Medial Tissue Research Articles

An anterior medial cell population with an apical-organ-like transcriptional profile that pioneers the central nervous system in the centipede Strigamia maritima

The apical plate of primary marine larvae is characterized by a common set of transcription factors comprising six3, rx, hbn, nk2.1 and FoxQ2. It harbours the apical organ, a neural and ciliary structure with neurosecretory properties. Recent studies in lophotrochozoans have found that apical organ cells form the anterior tip of the developing central nervous system.We identify an anterior medial tissue in the embryonic centipede head that shares the transcriptional profile of the apical plate of marine larvae, including nested domains of FoxQ2 and six3 expression. This domain gives rise to an anterior medial population of neural precursors distinct from those arising within the segmental neuroectoderm. These medial cells do not express achaete scute homologue in proneural clusters, but express collier, a marker for post mitotic cells committed to a neural fate, while they are still situated in the surface ectodermal layer. They then sink under the surface to form a compact cell cluster. Once internalized these cells extend axons that pioneer the primary axonal scaffold of the central nervous system. The same cells express phc2, a neural specific prohormone convertase, which suggests that they form an early active neurosecretory centre. Some also express markers of hypothalamic neurons, including otp, vtn and vax1.These medial neurosecretory cells of the centipede are distinct from those of the pars intercerebralis, the anterior neurosecretory part of the insect brain. The pars intercerebralis derives from vsx positive placodal-like invagination sites. In the centipede, vsx expressing invaginating ectoderm is situated bilaterally adjacent to the medial pioneer cell population. Hence the pars intercerebralis is present in both insect and centipede brains, whereas no prominent anterior medial cluster of pioneer neurons is present in insects. These observations suggest that the arthropod brain retained ancestrally an anterior medial population of neurosecretory cells homologous to those of the apical plate in other invertebrate phyla, but that this cell population has been lost or greatly reduced in insects.

H.M.'s contributions to neuroscience: A review and autopsy studies

H.M., Henry Molaison, was one of the world's most famous amnesic patients. His amnesia was caused by an experimental brain operation, bilateral medial temporal lobe resection, carried out in 1953 to relieve intractable epilepsy. He died on December 2, 2008, and that night we conducted a wide variety of in situ MRI scans in a 3 T scanner at the Massachusetts General Hospital (Mass General) Athinoula A. Martinos Center for Biomedical Imaging. For the in situ experiments, we acquired a full set of standard clinical scans, 1 mm isotropic anatomical scans, and multiple averages of 440 μm isotropic anatomical scans. The next morning, H.M.'s body was transported to the Mass General Morgue for autopsy. The photographs taken at that time provided the first documentation of H.M.'s lesions in his physical brain. After tissue fixation, we obtained ex vivo structural data at ultra-high resolution using 3 T and 7 T magnets. For the ex vivo acquisitions, the highest resolution images were 210 μm isotropic. Based on the MRI data, the anatomical areas removed during H.M.'s experimental operation were the medial temporopolar cortex, piriform cortex, virtually all of the entorhinal cortex, most of the perirhinal cortex and subiculum, the amygdala (except parts of the dorsal-most nuclei-central and medial), anterior half of the hippocampus, and the dentate gyrus (posterior head and body). The posterior parahippocampal gyrus and medial temporal stem were partially damaged. Spared medial temporal lobe tissue included the dorsal-most amygdala, the hippocampal-amygdalo-transition-area, ∼2 cm of the tail of the hippocampus, a small part of perirhinal cortex, a small portion of medial hippocampal tissue, and ∼2 cm of posterior parahippocampal gyrus. H.M.'s impact on the field of memory has been remarkable, and his contributions to neuroscience continue with a unique dataset that includes in vivo, in situ, and ex vivo high-resolution MRI.

A combined numerical and experimental framework for determining permeability properties of the arterial media.

