Mincheol Choi

Umbilical-cord-blood-derived mesenchymal stem cells seeded onto fibronectin-immobilized polycaprolactone nanofiber improve cardiac function

Stem cells seeded onto biofunctional materials have greater potency for therapeutic applications. We investigated whether umbilical-cord-blood-derived mesenchymal stem cell (UCB-MSC)-seeded fibronectin (FN)-immobilized polycaprolactone (PCL) nanofibers could improve cardiac function and inhibit left ventricle (LV) remodeling in a rat model of myocardial infarction (MI). Aligned nanofibers were uniformly coated with poly(glycidyl methacrylate) by initiated chemical vapor deposition followed by covalent immobilization of FN proteins. The degree of cell elongation and adhesion efficacy were improved by FN immobilization. Furthermore, genes related to angiogenesis and mesenchymal differentiations were up-regulated in the FN-immobilized PCL nanofibers in comparison to control PCL nanofibers in vitro. 4 weeks after the transplantation in the rat MI model, the echocardiogram showed that the UCB-MSC-seeded FN-immobilized PCL nanofiber group increased LV ejection fraction and fraction shortening as compared to the non-treated control and acellular FN-immobilized PCL nanofiber groups. Histological analysis indicated that the implantation of UCB-MSCs with FN-immobilized PCL nanofibers induced a decrease in MI size and fibrosis, and an increase in scar thickness. This study indicates that FN-immobilized biofunctional PCL nanofibers could be an effective carrier for UCB-MSC transplantation for the treatment of MI.

DETERMINATION OF NUMBER DENSITY, SIZE AND MORPHOLOGY OF AGGREGATES IN COFLOW DIFFUSION FLAMES USING LIGHT SCATTERING AND LOCAL SAMPLING

Abstract Number density, size, and morphology of silica aggregate particles in coflow diffusion flames have been determined experimentally using a novel method combining light scattering and local sampling techniques. The number density and volume fraction of both aggregates and spherical particles have been calculated from the scattering cross section measured from 90° light scattering with the combination of determining particle size and morphology from the localized sampling and TEM image analysis. Rayleigh–Debye–Gans and Mie theories have been applied to the calculations for fractal aggregates and spherical particles, respectively. Of particular interests are the effects of carrier gas flow rates, different carrier gases, and flame temperatures on the growth of silica particles and the roles of radial heat and H 2 O diffusion have been studied when using N 2 or O 2 as a carrier gas.

Ultrasound-guided mesenteric lymph node iohexol injection for thoracic duct computed tomographic lymphography in cats.

Computed tomographic (CT) lymphography was performed in cats using percutaneous ultrasound-guided injection of contrast medium into a mesenteric lymph node. The thoracic duct and its branches were clearly delineated in CT images of seven cats studied. The thoracic duct was characterized by anatomic variation and appeared as single or multiple branches. The thoracic duct and the cisterna chyli were identified along the ventral or left ventral aspect of the vertebrae from the level of the cranial lumbar to the caudal cervical vertebrae. The thoracic duct was identified in the central caudal mediastinum, deviated to the left in the cranial mediastinum, and finally moved toward the venous system. Small volumes of extranodal contrast medium leakage were identified in all cats. After injection, the mesenteric lymph nodes were cytologically normal. Ultrasound-guided CT lymphography via percutaneous mesenteric lymph node injection appears safe and effective in cats.

Targeted Genome Engineering to Control VEGF Expression in Human Umbilical Cord Blood‐Derived Mesenchymal Stem Cells: Potential Implications for the Treatment of Myocardial Infarction

