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Biomedical subjects

A Atala

Publications and source records attributed to A Atala.

At least 73 records · Page 4Linked to original sources
PubMed1998

Videofetoscopically assisted fetal tissue engineering: skin replacement.

BACKGROUND/PURPOSE: Treatment of several congenital anomalies is frequently hindered by lack of enough tissue for surgical reconstruction in the neonatal period. The purposes of this study were (1) introduction of a novel concept in perinatal surgery, involving minimally invasive harvest of fetal tissue, which is then processed through tissue engineering techniques in vitro while pregnancy is allowed to continue, so that, at delivery, the newborn can benefit from having autologous, expanded tissue promptly available for surgical implantation at birth; (2) analysis of the progress of an engineered fetal skin graft with time, after implantation in the neonate; and (3) study of the effects of current tissue engineering techniques on fetal keratinocytes and fetal dermal fibroblasts. METHODS: Ten 90- to 95-day-gestation fetal lambs underwent surgical creation of two large paramedian excisional skin defects on the posterior body wall. Subsequently, fetal skin specimens no larger than 1.5 x 1.5 cm were videofetoscopically harvested. Fetal keratinocytes and dermal fibroblasts were then separately cultivated and expanded in vitro for 45 to 50 days, resulting in a total of approximately 250 to 300 million cells. Seven to 10 days before fetal delivery, all cells were seeded in two layers on a 16 to 20-cm2, 3-mm thick biodegradable polyglycolic acid polymer matrix. One to 4 days after delivery, the autologous engineered skin was implanted over one of two previously created skin defects. The second skin defect region received an absorbable polymer scaffold without cells as a control. If necessary, the original skin wounds were further amplified before implantation. Each animal provided at least one time-point for histological analysis of both types of repair through excisional biopsies performed at weekly intervals, up to 8 weeks postimplantation. Normal skin specimens were also used as controls. RESULTS: Fetal and neonatal survival rates were 100%. Based on previous postnatal skin engineering studies, fetal dermal fibroblasts multiplied significantly faster in vitro (approximately fivefold) than expected. Fetal keratinocytes multiplied at expected postnatal rates. The engineered grafts induced faster epithelization of the wound (partial at 1 week and complete between 2 and 3 weeks postoperatively) than did the acellular ones (partial at 3 weeks and complete between 3 and 4 weeks postoperatively). Analysis of skin architecture showed a higher level of epidermal organization and less dermal scarring in the wounds that received the engineered, cell-implanted polymer scaffold. CONCLUSIONS: (1) Videofetoscopically assisted fetal tissue engineering is a viable method for obtaining expanded autologous tissue for prompt surgical reconstruction at birth. (2) Fetal skin can be expanded and engineered in vitro at faster rates than expected postnatally, with current tissue engineering techniques. (3) Engineered autologous fetal skin induces a faster and more organized healing of neonatal skin defects than that observed with second intention. This concept may prove useful for the treatment of certain human neonatal conditions such as giant neoplasias, ectopia cordis, and other body wall defects.

Animals↗
PubMed1998

Constant elevation in renal pelvic pressure induces an increase in urinary N-acetyl-beta-D-glucosaminidase in a nonobstructive porcine model.

PURPOSE: To clarify the physiological significance of renal pelvic pressure elevations encountered in the evaluation of hydronephrotic kidney we examined the effects of different levels of renal pelvic pressure on the induction of renal injury. MATERIALS AND METHODS: A nonobstructive porcine model was created in which the urine drained against a constant predetermined pressure gradient. Renal pelvic pressure of 10, 20 and 40 cm. was created in 2, 2 and 4 animals, respectively. During 18 to 23 hours serial urinary N-acetyl-beta-D-glucosaminidase levels were determined as an indicator of renal tubular injury. Tissue specimens were examined histologically and renal arterial blood flow was monitored. RESULTS: Urinary N-acetyl-beta-D-glucosaminidase levels in the kidneys subjected to 10 cm. water remained essentially unchanged. However, at 20 and 40 cm. water statistically significant increases were observed. Similarly, renal arterial blood flow was unchanged at 10 cm. water but it became significantly lower than in controls at 20 and 40 cm. water. Histological evaluation revealed mild to moderate tubular dilatation in the kidneys subjected to 20 and 40 cm. water. CONCLUSIONS: Excessively high collecting system pressure induced renal cellular injury, as reflected by an increase in urinary N-acetyl-beta-D-glucosaminidase levels. While renal pelvic pressure up to 10 cm. water appeared to be innocuous, renal cellular injury was evident within as little as 1 hour at renal pelvic pressures 20 cm. water or greater. The degree of N-acetyl-beta-D-glucosaminidase in the urine also correlated with a decrease in renal arterial blood flow.

