Tüm sıçanlar, Başkent Tıp Üniversitesi, Pekin Dostluk Hastanesi Hayvan Deney Komitesi tarafından onaylanan protokollere göre kullanıldı.
1. Cerrahi hazırlık, anestezi ve cerrahi teknikler
NOT: Bu çalışma için 200-220 g ağırlığında toplam 40 dişi Wistar sıçanı ticari olarak elde edilmiştir. 40 sıçandan 10'u rastgele kontrol grubu olarak seçildi ve geri kalanı test grubu olarak kabul edildi. Tüm hayvanlar, Başkent Tıp Üniversitesi, Pekin Dostluk Hastanesi'nin hayvan tesislerinde steril bir ortamda barındırıldı.
<ol><li>Sodyum pentobarbital intraperitoneal (40 mg/kg) uygulayarak genel anestezi uygulayın. Alternatif olarak,% 3-4% izofluran kullanarak anesteziyi indükleyin ve% 1 -% 3'te (solunan) tutun. Kuruluğu önlemek için gözlere oftalmik merhem uygulayın. Ardından, sıçanı cerrahi platforma yerleştirin ve termal destek sağlayın. NOT: İşlemin başında buprenorfin, 0.05 mg/kg, SC, 0.1-0.2 mL gibi analjezikler uygulayın.</li>
	<li>Ayak parmağı sıkışmasına yanıt vermemek suretiyle anestezi derinliğini kontrol edin. Kürkü tüm arka bölgeden tıraş bıçağı ile tıraş edin.</li>
	<li>Cerrahi bölgeyi, Klorheksidin veya Povidon iyot ve ardından İzopropil alkol gibi iki aşamalı bir ovmanın en az 3 döngüsü ile sterilize edin. Uzuvları cerrahi bantla sabitleyin ve cerrahi makasla sırtta yaklaşık 3 cm'lik bir medyan kesi yapın.</li>
	<li>Cerrahi makas kullanarak kesiyi deri altı dokulardan derinleştirin ve omurgaya bağlı kasları kesin.</li>
	<li>13. kaburgayı görsel olarak tanımlayın ve ortaya çıkarın (bu kaburgaya bağlı intervertebral boşluk T13\u2012L1 aralığıdır). 13. kaburgayı bir dikiş kullanarak işaretleyin.</li>
	<li>Tanımlamadan sonra, omurgaya bağlı kasları dikkatlice rezeke edin ve vertebral kolonu açığa çıkarın. Vertebral kolonun doğru tanımlanması için supraspinöz ligament ve interspinöz ligamenti rezeke edin. L4\u2012L5 seviyesini cerrahi makas ve forseps ile ortaya çıkarın. NOT: Supraspinöz ligament, ince deri altı dokusunun varlığı nedeniyle kolayca tanımlanabilir. Supraspinöz ligamentin rezeksiyonundan sonra, spinöz süreç arasındaki ligament interspinöz ligamenttir.</li>
	<li>L4\u2012L5 vertebral dikenli süreci ve omuriliği açığa çıkarmak için Kelly forseps kullanarak enine sürecin parçalarını dikkatlice inceleyin (Şekil 1).</li>
	<li>Konus medullarisin L4\u2012L5 seviyesinde tamamen açığa çıkarılması ve iridektomi makası ile konus medullarisin tamamen transekte edilmesi. Omuriliğin iyileşmesini engellemek için bazı doku ambalajları yerleştirin.</li>
	<li>Dış deri tabakasındaki üst kas ve cildi 4-0 emilemeyen dikiş kullanarak kapatın.</li>
	<li>Kontrol grubu için 1.1\u20121.7 adımlarını uygulayın ve conus medullaris'i olduğu gibi bırakın. İnsizyonu adım 1.9'a göre kapatın.</li>
