MICROSTRUCTURAL STUDY ON DETRUSOR MUSCLE AFTER BLADDER FUNCTIONAL RECONSTRUCTION FOR ATONIC BLADDER CAUSED BY MEDULLARY CONE INJURY IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Yuelei , SHENG Jun , HOU Chunlin , LIN Haodong.

Department of Orthopedics, Shanghai Changzheng Hospital, Shanghai, 200003, P.R.China. Corresponding author: HOU Chunlin, E-mail: chunlin _hou@yahoo.com.cn;

Corresponding author：

Keywords：

Medullary cone injury; Bladder functional reconstruction; Detrusor muscle; Neuromuscular junction; Rat

DOI：

10.7507/1002-1892.20130183

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Objective To study the microstructural change of detrusor muscle and neuromuscular junction (NMJ) after bladder functional reconstruction for atonic bladder caused by medullary cone injury and to discuss the feasibility of bladder functional reconstruction for improving the detrusor muscle degeneration. Methods A total of 104 adult female Sprague-Dawley rats (weighing, 200-250 g) were randomized divided into 3 groups: normal group (n=8), control group (n=48), and experimental group (n=48). No treatment was given in normal group; the medullary cone injury was established by sharp transection of spinal cord at L4, 5 levels in control group; and the anastomosis of bilateral L5 ventral root (VR)-S2 VR and L5 dorsal root (DR)-S2 DR was performed for bladder functional reconstruction after modeling of medullary cone injury in experimental group. After operation, the survival condition of rats was observed. At 3 days and 3 consecutive days before 1, 2, 3, 4, 5, and 6 months after operation, the residual urine volume was measured; at 1, 2, 3, 4, 5, and 6 months after operation, the detrusor muscle was harvested to measure the muscle fiber cross-sectional area by HE staining, to calculate the percentage of connective tissue by Masson trichrome staining, and to observe the ultrastructure of the detrusor muscle and the NMJ by transmission electron microscope (TEM). Results Eleven rats were supplemented because of death after operation. In control group, a significant increase of the residual urine volume was observed with the extension of time (P lt; 0.05); in experimental group, an increase was observed at the first 3 months after operation, and then gradually decreased, showing significant differences between the other time point (P lt; 0.05) except between at 3 days and at 5 months after operation (P gt; 0.05); there was significant difference between control and experimental groups at other each time point (P lt; 0.05) except at 3 days, 1 month, and 2 months (P gt; 0.05). HE staining and Masson trichrome staining indicated that the muscle fibers arranged in disorder with gradually aggravated atrophy and gradually increased connective tissue in control group, while the shape of the detrusor muscle recovered with no increased connective tissue at 4, 5, and 6 months after operation in experimental group; there was significant difference in cross-sectional area of detrusor muscle and percentage of connective tissue between normal group and experimental group, and between normal group and control group at each time point (P lt; 0.05). In control group, the cross-sectional area of detrusor muscle decreased and the percentage of connective tissue increased with the extension of time (P lt; 0.05). In experimental group, the cross-sectional area of detrusor muscle decreased at the first 3 months and then increased, and the percentage of connective tissue increased slowly with the extension of time. There was no significant difference of cross-sectional area of detrusor muscle at the first 3 months between control and experimental groups (P gt; 0.05), but the values in experimental group were significantly higher than those in control group at 4, 5, and 6 months after operation (P lt; 0.05). There were significant differences of the percentage of connective tissue between control and experimental groups at each time point (P lt; 0.05). In control group, the amount of synaptic vesicles decreased in the NMJ with time passing; vacuole like structure was observed in NMJ at 3 months; there was almost no nerve ending at 6 months. In experimental group, the amount of synaptic vesicles decreased at 1 and 3 months after operation, but obviously increased at 6 months. Conclusion The reconstruction of bladder function with L5 nerve roots above the paraplegic plane can effectively inhibit the degeneration of detrusor muscle and improve its microstructural changes after medullary cone injury.

Citation： ZHANG Yuelei,SHENG Jun,HOU Chunlin,LIN Haodong.. MICROSTRUCTURAL STUDY ON DETRUSOR MUSCLE AFTER BLADDER FUNCTIONAL RECONSTRUCTION FOR ATONIC BLADDER CAUSED BY MEDULLARY CONE INJURY IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(7): 836-842. doi: 10.7507/1002-1892.20130183 Copy