The medial layer of the arterial wall may play an important role in the regulation of water and solute transport across the wall. In particular, a high medial resistance to transport could cause accumulation of lipid-carrying molecules in the inner wall. In this study, the water transport properties of medial tissue were characterised in a numerical model, utilising experimentally obtained data for the medial microstructure and the relative permeability of different constituents. For the model, a new solver for flow in porous materials, based on a high-order splitting scheme, was implemented in the spectral/hp element library nektar++ and validated. The data were obtained by immersing excised aortic bifurcations in a solution of fluorescent protein tracer and subsequently imaging them with a confocal microscope. Cuboidal regions of interest were selected in which the microstructure and relative permeability of different structures were transformed to a computational mesh. Impermeable objects were treated fictitiously in the numerical scheme. On this cube, a pressure drop was applied in the three coordinate directions and the principal components of the permeability tensor were determined. The reconstructed images demonstrated the arrangement of elastic lamellae and interspersed smooth muscle cells in rat aortic media; the distribution and alignment of the smooth muscle cells varied spatially within the extracellular matrix. The numerical simulations highlighted that the heterogeneity of the medial structure is important in determining local water transport properties of the tissue, resulting in regional and directional variation of the permeability tensor. A major factor in this variation is the alignment and density of smooth muscle cells in the media, particularly adjacent to the adventitial layer.

Isolation and characterization of mesenchymal progenitor cells from human orbital adipose tissue.

Adipose-derived stem cells (ASCs) have gained importance due to their myriad potential clinical applications. We hypothesize that progenitor cells also exist besides those conventionally isolated from the stromal vascular fraction (SVF). Central and medial orbital adipose tissues obtained from patients during eyelid surgery were digested with collagenase for 3 or 16 hours at 37°C with or without shaking. After centrifugation, the remaining cell pellet was resuspended and filtered to yield flow through in SVF and retained cells (RC) on the filter. Single cells from RC and SVF were cultured on 5% coated Matrigel in serum-free modified embryonic stem cells medium (MESCM) for 10 passages. The progenitor status was evaluated by the expression of a number of markers by qPCR and immunofluorescence staining as well as their plasticity for endothelial and tri-lineage differentiation. Type I collagenase digestion for 3 hours under shaking was significantly less effective in releasing progenitor cells than collagenase A digestion for 16 hours without shaking. Following filtration, cells in SVF and RC, of which the latter were tangled in collagen IV-containing matrix, expressed different markers of progenitor cells. Cells from SVF and RC could be expanded for 10 passages on coated Matrigel in MESCM and exhibited similar or better potential to differentiate into vascular endothelial cells, chondrocytes, osteocytes, and adipocytes than SVF cells expanded on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). Different progenitor cells can be isolated and expanded from orbital adipose tissues. Further characterization of their mesodermal or neuroectodermal origin might enhance clinical outcome when used as a source of autologous stem cells for ocular surface regeneration.

MPFL reconstruction using a quadriceps tendon graft: Part 2: Operative technique and short term clinical results

BackgroundWe describe the preliminary clinical results of a new operative technique for MPFL reconstruction using a strip of quadriceps tendon (QT). MethodsPatients: 17 patients (7 male, 10 female; mean age 21.5years±3.9) have been operated on with this technique. All patients were evaluated clinically, radiologically and with subjective questionnaires (Tegner-, Lysholm-, Kujala Score) pre-operatively and post-operatively at 6 and 12months (m).Surgical technique: A 10 to 12mm wide, 3mm thick and 8 to 10cm long strip from the central aspect of quadriceps tendon is harvested subcutaneously. The tendon strip is then dissected distally on the patella, left attached, diverged 90° medially underneath the medial prepatellar tissue and fixed with 2 sutures. The graft is fixed in 20° of knee flexion with a bioabsorbable interference screw. ResultsLysholm score at 6m was 81.9±11.7 and at 12m 88.1±10.9, Kujala score at 12m was 89.2±7.1 and Tegner Score was 4.9±2.0 (6m) and 5.0±1.9 (12m). Two patients had a positive apprehension test at 12months. There was no re-dislocation during the follow-up period. ConclusionMPFL reconstruction with a strip of QT harvested in a minimal invasive technique was found to be associated with good short term clinical results. We think that this technique presents a valuable alternative to common hamstring techniques for primary MPFL reconstruction in children and adults, as well as for MPFL revision surgery. Level of evidenceIV, prospective case series.