</ol>2. Hayvan kurtarma
<ol><li>Sıçanları ameliyattan sonraki ilk saat boyunca sıcaklık kontrollü bir inkübatörde (37 ° C) tutun ve sternal veya aktif olarak hareket edene kadar izleyin. NOT: Tam kurtarma işlemi yaklaşık yarım saat sürer.</li>
	<li>Hayvanı yeterli yiyecek ve su içeren temiz bir kafese aktarın. Sıçanları ayrı kafeslerde tutun. NOT: Transeksiyonun başarısı, test grubundaki sıçanlar sadece ön ayakların yardımıyla hareket ederken, kontrol grubundaki sıçanlar normal şekilde yürüyebildiklerinde belirtilir.</li>
</ol>3. Operasyon sonrası yönetim
<ol><li>İntraperitoneal olarak bir antibiyotik olan Penisilin G'yi (hayvan başına 50.000 U / mL) enjekte edin. Buprenorfin, 0.05 mg / kg, SC, 0.1-0.2 mL gibi analjezikleri ameliyat sonrası 48 saat boyunca her 6-12 saatte bir uygulayın.</li>
	<li>İşemeye yardımcı olmak için idrar kesesini hipogastriyumda sıkıştırın. Bunu altı hafta boyunca günde iki kez aynı anda (sabah 8 ve akşam 8) gerçekleştirin. NOT: Detrusor'un normal daralmasının kaybı, DU'nun sembolüdür.</li>
	<li>Tüm sıçanları metabolik kafeslerde barındırın, her biri miksiyon ve inkontinansı izlemek için daha önce tartılmış emici bir kağıdın üzerine yerleştirilmiş bir idrar toplama hunisi içerir.</li>
	<li>İşemiş hacmi (VV) ve artık idrar hacmini ayrı ayrı gösteren emici kağıdın ağırlık değişimini toplayın ve not edin.</li>
</ol>4. Ürodinamik test
<ol><li>Ameliyattan altı hafta sonra, aşağıdaki gibi ürodinamik ölçüm ekipmanı kullanarak bir sistometrogram uygulayın.<ol><li>Sıçanları periton boşluğuna (3 mL/kg) %10 kloral hidrat enjekte ederek anestezi altına alın.</li>
		<li>İşeme için mesaneyi sıkıştırın, ardından sıçanı bir bant kullanarak cerrahi platforma sabitleyin.</li>
		<li>Epidural kateteri (3F) mesaneye yerleştirin ve ürodinamik ölçüm ekipmanını, epidural kateteri ve infüzyon pompasını üç uzuvlu tüp ile bağlayın.</li>
		<li>Ürodinamik ölçüm için fizyolojik salini 0,2 mL/dak hızında pompalayın (bakınız Malzeme Tablosu). MCC ve DOP'u ve mesanenin uygunluğunu kaydedin (δ mesane hacminin detrusörün δ basıncına bölünmesiyle hesaplanır).</li>
	</ol></li>
</ol>5. İstatistiksel analiz
<ol><li>Ticari olarak temin edilebilen yazılımları kullanarak istatistiksel analiz gerçekleştirin.</li>
	<li>Verilerin normalliğini test etmek için Kolmogorov-Smirnov testini kullanın.</li>
	<li>Normal olarak dağılmış değişkenleri standart sapmalarla ortalama değerler olarak ifade edin. Her iki gruptaki sistometrogram parametrelerini karşılaştırmak için iki kuyruklu eşleştirilmiş Öğrenci t-testlerini kullanın. NOT: p < 0,05, aradaki farkın istatistiksel anlamlılığa sahip olduğunu gösterir.</li>
</ol>

