Muscle flap mass preservation by sensory reinnervation with end-to-side neurorrhaphy: an experimental study in rats

The purpose of this study was to determine whether sensory reinnervation with end-to-side neurorrhaphy preserves muscle mass in pedicled muscle flaps. A new muscle flap model innervated by the common peroneal nerve (CPN) was tested in rats. Animals were divided into group 1 (CPN transected without repair), group 2 (CPN transected and immediately repaired by end-to-end neurorrhaphy), and groups 3A and 3B (CPN transected and repaired with the sural nerve, by end-to-end and end-to-side neurorrhaphy, respectively). We evaluated the muscle-preserving effect by measuring muscle weight and performed histological and morphometric analyses 3 months after the procedure. Sensory reinnervation significantly preserved the muscle mass, although less than motor reinnervation. There was no significant difference between the end-to-end and end-to-side procedures. Results of morphometric analysis in each group paralleled those of mean muscle weight. Sensory reinnervation with end-to-side neurorrhaphy appears to be useful in the preservation of muscle flap mass.     

周围神经端侧缝合后神经再生及其趋化性的实验研究

目的进一步验证周围神经端侧缝合的有效性,初步探讨端侧缝合后神经再生的趋化性问题。方法雌性清洁级SD大鼠10只20侧,实验分3组,分别为神经端侧缝合组、正常对照组、切断对照组。神经缝合后5个月采用电生理、组织学、电镜等方法,观测再生神经纤维及其靶器官结构功能的改变;乙酰胆碱酯酶染色观测再生运动神经纤维及比例。结果端侧组肱二头肌复合肌肉动作电位的(CMAP)潜伏期较正常组减慢64％,最大波幅为正常组的27％。肱二头肌肌湿重为正常组的72％,肌纤维横截面积与正常组元明显差异。端侧组肌皮神经有髓神经纤维计数占正常组的44．5％,有髓神经纤维髓鞘厚度、最大直径、最小直径分别占正常组的85．9％、77．3％和65．5％。乙酰胆碱酯酶染色示端侧组肌皮神经主干运动神经纤维比例为[(0．39±0．07)％,x±s,下同],与正常组的差异无统计学意义;肱二头肌肌支运动神经纤维比例为(0．38±0．07)％,与正常组比较差异有统计学意义(P>0．01)。结论周围神经端侧缝合后有相当数目的再生神经纤维,但以髓鞘薄,直径小的纤维为主,同时能有效防止肌肉萎缩。周围神经端侧缝合后神经侧方出芽对特定靶器官的趋化性不明显。     

螺旋式改良神经端侧缝合法修复周围神经损伤的实验研究

目的 对治疗周围神经损伤的常规端侧缝合方法进行螺旋式改良端侧缝合实验研究,观察神经再生修复效果,为周围神经损伤的治疗提供效果更好的修复方法.方法 选用60只健康SD大鼠,采用右侧腓总神经修复模型.术中根据手术方法的不同,随机分为A、B、C三组,每组20只.每组将右侧腓总神经在其坐骨神经分出后3mm处局部封闭,利刀切断.A组神经远断端切成90°断面,行腓总神经端端缝合;B组神经远断端切成45°斜面,同时供体神经干外束膜开窗行端侧缝合;C组神经远断端切成10°斜面,供体神经干外束膜开窗,以螺旋式改良端侧缝合法进行神经束膜及外膜缝合.术后第8周分别对各组大鼠进行组织学、肌湿重、神经电生理检测,有髓神经纤维计数及神经示踪法观察.结果 螺旋式改良缝合组(C组)观察指标均优于常规端侧缝合组(B组)(P＜0.05),与端端缝合组(A组)比较差异无统计学意义(P＞0.05).螺旋式改良端侧缝合法对促进神经纤维的再生明显优于常规端侧缝合法.结论 神经断端采用螺旋式改良缝合后,神经再生良好;螺旋式改良缝合法较常规端侧缝合法能获得更有效的神经再生;长入远端的神经纤维多少与受端缝合接触面积大小有关.当临床遇动力神经缺乏时,采用神经螺旋式改良端侧缝合法可获得更好的修复效果.     