Minimally Invasive Reconstruction of the Medial Patellofemoral Ligament Using Quadriceps Tendon

Reconstruction of the medial patellofemoral ligament (MPFL) for the treatment of patellar instability has received increased attention over the past few years. Most operative techniques use hamstring grafts fixed with bone tunnels and/or anchors on the patella. Despite good clinical results using these techniques, complications such as implant breakage, patellar fractures through bone tunnels, and loss of knee motion have occurred. We present a minimally invasive technique for MPFL reconstruction using a strip of quadriceps tendon. With the use of specially designed instruments, the graft is harvested through a 3-cm transverse incision at the proximal pole of the patella. The tendon strip is then dissected distally on the patella, left attached, and diverged 90° medially underneath the medial prepatellar tissue. The graft is fixed on the femur in 20° of knee flexion in a bone tunnel with a bioabsorbable interference screw (adults) or a bone anchor (children). We think that this technique presents a valuable alternative to common hamstring techniques for primary MPFL reconstruction in children and adults, as well as for MPFL revision surgery.

Structural reorganization of pyramidal neurons in the medial prefrontal cortex of alcohol dependent rats is associated with altered glial plasticity.

In rodents, chronic intermittent ethanol vapor exposure (CIE) produces alcohol dependence, alters the activity of pyramidal neurons and decreases the number of glial progenitors in the medial prefrontal cortex (mPFC). Adult male Wistar rats were exposed to CIE and were injected with mitotic markers to label and phenotype proliferating cells to test the hypothesis that CIE produces concurrent alterations in the structure of pyramidal neurons and the cell cycle kinetics and developmental stages of glial progenitors in the mPFC. Medial prefrontal cortical tissue was processed for Golgi-Cox staining, immunohistochemistry and Western blotting analysis. CIE increased dendritic arborization and spine densities within basal and apical dendrites of pyramidal neurons via aberrant reorganization of actin cytoskeleton-associated molecules. CIE concomitantly increased the expression of total NR2B subunits without affecting phosphorylation of NR2B at Tyr-1472 or levels of PSD-95. CIE reduced the length of S-phase of the cell cycle of glial progenitors and reduced proliferation and differentiation of progenitors into bHLH transcription factor Olig2-expressing premyelinating oligodendrocyte progenitor cells (OPCs). CIE also produced a corresponding hyperphosphorylation of Olig2, and reduced expression of myelin basic protein. Our findings demonstrate that CIE-induced alterations in OPCs and myelin-related proteins are associated with profound alterations in the structure of pyramidal neurons. In sum, our results not only provide evidence that alcohol dependence leads to pathological changes in the mPFC, which may in part define a cellular basis for cognitive impairments associated with alcoholism, but also show dependence-associated morphological changes in the PFC at the single neuron level.

Transcriptomic changes in the frontal cortex associated with paternal age

BackgroundAdvanced paternal age is robustly associated with several human neuropsychiatric disorders, particularly autism. The precise mechanism(s) mediating the paternal age effect are not known, but they are thought to involve the accumulation of de novo (epi)genomic alterations. In this study we investigate differences in the frontal cortex transcriptome in a mouse model of advanced paternal age.FindingsTranscriptomic profiling was undertaken for medial prefrontal cortex tissue dissected from the male offspring of young fathers (2 month old, 4 sires, n = 16 offspring) and old fathers (10 month old, 6 sires, n = 16 offspring) in a mouse model of advancing paternal age. We found a number of differentially expressed genes in the offspring of older fathers, many previously implicated in the aetiology of autism. Pathway analysis highlighted significant enrichment for changes in functional networks involved in inflammation and inflammatory disease, which are also implicated in autism.ConclusionsWe observed widespread alterations to the transcriptome associated with advanced paternal age with an enrichment of genes associated with inflammation, an interesting observation given previous evidence linking the immune system to several neuropsychiatric disorders including autism.