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Yazarların açıklayacak hiçbir şeyleri yoktur.

DÜ, hem erkeklerde hem de kadınlarda alt üriner sistem semptomlarının yaygın bir nedenidir. Etkilenenlerin yaşam kalitesini (Qol) önemli ölçüde azaltabilen az sayıda tedavi seçeneğine sahip karmaşık bir semptom takımyıldızıdır18. DÜ'nün multifaktöriyel olduğuna inanılmasına rağmen, patogenezinin anlaşılması ilkeldir. Çalışmalar, DÜ'nün patogenezinin miyojenik ve nörojenik faktörlerle ilişkili olabileceğini göstermiştir.
Miyojeni...

Defrüsör yetersizliği (DU), halen çalışılan tipik bir alt üriner sistem disfonksiyonudur. DU Uluslararası Kontinans Derneği (ICS)1 tarafından tanımlanmış olmasına rağmen, bu hastalığa atıfta bulunmak için çok sayıda farklı terminoloji kullanılmaktadır, örneğin, "detrusor yetmezliği", "akonttil mesane", "detrusor arefleksia"2. DU, 2002 yılında Uluslararası Kontinans Derneği (ICS) tarafından tanımlandığı gibi, mesane boşaltma için zamanın uzun süreli artmasına neden olan ve böylece normal bir süre içinde tam mesane boşalmasının sağlanamamasına neden olan, azaltılmış güç ve sürenin daralmasıdır.
DU, alt üriner sistem semptomları olan erkeklerin %48'ini ve kadınların (>70 yaş arası)3 %12'sini etkileyebilir. Multifaktöriyel gibi görünüyor ve etkili bir tedavi mevcut değil. Multipl skleroz4, diabetes mellitus5, Parkinson hastalığı6 veya serebral inme7 gibi nörojenik mesane disfonksiyonu olan hastalarda DU'nun her yerde olduğu bildirilmektedir. DU ayrıca laparoskopik histerektomi, prostatektomi veya küçük pelvis8'deki diğer cerrahi müdahaleler gibi iyatrojenik sinir hasarından da kaynaklanabilir. Patofizyoloji değişiklikleri ve DÜ'nün mevcut tedavileri, çalışma için uygun bir hayvan modelinin bulunmaması nedeniyle hala kafa karıştırıcıdır.
Miktüritasyon refleksi, pontin miktüritasyon merkezini, sakral parasempatik çekirdeği ve daha kıdemli korteks merkezlerini birleştiren spino-bulbospinal yollar tarafından kontrol edilir9. Miktürition refleksinin aktivasyonu ve sürdürülmesi esas olarak duyusal sinyallerin mesaneden daha kıdemli korteks merkezlerine düzenli olarak taşınmasına bağlıdır. Duyusal disfonksiyonun DU'ya katkıda bulunduğu varsayılabilir.
Alt üriner sistem disfonksiyonlarıyla ilgili deneysel hayvan çalışmalarının çoğu, aşırı aktif mesane (OAB) modellerine odaklanmıştır10. Bu modeller OAB patofizyolojisi ve prognozu hakkında makul bir anlayış sağlar. Bununla birlikte, sadece birkaç DU modeli bildirilmiştir, örneğin, supraspinal yaralanma (lokal lezyonlar, deserebrasyon ve orta serebral arter tıkanıklığı), omurilik transeksiyonu veya kontüzyon yaralanması, sistemik (örneğin, siklofosfamid) veya tahriş edici veya enflamatuar ajanların intravezikal uygulaması (örneğin, asit, akrolein ve lipopolisakkarit)11,12,13,14 . Bu yöntemler arasında DU13'ün hayvan modelinin oluşturulmasında sadece omurilik transeksiyonu veya kontüzyon yaralanması yöntemi kullanılabilir. Pontin miktüritasyon merkezinin ve yüksek korteks merkezlerinin yaralanmasını içeren girişimler ciddi travma nedeniyle terk edildi. Bu nedenle, DU'yu minimum yan etkilerle indüklemek için miksiyon refleks merkezinde doğru bir yer bulmak için daha fazla dikkat edilmektedir.
Daha önce de belirtildiği gibi, DU'yu indükleme mekanizmalarından biri, miksiyon refleksinin sinyal yoluna zarar vermek için omuriliğe zarar vermektir. Allen'ın ağırlık bırakma yöntemi, yaralı omurilikleri olan laboratuvar hayvanları oluşturmak için geliştirilmiştir15. Bununla birlikte, bu yöntem hakkında daha fazla deneysel veri bulunmamaktadır. Dahası, hayvanların bir kısmı DU olmadan inmeden sonra omurga fonksiyonunu geri kazandığından, bir DU hayvan modeli16 oluşturmak için mükemmel bir yöntem olarak düşünülemez.
1987'de Bregman, DU hayvan modelini üretmek için omuriliğin transeksiyonu sürecini çıkardı ve deneysel veriler elde etti17. Bununla birlikte, bu yöntem DU hayvan modelini oluşturmak için uygulanmamıştır. O zamanlar, araştırmacılar hala DU'nun patogenezi konusunda kafaları karışıktı. OAB veya DU indüksiyonu ile ilişkili omurilikteki yerler birbirine bitişik olduğundan, DU17'yi indüklemek için omuriliğe doğru hasar bölgesini bulamadılar. OAB ve DU bu yöntemle birlikte veya ayrı ayrı tanıtıldı. Bu nedenle, bu yöntem DU'yu tanıtmasına rağmen, kesin değildi ve DU'nun oluşumunun ve işlenmesinin anlaşılması için kullanılamazdı.
Yukarıda belirtildiği gibi, DU'nun uygun bir hayvan modelinin olmaması, DU'nun çalışmasının önündeki ana engellerden biridir. Araştırmacılar sürekli olarak DU'nun patolojisini simüle edebilecek doğru ve yönetilebilir bir model aramaktadır. DU için tedavi seçenekleri bile son 20 yılda önemli ölçüde iyileşmemiştir. Toplu olarak, DU'nun bir hayvan modelini oluşturmak için standart bir protokol tanımlamaya büyük ihtiyaç vardır.
Bu nedenle, bu yazıda, konus medullaris transeksiyonu ile DU'nun sıçan modelini başarılı bir şekilde oluşturmak için bir yöntem açıklanmaktadır. Konüs medullaris'i ayırmak için L4\u2012L5 seviyesinde transeksiyon yapıldı. Maksimum sistometrik kapasite (MCC), detrusor açma basıncı (DOP) ve mesanenin uyumu kaydedildi ve protokolü doğrulamak için analiz edildi. Aşağıda belirtilen protokol, DU hayvan modelini oluşturmak için hem fizibiliteyi hem de güvenilirliği standartlaştırılmış bir şekilde birleştirir ve DU'nun oluşumunu ve işlenmesini simüle eder. Protokol, DU'nun daha ileri çalışmaları için bir teknik olarak kullanılabilir.