Acetylglucosaminidase↗
PubMed1998

Bladder augmentation using allogenic bladder submucosa seeded with cells.

OBJECTIVES: The search for a suitable material to reconstruct the genitourinary tract has been a challenging task. Bowel has been widely used for urinary tract reconstruction, despite its subsequent complications. We investigated the possibility of using allogenic bladder submucosa, a tissue consisting of nonimmunogenic acellular collagen, either with or without cells, as a material for bladder augmentation. METHODS: Partial cystectomies were performed in 10 beagle dogs. Both urothelial and smooth muscle cells were harvested and expanded separately in 5 animals. The allogenic bladder submucosa obtained from sacrificed dogs was seeded with muscle cells on one side and urothelial cells on the opposite side. All beagles underwent cruciate cystotomies on the bladder dome. Augmentation cystoplasty was performed with the allogenic bladder submucosa seeded with cells in 5 animals and with the allogenic bladder submucosa without cells in 5. The augmented bladders were retrieved 2 and 3 months after augmentation. RESULTS: Bladders augmented with the allogenic bladder submucosa seeded with cells showed a 99% increase in capacity compared with bladders augmented with the cell-free allogenic bladder submucosa, which showed only a 30% increase in capacity. All dogs showed a normal bladder compliance, as evidenced by urodynamic studies. Histologically, all retrieved bladders contained a normal cellular organization consisting of a urothelial lined lumen surrounded by submucosal tissue and smooth muscle. Immunocytochemical analyses confirmed the urothelial and muscle cell phenotype and showed the presence of nerve fibers. CONCLUSIONS: These results show that allogenic bladder submucosa seeded with cells appears to be an excellent option as a biomaterial for bladder augmentation.

Animals↗
PubMed1998

Urachal anomalies: defining the best diagnostic modality.

OBJECTIVES: Urachal abnormalities are uncommon and the literature is primarily comprised of case reports. Conclusions regarding the presentation and diagnosis of these abnormalities may be elucidated by reviewing a large experience. METHODS: The records of 45 patients with urachal abnormalities in the pediatric age group were reviewed from 1970 to 1997. This included 24 boys and 21 girls with an age range from 1 day to 20 years (average 4.0 years). The presenting complaint was periumbilical discharge in 19 patients (42%), umbilical cyst or mass in 15 (33%), abdominal or periumbilical pain in 10 (22%), and dysuria in 1 (2%). The diagnosis consisted of a urachal sinus in 22 children (49%), a urachal cyst in 16 (36%), and a patent urachus in 7 (15%). Various radiographic studies were used to establish the diagnosis. RESULTS: Patients with a urachal sinus had 16 voiding cystourethrograms performed (only 1 diagnostic), 9 sinograms (all diagnostic), 8 ultrasounds (4 diagnostic), and 1 excretory urogram (normal). Those with a urachal cyst had 8 voiding cystourethrograms (1 diagnostic), 5 excretory urograms (all normal), 4 ultrasounds (all diagnostic), and 1 computed tomography scan (diagnostic). Children with a patent urachus had 2 excretory urograms (both diagnostic), 1 voiding cystourethrogram (diagnostic), and 2 ultrasounds (normal). One baby with a patent urachus was diagnosed prenatally during ultrasound screening. The diagnosis was made by history and physical examination alone in 5 children and at the time of surgery in 1. Treatment consisted of surgical excision of the urachal abnormality with a cuff of bladder in 22 children, surgical excision without a bladder cuff in 22, incision and drainage of a urachal cyst (1%), and laparoscopic excision of a patent urachus with a bladder cuff in another (1%). There were three wound infections postoperatively. None developed any long-term sequelae. CONCLUSIONS: The diagnosis of urachal abnormalities can be made with certainty if a good physical examination and the appropriate radiographic test are performed. A patient who presents with periumbilical drainage should have a sinogram performed, which should be diagnostic for both a urachal sinus and a patent urachus. Any child who presents with a periumbilical mass should have an ultrasound performed, which should be diagnostic for a urachal cyst.