Perivascular Delivery of Notch 1 siRNA Inhibits Injury-Induced Arterial Remodeling

ObjectivesTo determine the efficacy of perivascular delivery of Notch 1 siRNA in preventing injury-induced arterial remodeling.Methods and ResultsCarotid artery ligation was performed to induce arterial remodeling. After 14 days, morphometric analysis confirmed increased vSMC growth and subsequent media thickening and neointimal formation. Laser capture microdissection, quantitative qRT-PCR and immunoblot analysis of medial tissue revealed a significant increase in Notch1 receptor and notch target gene, Hrt 1 and 2 expression in the injured vessels. Perivascular delivery of Notch 1 siRNA by pluronic gel inhibited the injury-induced increase in Notch 1 receptor and target gene expression when compared to scrambled siRNA controls while concomitantly reducing media thickening and neointimal formation to pre-injury, sham-operated levels. Selective Notch 1 knockdown also reversed the injury-induced inhibition of pro-apoptotic Bax expression while decreasing injury-induced anti-apoptotic Bcl-XL expression to sham-operated control levels. In parallel experiments, proliferative cyclin levels, as measured by PCNA expression, were reversed to sham-operated control levels following selective Notch 1 knockdown.ConclusionThese results suggest that injury-induced arterial remodeling can be successfully inhibited by localized perivascular delivery of Notch 1 siRNA.

Fabrication Factory for Tubular Vascular Tissue Mimics based on Automated Rolling Manipulation and Thermo-Responsive Polymers

This study presents a novel methodology for fabrication of tubular vascular tissue mimics by an automated factory using thermoresponsive polymers which have temperature-induced reversible phase transition characteristics; Poly(N-Isopropylacrylamide)-Grafted Gelatin (PNIPAM-gelatin), PNIPAM and gelatin. The custom-designed factory consists of a moving tray, a roller unit driven by rack-and-pinion mechanics, and a local temperature-control unit. PNIPAM-gelatin serves as the matrix, providing temperature-dependent adhesion and detachment of cell sheets. We describe two prototypes of vascular tissue mimics: a tubular medial tissue mimic composed of randomly oriented Smooth Muscle Cells (SMCs), and a hierarchically structured vascular tissue mimic (intimal/medical tissue mimics) composed of longitudinally oriented Endothelial Cells (ECs) on the luminal surface and circumferentially oriented multilayered SMCs in the outer layer. We observed these prototypes under electron microscopes and a Confocal Laser-Scanning Microscope (CLSM). An advanced model of our prototype factory would have the potential to produce high-quality on-demand vascular grafts without incorporating any foreign materials.

Coronal plane socket stability during gait in persons with transfemoral amputation: Pilot study.

Little research describes which transfemoral socket design features are important for coronal plane stability, socket comfort, and gait. Our study objectives were to (1) relate socket comfort during gait to a rank order of changes in ischial containment (IC) and tissue loading and (2) compare socket comfort during gait when tissue loading and IC were systematically manipulated. Six randomly assigned socket conditions (IC and tissue compression) were assessed: (1) IC and high, (2) IC and medium, (3) IC and low, (4) no IC and high, (5) no IC and medium, and (6) no IC and low. For the six subjects in this study, there was a strong, negative relationship between comfort and changes in IC and tissue loading (rho = -0.89). With the ischium contained, tissue loading did not influence socket comfort (p = 0.47). With no IC, the socket was equally comfortable with high tissue loading (p = 0.36) but the medium (p = 0.04) and low (p = 0.02) tissue loading conditions decreased comfort significantly. Coronal plane hip moments, lateral trunk lean, step width, and walking speed were invariant to changes in IC and/or tissue loading. Our results suggest that in an IC socket, medial tissue loading mattered little in terms of comfort. Sockets without IC required high tissue loading to be as comfortable as those with IC, while suboptimal tissue loading compromised comfort.

Divergent palate morphology in turtles and birds correlates with differences in proliferation and BMP2 expression during embryonic development

During embryonic development, amniotes typically form outgrowths from the medial sides of the maxillary prominences called palatal shelves or palatine processes. In mammals the shelves fuse in the midline and form a bony hard palate that completely separates the nasal and oral cavities. In birds and lizards, palatine processes develop but remain unfused, leaving a natural cleft. Adult turtles do not possess palatine processes and unlike other amniotes, the internal nares open into the oral cavity. Here we investigate craniofacial ontogeny in the turtle, Emydura subglobosa to determine whether vestigial palatine processes develop and subsequently regress, or whether development fails entirely. We found that the primary palate in turtles develops similarly to other amniotes, but secondary palate ontogeny diverges. Using histology, cellular dynamics and in situ hybridization we found no evidence of palatine process development at any time during ontogeny of the face in the turtle. Furthermore, detailed comparisons with chicken embryos (the model organism most closely related to turtles from a molecular phylogeny perspective), we identified differences in proliferation and gene expression patterns that correlate with the differences in palate morphology. We propose that, in turtles, palatine process outgrowth is never initiated due to a lack of mesenchymal bone morphogenetic protein 2 (BMP2) expression in the maxillary mesenchyme, which in turn fails to induce the relatively higher cellular proliferation required for medial tissue outgrowth. It is likely that these differences between turtles and birds arose after the divergence of the lineage leading to modern turtles.