<table><tbody><tr><td>0.9% saline</td><td>Wuhan Prosai Company</td><td>EY-C1178</td><td>pump for urodynamic measurement</td></tr><tr><td>10% chloral hydrate</td><td>Shandong Yulong Co., Ltd</td><td>H37022673</td><td>3mL/kg, administered intraperitoneally</td></tr><tr><td>Buprenorphine Hydrochloride Injection</td><td>Tianjin Pharmaceutical Research Institute Pharmaceutical Co. LTD</td><td>H12020275</td><td>0.05mg/kg subcutaneously 24h and 48h postoperation</td></tr><tr><td>Epidural Catheter</td><td>Shandong Xinghua Co, Ltd</td><td>VABR3L</td><td>for urodynamic measurement</td></tr><tr><td>Penicillin G</td><td>Alta Technology Co., Ltd</td><td>1ST5637</td><td>50,000 unit/ml per animal</td></tr><tr><td>pentobarbital</td><td>Beijing solabo Technology Co., Ltd</td><td>NK-WF0001</td><td>40 mg/kg, administered intraperitoneally</td></tr><tr><td>Suture line(4-0)</td><td>ETHICON</td><td>VCP422H</td><td>suture the injury</td></tr><tr><td>Three-limb tube</td><td>Shandong Xinghua Co, Ltd</td><td>VAB3T</td><td>for urodynamic measurement</td></tr><tr><td>Trace infusion pump</td><td>Zhejiang Smith Medical Instrument Co., Ltd</td><td>20162540335</td><td>Pump the saline at a speed of 0.2ml/min for urodynamic measurement</td></tr><tr><td>Urodynamic measurement equipment</td><td>Medical Measurement SystemsB.V.</td><td>08-0467</td><td>urodynamic measurement equipment can not only help the diagnosis of dysuria, but also provide objective materials for treatment and therapeutic effect. It is the most commonly used examination method in clinical diagnosis and treatment of lower urinary tract functional diseases</td></tr><tr><td>Wistar Rats</td><td>HFK Biotechnology Co.Ltd,Beijing ,China</td><td>SCXK2012-0023</td><td>200-220g</td></tr></tbody></table>

