神经端—侧缝合治疗臂丛损伤的实验研究
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摘要
目的比较周围神经标准端-侧缝合方法与其他各种改良手术方式在修复效果上的差异。
方法SD大鼠126只,随机分为7组。将右侧上臂段肌皮神经切断作为受体神经,以同侧尺神经作为供体神经,按照(1)端-端缝合组(end-to-end group,E-E组)；(2)端-侧缝合组(end-to-side group, E-S组)；(3)侧-侧缝合外膜开窗组(side-to-side group, S-S组)；(4)侧-侧缝合外膜不开窗组(side-to-side'group. S-S')组；(5)螺旋缠绕外膜去除组(spiral group, Sp组)；(6)螺旋缠绕外膜保留组(spiral' group,Sp'组);(7)阳性对照组(controlling group, CG组)制作端-侧缝合修复肌皮神经的动物模型。各组分别于术后1、2、3月进行取材,行神经电生理,肌、神经组织病理学检测,比较上述指标的差异。
结果自术后1月起至3月止,各端-侧缝合组均有明显神经再生现象,但各项检测指标均不及端-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>0.05)；神经外膜保留的两组中,检测指标为各实验组中最差。
结论各种缝合方式间大部分检测指标没有显著差别；神经外膜对端-侧缝合后的效果产生一定的影响；依靠增加供、受体神经之间的接触面积并不能消除神经外膜对神经端-侧缝合后出芽的阻碍作用；单纯通过改变手术方式以提高端-侧缝合后神经再生效果的能力非常有限。
目的建立采用不同供体神经修复臂丛神经的可行的动物模型；比较不同供体神经端-侧缝合后修复效果上的差异,并分析可能的影响因素。
方法SD大鼠90只,随机分为5组,切断动物右侧颈5,6神经根,模拟臂丛上干损伤的病理情况；分别以实验侧膈神经及同侧颈7神经根作为供体神经,按照(1)膈神经-肌皮神经端-端缝合修复组(Phr-MC-E-E);(2)同侧C7-肌皮神经端-端缝合修复组(C7-MC-E-E)组；(3)膈神经-肌皮神经端-侧缝合修复组(Phr-MC-E-S);(4)同侧C7-肌皮神经端-侧缝合修复组(C7-MC-E-S)组；(5)阳性对照组(Controlling group, CG)进行模型制作。各组分别于术后1、2、3月取材,对所有动物进行神经行为学评分、神经电生理以及病理组织学检测。
结果术后1-3月,采用神经端-侧缝合方式修复的模型可以观察到功能学及组织形态学恢复的证据,但各项检测指标均不及对应的端-端缝合组。以膈神经为供体神经的端-侧缝合组,各项指标得到了一定程度的恢复,但同侧颈7神经根端-侧缝合组神经再生的效果较差,各项检测指标为各修复组最低,差异具有统计学意义。
结论在大鼠上干损伤模型中,利用膈神经及同侧颈7神经根进行端-侧缝合修复肌皮神经是有效的；采用不同的供体神经进行端-侧缝合效果存在一定差异,膈神经与同侧颈7神经根相比,是端-侧缝合相对较好的供体神经；利用神经端.侧缝合修复臂丛神经损伤的效果尚不能达到与端-端缝合相同的效果,临床使用需掌握严格的适应征。
目的应用荧光素逆行追踪法验证神经端-侧缝合修复臂丛损伤后,重建神经通路的实际功能,对再生神经进行脊髓平面的定位。
方法SD大鼠24只,随机分为4组,造成臂丛上干损伤模型,分别以膈神经、同侧颈7神经根为供体神经,按照端-侧和端-端两种缝合方式修复肌皮神经。在术后3个月进行二期手术,采用True Blue和Diamidino Yellow两种荧光素分别注入大鼠肌皮神经和相应供体神经,对再生神经纤维的来源进行逆行示踪。在二期手术后3、7、14天后对实验动物进行灌注固定,取颈段脊髓进行冰冻连续切片,以荧光显微镜进行观察,并分析不同脊髓截段荧光素标记细胞的分布特征。
结果自术后3天起,供体神经来源的背根节及脊髓前角均出现荧光标记细胞,并随时间的延长逐渐增多。以同侧颈7为供体神经组,标记细胞仅出现在该节段,而以膈神经为供体神经组,标记细胞出现在颈3-5节段。端-侧缝合组在相应脊髓前角或背根神经节出现同时具有两种荧光剂的双标细胞或在同一脊髓节段同时出现分别以两种荧光剂标记的单标细胞。
结论采用不同供体神经进行端-侧缝合联合神经移植修复臂丛可使神经再生,端-侧缝合后,供-受体神经间建立了新的神经投射联系,并具有物质转运的实际功能；荧光素逆性示踪法可以准确定位端-侧缝合后再生神经的来源。
目的比较不同时间延迟修复对神经端-侧缝合疗效的影响,对动物实验结果与临床疗效之间存在差异的原因进行初步的探讨。
方法清洁级SD成年雌性大鼠30只,随机等分为5组,将右侧肌皮神经切断作为受体神经,以同侧尺神经作为供体神经。将肌皮神经切断后旷置不同时间,分别按照：(1)损伤后即刻修复组(Od)；(2)伤后1周(7d)修复组；(3)伤后3周(21d)修复组；(4)伤后6周(42d)修复组；(5)对照组(Controlling group,CG),制作模型。饲养3月后,对实验动物进行神经电生理及病理组织学检测。
结果术后3月,延迟1周修复组在电生理、肌肉、神经组织病理学等各项指标均为各组中最佳,与其余各组相比,差异具有统计学意义；直接修复组与延迟3周修复组部分指标差异不显著；而延迟6周后进行修复组,各项指标的恢复程度较低,与其他各组具有显著性差异(p<0.05)。
结论神经损伤后间隔不同时间进行修复,会对端-侧缝合的疗效造成影响；神经预变性一周后进行端-侧缝合可以达到较好的效果,但随着预变性时间的延长,手术效果逐渐变差；失神经支配后靶肌肉萎缩程度加重,其对神经端-侧缝合后功能恢复的影响可能是造成实验结果与临床疗效之间差异的原因之一。
Objective To compare the curative effectiveness of standard end-to-side neurorrhaphy and other modifications.
Methods 126 SD rats were randomized into 7 groups. The right musculocutaneous nerves were transected in all animals. The ipsilateral ulnar nerve served as the donor nerve. In group 1 the ulnar nerve and musculocutaneous nerve were coaptated in end-to-end pattern. In group 2, the musculocutaneous nerve was coaptated with ulnar nerve in standard end-to-side pattern. Side-to-side neurorrhaphy with and without removal of the epineural sheath were undergone in group 3 and 4. In group 5 and 6, the recipient nerve was coaptated with the donor nerve in helicoid pattern, with and without removal of the Epineurium The musculocutaneous nerve in group 7 were not reconstructed, and served as controlling group. Electrophysiological and histological analyses were performed in the 1st,2nd and 3rd month postoperatively.
