逆行追踪神经端侧缝合后侧支生长的神经元胞体实验研究

目的 探讨神经端事后侧支生长的神经远胞体来源。方法 将实验组端侧缝合的腓总神经距吻合口２．５ｃｍ外切断,其近端植入有辣根过氧化酶（ｈｏｒｓｅｒａｄｉｓｈ ｐｅｒｏｘｉｄａｓｅ,ＨＲＰ）的硅胶囊中,同法标记正常腓总神经与正赏胫神经近端。观察脊髓前角Ｌ７￣Ｓ２段及相应疹神经节。结果 对照组为正常腓总神经伸肌运动神经元核群;实验组标记细胞会布于正常总神经伸肌运动神经元核群的复内侧,与下沉胫神经标记细胞分     

自体雪旺细胞促周围神经端侧吻合轴突侧支发芽的实验研究

目的：研究周围神经损伤行端侧吻合口自体雪旺细胞（Sc)移植对端侧吻合的作用，探讨改善端铁合质量的方法。方法：72只Wistar大鼠随机平均分为3组：A组，左腓神经端端吻合组；B组，左胫腓神经端侧吻合组；C组，左胫腓神经端侧吻合后再生小室内注自体雪旺细胞悬液组。于术后2、4、6个月每组分别取8只大鼠取材、检测，依次行大体观察、神经电生理检测、肌湿重、组织学及电镜观察等。结果：C组与B组各项指标比较：肌力、肌纤维截面积、有髓纤维截面积差异有显著性（P＜0．05）。神经电生理、肌湿重、有髓纤维数目差异有非常显著性（P＜0．01），显示恢复较好，某些指标（神经电生理、肌湿重、肌纤维截面积）在6个月时接近A组（P＞0．05）。结论：自体雪旺细胞移植可以促进周围神经端侧吻合轴突侧支发芽，改善端侧吻合口质量。     

Modified end-to-side neurorrhaphy enhances axonal sprouting: a motor functional and morphological study

This study evaluated the effect of modified end-to-side neurorrhaphy on functional recovery by modulating the epineurial window and contact areas, with a donor nerve, using a rat median nerve and finger flexors. Grasping testing, muscle contractility testing, and a histological study were performed 20 weeks after surgery. The modified end-to-side repair can enhance axonal sprouting from an intact nerve, and improve the functional recovery. Either the epineurial window surface area, or the contact configuration with the donor nerve is an important factors in an end-to-side coaptation model.     

Evaluation of collateral sprouting after end-to-side nerve coaptation using a fluorescent double-labeling technique.

The mechanism of end-to-side neurorrhaphy is believed to be by collateral sprouting, although evidence for this is lacking. This study validates whether axonal sprouting originates from the donor intact nerve by collateral sprouting with the use of a fluorescent double-labeling technique. End-to-side neurorrhaphy was performed on adult female Sprague-Dawley rats. Eight and 12 months postoperatively, animals were injected with true blue and diamidino yellow into the tibialis anterior and/or gastrocnemius muscles and were transcardially perfused with fixative after 7 days of retrograde transport. The lumbar enlargement and the dorsal root ganglia from L3 to L6 were harvested and serial sectioning and fluorescent microscopy were performed. No double-labeling neurons were observed in control animals, but a group of neurons that were greenish or yellowish in color were seen with single labeling. Double-labeling neurons, however, were seen in animals treated with end-to-side neurorrhaphy whether or not perineurotomy was performed. These results demonstrate that one parent nerve fiber can emanate another axon by collateral sprouting following end-to-side neurorrhaphy. We hypothesize that the causes of collateral sprouting might result from "switching signals" and/or "switching factors." Copyright 1999 Wiley-Liss, Inc.     

神经端侧吻合植入失神经皮瓣感觉功能研究

目的 探讨感觉神经端侧吻合后供神经的轴突长入神经移植体后,最终能否形成具有功能的感觉末梢。方法 用新西兰兔15只,将兔一条耳大神经做为供神经,在另侧耳取耳大神经移植体与供神经作端侧吻合后埋入失神经皮瓣,另设正常皮肤组和未植神经作对照组,每组动物5只。4个月后用神经单纤维放电技术观察皮瓣内再生神经纤维放电数量、 分布和类型。结果 对移植神经4个月的皮瓣进行神经电生理检查,见各类敏感纤维放电数量、分布和类型。结果 对移植神经4个月的皮瓣进行神经电生理检查,见各类敏感纤维放电均有出现,放电纤维总数在到正常皮肤经组的58％,而同期末植神经对照组,仅在皮瓣边缘靠近神经的一侧有极少放电。结论 神经端侧吻合方法可以重建皮瓣感觉功能。     

Muscle flap mass preservation with end-to-side neurorrhaphy: an experimental study

