Avaliação morfofuncional do enxerto de túnica albugínea suína na cistoplastia em ratos

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Abstract

RESUMO O objetivo deste trabalho é avaliar o uso da túnica albugínea suína na cistoplastia em ratos, avaliando funcionalidade, capacidade de reparação do órgão e possibilidades de complicações. Foram selecionados 30 ratos Wistar, machos, de seis meses de idade, divididos em: um grupo teste (TA), em que os animais receberam o enxerto de túnica albugínea suína após a cistectomia parcial e um grupo controle (C), em que os animais sofreram somente a cistectomia parcial. Os animais pertencentes a ambos os grupos foram divididos igualmente em subgrupos de cinco animais cada, que sofreram eutanásia em sete, 28 e 42 dias de pós-operatório. Foi realizada uma análise macroscópica e, posteriormente, uma análise histopatológica da região da ferida cirúrgica. Aos sete e 28 dias, os animais pertencentes ao grupo C e ao grupo TA apresentaram urotelização, regeneração da lâmina própria e da musculatura, porém o grupo TA apresentou menores sinais inflamatórios e maior organização tecidual, principalmente com relação à formação das fibras musculares. Aos 42 dias de pós-operatório, ambos os grupos já apresentavam características histológicas normais. Concluiu-se que o enxerto de túnica albugínea suína obteve sucesso na regeneração da bexiga de ratos, mantendo a funcionalidade do órgão, sem rejeição, e favorecendo a migração celular.

Translated abstract

ABSTRACT The aim of this study is to evaluate porcine tunica albuginea as a graft for cystoplasty in rats, regarding bladder function, capacity and possible complications. 30 male Wistar rats with six monthes of age have been selected and separated into two different groups: A test group (TA) in which the animals received a tunica albuginea graft after partial cystectomy and a control group (C) in which partial cystectomy was performed, followed by bladder suture. In each group the animals were euthanized at seven, 28 and 42 days after surgery. Macroscopic and Histological analysis have been performed. At seven and 28 days after surgery the samples from both groups had urothelial lining upon a lamina propria and smooth muscle fibers in regeneration process. However, the TA group showed less inflammatory signs and more organized structure, mainly regarding the smooth muscle formation. At 42 days after surgery all groups showed a bladder wall structure qualitatively identical to the normal tissue. We could conclude that tunica albuginea graft is able to maintain bladder function and support cellular migration without any kind of rejection.

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Challenges in a larger bladder replacement with cell-seeded and unseeded small intestinal submucosa grafts in a subtotal cystectomy model.

Dominic Frimberger, Yuanyuan Zhang, Y. Cheng … (2006)

To evaluate small intestinal submucosa (SIS), unseeded or seeded, as a possible augmentation material in a canine model of subtotal cystectomy. In all, 22 male dogs had a 90% partial cystectomy and were then divided into three groups. At 1 month after the initial cystectomy, dogs in group 1 (unseeded, six) and group 2 (seeded, six) received a bladder augmentation with a corresponding SIS graft. The dogs in group 3 (ten) received no further surgery and were considered the surgical control group. All dogs were evaluated before and after surgery with blood chemistry, urine culture, intravenous urography, cystograms and cystometrograms. After surgery (at 1, 5 and 9 months), the bladders were examined using routine histology and immunohistochemistry. All 22 dogs survived the subtotal cystectomy, and 18 survived their intended survival period. One dog, in group 2 (seeded), was killed at 1 month after augmentation due to bladder perforation caused by a large piece of incompletely absorbed SIS. Three other dogs (group 1, two; and group 2, one) were killed within 2 months after augmentation due to bladder obstruction by stones. Group 1 and group 2 SIS grafts had moderate to heavy adhesion, graft shrinkage, and some had bone and calcification at the graft site. Histologically, there was limited bladder regeneration in both groups. Interestingly, dogs in group 3 at 1 month after cystectomy (when group 1 and 2 received their augmentations) had severely shrunken bladders and histologically had severe inflammation, fibroblast infiltration and muscle hypertrophy. These results verify the subtotal cystectomy model. The use of seeded or unseeded SIS in a subtotal cystectomy model does not induce the same quality and quantity of bladder regeneration that is seen in the 40% non-inflammatory cystectomy model. This study provides important insights into the process of regeneration in a severely damaged bladder. The results led us to re-evaluate the critical elements required for a complete bladder replacement using tissue-engineering techniques.

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Biomatrices for bladder reconstruction.

Hsueh-Kung Lin, Sundar Madihally, Blake W. Palmer … (2015)

There is a demand for tissue engineering of the bladder needed by patients who experience a neurogenic bladder or idiopathic detrusor overactivity. To avoid complications from augmentation cystoplasty, the field of tissue engineering seeks optimal scaffolds for bladder reconstruction. Naturally derived biomaterials as well as synthetic and natural polymers have been explored as bladder substitutes. To improve regenerative properties, these biomaterials have been conjugated with functional molecules, combined with nanotechology, or seeded with exogenous cells. Although most studies reported complete and functional bladder regeneration in small-animal models, results from large-animal models and human clinical trials varied. For functional bladder regeneration, procedures for biomaterial fabrication, incorporation of biologically active agents, introduction of nanotechnology, and application of stem-cell technology need to be standardized. Advanced molecular and medical technologies such as next generation sequencing and magnetic resonance imaging can be introduced for mechanistic understanding and non-invasive monitoring of regeneration processes, respectively.

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In vivo bladder regeneration using small intestinal submucosa: experimental study.

N Capozza, D Zavaglia, G Palombaro … (2006)

Significant side effects are correlated with bladder augmentation. Recently, small intestinal submucosa (SIS) has been proposed for clinical use. The efficacy of SIS bladder regeneration was studied in a porcine experimental model. Partial cystectomy (40-60% of bladder wall) was performed and replaced by SIS graft. Animals were planned to be killed at 2 weeks, 5 weeks and 3 months. Bladder capacity at 40 cmH(2)O pressure and macroscopic graft morphology were assessed before and after SIS implant. Histological examination was carried out with computer assisted morphometric analysis for collagen/smooth muscle ratio. Student's t test was adopted for statistical analysis. Two piglets died on the 9th and 10th post-operative day due to urinary peritonitis. The remaining piglets were killed after uneventful post-operative period at 5 weeks (two animals) and 3 months (two animals). The bladder capacity was reduced (-18%) at the 5 week follow-up and quite similar to the pre-operative volume (+2.5%) at the 3 months control. No diverticular formation, bladder calculi, mucus and urinary infection were found. The SIS graft resulted not significantly contracted. Histology at 10 days showed SIS membrane lined by transitional epithelium islands with some capillaries. At 5 weeks, transitional epithelium was fully covering the graft; new blood vessels and fibroblasts with smooth muscle cells were observed. At 3 months, the SIS was not evident. Two layers were defined: inner transitional epithelium, outer collagen with fibroblasts and muscular bundles. Computer assisted morphometric analysis showed collagen/muscle ratio 70/30% (normal bladder=56/44%, P<0.05). The SIS was effective as a scaffold for bladder wall regeneration in four out of six animals. Long-term studies are required to confirm the efficacy of the newly developed wall and for eventual clinical use.