神经高位端端与低位端侧或侧侧缝合相结合提高神经修复能力的实验研究

目的对无缺损的周围神经高位损伤，提出高位端端与低位端侧或侧侧缝合相结合的新方法，观察神经再生和靶器官的恢复情况。方法SD大鼠80只，高位切断左侧胫神经。随机分为5组：A组：胫神经两断端行端端缝合，远端于膝关节水平与腓神经干行侧侧缝合。B组：断端处理同A组，远端移植正中神经作胫腓神经干之间的端侧桥接缝合。C组：单纯作断端的端端吻合。D组：胫神经干近端结扎并固定，远端与腓神经干行侧侧缝合。E组：近端处理同D组，远端切除部分神经段后，与腓神经干行端侧缝合。术后行肌电图检查及组织学观察并作统计学分析。结果术后早期（4周）D、E组有神经再生，术后12周A、B组的神经再生、传导功能及靶肌肉和运动终板的恢复情况均优于C、D、E组。结论高位端端与低位端侧或侧侧缝合相结合的方法，可尽早恢复对靶组织的营养和神经再支配，为高位缝合处高质量神经的长入赢得时间，提高了有效功能的恢复。     

Repair of the main nerve trunk of the upper limb with end-to-side neurorrhaphy: an experimental study in rabbits

The aim of this study was to assess the effectiveness of reinnervation using end-to-side neurorrhaphy in the upper extremity of the rabbit. The cut right ulnar nerve was repaired and sutured to the side of the median nerve about 3 cm above the elbow joint. The extent of reinnervation was quantitatively evaluated, as well as the integrity of the intact donor nerve in 36 rabbits randomly treated with fresh or delayed nerve repair with or without perineurotomy. Evaluations included nerve conduction velocity (NCV) of both the ulnar and medial nerves, dry muscle weight, and histologic examination (neurofilament stain and morphometric assessment) at 3 and 6 months postoperatively. NCV recovery rates were 79% and 87% for the ulnar nerve, and 89% and 94% for the median nerve compared to contralateral intact nerves, at 3 and 6 months, respectively. Flexor carpi ulnaris muscle mass measurements revealed a recovery in dry muscle weight of about 81% and 88% at 3 and 6 months, respectively, compared to the intact contralateral flexor carpi ulnaris. Histologic studies with neurofilament staining reveal numerous axonal sprouts at the distal end of the median nerve, indicative of myelinated axonal regeneration. Morphometric analysis demonstrated no difference between fresh and delayed repairs. These results indicate that in the upper extremity of rabbits, end-to-side neurorrhaphy permits axonal regeneration from the intact donor nerve, and is associated with satisfactory recovery. The effect of the procedure on the donor nerve was negligible.     

端侧神经吻合后“再生”神经纤维的逆行追踪实验研究

目的研究兔坐骨神经模型端侧神经吻合术后神经“再生”和功能恢复状况。方法20只健康白兔,随机分为2组:实验组右侧腓总神经切断,远断端吻合至去除外膜的胫神经外侧部,近断端包埋入股收肌;对照组右侧腓总神经切断,并于胫神经外侧部作外膜开窗,两断端包埋入邻近肌肉。16周后,观察展趾反射、针刺试验,辣根过氧化物酶逆行标记,胫前肌、腓肠肌湿重测量。结果针刺试验、展趾反射提示有感觉和运动功能恢复,但较差;湿重比率提示肌肉功能无恢复。辣根过氧化物酶标记结果显示标记感觉神经元虽比运动神经元多,但后者“再生”比率为(67.72±20.18)%,感觉“再生”比率仅为(8.81±2.90)%,这与功能恢复程度不一致。结论无效“再生”支配是该坐骨神经模型功能恢复差的主要原因。     

Ulnar nerve repair by end-to-side neurorrhaphy on the median nerve with interposition of a vein: an experimental study

Our study used a rat animal model to verify that Schwann cell migration, collateral axonal sprouting, and regeneration were not hindered by interposition of a 10-mm vein segment between the distal stump of the transected ulnar nerve and the donor median nerve in end-to-side reconstruction. Reconstructed nerves were withdrawn at 7, 14, 30, and 60 days after surgery, then analyzed by confocal laser microscopy after immunolabeling with anti-neurofilament-200 kD (an axonal marker) and anti-S100 (a glial marker) antibodies. Results are reported at 7, 14, 30, and 60 days after surgery. Our observations indicate that chemotactic factors that stimulate end-to-side nerve regeneration can exert their action at this distance. These findings suggest a possible clinical application for this surgical technique in cases when the severed nerve stump is far from the potential donor nerve.     