Human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSCs) exhibit potency for the regeneration of infarcted hearts. Vascular endothelial growth factor (VEGF) is capable of inducing angiogenesis and can boost stem cell‐based therapeutic effects. However, high levels of VEGF can cause abnormal blood vessel growth and hemangiomas. Thus, a controllable system to induce therapeutic levels of VEGF is required for cell therapy. We generated an inducible VEGF‐secreting stem cell (VEGF/hUCB‐MSC) that controls the expression of VEGF and tested the therapeutic efficacy in rat myocardial infarction (MI) model to apply functional stem cells to MI. To introduce the inducible VEGF gene cassette into a safe harbor site of the hUCB‐MSC chromosome, the transcription activator‐like effector nucleases system was used. After confirming the integration of the cassette into the locus, VEGF secretion in physiological concentration from VEGF/hUCB‐MSCs after doxycycline (Dox) induction was proved in conditioned media. VEGF secretion was detected in mice implanted with VEGF/hUCB‐MSCs grown via a cell sheet system. Vessel formation was induced in mice transplanted with Matrigel containing VEGF/hUCB‐MSCs treated with Dox. Moreover, seeding of the VEGF/hUCB‐MSCs onto the cardiac patch significantly improved the left ventricle ejection fraction and fractional shortening in a rat MI model upon VEGF induction. Induced VEGF/hUCB‐MSC patches significantly decreased the MI size and fibrosis and increased muscle thickness, suggesting improved survival of cardiomyocytes and protection from MI damage. These results suggest that our inducible VEGF‐secreting stem cell system is an effective therapeutic approach for the treatment of MI. Stem Cells Translational Medicine 2017;6:1040–1051

Assessment of glomerular filtration rate with dynamic computed tomography in normal Beagle dogs.

The objective of our study was to determine individual and global glomerular filtration rates (GFRs) using dynamic renal computed tomography (CT) in Beagle dogs. Twenty-four healthy Beagle dogs were included in the experiment. Anesthesia was induced in all dogs by using propofol and isoflurane prior to CT examination. A single slice of the kidney was sequentially scanned after a bolus intravenous injection of contrast material (iohexol, 1 mL/kg, 300 mgI/mL). Time attenuation curves were created and contrast clearance per unit volume was calculated using a Patlak plot analysis. The CT-GFR was then determined based on the conversion of contrast clearance per unit volume to contrast clearance per body weight. At the renal hilum, CT-GFR values per unit renal volume (mL/min/mL) of the right and left kidneys were 0.69 ± 0.04 and 0.57 ± 0.05, respectively. No significant differences were found between the weight-adjusted CT-GFRs in either kidney at the same renal hilum (p = 0.747). The average global GFR was 4.21 ± 0.25 mL/min/kg and the whole kidney GFR was 33.43 ± 9.20 mL/min. CT-GFR techniques could be a practical way to separately measure GFR in each kidney for clinical and research purposes.

Computed tomographic evaluation of cervical vertebral canal and spinal cord morphometry in normal dogs.

The height, width, and cross-sectional area of the vertebral canal and spinal cord along with the area ratio of spinal cord to vertebral canal in the cervical vertebra were evaluated in images obtained using computed tomography (CT). Measurements were taken at the cranial, middle, and caudal point of each cervical vertebra in eight clinically normal small breed dogs (two shih tzu, two miniature schnauzers, and four mixed breed), 10 beagles, and four German shepherds. CT myelography facilitated the delineation of the epidural space, subarachnoid space, and spinal cord except at the caudal portion of the 7th cervical vertebra. The spinal cord had a tendency to have a clear ventral border in the middle portion of the vertebral canal and lateral borders near both end plates. The height, width, and area of the vertebral canal and spinal cord in the cervical vertebra were increased as the size of dog increased. However, the ratio of the spinal cord area to vertebral canal area in the small dogs was higher than that of the larger dogs. Results of the present study could provide basic and quantitative information for CT evaluation of pathologic lesions in the cervical vertebra and spinal cord.

Computed tomographic evaluation of abdominal fat in minipigs

Computed tomography (CT) exams were conducted to determine the distribution of abdominal fat identified based on the CT number measured in Hounsfield Units (HU) and to measure the volume of the abdominal visceral and subcutaneous fat in minipigs. The relationship between the CT-based fat volumes of several vertebral levels and the entire abdomen and anthropometric data including the sagittal abdominal diameter and waist circumference were evaluated. Moreover, the total fat volumes at the T11, T13, L3, and L5 levels were compared with the total fat volume of the entire abdomen to define the landmark of abdominal fat distribution. Using a single-detector CT, six 6-month-old male minipigs were scanned under general anesthesia. Three radiologists then assessed the HU value of visceral and subcutaneous abdominal fat by drawing the region of interest manually at the T11, T13, L1, L3, and L5 levels. The CT number and abdominal fat determined in this way by the three radiologists was found to be correlated (intra-class coefficient = 0.9). The overall HU ranges for the visceral and subcutaneous fat depots were -147.47 to -83.46 and -131.62 to -90.97, respectively. The total fat volume of the entire abdomen was highly correlated with the volume of abdominal fat at the T13 level (r = 0.97, p < 0.0001). These findings demonstrate that the volume of abdominal adipose tissue measured at the T13 level using CT is a strong and reliable predictor of total abdominal adipose volume.