detrusor underactivity model in rats by conus medullaris transection

Sunulan protokolün amacı, konus medullaris transeksiyonu yoluyla sıçanda bir detrusor underactivity (DU) modeli oluşturmaktır. 200-220 g ağırlığındaki toplam 40 dişi Wistar sıçanına (kontrol grubu: 10 sıçan; test grubu: 30 sıçan) laminektomi uygulandı ve test grubunda konus medullaris L4\u2012L5 düzeyinde transekte edildi. Tüm sıçanlar altı hafta boyunca aynı çevre koşulları altında barındırıldı ve beslendi. Test grubunda, altı hafta boyunca günde iki kez idrar işemesi yapıldı ve ortalama rezidüel idrar hacmi kaydedildi. Her iki gruba da sistometrogram yapıldı. Mesanenin maksimum sistometrik kapasitesi (MCC), detrusor açma basıncı (DOP) ve uyumu kaydedildi ve hesaplandı. Test grubu, ameliyat sonrasında, spinal şok sırasında ve sonrasında önemli idrar retansiyonu gösterdi. Bununla birlikte, kontrol grubunda herhangi bir anormallik gözlenmedi. Kontrol grubuyla karşılaştırıldığında, MCC ve test grubundaki mesanenin uyumu test grubundan anlamlı derecede yüksekti (3.24 ± 2.261 mL'ye karşı 1.04 ± 0.571 mL; 0.43 ± 0.578 mL / cmH 2 O karşı 0.032 ± 0.016 mL / cmH 2 O), test grubundaki DOP kontrolden daha düşüktü (20.28 ± 14.022 cmH 2 O'ya karşı 35 ± 13.258 cmH2 Conus medullaris transeksiyonu ile DU'nun hayvan modelini oluşturmanın bu yöntemi, DU'nun patofizyolojisini daha iyi anlamak için mükemmel bir fırsat sunmaktadır.

Conus medullaris transeksiyonunun tüm prosedürü deneyimli cerrahlar tarafından 45 dakika içinde tamamlanabilir. Laboratuvarımızda 100'ün üzerinde konus medullaris transeksiyon ameliyatı gerçekleştirilmiştir. Başarı oranı, sıçanların sağkalımı ve DU'nun başarılı indüksiyonu ile tanımlandığı gibi% 95'in üzerindedir. Ürodinamik test DU indüksiyonunu doğruladı.
Deneyimlerimize dayanarak, DU indüksiyonu rezidüel idrar hacmi ile önceden değerlendirilebilir. İd...

Watch this Scientific Journal Video about Detrusor Underactivity Model in Rats by Conus Medullaris Transection at JoVE.com

Detrusor Underactivity Model in Rats by Conus Medullaris Transection

Bu yazıda sıçanlarda conus medullaris transeksiyonu ile detrusor az aktivite modeli oluşturmak için bir yöntem sunulmaktadır. Detrusor az aktivitesi bu hayvanlarda başarıyla uyarıldı. Model, idrar yolu fonksiyonunu incelemek için kullanılabilir.

Conus Medullaris Transeksiyonu ile Sıçanlarda Detrusor Düşük Aktivite Modeli

The goal of the presented protocol was to establish a detrusor underactivity (DU) model in the rat through conus medullaris transection. Laminectomy was ...

detrusor-underactivity-model-in-rats-by-conus-medullaris-transection

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

2.0K Görüntüleme.  Capital Medical University. Bu yazıda sıçanlarda conus medullaris transeksiyonu ile detrusor az aktivite modeli oluşturmak için bir yöntem sunulmaktadır. Detrusor az aktivitesi bu hayvanlarda başarıyla uyarıldı. Model, idrar yolu fonksiyonunu incelemek için kullanılabilir.

Video: Detrusor Underactivity Model in Rats by Conus Medullaris Transection

Evaluation of Biomaterials for Bladder Augmentation using Cystometric Analyses in Various Rodent Models