TCT-122 Electrode irrigation alters RF ablation treatment zone geometry and preserves medial and adventitial tissue while maintaining injury to renal nerves

We quantitatively compared lesion geometry and artery and nerve injury with and without irrigation during RF ablation to enable optimization of catheter design and understanding of temperature gradients. RF ablations, using spatially discrete electrodes, were conducted in swine using THERMOCOOL(r)

Extent and severity of coronary artery disease by coronary CT angiography is associated with elevated left ventricular diastolic pressures and worsening diastolic function

BackgroundPatients with flow-limiting coronary stenoses exhibit elevated left ventricular end-diastolic pressure (LVEDP) and abnormal left ventricular (LV) relaxation. ObjectiveWe investigated the relationship of extent and severity of coronary artery disease (CAD) by coronary CT angiography (CTA) to LVEDP and measures of LV diastolic dysfunction. MethodsWe identified consecutive patients undergoing coronary CTA and transthoracic echocardiography who were assessed for diastolic function. CAD was evaluated on a per-patient, per-vessel, and per-segment basis for intraluminal diameter stenosis by using an 18-segment model (0 = none, 1 = 1%–49%, 2 = 50%–69%, and 3 = 70%–100%) and summed over segments to obtain overall coronary plaque burden (segment stenosis score [SSS]; maximum = 54). Transthoracic echocardiography evaluated mitral inflow E wave-to-A wave ratio, tissue Doppler early mitral annual tissue velocity axial excursion, stage of diastolic dysfunction, and LV dimensions and estimated LVEDP from the ratio of mitral inflow velocity to early mitral annular (medial) tissue velocity. ResultsFour hundred seventy-eight patients (57% women; mean age, 57.9 ± 14.6 years; 24.9% prior CAD) comprised the study population. Increasing per-patient maximal coronary stenosis, number of vessels with obstructive stenosis, and SSS were associated with increased LVEDP. The prevalence of advanced diastolic dysfunction increased with greater number of obstructive vessels. In multivariable analyses, SSS was associated with increased LVEDP (0.8 mm Hg per tertile increase in SSS, 0.5–1.1; P < .001); reduced E′ axial excursion (−0.3; 95% confidence interval [CI], −0.5 to −0.1; P = .001), increased LV mass index (1.6 g/m2 per tertile increase in SSS; P = .04), and increased relative wall thickness (0.005; 95% CI, 0.004–0.009; P = .03), with consistent relationships persisting even among persons with per-patient maximal stenosis <50% and LV ejection fraction ≥55%. ConclusionsExtent and severity of obstructive as well as nonobstructive CAD by coronary CTA are associated with increased LVEDP and measures of diastolic dysfunction.

Calmodulin Expression Distinguishes the Smooth Muscle Cell Population of Human Carotid Plaque

Several observations suggest the expansion of a distinct medial smooth muscle cell (SMC) subset in atherosclerosis and restenosis. We characterized the phenotypic features of SMC subsets in cultures derived from human carotid endarterectomy specimens. Specimens comprised an undiseased portion (thin intimal thickening with the underlying media) and a diseased portion (atherosclerotic plaque with the underlying media). From plaque tissues of the diseased portion, only macrophage-derived foam cells were retrieved. From medial tissues, two SMC phenotypes were isolated: large SMCs (flat with a monolayered growth pattern, from the undiseased portion) and small SMCs (fusiform and growing in multilayers, from the undiseased and diseased portions after co-culture with macrophage-derived foam cells). Small SMCs displayed higher proliferative and migratory activities and were less differentiated than large SMCs. Proteomic analysis showed that calmodulin was predominant in small SMCs. Co-culture of large SMCs with macrophage-derived foam cells induced a transition to the small phenotype with increased calmodulin expression. The calmodulin inhibitor W-7 decreased the proliferation of small SMCs and prevented the large to small phenotypic transition. Invivo, calmodulin was markedly expressed in SMCs of atherosclerotic plaques and was barely detectable in the media. Macrophage-derived foam cells promote selective migration from the media of atheroma-prone SMCs characterized by calmodulin overexpression. Further studies of small SMCs could be instrumental in understanding atherosclerosis pathogenesis and in planning therapeutic strategies.

