From stem cells to viable autologous semilunar heart valve.

BACKGROUND: An estimated 275,000 patients undergo heart valve replacement each year. However, existing solutions for valve replacement are complicated by the morbidity associated with lifelong anticoagulation of mechanical valves and the limited durability of bioprostheses. Recent advances in tissue engineering and our understanding of stem cell biology may provide a lifelong solution to these problems. METHODS AND RESULTS: Mesenchymal stem cells were isolated from ovine bone marrow and characterized by their morphology and antigen expression through immunocytochemistry, flow cytometry, and capacity to differentiate into multiple cell lineages. A biodegradable scaffold was developed and characterized by its tensile strength and stiffness as a function of time in cell-conditioned medium. Autologous semilunar heart valves were then created in vitro using mesenchymal stem cells and the biodegradable scaffold and were implanted into the pulmonary position of sheep on cardiopulmonary bypass. The valves were evaluated by echocardiography at implantation and after 4 months in vivo. Valves were explanted at 4 and 8 months and examined by histology and immunohistochemistry. Valves displayed a maximum instantaneous gradient of 17.2+/-1.33 mm Hg, a mean gradient of 9.7+/-1.3 mm Hg, an effective orifice area of 1.35+/-0.17 cm2, and trivial or mild regurgitation at implantation. Gradients changed little over 4 months of follow-up. Histology showed disposition of extracellular matrix and distribution of cell phenotypes in the engineered valves reminiscent of that in native pulmonary valves. CONCLUSIONS: Stem-cell tissue-engineered heart valves can be created from mesenchymal stem cells in combination with a biodegradable scaffold and function satisfactorily in vivo for periods of >4 months. Furthermore, such valves undergo extensive remodeling in vivo to resemble native heart valves.

A nuclear form of the heparin-binding epidermal growth factor-like growth factor precursor is a feature of aggressive transitional cell carcinoma.

Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a member of the ErbB receptor ligand family, exists in distinct molecular forms with disparate biological activities. Previous studies have shown that the HB-EGF precursor, proHB-EGF, localizes to the cytoplasm of transitional cells of the human bladder urothelium and that the soluble form of the growth factor is an autocrine urothelial cell mitogen. In this study, we identify a potential role for proHB-EGF in transitional cell carcinoma (TCC) of the bladder. In an analysis of 33 TCC specimens and 8 normal controls, proHB-EGF, identified using an antibody directed against the cytoplasmic tail domain, localized to cell nuclei in a manner that correlated positively with tumor stage and grade (P < 0.001). The ability of proHB-EGF to localize to the nucleus was independently confirmed in a TCC cell line (TCCSUP), in which approximately 40% of transfected proHB-EGF was found to reside in the nuclear compartment. In Kaplan-Meier survival analysis, TCC patients with >20% proHB-EGF-positive cell nuclei demonstrated markedly reduced survival compared with patients with <20% proHB-EGF-positive nuclei (P < 0.005, log-rank test). In multivariate analysis, nuclear localization of proHB-EGF of >20% was an independent prognostic indicator of disease-specific mortality. This is the first report in any cell type that HB-EGF is capable of translocating to the cell nucleus. In addition, our findings suggest that nuclear proHB-EGF may play a role in disease progression in bladder cancer and possibly other cancers.

Radiation exposure reduction during voiding cystourethrography in a pediatric porcine model of vesicoureteral reflux.

