investigation for adequate assessment. Problems of differential diagnosis between anatomical structures (normal or variant) and pathological findings also frequently arise [1]. In the present work, we focused on anatomical bases of spinal lesions in two autoptical cases of abusive head trauma, with particular reference to methodological issues. Both cases presented brain subdural haemorrhage and multiple bilateral retinal haemorrhages. In both cases, the spinal cord was sampled in continuity with the dura mater and it was subjected to complete sectioning. Spinal subdural haemorrhages were found along all the spinal levels. The histopathological characteristics of these haemorrhages also permitted to reveal different chronologies of the lesions, with consequent forensic implications. Hypoxic-ischaemic damages coexisted, mainly at the level of cervical and lumbar spinal cord, together with glio-mesodermic response. On the basis of in vivo imaging suggesting cervical sub-dislocations, portions of the vertebral column were also sampled and subjected to postmortem imaging before further hystopathological sampling. In one case, postmortem imaging permitted to confirm anterolisthesis of the second vertebral body over the third one. Histopathological analysis also showed the presence of haemorrhagic infiltrations of the epidural adipose tissue at the level of the atlanto-axial joints. A consistent methodology of analysis of the spinal structures should involve integration of postmortem imaging with detailed and exhaustive histopathological study

High-quality digital 3D reconstruction of the terminal pathway of a heart stab wound

High-quality digital 3D reconstructions of microscopic findings have been involved in anatomical and histopathological research, but their potentialities in forensic pathology may also be of particular interest. We here present the use of such methods to furtherly analyse a heart stab wound in a suicidal case. External examination revealed multiple incised wounds on the wrists, three stab wounds at the neck and a single stab wound on the chest. At autopsy, injuries to the neck and wrists were superficial whereas the thoracic stab wound penetrated the chest wall and pericardium. Heart examination showed an 8-mm-long stab wound on the anterior surface of the left ventricle. Heart sectioning and inspection of the correspondent internal aspect of the left ventricle did not show a macroscopic injury, but the irregularity of the trabeculae carneae did not permit to exclude a microscopic pathway. Thus, the heart wall including the stab wound was paraffin-embedded and subjected to complete sectioning for microscopic analysis along all the wound extension. Every 10th section was stained with haematoxylin-eosin and was acquired by using a Leica DMR microscope and a high resolution digital camera. The three-dimensional aspect of the lesion was reconstructed with a software system for 3D computer graphics. Microscopic examination and 3D reconstruction demonstrated that the lesion extended to the internal surface of the ventricle wall, although for a very limited extension. 3D reconstruction also showed a certain curvilinear pattern of the lesion in the myocardium, consistent with myocardial contraction at the moment of injury and consequent vitality of the lesion. Moreover, 3D reconstruction permitted to obtain the dimensions of the intra-myocardial injury (corrected for shrinkage and evaluated in the context of muscle contraction), also permitting to furtherly confirm the identification of the knife involved. In conclusion, the present case is indicative of how complete microscopic sectioning and 3D reconstruction may add further information about characteristics of injuries of forensic interest

Hyperoxia-induced changes in morphometric parameters of postnatal neurogenic sites in rat

In literature many works address the effects of hypoxia exposure on postnatal neurogenesis but few data are available about hyperoxia effects, although high oxygen concentrations are frequently used for ventilation of premature newborns. Thus, the aim of the present study was to compare with controls the morphometrical parameters of the main neurogenic sites (subventricular zone and dentate gyrus) in newborn Sprague-Dawley rats exposed to 60% or 95% oxygen for the first 14 postnatal days. Six rats were studied for each of the three groups. The unbiased quantitative method of the optical disector was applied to analyze neuronal densities, nuclear volumes, and total neuron numbers of the subventricular zone and hippocampal dentate gyrus. Apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling, TUNEL) and proliferation (Ki67) were also studied. The subventricular zone of newborn rats exposed to 95% hyperoxia showed statistically significant higher volume (mean value ± coefficient of variation: 0.40 ± 0.20 mm3) than subventricular zone of rats raised in normoxia (0.20 ± 0.11 mm3) or 60% hyperoxia (0.26 ± 0.18 mm3). Total neuron number was also significantly higher in 95% hyperoxia while neuronal densities did not reach statistically significant differences. TUNEL showed increased apoptotic indexes in hyperoxic rats. The percentage of proliferating KI67 positive cells was also higher in hyperoxia. The dentate gyrus granular layer of the normoxic rats showed higher volume (0.65 ± 0.11 mm3) than both the hyperoxic groups (60% hyperoxia: 0.39 ± 0.14 mm3; 95% hyperoxia: 0.36 ± 0.16 mm3). Total neuron numbers of hyperoxic dentate gyrus were also significantly reduced; neuronal densities were not modified. Hyperoxia-exposed rats also showed higher apoptotic and proliferating indexes in the dentate gyrus. Hyperoxia exposure in the first postnatal period may affect the main neurogenic areas (subventricular zone and dentate gyrus) increasing apoptosis but also inducing a certain reparative response consisting of increased proliferation. In particular, the increased volume of the subventricular zone may be ascribed to compensatory neurogenic response to the hyperoxic damage. Conversely, the decreased volume of the dentate gyrus granular layer could derive by a non sufficient neurogenic response to counterbalance the hyperoxic neuronal injury