Modifications of Chromatin Dynamics Control Smad2 Pathway Activation in Aneurysmal Smooth Muscle Cells

The activation of the Smad2 signaling pathway is thought to play an important role in human aneurysmal diseases as described by an important body of research. We previously showed that constitutive Smad2 activation is associated with Smad2 mRNA overexpression in aneurysmal vascular smooth muscle cells (VSMCs), which is dependent on epigenetic regulation of the SMAD2 promoter involving histone modifications. However, the underlying molecular mechanisms controlling Smad2 overexpression are currently unknown. The aim of the present study is to understand the mechanisms regulating the constitutive Smad2 overexpression in VSMCs by identification of the histone-modifying enzymes, transcription factors, and cofactors responsible for Smad2 promoter activation in aneurysmal disease. This study was performed on medial tissue extracts and primary cultures of VSMCs of human thoracic aneurysms (n=17) and normal thoracic aortas (n=10). Here, we demonstrate that the activation of SMAD2 promoter is driven by the recruitment of a multipartner complex, including the transcription factor p53 and histone acetyltransferases. Remarkably, the transcriptional regulatory network of the SMAD2 promoter is dramatically altered in human aneurysmal VSMCs in vitro and in situ with a switch from Myc-dependent repression of SMAD2 in normal vessel to a p53-dependent activation of SMAD2 in aneurysms. Furthermore, histone acetyltransferases p300 and P300/CBP-associated protein play a major role in SMAD2 promoter activation by acting on histone acetylation, p53 recruitment, and acetylation. These results provide evidence for a major role of p53 and the complex composed of p300 and p300/CBP-associated protein in Smad2 activation in human aneurysmal VSMCs.

Smad2-Dependent Protease Nexin-1 Overexpression Differentiates Chronic Aneurysms From Acute Dissections of Human Ascending Aorta

Tissue activation of proteolysis is involved in acute intramural rupture (dissections, acute ascending aortic dissection) and in progressive dilation (aneurysms, thoracic aneurysm of the ascending aorta) of human ascending aorta. The translational aim of this study was to characterize the regulation of antiproteolytic serpin expression in normal, aneurysmal, and dissecting aorta. We explored expression of protease nexin-1 (PN-1) and plasminogen activator inhibitor-1 and their regulation by the Smad2 signaling pathway in human tissue and cultured vascular smooth muscle cells (VSMCs) of aneurysms (thoracic aneurysm of the ascending aorta; n=46) and acute dissections (acute ascending aortic dissection; n=10) of the ascending aorta compared with healthy aortas (n=10). Both PN-1 and plasminogen activator inhibitor-1 mRNA and proteins were overexpressed in medial tissue extracts and primary VSMC cultures from thoracic aneurysm of the ascending aorta compared with acute ascending aortic dissection and controls. Transforming growth factor-β induced increased PN-1 expression in control but not in aneurysmal VSMCs. PN-1 and plasminogen activator inhibitor-1 overexpression by aneurysmal VSMCs was associated with increased Smad2 binding on their promoters and, functionally, resulted in VSMC self-protection from plasmin-induced detachment and death. This phenomenon was restricted to aneurysms and not observed in acute dissections. These results demonstrate that epigenetically regulated PN-1 overexpression promotes development of an antiproteolytic VSMC phenotype and might favor progressive aneurysmal dilation, whereas absence of this counter-regulation in dissections would lead to acute wall rupture.