1.	Heary RF, Salas S, Bono CM, et al. Complication avoidance: thoracolumber and lumbar burst fractures. Neurosurg Clin N Am, 2006, 17(3): 377-388.
2.	Benevento BT, Sipski ML. Neurogenic bladder, neurogenic bowel, and sexual dysfunction in people with spinal cord injury. Phys Ther, 2002, 82(6): 601-612.
3.	Jeong SJ, Cho SY, Oh SJ. Spinal cord/brain injury and the neurogenic bladder. Urol Clin North Am, 2010, 37(4): 537-546.
4.	de Groat WC, Yoshimura N. Plasticity in reflex pathways to the lower urinary tract following spinal cord injury. Exp Neurol, 2012, 235(1): 123-132.
5.	林浩东, 侯春林, 郑宪友, 等. 大鼠脊髓损伤后膀胱生理反射弧重建的实验研究. 中国修复重建外科杂志, 2008, 22(6): 719-723.
6.	Lin H, Hou C. Transfer of normal S1 nerve root to reinnervate atonic bladder due to conus medullaris injury. Muscle Nerve, 2013, 47(2): 241-245.
7.	Zheng XY, Hou CL, Chen AM, et al. Optimal timing of operation for repairing atonic bladder after medullary cone injury: an experimental study in rats. Spinal Cord, 2008, 46(8): 574-581.
8.	王金武, 侯春林, 卢宁, 等. 利用腹壁反射重建膀胱反射弧的远期功能性研究. 中华显微外科杂志, 2002, 25(4): 280-284.
9.	胥维勇, 杨群, 范小莉. 介绍一种改良Masson三色染色法. 诊断病理学杂志, 2005, 12(1): 63．.
10.	de Groat WC, Wickens C. Organization of the neural switching circuitry underlying reflex micturition. Acta Physiol, 2013, 207(1): 66-84.
11.	Hoang TX, Pikov V, Havton LA, et al. Functional reinnervation of the rat lower urinary tract after cauda equina injury and repair. J Neurosci, 2006, 26(34): 8672-8679.
12.	赵继懋, 张玉海, 杜林栋. 神经原性膀胱逼尿肌超微结构的变化. 中华泌尿外科杂志, 2004, 25(10): 682-685.
13.	Hindley RG, Brierly RD, McLarty E, et al. A qualitative ultrastructural study of the hypocontractile detrusor. J Urol, 2002, 168(1): 126-131.
14.	Miyazato M, Yoshimura N, Chancellor MB, et al. The other bladder syndrome: underactive bladder. Rev Urol, 2013, 15(1): 11-22.
15.	Chartier-Kastler E, Denys P. Intermittent catheterization with hydrophilic catheters as a treatment of chronic neurogenic urinary retention. Neurourol Urodyn, 2011, 30(1): 21-31.
16.	Pannek J, Bertschy S. Mission impossible? Urological management of patients with spinal cord injury during pregnancy: a systematic review. Spinal Cord, 2011, 49(10): 1028-1032.
17.	Jean-Jacques Wyndaele. Conservative treatment of patients with neurogenic bladder. European Urology Supplements, 2008, 7: 557-565.
18.	Lin H, Hou C, Chen A, et al. Reinnervation of atonic bladder after conus medullaris injury using a modified nerve crossover technique in canines. World Neurosurg, 2010, 73(5): 582-586.
19.	Lin H, Hou CL, Zhong G, et al. Reconstruction of reflex pathways to the atonic bladder after conus medullaris injury: Preliminary clinical result. Microsurgery, 2008, 28(6): 429-435.
20.	Lin H, Hou C, Zhen X. Bypassing spinal cord injury: surgical reconstruction of afferent and efferent pathways to the urinary bladder after conus medullaris injury in a rat model. J Reconstr Microsurg, 2008, 24(8): 575-581.
21.	Choi YS, Kim JC, Lee KS, et al. Analysis of female voiding dysfunction: a prospective, multi-center study. Int Urol Nephrol, 2013. [Epub ahead of print].
22.	侯春林, 郑宪友, 陈爱民, 等. 大鼠脊髓圆锥损伤后膀胱逼尿肌及其神经肌肉接头形态学变化的研究. 中国修复重建外科杂志, 2007, 21(5): 473-477.
23.	Smith PP, DeAngelis A, Kuchel GA. Detrusor expulsive strength is preserved, but responsiveness to bladder filling and urinary sensitivity is diminished in the aging mouse. Am J Physiol Regul Integr Comp Physiol, 2012, 302(5): R577-586.
24.	Pisarska-Krawczyk M, Urbaniak T, Klejewski A. Overactive bladder—diagnosis et treatment. Przegl Lek, 2012, 69(10): 1156-1159.
25.	Säily M, Vasarainen H, Sairanen J, et al. Urinary retention in women. Duodecim, 2012, 128(21): 2244-2250.