Evaluation of triphasic helical computed tomography of the kidneys in clinically normal dogs

OBJECTIVE To determine computed tomography (CT) delay times by use of a sequential scan and identify the normal enhancement pattern in each phase of a triphasic CT scan of the kidneys in dogs. ANIMALS 14 healthy Beagles. PROCEDURES A sequential CT scan was used for investigating delay time, and a triphasic helical CT scan was used for identifying the normal enhancement pattern and determining Hounsfield unit values in the kidneys of dogs. RESULTS In the cine scan (single-slice dynamic scan), the optimal delay times were 10 seconds in the corticomedullary phase and 44 seconds in the nephrographic phase, after contrast medium injection. By use of triphasic CT images, Hounsfield unit values were acquired in each phase. CONCLUSIONS AND CLINICAL RELEVANCE Triphasic CT of the kidneys in clinically normal dogs was established by acquisition of delay times in a cine scan and may become an important imaging modality in the diagnosis of renal diseases and in treatment planning in dogs.

CT THORACIC DUCT LYMPHOGRAPHY IN CATS BY POPLITEAL LYMPH NODE IOHEXOL INJECTION

Three different doses (1.0, 1.5, and 2.0 ml) of iohexol (300 mgl/ml) were injected percutaneously into the popliteal lymph node of eight adult cats under ultrasound guidance. Serial transverse CT images of five regions of interest (L3, T13, T8, T4, and T1 level) were performed at 2-min intervals, and the attenuation in Hounsfield Units (HU) of the lymphatic vessels was measured for determination of the optimal dose of iohexol and CT scan parameters. The optimal dose was 1.5 ml and helical CT acquisition is recommended to be performed as soon as possible after iohexol injection. In helical scans, the thoracic duct was characterized by variable branch numbers that formed a single trunk and entered the venous system at variable levels. CT lymphography using this protocol was performed in a cat with chylothorax. The thoracic duct was tortuous and focally dilated, and leakage of contrast medium was observed. Percutaneous CT lymphography using ultrasound-guided administration of iohexol into the popliteal lymph node appears reliable for delineation of the thoracic duct in cats.

Cranial epidural spread of contrast medium and new methylene blue dye in sternally recumbent anaesthetized dogs

OBJECTIVE To examine the spread of solution in the epidural space of sternally recumbent dogs. STUDY DESIGN Prospective experimental trial. Animals  Ten healthy adult Beagle dogs weighing 7.6 ± 1.1 kg. METHODS Dogs were anaesthetized with total intravenous propofol infusion, and placed in sternal recumbency. A volume of 0.2 mL kg(-1) contrast medium (CM) containing 1% new methylene blue (MB) dye was administered into the lumbosacral epidural space. Left to right lateral radiographs using a horizontal beam were taken every 5 minutes for 45 minutes. The perpendicular height (PH) between floor of the epidural canal of the highest vertebra and that of lumbosacral spinal canal was measured on radiographs. The angle of slope from the injection point toward the highest vertebral floor was measured. Immediately after taking the last radiographic image, dogs were euthanized and a laminectomy was performed from the cervical to lumbar vertebrae for visual evaluation of MB spread. The spread of CM and of MB as counted in number of stained vertebra were compared, and each of these data sets were further compared to PH and angle, using linear regression analyses. RESULTS The PH and angle were (mean ± SD) 3.8 ± 0.8 cm and 14.8 ± 2.8° respectively. The most cranial spread of CM was at 12.7 ± 5.7 (range: C6-L3) vertebrae, and at 14.0 ± 5.4 (range: C6-L2) vertebrae for MB staining. There were no significant correlations between PH and spread of CM (R(2)  = 0.08) or MB (R(2)  = 0.13), between angle and spread of CM (R(2)  = 0.05) or MB (R(2)  = 0.02), respectively. CM and MB demonstrated proportional relationship (R(2)  = 0.82, p < 0.001). CONCLUSIONS No significant inhibitory effect of upward slope on cranial epidural spread of the solution was observed. Other factors may have greater effect on epidural spread in sternally recumbent dogs.