The Effect of Pulsatile Loading and Scaffold Structure for the Generation of a Medial Equivalent Tissue Engineered Vascular Graft

A reliable and cost-effective scaffold for tissue-engineered vascular graft that would not only support cell proliferation and growth but also maintain cell phenotype has been a long-term challenge. In this study, we propose a biodegradable and biomimetic copolymer of gelatin with vinyl acetate synthesized via a graft copolymerization technique to generate tubular scaffolds for vascular tissue engineering. Two fabrication techniques, freeze drying and electrospinning, were used to generate the differing architectures for the scaffolds and characterized. The electrospun scaffolds were found to have a faster rate of mass loss in physiological saline of 81.72% within 4 months compared with 60% mass loss for the freeze-dried samples, though the materials were more crystalline. Vascular (v) smooth muscle cells (SMCs) were seeded on these tubes, which were then subjected to dynamic pulsatile stimulation on a vascular bioreactor for a week. Gross examination of the tissue-engineered constructs revealed that the cells secreted extensive extracellular matrix, with the dynamically conditioned samples exhibiting well-orientated SMCs and collagenous fibers in comparison with growth in static conditions. In addition, the alignment of cells in the direction of strain was greater in the electrospun constructs. The electrospun scaffolds maintained the characteristic contractile phenotype of SMCs, which was confirmed by higher gene expression rates of contractile protein markers like SM22α and calponin. A significant increase in the total matrix components (collagen and elastin) in the electrospun constructs compared with the freeze-dried samples was confirmed by biochemical analysis. The results of this study indicate that a combination approach involving a biomimetic scaffold with the nanofibrillar architecture and good mechanical strength conducive to the growth of SMCs and the use of the pulsatile forces to modulate the cell morphology and phenotypic plasticity of vSMCs helps in the successful engineering of a medial layer of blood vessel.

Psychological stress, cocaine and natural reward each induce endoplasmic reticulum stress genes in rat brain

Our prior research has shown that the transcription of endoplasmic reticulum (ER) stress transcription factors activating transcription factor 3 (ATF3) and ATF4 are induced by amphetamine and restraint stress in rat striatum. However, presently the full extent of ER stress responses to psychological stress or cocaine, and which of the three ER stress pathways is activated is unknown. The current study examines transcriptional responses of key ER stress target genes subsequent to psychological stress or cocaine. Rats were subjected to acute or repeated restraint stress or cocaine treatment and mRNA was isolated from dorsal striatum, medial prefrontal cortex and nucleus accumbens brain tissue. ER stress gene mRNA expression was measured using quantitative polymerase chain reaction (PCR) and RNA sequencing. Restraint stress and cocaine-induced transcription of the classic ER stress-induced genes (BIP, CHOP, ATF3 and GADD34) and of two other ER stress components x-box binding protein 1 (XBP1) and ATF6. In addition, rats living in an enriched environment (large group cage with novel toys changed daily) exhibited rapid induction of GADD34 and ATF3 after 30min of exploring novel toys, suggesting these genes are also involved in normal non-pathological signaling. However, environmental enrichment, a paradigm that produces protective addiction and depression phenotypes in rats, attenuated the rapid induction of ATF3 and GADD34 after restraint stress. These experiments provide a sensitive measure of ER stress and, more importantly, these results offer good evidence of the activation of ER stress mechanisms from psychological stress, cocaine and natural reward. Thus, ER stress genes may be targets for novel therapeutic targets for depression and addiction.

A case report of semitendinosus tendon autograft for reconstruction of the meniscal wall supporting a collagen implant

PurposeDescribe the evolution of the reconstruction of meniscal rim with semitendinosus tendon in a patient with knee pain after a subtotal meniscectomy and absence of meniscal wall.Method32 years old male with a six-month history of the left knee pain after a subtotal meniscectomy. The MRI indicated a small internal meniscal remainder without posterior horn attachment. Taking this absence as a relative contraindication for implant and meniscal transplantation, the reconstruction of a new meniscal wall with semitendinosus tendon autograft was considered. A collagen meniscal implant was attached to the new wall five months later.ResultsAfter two years the patient referred only non specific discomfort with full pain relief in the medial compartment. The MRI revealed integration of implants without significant degenerative changes compared to previous images.ConclusionsThis staged technique was designed to restore medial meniscus-like biologic tissue in a symptomatic patient following arthroscopic subtotal meniscectomy with a significant loss of the peripheral meniscus rim. Symptomatic improvement was obtained at two years follow-up.