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and expels it with a precisely orchestrated contraction. A number of congenital and acquired urological anomalies including posterior urethral valves, benign prostatic hyperplasia, and neurogenic bladder secondary to spina bifida/spinal cord injury can result in pathologic tissue remodeling leading to impaired compliance and reduced capacity1. Functional or anatomical obstruction of the urinary tract is frequently associated with these conditions, and can lead to urinary incontinence and kidney damage from increased storage and voiding pressures2. Surgical implantation of gastrointestinal segments to expand organ capacity and reduce intravesical pressures represents the primary surgical treatment option for these disorders when medical management fails3. However, this approach is hampered by the limitation of available donor tissue, and is associated with significant complications including chronic urinary tract infection, metabolic perturbation, urinary stone formation, and secondary malignancy4,5.
Current research in bladder tissue engineering is heavily focused on identifying biomaterial configurations which can support regeneration of tissues at defect sites. Conventional 3-D scaffolds derived from natural and synthetic polymers such as small intestinal submucosa and poly-glycolic acid have shown some short-term success in supporting urothelial and smooth muscle regeneration as well as facilitating increased organ storage capacity in both animal models and in the clinic6,7. However, deficiencies in scaffold mechanical integrity and biocompatibility often result in deleterious fibrosis8, graft contracture9, and calcification10, thus increasing the risk of implant failure and need for secondary surgical procedures. In addition, restoration of normal voiding characteristics utilizing standard biomaterial constructs for augmentation cystoplasty has yet to be achieved, and therefore research and development of novel matrices which can fulfill this role is needed.
In order to successfully develop and evaluate optimal biomaterials for clinical bladder augmentation, efficacy research must first be performed in standardized animal models using detailed surgical methods and functional outcome assessments. We have previously reported the use of a bladder augmentation model in mice to determine the potential of silk fibroin-based scaffolds to mediate tissue regeneration and functional voiding characteristics.11,12 Cystometric analyses of this model have shown that variations in structural and mechanical implant properties can influence the resulting urodynamic features of the tissue engineered bladders11,12. Positive correlations between the degree of matrix-mediated tissue regeneration determined histologically and functional compliance and capacity evaluated by cystometry were demonstrated in this model11,12. These results therefore suggest that functional evaluations of biomaterial configurations in rodent bladder augmentation systems may be a useful format for assessing scaffold properties and establishing in vivo feasibility prior to large animal studies and clinical deployment. In the current study, we will present various surgical stages of bladder augmentation in both mice and rats using silk scaffolds and demonstrate techniques for awake and anesthetized cystometry.

Surgical stages of bladder augmentation are described using 3-D scaffolds in murine and rat models. To test the efficacy of biomaterial configurations for use in bladder augmentation, techniques for both awake and anesthetized cystometry are presented.

Çeşitli Kemirgen Modeller sistometrik Analizleri kullanarak Mesane Büyütme için Biyomalzeme Değerlendirilmesi

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and ...

Biyomühendislik

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

Küçük Kemirgenler sistometri ve Kullanımı: Mesane Chemosensation Çalışması

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

Tıp

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

Farelerde Mesane Ağrı için bir model olarak Mesane Distansiyonunun Uyarılmış Visceromotor Cevapları

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology

We describe an in vitro method to measure bladder smooth muscle contractility, and its use for investigating physiological and pharmacological properties of the smooth muscle as well as changes induced by pathology. This method provides critical information for understanding bladder function while overcoming major methodological difficulties encountered in in vivo experiments, such as surgical and pharmacological manipulations that affect stability and survival of the preparations, the use of human tissue, and/or the use of expensive chemicals. It also provides a way to investigate the properties of each bladder component (i.e. smooth muscle, mucosa, nerves) in healthy and pathological conditions.
The urinary bladder is removed from an anesthetized animal, placed in Krebs solution and cut into strips. Strips are placed into a chamber filled with warm Krebs solution. One end is attached to an isometric tension transducer to measure contraction force, the other end is attached to a fixed rod. Tissue is stimulated by directly adding compounds to the bath or by electric field stimulation electrodes that activate nerves, similar to triggering bladder contractions in vivo. We demonstrate the use of this method to evaluate spontaneous smooth muscle contractility during development and after an experimental spinal cord injury, the nature of neurotransmission (transmitters and receptors involved), factors involved in modulation of smooth muscle activity, the role of individual bladder components, and species and organ differences in response to pharmacological agents. Additionally, it could be used for investigating intracellular pathways involved in contraction and/or relaxation of the smooth muscle, drug structure-activity relationships and evaluation of transmitter release.
The in vitro smooth muscle contractility method has been used extensively for over 50 years, and has provided data that significantly contributed to our understanding of bladder function as well as to pharmaceutical development of compounds currently used clinically for bladder management.

This manuscript presents a simple, yet powerful, in vitro method for evaluating smooth muscle contractility in response to pharmacological agents or nerve stimulation. Main applications are drug screening and understanding tissue physiology, pharmacology,&#160;and pathology.

Alt Üriner Sistem Farmakoloji değerlendirin bir Yöntem olarak mesane düz kas Şerit Kasılabilme

We describe an in vitro method to measure bladder smooth muscle contractility, and its use for investigating physiological and pharmacological properties ...

