Micro‐computed tomography assessment of vertebral column defects in retinoic acid‐Induced rat model of myelomeningocele

Barbe, Mary F.; Adiga, Radhika; Gordiienko, Oleg; Pleshko, Nancy; Selzer, Michael E.; Krynska, Barbara

doi:10.1002/bdra.23254

Cited by 11 publications

(12 citation statements)

References 30 publications

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“…Time-dated Sprague-Dawley pregnant rats obtained from Charles River Laboratories (Wilmington, MA) were placed on a standard dark: light schedule. An MMC defect was induced in the fetuses of time-dated pregnant rats by gavage of a single dose of 50 mg/kg of all trans-retinoic acid (Sigma-Aldrich Chemical, Saint Louis, MO) dissolved in olive oil on embryonic day 10 (E10), as described previously [ 32 , 34 ]. Normal control mothers were gavage fed with olive oil.…”

Section: Methodsmentioning

confidence: 99%

Clusters of amniotic fluid cells and their associated early neuroepithelial markers in experimental myelomeningocele: Correlation with astrogliosis

et al. 2017

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Myelomeningocele (MMC) is the most common and severe disabling type of spina bifida resulting in the exposure of vulnerable spinal cord to the hostile intrauterine environment. In this study, we sought to examine the cellular content of fetal amniotic fluid (AF) in MMC and explore a correlation between these cells and pathological development of MMC. MMC was induced in fetal rats by exposing pregnant mothers to all-trans retinoic acid and AF samples were collected before term. Cells were isolated from AF samples and morphologically and phenotypically characterized in short-term cultures. In addition, the spinal cord injury in MMC fetuses was assessed by immunohistochemical examination of astrogliosis. We identified a population of cells from the AF of MMC fetuses (MMC-AF) that formed adherent clusters of tightly packed cells, which were absent from the AF of normal control fetuses (norm-AF). MMC-AF clusters contained cells co-expressing adherens junction associated proteins (ZO-1), N-cadherin and F-actin at sites of cell-cell contacts. In addition, they expressed markers of early neuroepithelial cells such as SOX-1 and Pax-6 along with other stem/progenitor cell markers such as SOX-2 and nestin. Subpopulations of cells in MMC-AF clusters also expressed more advanced differentiation markers such as doublecortin and GFAP. We found that the appearance of cluster forming cells in cultures from MMC-AF correlated with activation of astrogliosis associated with the spinal cord injury in MMC fetuses. In summary, we identified a neuroepithelial cell population in the AF of MMC fetuses that formed adherent clusters in culture and we characterized cellular markers of these cells. Our data suggests that the phase of the disease is a crucial factor in the emergence of these cells into the AF and that these cells may provide a new and important platform for studying the progression of MMC and development of improved strategies for the repair and diagnosis of MMC prenatally.

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Section: Methodsmentioning

confidence: 99%

Clusters of amniotic fluid cells and their associated early neuroepithelial markers in experimental myelomeningocele: Correlation with astrogliosis

et al. 2017

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“…The 3D-printed IA model is coated with paint comprised of 0.002×0.008μm hex-cut metallic flakes (KUS MSF15-2Z, TCP Global Corporation) to distinguish the inner tissue surface from the 3D-printed model in the micro-CT images 1 . Mounting Clay (Renishaw Inc.) is used to fix the model to a brass mount compatible with CT scanning, Figure 2F.…”

Section: Methodology For Mapping Resected Ia Tissue To Reconstructmentioning

confidence: 99%

Regional Mapping of Flow and Wall Characteristics of Intracranial Aneurysms

et al. 2016

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The evolution of intracranial aneurysms (IAs) is thought to be driven by progressive wall degradation in response to abnormal hemodynamics. Previous studies focused on the relationship between global hemodynamics and wall properties. However, hemodynamics, wall structure and mechanical properties of cerebral aneurysms can be non-uniform across the aneurysm wall. Therefore, the aim of this work is to introduce a methodology for mapping local hemodynamics to local wall structure in resected aneurysm specimens. This methodology combines image-based computational fluid dynamics, tissue resection, micro-CT imaging of resected specimens mounted on 3D-printed aneurysm models, alignment to 3D vascular models, multi-photon microscopy of the wall, and regional mapping of hemodynamics and wall properties. This approach employs a new 3D virtual marking tool for surgeons to delineate the location of the resected specimen directly on the 3D model, while in the surgical suite. The case of a middle cerebral artery aneurysm is used to illustrate the application of this methodology to the assessment of the relationship between local wall shear stress and local wall properties including collagen fiber organization and wall geometry. This methodology can similarly be used to study the relationship between local intramural stresses and local wall structure.

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“…Alternatively, if the sample is very small, cling wrap (Glad ®, CA, USA) can be used to mount the sample on the stub (Barbe, 2014). This particular brand of wrap was found to avoid substantial x-ray attenuation.…”

Section: Use Of Micro-computed Tomography For a Priori Selection Of Imentioning

confidence: 99%

“…He, along with his collaborators, conceived the method of using the tracer for ex vivo calcification imaging under a confocal microscope (Hutcheson et al., ). We appreciate insights from Dr. Mary F. Barbe of Temple University School of Medicine, who shared methods for using plastic wrap to keep the samples hydrated during micro‐CT scanning (Barbe et al., ). We would also like to thank undergraduate summer intern Thomas Bina for manually tracing fibers for the collagen images in Figure .…”

Section: Commentarymentioning

confidence: 99%

“…To ensure maximum field of view, place the sample in the center of tube, symmetrically located with respect to the vertical axis. Alternatively, if the sample is very small, cling wrap (Glad ®, CA, USA) can be used to mount the sample on the stub (Barbe, 2014). This particular brand of wrap was found to avoid substantial x-ray attenuation.…”

Section: Use Of Micro-computed Tomography For a Priori Selection Of Imentioning

confidence: 99%

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Multiphoton Imaging of Collagen, Elastin, and Calcification in Intact Soft‐Tissue Samples

2018

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Multiphoton induced second harmonic generation and two photon excitation enables imaging of collagen and elastin fibers at micron level resolution to depths of hundreds of microns, without exogenous stains. These attributes can be leveraged for quantitative analysis of the 3D architecture of collagen and elastin fibers within intact, soft tissue specimens such as the artery and bladder wall. This architecture influences the function of intramural cells and also plays a primary role in determining tissue passive mechanical properties. Calcification deposition in soft tissues is a highly prevalent pathology in both older and diseased populations that can alter tissue properties. In this unit, we provide a protocol for simultaneous multiphoton microscopy (MPM) imaging and analysis of the 3D collagen and elastin structure with calcification, effective for fixed and fresh intact samples. We also provide an associated micro-CT protocol to identify regions of interest in the samples to target the MPM imaging.

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Micro‐computed tomography assessment of vertebral column defects in retinoic acid‐Induced rat model of myelomeningocele

Abstract: High-resolution micro-CT combined with digital quantification methods is a powerful technique ideally suited for precise assessment of complex congenital skeletal abnormalities such as examined in this rodent model of MMC.

Cited by 11 publications

References 30 publications

Clusters of amniotic fluid cells and their associated early neuroepithelial markers in experimental myelomeningocele: Correlation with astrogliosis

Clusters of amniotic fluid cells and their associated early neuroepithelial markers in experimental myelomeningocele: Correlation with astrogliosis

Regional Mapping of Flow and Wall Characteristics of Intracranial Aneurysms

Multiphoton Imaging of Collagen, Elastin, and Calcification in Intact Soft‐Tissue Samples

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