The authors examined the preservation of rat gracilis muscle flap mass after motor and sensory end-to-side neurorrhaphy. The rat gracilis muscle flap model was designed based on a previous study. Twenty-four Sprague-Dawley rats were divided into three groups. In Group 1 (n = 8), the flap was denervated by transecting the obturator nerve. In Group 2 (n = 8), the flap was reinnervated by coapting the proximal saphenous nerve to the distal obturator nerve. In Group 3 (n = 8), the flap was reinnervated by coapting the motor branch of the femoral nerve to the distal stump of the obturator nerve. At 6 months postoperatively, the gracilis muscle flaps were examined, harvested, and weighed individually. Results showed that the flaps with motor nerve reinnervation retained good bulk, with a weight of 634.0 +/- 65.1 gm, which was statistically significantly higher than the denervated group (457.5 +/- 125.3 gm, p < 0.01). However, muscle mass preservation in the sensory reinnervated group (606.9 +/- 209.1 gm) was not significantly different, compared to the denervated group. Histology revealed atrophic changes in the denervated group, compared to the sensory and motor-reinnervated groups. The authors concluded that muscle mass can be preserved by end-to-side nerve repair. Motor nerve reinnervation is able to better arrest atrophic changes of the muscle flaps.     

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。     

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.     

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.     

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.     

不同端-侧缝合方式修复周围神经的比较研究

目的 比较周围神经标准端-侧缝合方式与其他变体在修复效果上的差异.方法 SD大鼠126只,随机分为7组,将右侧肌皮神经切断,以同侧尺神经作为供体神经,按照端-端缝合组(end-to-end,E-E组)、端-侧缝合组(end-to-side,E-S组)、侧-侧缝合外膜开窗组(side-to-side,S-S组)、侧-侧缝合外膜不开窗组(side-to-side,S-S'组)、螺旋缠绕外膜去除组(spiral,Sp组)、螺旋缠绕外膜保留组(spiral,Sp'组)、阳性对照组(controlling group,CG组)制作模型,各组分别于术后1、2、3个月取材,进行神经电生理、病理组织学检测.结果 术后3个月,各端-侧缝合组均有明显神经再生现象,但效果均不及端-端缝合组,各种端-侧缝合术式的变体与标准端-侧缝合方式相比差异不明显(P＞O.05),保留神经外膜的两组为各组中效果最差.结论 神经外膜对端-侧缝合效果有影响,各种缝合方式差异无统计学意义,单纯通过手术方式提高神经端-侧缝合后神经再生能力的效果非常有限.     

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.     

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。     

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.     

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.     

Imaging of experimental end-to-side anastomoses in a cast model.

OBJECTIVE: To evaluate cast models in the imaging of experimental end-to-side anastomoses. DESIGN: Experimental study. SETTING: Teaching hospital, Finland. INTERVENTIONS: 20 non-stenosed and 10 stenosed end-to-side anastomoses were constructed using a Dacron prosthesis 8 mm in diameter. Cast models were made of all anastomoses, and anteroposterior and oblique projection radiographs were taken of them. Cross-sectional radiographs were also taken of the stenosed anastomoses. MAIN OUTCOME MEASUREMENTS: The diameters and areas of anastomoses were measured on the radiographs and compared with the corresponding measurements of the casts. RESULTS: No filling defects were observed in any of the cast models and only minor differences were found between the corresponding measurements of the anteroposterior and oblique projection radiographs and casts. The lower density of the contrast medium in the stenotic anastomoses indicated a stenosis. In the cross-sectional radiographs of the stenotic anastomoses there was a significant reduction (p < 0.001) both in the diameter and the area, the mean of which was 63% and the median 61%. CONCLUSIONS: Radiographs of cast models look promising in imaging of experimental end-to-side anastomoses when cross-sectional projections are also used.     

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.     

Low-power laser biostimulation enhances nerve repair after end-to-side neurorrhaphy: a double-blind randomized study in the rat median nerve model

Previous studies have shown that low-power laser biostimulation (lasertherapy) promotes posttraumatic nerve regeneration. The objective of the present study was to investigate the effects of postoperative lasertherapy on nerve regeneration after end-to-side neurorrhaphy, an innovative technique for peripheral nerve repair. After complete transection, the left median nerve was repaired by end-to-side neurorrhaphy on the ulnar donor nerve. The animals were then divided into four groups: one placebo group, and three laser-treated groups that received lasertherapy three times a week for 3 weeks starting from postoperative day 1. Three different types of laser emission were used: continuous (808 nm), pulsed (905 nm), and a combination of the two. Functional testing was carried out every 2 weeks after surgery by means of the grasping test. At the time of withdrawal 16 weeks postoperatively, muscle mass recovery was assessed by weighing the muscles innervated by the median nerve. Finally, the repaired nerves were withdrawn, embedded in resin and analyzed by light and electron microscopy. Results showed that laser biostimulation induces: (1) a statistically significant faster recovery of the lesioned function; (2) a statistically significant faster recovery of muscle mass; (3) a statistically significant faster myelination of the regenerated nerve fibers. From comparison of the three different types of laser emissions, it turned out that the best functional outcome was obtained by means of pulsed-continuous-combined laser biostimulation. Taken together, the results of the present study confirm previous experimental data on the effectiveness of lasertherapy for the promotion of peripheral nerve regeneration and suggest that early postoperative lasertherapy should be considered as a very promising physiotherapeutic tool for rehabilitation after end-to-side neurorrhaphy.