Adolescent↗
PubMed1998

Tissue engineering in urologic surgery.

The use of nonurologic tissues in the genitourinary tract is common (owing to a lack of a better alternative) despite the known possible adverse effects. Selective cell transplantation is providing a means to engineer genitourinary tissues that may be used for reconstruction. This novel technology involves an interdisciplinary approach, combining techniques of cell biology and materials sciences towards the development of functional tissues or organs. Tissues associated with urology, such as clitoral, cavernosal, urethral, vesical, ureteral, and renal have been created in the laboratory, with varying degrees of function. Cells have also been recently used in patients as bulking agents for the treatment of vesicoureteral reflux and urinary incontinence. As the science of tissue engineering evolves, one can expect a wider application of this technology to the armamentarium of urologic surgery.

Cell Transplantation↗
PubMed1998

Cartilage rods as a potential material for penile reconstruction.

PURPOSE: Sex assignment is made in patients with ambiguous genitalia, genital trauma or iatrogenic injury after a thorough diagnostic evaluation and careful consultation with the family. In numerous instances a decision is made to rear the child as the female gender due to inadequate genitalia regardless of karyotype. Although a silicone penile prosthesis is accepted treatment in adults who require penile reconstruction, it has not been generally used in the pediatric population, mainly due to associated long-term problems. We determine the feasibility of creating natural penile prostheses of cartilage which, if biocompatible and elastic, may be used in patients who require genital reconstruction. MATERIALS AND METHODS: Cartilage was harvested from the articular surface of calf shoulders. Chondrocytes were isolated, grown and expanded in vitro. Cells were seeded onto preformed cylindrical polyglycolic acid polymer rods 1 cm. in diameter and 3 cm. long at a concentration of 50 x 10(6) chondrocytes per cm.3. A total of 40 polymer scaffolds were implanted in the subcutaneous space of 20 athymic mice. In each mouse 2 implantation sites consisted of a polymer scaffold seeded with chondrocytes and a control (polymer alone). Mice were sacrificed 1, 2, 4 and 6 months after implantation, respectively. Stress relaxation studies to measure biomechanical properties, including compression, tension and bending, were performed on the retrieved structures. Histological analyses were done with hematoxylin and eosin, aldehyde fuchsin-alcian blue and toluidine blue staining. RESULTS: Gross examination revealed well formed, milk-white rod-shaped solid cartilaginous structures the same size as the initial implant. Compression, tension and bending studies demonstrated that the cartilaginous structures were readily elastic and withstood high degrees of pressure. Histochemical analyses showed mature, well formed chondrocytes in all implants. There was no evidence of cartilage formation in the controls. CONCLUSIONS: Chondrocytes seeded on preformed biodegradable polymer structures form cartilage rods. The use of an entirely autologous system composed of biodegradable polymers and chondrocytes precludes an immunological reaction. This technology appears to be useful for the creation of a biocompatible malleable penile prosthesis, which may be useful in children with ambiguous genitalia and patients undergoing penile reconstruction.

Animals↗
PubMed1998

Acute scrotal symptoms in boys with an indeterminate clinical presentation: comparison of color Doppler sonography and scintigraphy.

PURPOSE: To compare the performance of color Doppler ultrasonography (US) and scintigraphy in assessing testicular perfusion in boys with clinically equivocal presentations. MATERIALS AND METHODS: Forty-one boys with clinically equivocal testicular perfusion underwent color Doppler US and scintigraphy. Studies were retrospectively classified as consistent with torsion, consistent with nontorsion, or indeterminate. Sensitivity and specificity were determined with alternate positivity criteria (indeterminate studies first considered positive and then negative for torsion). RESULTS: Color Doppler US demonstrated nine of 11 cases of torsion and 23 of 30 cases of nontorsion, with one false-positive and eight indeterminate studies. Scintigraphy demonstrated 10 of 11 cases of torsion and 29 of 30 cases of nontorsion, with two indeterminate studies (both in patients with inguinal testis). When indeterminate studies were considered positive for torsion, specificity was 77% for color Doppler US versus 97% for scintigraphy (P = .05). There were no other statistically significant differences between the sensitivities and specificities. CONCLUSION: Color Doppler US and scintigraphy demonstrate no statistically significant difference in ability to demonstrate testicular torsion in boys with acute scrotal symptoms and indeterminate clinical presentations. Owing to its greater specificity, scintigraphy may help prevent unnecessary surgery when color Doppler US shows equivocal flow.

