Computed Tomography Volumetry of Bone Cement: Retrospective Blinded Validation of Commercially Available Semi-automated Edge Detection Software.

OBJECTIVE: Cement volumes are increasingly linked to orthopedic oncology and neurosurgical outcomes (construct durability, adjacent fracture), but manual cement volumetry remains time prohibitive. The authors aim to report performance of PACS-integrated volumetric software specifically for barium-enhanced polymethylmethacrylate cement. METHODS: Institutional review board-approved single-institution retrospective review of patients from 2019-2022 undergoing kyphoplasty for pathological compression fractures with a quantitative cement infuser providing true cement volume. An operator blinded to true cement volumes retrospectively performed software-assisted volumetry on follow-up computed tomography scans. RESULTS: Included were 91 kyphoplasty levels in 56 patients: mean age, 62 years (range, 34-85 years), 73% female. True cement volume (available for 44 of 66 procedures) was mean 4.5 mL per level (range, 1.2-15.6 mL). Measured cement volume (available for all procedures) yielded a mean of 6.1 mL per level (range, 1.5-27.9 mL). For the 57 levels (39 patients) where both true and measured cement volumes were available, linear regression intercept and slope were 1.46 (95% CI = 0.97-1.95, P < 0.001) and 0.52 (CI = 0.47-0.57, P < 0.001), respectively, suggesting measured volume averaged 1.46 mL greater than true volume, with each additional milliliter of measured volume corresponding to approximately 0.52 mL of true volume. There was no significant difference in the relationship between estimated and actual cement volume in thoracic levels (intercept = -0.24, CI = -1.13 to 0.66, P = 0.61; slope = 0.03, CI = -0.14 to 0.19, P = 0.73) compared with lumbar levels. The goodness-of-fit of the regression model was strong ( R2 = 0.81). Discrepancies ranged from 90% underestimation to 52% overestimation; average, 17% overestimation. CONCLUSIONS: Semi-automated volumetry maintained a strong correlation with true volumes across the thoracic and lumbar curvatures, overestimating cement volume by a mean of 17% or 1.46 mL.

Intranodal Lymphangiography with Thoracic Duct Embolization for Treatment of Chyle Leak after Thoracic Outlet Decompression Surgery.

From 2015 to 2019, 9 patients underwent ultrasound-guided intranodal lymphangiography for the treatment of a chyle leak following thoracic outlet decompression surgery. Chyle leaks were identified by Lipiodol (Guerbet, Roissy, France) extravasation near the left supraclavicular surgical bed in all patients. The technical success rate of thoracic duct embolization was 67% (6 of 9), including fluoroscopic transabdominal antegrade access (n = 4) and ultrasound-guided retrograde access in the left neck (n = 2). Clinical success was achieved in 89% of patients (8 of 9). The mean interval from lymphangiography to drain removal was 6.6 days (range, 4-18 d). No patients had a chyle leak recurrence during clinical follow-up (mean, 304 d).

Effect of hyaluronic acid hydrogels containing astrocyte-derived extracellular matrix and/or V2a interneurons on histologic outcomes following spinal cord injury.

One reason for the lack of regeneration, and poor clinical outcomes, following central nervous system (CNS) injury is the formation of a glial scar that inhibits new axon growth. In addition to forming the glial scar, astrocytes have been shown to be important for spontaneous SCI recovery in rodents, suggesting some astrocyte populations are pro-regenerative, while others are inhibitory following injury. In this work, the effect of implanting hyaluronic acid (HA) hydrogels containing extracellular matrix (ECM) harvested from mouse embryonic stem cell (mESC)-derived astrocytes on histologic outcomes following SCI in rats was explored. In addition, the ability of HA hydrogels with and without ECM to support the transplantation of mESC-derived V2a interneurons was tested. The incorporation of ECM harvested from protoplasmic (grey matter) astrocytes, but not ECM harvested from fibrous (white matter) astrocytes, into hydrogels was found to reduce the size of the glial scar, increase axon penetration into the lesion, and reduce macrophage/microglia staining two weeks after implantation. HA hydrogels were also found to support transplantation of V2a interneurons and the presence of these cells caused an increase in neuronal processes both within the lesion and in the 500 µm surrounding the lesion. Overall, protoplasmic mESC-derived astrocyte ECM showed potential to treat CNS injury. In addition, ECM:HA hydrogels represent a novel scaffold with beneficial effects on histologic outcomes after SCI both with and without cells.