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

Kadın Küçük Kemirgenler Sinir koruyucu Orta üretral Tıkanıklığı (Nemo)

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model

Awake filling cystometry has been used for a long time to evaluate bladder function in freely moving mice, however, the specific methods used, vary among laboratories. The goal of this study was to describe the microsurgical procedure used to implant an intravesical tube and the experimental technique for recording urinary bladder pressure in an awake, freely moving mouse. In addition, experimental data is presented to show how surgery, as well as tubing type and size, affect lower urinary tract function and recording sensitivity. The effect of tube diameter on pressure recording was assessed in both polyethylene and polyurethane tubing with different internal diameters. Subsequently, the best performing tube from both materials was surgically implanted into the dome of the urinary bladder of male C57BL/6 mice. Twelve-hour, overnight micturition frequency was recorded in healthy, intact animals and animals 2, 3, 5, and 7 days post-surgery. At harvest, bladders were assessed for signs of swelling using gross observation and were subsequently processed for pathological analysis. The greatest extent of bladder swelling was observed on day 2 and 3, which correlated with behavioral voiding data showing significantly impaired bladder function. By day 5, bladder histology and voiding frequency had normalized. Based on the literature and evidence provided by our studies, we propose the following steps for in vivo recording of intravesical pressure and voided volume in an awake mouse: 1) Perform the surgery using an operating microscope and microsurgical tools, 2) Use polyethylene-10 tubing to minimize movement artifacts, and 3) Perform cystometry on post-operative day 5, when bladder swelling resolves.

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

Bir Fare Modelinde Seyrek Sistometri Uygulamasının Değerlendirilmesi

Awake filling cystometry has been used for a long time to evaluate bladder function in freely moving mice, however, the specific methods used, vary among ...

A Decentralized (Ex Vivo) Murine Bladder Model with the Detrusor Muscle Removed for Direct Access to the Suburothelium during Bladder Filling

Previous studies have established the release of chemical substances from flat bladder mucosa sheets affixed in Ussing chambers and exposed to changes in hydrostatic pressure or mechanical stretch and from cultured urothelial cells upon hydrostatic pressure changes, stretch, cell swelling, or drag forces, and in bladder lumen at end of filling. Such findings led to the assumption that these mediators are also released in suburothelium (SubU)/lamina propria (LP) during bladder filling, where they affect cells deep in the bladder wall to ultimately regulate bladder excitability. There are at least two obvious limitations in such studies: 1) none of these approaches provide direct information about the presence of mediators in SubU/LP, and 2) the stimuli used are not physiological and do not recapitulate authentic filling of the bladder. Here, we discuss a procedure that enables direct access to the suburothelial surface of the bladder mucosa in the course of bladder filling. The murine detrusor-free preparation we created closely resembles filling of the intact bladder and allows pressure-volume studies to be performed on the bladder in the absence of confounding signaling from spinal reflexes and detrusor smooth muscle. Using the novel detrusor-free bladder model, we recently demonstrated that intravesical measurements of mediators cannot be used as a proxy to what has been released or present in the SubU/LP during bladder filling. The model enables examination of urothelium-derived signaling molecules that are released, generated by metabolism and/or transported into the SubU/LP during the course of bladder filling to transmit information to neurons and smooth muscle of the bladder and regulate its excitability during continence and micturition.

A Decentralized Ex Vivo Murine Bladder Model with the Detrusor Muscle Removed for Direct Access to the Suburothelium during Bladder Filling

The detrusor-free bladder model enables direct access to the suburothelium to study local mechanisms for regulation of biologically active mediator availability in suburothelium/lamina propria during storage and voiding of urine. The preparation closely resembles filling of an intact bladder and allows pressure-volume studies to be performed without systemic influences.

Mesane Dolgusu Sırasında Suburothelium Doğrudan Erişim için Kaldırılan Detrusor Kas ile Ademi Merkeziyet (Ex Vivo) Murine Mesane Modeli

Previous studies have established the release of chemical substances from flat bladder mucosa sheets affixed in Ussing chambers and exposed to changes in ...

Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually performed as terminal analysis under urethane anesthesia. It is well known that anesthetic drugs can influence bladder function. Hence, the aim of this technique is to perform cystometric measurements of the urinary bladder and external urethral sphincter in lightly restrained awake rats. For this purpose, a bladder catheter is implanted into the bladder dome. Subsequently, two electrodes are implanted bilateral to the external urethral sphincter and a ground electrode is sutured to a non-responsive skeletal muscle. The bladder catheter and the three electrodes are finally tunneled subcutaneously to the neck region and affixed to a harness. With this technique, the lower urinary tract can be measured at multiple time points in the same animal to assess lower urinary tract function. The main application of this technique is the follow-up of simultaneous urinary bladder and external urethral sphincter function in awake healthy rats and after induction of a disease or injury. Moreover, subsequent lower urinary tract monitoring can be performed during evaluation of the disease/injury and to monitor treatment efficacy.

This protocol first describes the surgical procedure of the permanent implantation of a urinary bladder catheter combined with external urethral sphincter electrodes, and second, the measurement of the function of the urinary bladder and external urethral sphincter in implanted awake animals.

Cystometric ve dış üretral sfinkter ölçümleri implant kateter ve tekrarlanan ölçülerini izin elektrotları ile uyanık Sıçanlarda

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually ...

Biyoloji

Urethral Stricture Induction Followed by Buccal Mucosa Graft Urethroplasty in a Rat Model

Urethral reconstruction is an important area of expertise for urologists. The buccal mucosa is considered the best option when urethral grafting is necessary, although in some cases, it is inappropriate or needs to be optimized to repair a given stricture. Therefore, developing innovative procedures and evaluating their putative success in experimental models is crucial to fit the clinical need. With this goal, this study describes a protocol in which urethral stricture was induced by electrocautery in Wistar rats. Urethral reconstruction was performed 1 week later with a buccal mucosa graft, harvested from the lower lip, and placed in a ventral onlay fashion. A retrograde urethrogram showed a significant improvement in urethral diameter after urethroplasty compared to the respective value after stricture induction. Additionally, the graft placement was assessed by blood perfusion analysis using laser Doppler. As expected, a dark blue area corresponded to the non-vascularized buccal mucosa graft. This procedure can successfully simulate the normal pathophysiological process of urethral injury and tissue modulation, as well as urethral reconstruction using a buccal mucosa graft in a reproducible manner, and serve as the basis for future research based on tissue engineering or urethral grafts.

In the present protocol, a urethral stricture induction was developed in Wistar rats, followed by urethral reconstruction with a buccal mucosa graft. A retrograde urethrogram and laser Doppler assessment were performed, validating urethral reconstruction (after stricture formation) and graft placement.

Üretra darlığı indüksiyonunu takiben sıçan modelinde bukkal mukoza greft üretroplastisi

Urethral reconstruction is an important area of expertise for urologists. The buccal mucosa is considered the best option when urethral grafting is ...

Efficient Rat Model for Urethral Stricture: Electric Knife Method

Iatrogenic Injury Recapitulated: Electroexcision Technique for Urethral Stricture Modeling in Rats

Urethral stricture (US) is a common clinical condition in urology, characterized by high prevalence and morbidity across all ages. Current treatments for US, such as urethral dilatation and internal urethrotomy, fail to fully resolve the condition and are associated with high rates of recurrence and complications. 
Additionally, the pathogenesis of US is not well understood. To explore the pathogenesis of US and develop new therapeutic strategies, it is crucial to establish a standardized rat model that accurately reflects the clinical manifestations. This study outlines a straightforward and repeatable method for inducing US in rats using a high-frequency electric knife. The method involves making a longitudinal incision with the electric knife set to a unipolar mixed cutting mode at 4 W, which inflicts significant urethral damage. Histopathological analysis shows thickening of the urothelium, inflammatory infiltration, and disorganized collagen fibers. This model effectively replicates iatrogenic injury through electroexcision in the rat urethra. In summary, this study successfully establishes a new, efficient, and stable rat model of US that closely mimics the clinical scenario, providing a valuable tool for further research into the mechanisms and novel treatments for US.

In this study, we present a novel, efficient, and stable rat model of urethral stricture created through electroexcision of the rat urethra, which effectively simulates iatrogenic injury observed in clinical settings.

Urethral stricture (US) is a common clinical condition in urology, characterized by high prevalence and morbidity across all ages. Current treatments for ...