Acute Disease↗
PubMed1998

Current findings in diagnostic laparoscopic evaluation of the nonpalpable testis.

PURPOSE: We delineate the current findings and contribution of diagnostic laparoscopic evaluation in the management of nonpalpable testis. MATERIALS AND METHODS: We reviewed all cases in which laparoscopy was considered the management associated with a nonpalpable testis in a 4-year period. Since our previous series, we have performed a careful examination for the testis after induction of anesthesia but before committing to laparoscopy. We recorded testis position and quality, character of the vas deferens and spermatic vessels, type of management and contribution of laparoscopy. We also reviewed contemporary published series and collated the findings of studies performed elsewhere. RESULTS: We identified 263 nonpalpable testes in 225 patients between September 1992 and 1996. In 40 patients 46 testes (18%) were found during physical examination under anesthesia. Of the remaining cases considered appropriate for laparoscopy 215 with complete records were further analyzed. Only 12.6% could be considered missed on examination due to a viable testis distal to the inguinal ring. Of the testes 45.7% would have been found during inguinal exploration alone. In 9.8% of the patients there were intra-abdominal vanishing testes, while 4.2% had indeterminate cord structures on inguinal exploration that would have prompted abdominal extension without a laparoscopic demonstration that the vas and vessels entered the canal. A conventional inguinal incision would have provided optimal exposure for operative management in 34% of the testes. For testes distal to the internal ring when the vas and vessels were distinctly atretic we never identified a viable testis, while a normal appearing vas and vessel were associated with a 45% chance of a salvageable testis. Laparoscopy was informative regarding testis position in all cases in which it was performed. CONCLUSIONS: In 13.2% of the cases laparoscopic findings precluded unnecessary abdominal exploration. The typical surgical incision for inguinal exploration would have left the surgeon compromised in 66% of the cases compared to the approach optimized as a result of laparoscopic testicular localization. Of the patients 34% arguably did not benefit from laparoscopy versus inguinal exploration. A simple examination under anesthesia significantly decreases the number of uninformative laparoscopic evaluations, and it is well worth the cost of a few minutes of operative time.

Adolescent↗
PubMed1998

Comparison of sonicated albumin enhanced sonography to fluoroscopic and radionuclide voiding cystography for detecting vesicoureteral reflux.

PURPOSE: We compared sonicated albumin enhanced sonography to fluoroscopic and radionuclide voiding cystography for detecting vesicoureteral reflux. MATERIALS AND METHODS: After obtaining informed consent we enrolled in our study 20 patients with known or suspected vesicoureteral reflux and no contraindications to intravesical sonicated albumin. All patients underwent albumin enhanced sonography, following which 10 patients each underwent radionuclide and fluoroscopic voiding cystography. Reflux was graded by the observing radiologist and urologist. RESULTS: In 10 patients albumin enhanced sonography demonstrated reflux in 6 of the 7 (83%) ureters in which radionuclide cystography identified reflux. In 2 patients ultrasound studies were inadequate due to excessive movement during the procedure, and the patients were classified as unevaluable. In the remaining 10 patients 12 of 20 ureters (60%) were equal in the absence or presence of and degree of reflux on enhanced sonography and voiding cystourethrography. In 6 ureters voiding cystourethrography detected reflux more readily or revealed a higher grade of reflux. Two ureters had a higher reflux grade on enhanced sonography. No adverse effects were associated with intravesical sonicated albumin. CONCLUSIONS: In experienced hands sonicated albumin enhanced sonography is safe for evaluating vesicoureteral reflux. It provides the simultaneous evaluation of renal contours, parenchyma and size in addition to bladder visualization. This new technique may prove to be useful as a followup study in patients with previously documented reflux or as a primary study for sibling screening.

Adolescent↗
PubMed1998

Reservoir calculi: a comparison of reservoirs constructed from stomach and other enteric segments.