Different Mixed Astrocyte Populations Derived from Embryonic Stem Cells Have Variable Neuronal Growth Support Capacities.

Central nervous system injury often leads to functional impairment due, in part, to the formation of an inhibitory glial scar following injury that contributes to poor regeneration. Astrocytes are the major cellular components of the glial scar, which has led to the belief that they are primarily inhibitory following injury. Recent work has challenged this by demonstrating that some astrocytes are required for spinal cord regeneration and astrocytic roles in recovery depend on their phenotype. In this work, two mixed populations containing primarily either fibrous or protoplasmic astrocytes were derived from mouse embryonic stem cells (mESCs). Motoneuron and V2a interneuron growth on live cultures, freeze-lysed cultures, or decellularized extracellular matrix (ECM) from astrocytes were assessed. Both neuronal populations were found to extend significantly longer neurites on protoplasmic-derived substrates than fibrous-derived substrates. Interestingly, neurons extended longer neurites on protoplasmic-derived ECM than fibrous-derived ECM. ECM proteins were compared with in vivo astrocyte expression profiles, and it was found that the ESC-derived ECMs were enriched for astrocyte-specific proteins. Further characterization revealed that protoplasmic ECM had significantly higher levels of axon growth promoting proteins, while fibrous ECM had significantly higher levels of proteins that inhibit axon growth. Supporting this observation, knockdown of spondin-1 improved neurite growth on fibrous ECM, while laminin α5 and γ1 knockdown decreased neurite growth on protoplasmic ECM. These methods allow for scalable production of specific astrocyte subtype-containing populations with different neuronal growth support capacities, and can be used for further studies of the functional importance of astrocyte heterogeneity.

Three-dimensional cell culture environment promotes partial recovery of the native corneal keratocyte phenotype from a subcultured population.

Corneal disease is the fourth leading cause of blindness. According to the World Health Organization, roughly 1.6 million people globally are blind as a result of this disease. The only current treatment for corneal opacity is a corneal tissue transplant. Unfortunately, the demand for tissue exceeds supply, making a tissue-engineered in vitro cornea highly desirable. For an in vitro cornea to be useful, it must be transparent, which requires downregulation of the light-scattering intracellular protein alpha-smooth muscle actin (αSMA) and upregulation of the native corneal marker, aldehyde dehydrogenase 1A1 (ALDH1A1). This study focuses on the effects of a three-dimensional (3D) matrix on the expression levels of αSMA and ALDH1A1 by a subcultured population of rabbit corneal keratocytes and the comparison of the 3D matrix effects to other culture conditions. We show that, through western blot and quantitative real-time PCR, the presence of collagen strongly downregulates αSMA. Further, 3D cultures maintain low actin expression even in the presence of a proinflammatory cytokine, transforming growth factor-beta (TGF-ß). Finally, 3D culture conditions show a partial recovery of ALDH1A1 expression, which has never been previously observed in a serum-exposed subcultured cell population. Overall, this study suggests that 3D culture is not only a relatively stronger signal than both collagen and TGF-ß, it is also sufficient to induce some recovery of ALDH1A1 and the native corneal phenotype despite the presence of serum.

Precise nanometer localization analysis for individual fluorescent probes.

Calculation of the centroid of the images of individual fluorescent particles and molecules allows localization and tracking in light microscopes to a precision about an order of magnitude greater than the microscope resolution. The factors that limit the precision of these techniques are examined and a simple equation derived that describes the precision of localization over a wide range of conditions. In addition, a localization algorithm motivated from least-squares fitting theory is constructed and tested both on image stacks of 30-nm fluorescent beads and on computer-generated images (Monte Carlo simulations). Results from the algorithm show good agreement with the derived precision equation for both the simulations and actual images. The availability of a simple equation to describe localization precision helps investigators both in assessing the quality of an experimental apparatus and in directing attention to the factors that limit further improvement. The precision of localization scales as the inverse square root of the number of photons in the spot for the shot noise limited case and as the inverse of the number of photons for the background noise limited case. The optimal image magnification depends on the expected number of photons and background noise, but, for most cases of interest, the pixel size should be about equal to the standard deviation of the point spread function.