1. Heary RF, Salas S, Bono CM, et al. Complication avoidance: thoracolumber and lumbar burst fractures. Neurosurg Clin N Am, 2006, 17(3): 377-388.
2. Benevento BT, Sipski ML. Neurogenic bladder, neurogenic bowel, and sexual dysfunction in people with spinal cord injury. Phys Ther, 2002, 82(6): 601-612.
3. Jeong SJ, Cho SY, Oh SJ. Spinal cord/brain injury and the neurogenic bladder. Urol Clin North Am, 2010, 37(4): 537-546.
4. de Groat WC, Yoshimura N. Plasticity in reflex pathways to the lower urinary tract following spinal cord injury. Exp Neurol, 2012, 235(1): 123-132.
5. 林浩东, 侯春林, 郑宪友, 等. 大鼠脊髓损伤后膀胱生理反射弧重建的实验研究. 中国修复重建外科杂志, 2008, 22(6): 719-723.
6. Lin H, Hou C. Transfer of normal S1 nerve root to reinnervate atonic bladder due to conus medullaris injury. Muscle Nerve, 2013, 47(2): 241-245.
7. Zheng XY, Hou CL, Chen AM, et al. Optimal timing of operation for repairing atonic bladder after medullary cone injury: an experimental study in rats. Spinal Cord, 2008, 46(8): 574-581.
8. 王金武, 侯春林, 卢宁, 等. 利用腹壁反射重建膀胱反射弧的远期功能性研究. 中华显微外科杂志, 2002, 25(4): 280-284.
9. 胥维勇, 杨群, 范小莉. 介绍一种改良Masson三色染色法. 诊断病理学杂志, 2005, 12(1): 63．.
10. de Groat WC, Wickens C. Organization of the neural switching circuitry underlying reflex micturition. Acta Physiol, 2013, 207(1): 66-84.
11. Hoang TX, Pikov V, Havton LA, et al. Functional reinnervation of the rat lower urinary tract after cauda equina injury and repair. J Neurosci, 2006, 26(34): 8672-8679.
12. 赵继懋, 张玉海, 杜林栋. 神经原性膀胱逼尿肌超微结构的变化. 中华泌尿外科杂志, 2004, 25(10): 682-685.
13. Hindley RG, Brierly RD, McLarty E, et al. A qualitative ultrastructural study of the hypocontractile detrusor. J Urol, 2002, 168(1): 126-131.
14. Miyazato M, Yoshimura N, Chancellor MB, et al. The other bladder syndrome: underactive bladder. Rev Urol, 2013, 15(1): 11-22.
15. Chartier-Kastler E, Denys P. Intermittent catheterization with hydrophilic catheters as a treatment of chronic neurogenic urinary retention. Neurourol Urodyn, 2011, 30(1): 21-31.
16. Pannek J, Bertschy S. Mission impossible? Urological management of patients with spinal cord injury during pregnancy: a systematic review. Spinal Cord, 2011, 49(10): 1028-1032.
17. Jean-Jacques Wyndaele. Conservative treatment of patients with neurogenic bladder. European Urology Supplements, 2008, 7: 557-565.
18. Lin H, Hou C, Chen A, et al. Reinnervation of atonic bladder after conus medullaris injury using a modified nerve crossover technique in canines. World Neurosurg, 2010, 73(5): 582-586.
19. Lin H, Hou CL, Zhong G, et al. Reconstruction of reflex pathways to the atonic bladder after conus medullaris injury: Preliminary clinical result. Microsurgery, 2008, 28(6): 429-435.
20. Lin H, Hou C, Zhen X. Bypassing spinal cord injury: surgical reconstruction of afferent and efferent pathways to the urinary bladder after conus medullaris injury in a rat model. J Reconstr Microsurg, 2008, 24(8): 575-581.
21. Choi YS, Kim JC, Lee KS, et al. Analysis of female voiding dysfunction: a prospective, multi-center study. Int Urol Nephrol, 2013. [Epub ahead of print].
22. 侯春林, 郑宪友, 陈爱民, 等. 大鼠脊髓圆锥损伤后膀胱逼尿肌及其神经肌肉接头形态学变化的研究. 中国修复重建外科杂志, 2007, 21(5): 473-477.
23. Smith PP, DeAngelis A, Kuchel GA. Detrusor expulsive strength is preserved, but responsiveness to bladder filling and urinary sensitivity is diminished in the aging mouse. Am J Physiol Regul Integr Comp Physiol, 2012, 302(5): R577-586.
24. Pisarska-Krawczyk M, Urbaniak T, Klejewski A. Overactive bladder—diagnosis et treatment. Przegl Lek, 2012, 69(10): 1156-1159.
25. Säily M, Vasarainen H, Sairanen J, et al. Urinary retention in women. Duodecim, 2012, 128(21): 2244-2250.

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MICROSTRUCTURAL STUDY ON DETRUSOR MUSCLE AFTER BLADDER FUNCTIONAL RECONSTRUCTION FOR ATONIC BLADDER CAUSED BY MEDULLARY CONE INJURY IN RATS

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