PURPOSE: The intestinal augmented bladder has an increased propensity to form urinary calculi. Predisposing risk factors include chronic bacteriuria, urinary stasis and mucus production. Gastric reservoirs have negligible mucus production and the ability to acidify the urine. We determined whether they also have a decreased incidence of stones. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients undergoing augmentation cyptoplasty (215) or creation of a freestanding reservoir (44) between May 1976 and March 1996. Of these cases 83 were augmented with stomach and 179 were constructed from other intestinal segments, (that is ileal, ileocecal, sigmoid and/or a combination of these). Presenting diagnosis, patient age, gender, additional surgical procedures, interval to stone formation and calculous composition were recorded. Patients were excluded from study when there was less than 1 year of followup, as were those with renal or pre-augmentation bladder calculi. RESULTS: Stones formed in 32 of 207 patients (15%) after an average interval of 3.6 years (range 0.5 to 8.6). The majority of stones were composed of struvite (magnesium ammonium phosphate). Reservoirs containing gastric segments were significantly less likely to form calculi than those augmented with other intestinal segments (2 of 70 versus 30 of 137, p <0.0001 Fisher's exact test). Seven additional patients with pre-augmentation bladder stones who subsequently underwent gastrocystoplasty remain stone-free at an average followup of 3.0 years. CONCLUSIONS: Calculous formation is rare in reservoirs that incorporate a gastric segment. In patients with gastric augmentation stones appear only to develop when the patient is on histamine blockade or has a propensity to form stones that only favor an acidic environment, such as uric acid.

Adolescent↗
PubMed1997

Heparin-binding EGF-like growth factor is an autocrine growth factor for human urothelial cells and is synthesized by epithelial and smooth muscle cells in the human bladder.

The epidermal growth factor receptor (HER1) has been implicated in regenerative growth and proliferative diseases of the human bladder epithelium (urothelium), however a cognate HER1 ligand that can act as a growth factor for normal human urothelial cells (HUC) has not been identified. Here we show that heparin-binding EGF-like growth factor (HB-EGF), an activating HER1 ligand, is an autocrine regulator of HUC growth. This conclusion is based on demonstration of HB-EGF synthesis and secretion by primary culture HUC, identification of HER1 as an activatable HB-EGF receptor on HUC surfaces, stimulation of HUC clonal growth by HB-EGF, inhibition of HB-EGF-stimulated growth by heparin and of log-phase growth by CRM 197, a specific inhibitor of HB-EGF/HER1 interaction, and identification of human urothelium as a site of HB-EGF precursor (proHB-EGF) synthesis in vivo. ProHB-EGF expression was also detected in the vascular and detrusor smooth muscle of the human bladder. These data suggest a physiologic role for HB-EGF in the regulation of urothelial proliferation and regeneration subsequent to mucosal injury. Expression of proHB-EGF is also a feature of differentiated vascular and detrusor smooth muscle in the bladder. Because proHB-EGF is known to be the high affinity diphtheria toxin (DT) receptor in human cells, synthesis of the HB-EGF precursor by human urothelium also suggests the possibility of using the DT-binding sites of proHB-EGF as an in vivo target for the intraluminal treatment of urothelial diseases.

Bacterial Proteins↗
PubMed1997

Continent urinary diversion: the Children's Hospital experience.

PURPOSE: Continent urinary diversion has become increasingly important for treating childhood urinary tract pathology that cannot be managed by direct reconstructive techniques. We review our 9-year experience with continent diversion. MATERIALS AND METHODS: Since 1986 continent diversions were created in 74 patients 3 to 38 years old (mean age 13.7). The underlying pathological condition was the exstrophy/epispadias complex in 34 patients, neurological disorders in 23, malignancy in 13 and other congenital anomalies in 4. Followup averaged 5.2 years after the last procedure. Nonbladder reservoirs in 39 patients (53%) were fashioned from ileocolic (17), colic (7), gastrocolic (6), sigmoid (3), gastrosigmoid (2), ileosigmoid (2), ileal (1) and gastroileac (1) segments. When possible, the native bladder was incorporated into the reconstructive strategy. A total of 26 patients underwent bladder augmentation with intestine or stomach, including ileal (11), gastric (8), sigmoid (3), gastroileac (2) and ileocolic (2) segments. Nine other patients did not require bladder augmentation. Continence mechanisms were a flap valve (Mitrofanoff principle) in 50 patients, nipple valve in 15 and ileal plication (Indiana pouch) in 9. When the Mitrofanoff principle was used with a native bladder reservoir in 30 cases, outlet resistance was altered by bladder neck division (15), fascial sling placement (6) or Young-Dees-Leadbetter bladder neck reconstruction (2). In the remaining 7 patients the bladder neck remained intact. RESULTS: Excellent continence was obtained. The Mitrofanoff principle initially provided continence in 41 patients (82%). Six of the 9 incontinent patients were dry after a single revision. A total of 13 patients (87%) with nipple valves and 7 (78%) with Indiana pouches were dry, and the remaining 5 were cured after a single revision. Ultimately continence was achieved in 71 of the 74 patients (96%) after a maximum of 2 operations. Of the 48 complications in 29 patients the most common were difficulty in catheterizing (11), stones (11), infection (8) and upper tract deterioration (4). CONCLUSIONS: Many options exist for reconstructing complex anomalies. Choices must be individualized based on patient anatomy. The dry state may be achieved in most cases without resorting to a bag on the abdomen.