Results By the end of the experiment, regenerative signs were observed in all end-to-side groups. The curative effectiveness of end-to-side coaptation was not as satisfactory as that of the end-to-end neurorrhaphy. There was no significant difference in end-to-side groups (p>0.05).The most unsatisfactory result was observed in groups without removal of the epineural sheath.
Conclusion There was no significant difference in end-to-side coaptation patterns. Epineurium had negative impact to nerve regeneration after end-to-side neurorraphy. The effectiveness of end-to-side operative modification to improve the nerve regeneration was limited
Objective To evaluate the regenerate efficacy of different donor nerves in reconstructing brachial plexus injury by end-to-side coaptation
Methods 90 SD rats were subjected to brachial plexus upper thunk injury and randomized into 5 groups. The right musculocutaneous nerves of 4 groups were reconstructed by both end-to-side and end-to-end neurorraphy using phrenic nerve and ipsilateral C7 root respectively. Another group served as controlling group without treatment. Electrophysiologic and histologic examinations, accompanied with behavioral tests were performed in the 1st,2nd and 3rd month postoperatively.
Results By the end of the experiment, regenerative signs indicate that reinnervation was possible by end-to-side neurotization. But the outcomes were only passable. The application of the ipsilateral C7 root resulted unsatisfactorily.
Conclusion It is available to reconstruct the musculocutaneous nerves in rats of brachial plexus injury by end-to-side neurorraphy. The efficacy of various donor nerves is distinct. The phrenic nerve served as a better motor nerve in end-to-side coaptation, comparing with the ipsilateral C7 root. Unless there is no choice of end-to-end neurorraphy, the end-to-side coaptation would be an alternative for brachial plexus reconstruction.
Objective To explore the effectiveness of end-to-side neurorraphy for reconstruction in brachial plexus injury and the spinal localization of regenerating axons using flurorescence retrograde tracing.
Methods 24 female SD rats were subjected to brachial plexus superior trunk injurys and randomized into 4 groups:phrenic nerve to musculocutaneous nerve end-to-end transfer, phrenic nerve to musculocutaneous nerve end-to-side transfer ipsilateral cervical 7 root to musculocutaneous nerve end-to-end transfer and ipsilateral cervical 7 root to musculocutaneous nerve end-to-side transfer.3 months after surgery, the musculocutaneous nerve and the donor nerves were retrograde neurotraced with True Blue and Diamidino Yellow respectively. C2 to T2 dorsal root ganglion(DRG) and spinal cord were harvested after perfusion fixation 3,7 and 14 days later.30μm-thinkness slices were observed under the fluromicroscope.