End-to-side neurorrhaphy: an experimental study in rabbits

The concept of end-to-side nerve repair was recently introduced; however, most authors have reported conflicting results with this technique. This study was conducted to assess the effectiveness of end-to-side nerve repair in both fresh and predegenerated specimens by histological evaluation in an animal study in rabbits. Thirty male rabbits were divided into three groups. In group 1 (n = 14), the peroneal nerve was divided and sutured end-to-side to the tibial nerve via an epineurial window. In group 2 (n = 13), the peroneal nerve was divided and sutured end-to-side to the tibial nerve after a 1-week "predegeneration period." In group 3 (n = 3), which was considered the control group, the peroneal nerve was divided and sutured to the adjacent soft tissues. After 3 months, specimens were harvested for histological evaluation. Nerve fiber count, in normal peroneal nerves, averaged 532/cross section. In groups 1 and 2, average nerve fiber count in implanted peroneal nerves was 6.24 and 7.00/cross section, respectively. No significant statistical difference was observed between fresh and "predegenerated" groups (P = 0.90). These data suggest that collateral sprouting of donor nerves is possible after end-to-side neurorrhaphy through an epineurial window, but the number of nerve fibers in recipient nerves is too low to result in any functional recovery in the target organ.     

Axonal supercharging technique using reverse end-to-side neurorrhaphy in peripheral nerve repair: an experimental study in the rat model

OBJECT: In an attempt to improve peripheral nerve repair, the influence of the addition of reverse end-to-side neurorrhaphy for an injured peripheral nerve was investigated in the rat sciatic nerve transection model. METHODS: Twelve Sprague-Dawley rats were divided into two groups (six rats in each group). In Group I, the right sciatic nerve was cut at a point distal to the gluteal notch and repaired using end-to-end neurorrhaphy with four 10-0 nylon epineurial sutures. In Group II, after performing the same procedure as in Group I, the left sciatic nerve was cut distally and passed through a subcutaneous tunnel to the right side. The proximal stump of the left sciatic nerve was coapted to the epineurial window of the right sciatic nerve distal to the injured point in an end-to-side fashion using 10-0 nylon epineurial sutures. The effects were evaluated using analgesimeter recordings for the hind paw, electrophysiological tests, measurement of the muscle contraction force, a double-labeling technique, weight measurement and histological examination of the gastrocnemius muscle, histological examination of the bilateral sciatic nerves, and immunofluorescent staining. RESULTS: Results from the many tests used to evaluate the reverse end-to-side neurorrhaphy technique indicated that functional recovery of the denervated target organs was promoted by axonal augmentation. CONCLUSIONS: The reverse end-to-side neurorrhaphy technique could be useful in peripheral nerve repair.     

New pure motor nerve experimental model for the comparative study between end-to-end and end-to-side neurorrhaphy in free muscle flap neurotization

The present study demonstrates a new experimental model to compare the efficacy of end-to-end and end-to-end neurorrhaphy in free muscle flap neurotization. Forty animals were used, divided into four equal groups named A, B, C and D. The peripheral stump of the thoracodorsal nerve was stitched end-to-end in groups A and C and end-to-side in groups B and D to the long thoracic nerve. Free functional muscle transfer was simulated by putting vascular clamps to the thoracodorsal artery (FD SS8R, F: 15 to 20 g) and vein (FD SS6R, F: 10 to 15 g) for 60 minutes and transecting and then restitching the origin and insertion of the latissimus dorsi muscle. Electromyographic and histological studies were performed 150 days following completion of the experiment. The results could indicate the possibility that end-to-side neurorrhaphy might be used in free functional muscle transfer as an alternative to end-to-end neurorrhaphy. We believe that the proposed experimental model is useful for the comparative study between end-to-end and end-to-side neurorrhaphy in free muscle flap neurotization, as these are pure motor nerves and innervate synergistic muscles, are in close approximation, and have similar diameters.     

周围神经端侧缝合的研究进展

神经损伤的修复通常采用两神经断端的端端缝合法 ,直至今日仍然如此。神经端侧缝合是将损伤神经的远断端缝合到相邻健康的神经干上 ,或者取一段神经段以端侧缝合的方式桥接于正常神经和损伤神经之间 ,使损伤的神经功能得到一定的恢复。神经端侧缝合术在 2 0世纪初就已开展。 190 3年Ｂａｌａｎｃｅ应用神经端侧缝合术治疗面瘫 ,与此同时 ,Ｈａｍ和Ｌｏｗ将臂丛神经上干植入颈7神经根治疗臂丛上干损伤。在此后的90年 ,再没有学者进行过神经端侧缝合术的研究。1992年巴西学者Ｖｉｔｅｒｂｏ进行了一系列神经端侧缝合的实验研究及临床应用研究…     