Adolescent↗
PubMed1997

Estimating normal bladder capacity in children.

PURPOSE: An accurate estimation of normal bladder capacity can be helpful in evaluating the patient with genitourinary disease and in interpreting urodynamic data. Prior studies have provided initial estimates. We propose 2 new equations that are practical, easy to use and more accurate than those previously published. MATERIALS AND METHODS: We retrospectively reviewed the records of more than 5,000 children undergoing radionuclide cystography at our institution. Radionuclide cystography was conducted by instilling (99m)technetium pertechnetate via gravity drip in awake children. Bladder capacity was believed to be achieved when rate of inflow diminished to a minimal rate, initiation of voiding occurred or significant discomfort was indicated. Patients with vesicoureteral reflux, infravesical obstruction, urinary tract infection, dysfunctional voiding or other lower urinary tract pathology were excluded from the study. Linear and nonlinear regression modeling established the relationship between age and bladder capacity. RESULTS: A total of 2,066 children (598 boys and 1,468 girls) had normal radionuclide cystography and were included in the analysis. Analysis of variance demonstrated that increasing age was strongly predictive of bladder capacity (p <0.0001). Because a nonlinear model was the most accurate formula for all ages (4.5 x age(0.40) = capacity [ounces]), 2 practical linear equations were determined: 2 x age (years) + 2 = capacity (ounces) for children less than 2 years old, and age (years) divided by 2 + 6 = capacity (ounces) for those 2 years old or older. Although girls had larger capacities than boys, the rate of increase was not significantly different between them. CONCLUSIONS: The relationship between normal bladder capacity and age in children follows a nonlinear curve. This nonlinear relationship can be approximated by 2 practical linear formulas that are easy to remember and are derived from a larger population than any prior study. These formulas provided accurate estimations of bladder capacity when prospectively applied to normal patients.

Adolescent↗
PubMed1997

Endoscopic substances for the treatment of vesicoureteral reflux.

The endoscopic treatment of reflux, like that of urinary incontinence, is effective. Several materials and endoscopic delivery systems are currently under evaluation for the treatment of reflux and incontinence. These include silicone microimplants, glass particles, collagen, dextranomer microspheres, a detachable balloon system, chondrocytes, and muscle cells. Although the endoscopic treatment is rendered in a similar fashion for both urinary incontinence and reflux, the acceptable safety and efficacy parameters of the bulking agents may differ, depending on the condition treated and the age of the patient. A review of the endoscopic bulking substances and systems currently available is presented.

Biocompatible Materials↗
PubMed1997

Detachable self-sealing membrane system for the endoscopic treatment of incontinence.

PURPOSE: The endoscopic treatment of urinary incontinence is limited by the injectable substances currently available. The ideal injectable material should be able to conserve its volume, be nonmigratory and nonantigenic. Toward this goal we developed a system for the treatment of incontinence in which a catheter with an inflatable and detachable self-sealing silicone membrane fits through a standard cystoscopic needle. We present our experience with this system. MATERIALS AND METHODS: A total of 20 female beagle dogs underwent cystoscopy and in 15 a self-sealing membrane was placed endoscopically in the submucosal region of the proximal urethra at the 3 and 9 o'clock positions. Each membrane was inflated through the delivery catheter with 0.2 cc povidone. Coaptation of the urethra due to the relative bulking effect of the inflated membrane was confirmed endoscopically in each animal. Five control animals received only an injection of saline. Four animals were sacrificed at 1, 3, 6, 12 and 18 months, respectively. At sacrifice gross and histological examinations were performed. RESULTS: At retrieval up to 18 months after implantation the membrane remained inflated and at the same position at which it had been initially placed. There was no evidence of povidone volume loss or extravasation. Membranes were encapsulated by a fibrous capsule. Only a few inflammatory cells surrounded the capsule by month 1 and there were none by month 3. Histological examination of periurethral tissue and distant organs showed no evidence of particle migration. CONCLUSIONS: The detachable self-sealing membrane system is easily implantable, nonmigratory, nonantigenic and able to conserve its volume. These studies indicate that this system may be effective for the endoscopic treatment of incontinence.

