1.Tissue-engineered graft constructed by self-derived cells and heterogeneous acellular matrix.
Hui-min HUANG ; Shao-feng WU ; Hong REN
Journal of Zhejiang University. Science. B 2006;7(5):351-356
BACKGROUNDEndothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft.
METHODSA 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique. An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step decellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery.
RESULTSThe cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen alpha-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant.
CONCLUSIONCultured self-derived endothelial and smooth muscle cells could be used as seeding cells and heterogeneous acellularized matrix could be used as scaffold in constructing tissue-engineered graft.
Animals ; Cell Differentiation ; Cell Transplantation ; Cells, Cultured ; Graft Survival ; Sheep ; Swine ; Tissue Engineering ; instrumentation ; methods ; Transplantation, Heterologous
2.Contribution of the xenograft bone plate-screw system in lumbar transpedicular stabilization of dogs: an in-vitro study.
Hakan SALCI ; Sani SARIGUL ; Seref DOGAN ; Huseyin LEKESIZ ; Resat OZCAN ; O Sacit GORGUL ; Kaya AKSOY
Journal of Veterinary Science 2008;9(2):193-196
We performed biomechanical comparison of a xenograft bone plate-screw (XBPS) system for achieving cadaveric lumbar transpedicular stabilization (TS) in dogs. Twenty dogs' cadaveric L2-4 lumbar specimens were harvested and their muscles were removed, but the discs and ligaments were left intact. These specimens were separated to four groups: the L2-4 intact group as control (group I, n = 5), the L3 laminectomy and bilateral facetectomy group (LBF) (group II, n = 5), the LBF plus TS with metal plate-screw group (group III, n = 5) and the LBF plus TS with XBPS group (group IV, n = 5). Five kinds of biomechanical tests were applied to the specimens: flexion, extension, left-right bending and rotation. The averages of the 16 stiffness values were calculated and then these were statistically analyzed. The statistical results show that the XBPS system contributes spinal stability and this system can be a good choice for achieving TS.
Animals
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Biomechanics
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Bone Plates/*veterinary
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Bone Screws/*veterinary
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Cadaver
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Dogs
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Lumbar Vertebrae/physiology/*surgery
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Range of Motion, Articular/physiology
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Spinal Fusion/instrumentation/*methods
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Transplantation, Heterologous/instrumentation/*methods
3.Diffusion-Weighted Imaging of a Prostate Cancer Xenograft Model Seen on a 7 Tesla Animal MR Scanner: Comparison of ADC Values and Pathologic Findings.
Dae Chul JUNG ; Hak Jong LEE ; Jin Won SEO ; So Yeon PARK ; Sang Jin LEE ; Joo Hyuk LEE ; In Hoo KIM
Korean Journal of Radiology 2012;13(1):82-89
OBJECTIVE: To assess the relationship between apparent diffusion coefficient (ADC) values on diffusion-weighted magnetic resonance (MR) imaging and pathologic measures of a tumor using a prostate cancer xenograft model. MATERIALS AND METHODS: Eighteen athymic nude mice with 36 PC-3-induced tumors were sacrificed to obtain specimens immediately after MR imaging in order to compare the findings on MR images with those seen on pathological specimens. Using a high-field small-animal MR scanner, T1- and T2-weighted imaging and DW MR imaging was performed. Tumors were then processed for Hematoxylin and Eosin staining to evaluate tumor cellularity, intratumoral necrosis and immunostaining using antibodies directed against CD31 and vascular endothelial growth factor (VEGF) to determine the levels of microvessel density (MVD). Mean ADC values that were measured on the solid portion within each tumor were compared with tumor volume, cellularity, degree of necrosis, VEGF expression, and MVD in the corresponding section of the pathological specimen. RESULTS: Mean ADC values of the solid portion within the PC-3-induced high-grade tumors were significantly correlated with the degree of intratumoral necrosis (r = 0.63, p < 0.0001) and MVD (r = -0.44, p = 0.008) on pathologic slides. The ADC values were not significantly correlated with tumor cellularity, VEGF expression, or tumor volume in high-grade prostate cancer tissues. CONCLUSION: In the xenografted prostate cancer model, the ADC values of the solid portion of the tumors are significantly correlated with tumor necrosis and MVD of the pathologic specimens. The ADC values may be utilized as surrogate markers for the noninvasive assessment of tumor necrosis and MVD in high-grade prostate cancer.
Animals
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Diffusion Magnetic Resonance Imaging/instrumentation/*methods
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Male
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Mice
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Mice, Nude
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Prostatic Neoplasms/*pathology
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Transplantation, Heterologous
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Vascular Endothelial Growth Factor A/metabolism