端侧神经吻合后再生状况的实验研究

目的研究端侧神经吻合后,神经再生的可能性和再生纤维的类型.方法将失神经后的兔耳大神经制成端侧吻合模型,12只动物被分成2、4、6、8和12周5个实验组和1个正常对照组,在受神经侧注入HRP逆行示踪,实验到期后对耳大神经纤维和相应的神经节进行HRP染色和CGRP的免疫组化染色,光镜下观察结果.结果逆行示踪显示,神经端侧吻合2～4周时,供神经侧C2、C3背根神经节内无HRP标记阳性的细胞,至第6周,背根神经节内开始出现HRP阳性细胞,8～12周阳性细胞的数量更多,尤其是胞体直径在50μm以上的阳性细胞数显著增加;2～4周时CGRP免疫组化的染色显示,背根神经节内的CGRP阳性标记细胞逐步增强,6周以后阳性标记的细胞数量增加,阳性细胞的胞体直径在12～40 μm范围内,属于中小直径的细胞胞体,同时观察到供神经纤维内的CGRP阳性纤维逐步增多,并跨越神经吻合口长入受神经体内,8～12周背根神经节内的CGRP阳性标记细胞的增加不显著,吻合口及受神经体内的神经纤维也增加不明显.结论端侧神经吻合后再生纤维可长入移植体,纤维再生的顺序是细小类的纤维首先再生.     

神经端侧吻合的实验研究

目的：研究神经多个端侧吻合后远端神经的再生情况,并与一干单一神经端侧吻合进行比较,探讨其可行性及临床应用价值。方法：选用Wistar雄性大鼠36只,随机分成A、B组。A组：于右侧锁骨下将肌皮神经、尺神经、桡神经切断,并在正中神经干侧方的外膜上分别开3个1mm“小窗”。将离断神经的远端依次端侧吻合到正中神经的“开窗”处,B组：于右侧锁骨下将尺神经切断,远端端侧吻合到正中神经侧方的“开窗”处,两组左侧均不予处置,为对照侧,术后行神经电生、组织学、电镜及神经纤维计数等检查。结果：神经端侧吻合后,肌皮神经、尺神经、尺神经、桡神经远端均有不同数量的有髓神经纤维再生,且A、B组的尺神经远端神经再生数量的差异显著性。两组的正中神经主干功能的未受到明显影响。结论鼠类动物神经端侧吻合后,远端神经可再生。     

FK506缓释膜片促进外周神经端-侧吻合后神经再生的研究

目的 探讨免疫抑制剂FK506缓释膜片局部应用对周围神经端-侧吻合后神经再生的影响。方法 选用SD大白鼠40只，随机平均分成实验组和对照组。实验组：切断腓总神经，在邻近的胫神经干外膜上开一1mm窗口，将腓总冲经远端与胫神经干做端-侧神经吻合后在吻合口周围放置含有FK506分子可降解缓释膜片。对照组：单纯行神经端-侧吻合：两组分别于术后2、4、8、12周取材，对腓总神经和胫神经干用快蓝（FB）和荧光金（FG）注射标记，取背根神经节（DRG）和脊髓在荧光显微镜下观察被标记细胞。结果 实验组背根冲经节和脊髓内FB标记细胞明显多于对照组。结论 免疫抑制剂FK506能促进外周神经端-侧吻合后神经再生的速度和质量。     

终末器官对端侧神经吻合后轴突再生影响的实验研究

目的探讨受神经远端终末器官对端侧吻合后神经再生的影响.方法 SD雌性大鼠10只,动物随机分为A、B2组.A组在近端切断腓总神经,其远断端与胫神经外膜开窗处进行端侧吻合; B组同 A组两神经端侧吻合后,在距吻合口1.5cm处切断其与终末器官的联系,并将连接肌肉的腓总神经远端反折固定在邻近的肌肉组织.术后12周取吻合口处的神经标本,甲苯胺蓝和抗神经微丝抗体免疫组化染色、光镜检查及图像分析.结果甲苯胺蓝染色及神经微丝免疫组化染色结果说明 A、B2组再生神经纤维数目、髓鞘厚度、纤维结缔组织含量及神经纤维丝排列,均有明显差异.结论终末器官对促进端侧神经吻合后神经再生具有重要作用.     