Animals↗
PubMed1997

A novel gene delivery system using urothelial tissue engineered neo-organs.

PURPOSE: Presently gene delivery is most effectively achieved by ex vivo gene transfer, which includes removal of the target tissue, in vitro gene delivery to the target cells, possible selection to enhance the proportion of transfected cells and reintroduction of the gene modified cells. Reintroduction of transformed cells in vivo has been a challenging task. Based on the feasibility of tissue engineering techniques in which cells seeded on biodegradable polymer scaffolds form tissue when implanted in vivo, we explored the possibility of developing a neo-organ system for in vivo gene therapy. MATERIALS AND METHODS: Normal human urothelial cells were harvested, expanded in vitro and seeded on biodegradable polymer scaffolds. The cell-polymer complex was then transfected with PGL3-luc, pCMV-luc and pCMV beta-gal promoter reporter gene constructs. The transfected cell-polymer scaffolds were then implanted in athymic mice and the engineered tissue was retrieved 0, 1, 3, 5 and 7 days after implantation. RESULTS: The reporter gene assay demonstrated an expression of luciferase activity at days 1, 3, 5 and 7 with the peak at day 5. X-gal and beta-galactosidase antibody assays stained positive on the deoxyribonucleic acid treated transfection. CONCLUSIONS: Successful gene transfer can be achieved using biodegradable polymer scaffolds as a urothelial cell delivery vehicle. The transfected cell-polymer scaffold forms an organ-like structure with functional expression of the transfected genes. This study demonstrates that urothelial tissue engineered gene transfer is safe and effective.

Animals↗
PubMed1996

Human albumin solder supplemented with TGF-beta 1 accelerates healing following laser welded wound closure.

BACKGROUND AND OBJECTIVE: We examined the possibility that human albumin solder can be used as a vehicle for site specific delivery of growth factors for the purpose of accelerating tissue repair following laser welded wound closure. Certain human recombinant growth factors have been shown to accelerate wound healing in model systems. Pilot in vitro studies have established that several growth factors, including TGF-beta 1, maintain bioactivity following exposure to temperatures achieved during laser tissue welding. Using a temperature controlled laser delivery system (TCL) to precisely maintain welding temperatures, it is now possible to avoid thermal denaturation of exogenous bioactive molecules such as growth factors. STUDY DESIGN/MATERIALS AND METHODS: HB-EGF, bFGF, and TGF-beta 1 were tested in vitro for maintenance of bioactivity after exposure to 80 degrees C. In vivo experiments using porcine skin determined the efficacy of solders augmented with growth factors. Incisions were repaired using human albumin alone or supplemented with HB-EGF (2 micrograms), bFGF (10 micrograms), or TGF-beta 1 (1 microgram). Wounds were excised at 3, 5, and 7 days post-operatively. Tensile strength, total collagen content, and histology were performed. RESULTS: At 3 days, tensile strength (TS) of TGF-beta 1 wounds were 36% (P < 0.05) and 20% (n.s.) stronger than laser alone and suture closures, respectively. By 5 days the TS of the TGF-beta 1 group increased by 50% (P < 0.05) and 59% (P < 0.02) over laser alone and suture groups, respectively. At 7 days the TGF-beta 1 group was 50% (P < 0.05) and 79% (P < 0.01) stronger than laser solder alone or suture, respectively. The HB-EGF and bFGF groups were equivalent to the laser solder group at all time points. Total collagen TGF-beta 1 Accelerates Healing Following Laser Welding content at 7 days increased in the TGF-beta 1 group by 7% (n.s.) over the suture group and 21% (P < 0.05) in the laser group. CONCLUSION: Human albumin solder supplemented with TGF-beta 1 increases the early post-operative strength of laser welded wounds. This novel application of laser tissue soldering augmented with a growth factor has the potential to bring about immediate fluid tight seals while providing site specific delivery of biological modifiers. This may lead to an overall improvement in post-operative convalescence, wound infections, and hospital costs.

Albumins↗