Short Bowel Syndrome and Tissue Engineering: a preliminary study towards the development of a new regenerative approach in paediatric patients

Pediatric Short Bowel Syndrome (SBS) is a malabsorption state following massive surgical resections of the small intestine. The current therapeutic options issues (i.e. parental nutrition, surgical lengthening, transplantation) have prompt the research in Tissue Engineering. Thus, our aim was to preliminary investigate in vitro/in vivo two composite scaffolds for engineering the small intestine without resorting to autologous intestinal epithelial organoid units which, to date, are the cell source mainly considered for this purpose. In particular, we developed composite supports consisting of a novel biocompatible/resorbable cryogel that is oxidized polyvinyl alcohol (OxPVA) [1] crosslinked with intestinal mucosa whole (wIM/OxPVA) or homogenized (hIM/OxPVA). After evaluating the scaffolds by histology and Scanning Electron Microscopy (SEM), we assessed their interaction with adipose mesenchymal stem cells. Thereafter, the in vivo behavior of scaffolds was studied implanting them in the omentum of Sprague Dawley rats. At 4 weeks, explants were processed by histology and immunohistochemistry (CD3; F4/80; Ki-67; desmin; α-SMA; MNF116). Considering the in vitro evidence, both histological and SEM results proved the effectiveness of the decellularization, and allowed to appreciate the preservation of intestinal villi of the wIM as well as the characteristic features of the hIM. At 7 days from cell seeding, MTT assay showed that hIM/OxPVA scaffolds could support cell adhesion/proliferation even if the wIM/OxPVA ones seem to significantly increase cell growth (

Decellularized omentum as novel biologic scaffold for reconstructive surgery and regenerative medicine

Homologous tissues, such as adipose tissue, may be an interesting source of acellular scaffolds, maintaining a complex physiological three-dimensional (3D) structure, to be recellularized with autologous cells. The aim of the present work is to evaluate the possibility of obtaining homologous acellular scaffolds from decellularization of the omentum, which is known to have a complex vascular network. Adult rat and human omenta were treated with an adapted decellularization protocol involving mechanical rupture (freeze-thaw cycles), enzymatic digestion (trypsin, lipase, deoxyribonuclease, ribonuclease) and lipid extraction (2-propanol). Histological staining confirmed the effectiveness of decellularization, resulting in cell-free scaffolds with no residual cells in the matrix. The complex 3D networks of collagen (azan-Mallory), elastic fibers (Van Gieson), reticular fibers and glycosaminoglycans (PAS) were maintained, whereas Oil Red and Sudan stains showed the loss of lipids in the decellularized tissue. The vascular structures in the tissue were still visible, with preservation of collagen and elastic wall components and loss of endothelial (anti-CD31 and -CD34 immunohistochemistry) and smooth muscle (anti-alpha smooth muscle actin) cells. Fat-rich and well vascularized omental tissue may be decellularized to obtain complex 3D scaffolds preserving tissue architecture potentially suitable for recellularization. Further analyses are necessary to verify the possibility of recolonization of the scaffold by adipose-derived stem cells in vitro and then in vivo after re implantation, as already known for homologus implants in regenerative processes

Double origin of the superior cerebellar artery associated with homolateral haemorrhagic infarction of cerebellum

The superior cerebellar artery (SCA) shows the least variable course and the lowest incidence of anatomical variations among cerebellar arteries. In the present study, an 84-year-old woman was affected by a cerebellar infarction which underwent haemorrhagic evolution in the following days. Neuroimaging investigations also showed a probable double origin of the left SCA. Neuropathological examination confirmed the presence of a large haemorrhagic infarction at the level of the superior portion of the left cerebellar hemisphere and vermis. The left SCA arose from two small arteries arising from the left aspect of the basilar artery and joining together after a course of 9 mm. Previous studies have reported the association of cerebrovascular pathologies, such as intracranial aneurysms, with fenestrations and double origins of the posterior inferior cerebellar artery. In the present case, the occurrence of an haemorrhagic infarction in the vascular field of an SCA with double origin is intriguing in suggesting a possible pathophysiological association