神经端侧吻合术与侧侧吻合术对神经干损伤后功能恢复作用的比较

目的　比较神经端侧吻合与神经侧侧吻合后功能恢复作用的差异。方法　将Wistar大鼠 18只分成A、B两组 ,每组 9只。端侧吻合组 :左侧腓总神经切断并在胫神经干上外膜开窗后 ,与胫神经主干作端侧吻合 ;侧侧吻合组 :左侧腓总神经切断后与胫神经作侧侧吻合。术后 3个月 ,行形态学检查、电生理检查、肌湿重检查及图像分析 ,评价神经再生及肌肉功能恢复的情况。结果　神经端侧吻合组和神经侧侧吻合组神经纤维的数目分别为 73 2 .8± 5 3 .4、712 .5± 5 8.4,其截面积分别为 (712 .5± 5 8.4) μm2 、(182 .3± 3 9.7) μm2 ,神经的传导速度分别为 (2 5 .6± 8.3 )m /s、(18.5± 9.8)m/s ,胫前肌肉的湿重两组分别为 (2 0 5 .6± 2 0 .7)mg、(2 0 1.8± 2 1.3 )mg ,肌纤维的面积分别为 (13 2 0 .6± 10 3 .9) μm2 、(14 0 3 .7± 13 8.4) μm2 ,再生的运动终板面积分别为 (15 3 7.5±5 2 7.8) μm2 、(1774.2± 45 8.7) μm2 ,吸光度值为 0 .42± 0 .15、0 .5 5± 0 .17,所有测量值差异无显著性 (P <0 .0 5 )。结论　神经端侧吻合术与神经侧侧吻合术的功能恢复差异无显著性     

终末器官对端侧神经吻合后轴突再生影响的实验研究

目的　探讨受神经远端终末器官对端侧吻合后神经再生的影响。方法　SD雌性大鼠 1 0只 ,动物随机分为A、B2组。A组 :在近端切断腓总神经 ,其远断端与胫神经外膜开窗处进行端侧吻合 ;B组 :同A组两神经端侧吻合后 ,在距吻合口 1 .5cm处切断其与终末器官的联系 ,并将连接肌肉的腓总神经远端反折固定在邻近的肌肉组织。术后 1 2周取吻合口处的神经标本 ,甲苯胺蓝和抗神经微丝抗体免疫组化染色、光镜检查及图像分析。结果　甲苯胺蓝染色及神经微丝免疫组化染色结果说明A、B2组再生神经纤维数目、髓鞘厚度、纤维结缔组织含量及神经纤维丝排列 ,均有明显差异。结论　终末器官对促进端侧神经吻合后神经再生具有重要作用     

Ultrastructure of early axonal regeneration in an end-to-side neurorrhaphy model

The ultrastructure of the early regenerative response in an end-to-side neurorrhaphy rat model was studied using transmission electron microscopy. The ipsilateral saphenous nerve was grafted to the sciatic nerve under the following conditions: Group 1, the epineurium and perineurium of the sciatic nerve remained intact; Group 2, an epineurial and perineurial window was created at the site of the lateral neurorrhaphy; Group 3, the same as in Group 2 and, in addition, the sciatic nerve sustained a partial neurectomy. Rats were perfused through the heart with fixative containing 2 percent paraformaldehyde and 2.5 percent glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) at 4, 8, 12, 24 and 48 hr after surgery. In Group 1, no regenerating axons were observed and the myelin sheath in the donor nerve did not demonstrate any degenerative changes through 48 hr. In Group 2, an increased diameter of the unmyelinated axons and growth cones was observed in the donor nerve proximal to the coaptation site after 12 hr. Degenerative changes in the myelin sheath were observed after 12 hr within the several layers under the coaptation site. In Group 3, many growth cone-like structures were observed in the area proximal to the coaptation site after 12 hr. After 24 hr, proximal regenerating axons elongated to the coaptation site and, at 48 hr, many regenerating nerves grew inside the Schwann cell basement membrane of the graft nerve. These results indicate that the perineurial window and nerve graft are the critical conditions for inciting nerve regeneration in the donor nerve.     

肌皮神经与尺神经端侧缝合重建屈肘功能的实验研究

目的 研究周围神经端侧缝合后侧支发芽再生能力及用于臂丛上干撕脱伤重建屈肘功能的可行性.方法 选用24只Wistar大鼠.左前肢为实验侧:于距肱二头肌入肌点12mm处切断肌皮神经,远断与外膜开窗1.0mm×0.5mm的尺神经做端侧缝合.右前肢为对照侧:切断1/2尺神经束,近端与肌皮神经远断端做端端外膜缝合.按取材时间随机分为A、B、C三个时间组,每组8只,分别于术后3、6、12周取材,进行大体观察.神经组织学、电生理、肱二头肌收缩力和肌湿重、辣根过氧化物酶(HRP)示踪检测.结果A组实验侧再生纤维数目和B组实验侧动作电位振幅、潜伏期与对照例相比有显著性差异(P<0.01),实验侧均低于对照例.随术后时间的延长,其余各项指标无显著性差异(P>0.05).结论 肌皮神经与尺神经端侧缝合后确实存在侧支发芽能力,并获得部分神经肌肉功能恢复,为临床治疗臂丛神经上干撕脱伤重建屈肘功能提供了新思路.