PURPOSE: To compare grid-controlled variable-rate pulsed fluoroscopy (GCPFL) and continuous fluoroscopy (CFL) for the reduction of radiation exposure during voiding cystourethrography (VCUG) in a pediatric porcine model of vesicoureteral reflux. MATERIALS AND METHODS: Institutional animal care and use committee approval was obtained. Vesicoureteral reflux was simulated in four pigs, and 48 VCUG studies were performed (24 with GCPFL, 24 with CFL). VCUG was performed at abdominal girths of 8-10 cm (group 1, simulates human newborn to 6-month-old infant), 12-13 cm (group 2, simulates 2-3-year-old child), and 15-17 cm (group 3, simulates 10-year-old child). An electronic device calculated total radiation exposure during fluoroscopy and image recording. With five-point ordinal scales, VCUG images were scored independently for anatomic conspicuity and overall diagnostic quality by two radiologists (radiologists A and B). An analysis of variance was used to compare radiation exposures and fluoroscopy times between GCPFL and CFL and to determine whether radiation exposure and fluoroscopy time were dependent on the pig's abdominal girth. The Pearson product-moment correlation coefficient was used to assess whether fluoroscopy time was correlated with radiation exposure. Anatomic conspicuity and diagnostic quality scores were compared by means of the Wilcoxon signed rank test. RESULTS: Results of analysis of variance revealed that GCPFL resulted in a significant reduction in total radiation exposure compared with CFL for each of the three groups (P < .05 for each comparison), and this reduction was most marked in the larger animals. There were no significant differences in diagnostic quality of the recorded VCUG images (P > .05). Anatomic conspicuity was not significantly different for groups 2 and 3, but there was a significantly higher score for GCPFL in group 1 for radiologist A (P = .04). CONCLUSION: By using GCPFL in the performance of VCUG in a pediatric porcine model of vesicoureteral reflux, total radiation exposure can be reduced by a factor of 4.6-7.5 lower than with CFL, and diagnostic-quality images can be obtained.

Rate and predictors of spontaneous resolution of prenatally diagnosed primary nonrefluxing megaureter.

PURPOSE: We assessed clinical and ultrasound predictors of the spontaneous resolution of prenatally diagnosed, primary nonrefluxing megaureter. MATERIALS AND METHODS: Primary nonrefluxing megaureter was prenatally diagnosed in 54 neonates from 1993 to 1998, including 15 in whom it was bilateral (69 megaureters). Median followup period from birth to the last radiology examination was 25.8 months (range 2.3 to 72). RESULTS: Of the patients 10 underwent surgical repair for severe hydroureteronephrosis, including decreasing renal function on diuretic renography in 1, and an episode of bilateral obstruction and acute renal failure in 1. Hydroureteronephrosis resolved in 39 cases (72%) and persisted in 5 at 30 to 72 months of followup. Laterality, gender and retrovesical ureteral diameter had no significant effect on the resolution rate (p >0.05). Mean initial ureteral diameter in patients with resolution was less than in those without resolution and in those who underwent surgery (0.8 versus 1.15 and 1.32 cm., respectively). Presenting hydronephrosis grade was a significant predictor of the resolution rate (p = 0.03). Grades 1 to 3 hydronephrosis resolved at a median age of 12.9, 23.9 and 34.6 months, respectively (range 4.1 to 66). In patients in whom grades 4 and 5 hydronephrosis resolved the median age at resolution was 48.5 months. CONCLUSIONS: Most cases of prenatally diagnosed, primary nonrefluxing megaureter resolve spontaneously. Grades 1 to 3 hydronephrosis tend to resolve between ages 12 and 36 months. For these grades followup intervals may be progressively elongated after a pattern of improving hydronephrosis has been established. In children with grade 4 or 5 hydronephrosis, or a retrovesical ureteral diameter of greater than 1 cm. the condition may resolve slowly and require surgery.

Anterior urethral valves and diverticula in children: a result of ruptured Cowper's duct cyst?

OBJECTIVE: To review a series of children with anterior urethral valves and diverticula, to elucidate the pathophysiology and optimal management of this entity. PATIENTS AND METHODS: Nine cases (all boys; 1963 to 2003) were reviewed retrospectively. RESULTS: Seven of nine boys had bulbar diverticula. Continuity between Cowper's duct and the diverticulum was noted endoscopically in two and confirmed radiographically in one. Initially, open surgery was curative but more recently endoscopic management has been the procedure of choice. CONCLUSION: This series indicates that the distal lip of a ruptured syringocele may function as a flap-valve, leading to anterior urethral obstruction. Advances in imaging and endoscopic instruments have altered the mode of presentation and management of this entity.