Results Fluorescence labeled neurons were seen both in DRG and anterior horn of the spinal cord. There were more and more positively labeled neurons with pass of time. The fluorescence labeled neurons of C7 group were observed only in C7-DRG and anterior corner in correspond spinal cord.The ones of phrenic nerve were found in C3 to C5-DRG and anterior corner in. correspond spinal cord. Two kinds of single labeled neurons or double labeled neurons were observed in the same site in the groups of end-to-side neurorraphy
Conclusion Nerve regeneration is possible in an end-to-side neurorraphy brachial plexus reconstruction model. Fluorescence retrograde tracing can be used for precise neuron locating.
Objective To study the influence of different delayed-repairing time on end-to-side neurorraphy and to discuss the possible reasons of the disparity between the experimental studies and the clinical applications.
Methods 30 female SD rats were subjected to musculocutaneous nerve transsection and randomized into 5 groups. The musculocutaneous nerve were reconstructed in the end-to-side way with the ipsilateral ulnar nerve in different time. 1 week,3 weeks and 6 weeks were passed before the reconstructive surgery in delayed-reconstructed groups together with the immediately reconstructed group and the controlling group, in which the repair was abandoned.3 months after surgery, electrophysiologic and histologic examinations were taken.among the models.
Results 3 months later,1-week delayed group performed significantly better in electrophysiologic and histologic examinations than the other groups. There was no noticeable difference between immediately reconstructed group and 3-week delayed group in some tests. The outcomes were decreased in 6-week delayed group, which was significantly different with other groups (p<0.05).
Conclusion Delayed-rerairing time can influence the effectiveness of end-to-side neurorraphy after nerve injuries. The outcomes of the surgery were better after 1-week nerve pre-degeneration, but the therapeutic effect of end-to-side coaptation decreased with pass of time. Aggravation of myatrophy meight be one of the possible reasons to explain the imcompatible results between the experimental studies and the clinical applications.
方法SD大鼠126只,随机分为7组。将右侧上臂段肌皮神经切断作为受体神经,以同侧尺神经作为供体神经,按照(1)端-端缝合组(end-to-end group,E-E组)；(2)端-侧缝合组(end-to-side group, E-S组)；(3)侧-侧缝合外膜开窗组(side-to-side group, S-S组)；(4)侧-侧缝合外膜不开窗组(side-to-side'group. S-S')组；(5)螺旋缠绕外膜去除组(spiral group, Sp组)；(6)螺旋缠绕外膜保留组(spiral' group,Sp'组);(7)阳性对照组(controlling group, CG组)制作端-侧缝合修复肌皮神经的动物模型。各组分别于术后1、2、3月进行取材,行神经电生理,肌、神经组织病理学检测,比较上述指标的差异。
结果自术后1月起至3月止,各端-侧缝合组均有明显神经再生现象,但各项检测指标均不及端-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>0.05)；神经外膜保留的两组中,检测指标为各实验组中最差。
结论各种缝合方式间大部分检测指标没有显著差别；神经外膜对端-侧缝合后的效果产生一定的影响；依靠增加供、受体神经之间的接触面积并不能消除神经外膜对神经端-侧缝合后出芽的阻碍作用；单纯通过改变手术方式以提高端-侧缝合后神经再生效果的能力非常有限。
目的建立采用不同供体神经修复臂丛神经的可行的动物模型；比较不同供体神经端-侧缝合后修复效果上的差异,并分析可能的影响因素。
方法SD大鼠90只,随机分为5组,切断动物右侧颈5,6神经根,模拟臂丛上干损伤的病理情况；分别以实验侧膈神经及同侧颈7神经根作为供体神经,按照(1)膈神经-肌皮神经端-端缝合修复组(Phr-MC-E-E);(2)同侧C7-肌皮神经端-端缝合修复组(C7-MC-E-E)组；(3)膈神经-肌皮神经端-侧缝合修复组(Phr-MC-E-S);(4)同侧C7-肌皮神经端-侧缝合修复组(C7-MC-E-S)组；(5)阳性对照组(Controlling group, CG)进行模型制作。各组分别于术后1、2、3月取材,对所有动物进行神经行为学评分、神经电生理以及病理组织学检测。
结果术后1-3月,采用神经端-侧缝合方式修复的模型可以观察到功能学及组织形态学恢复的证据,但各项检测指标均不及对应的端-端缝合组。以膈神经为供体神经的端-侧缝合组,各项指标得到了一定程度的恢复,但同侧颈7神经根端-侧缝合组神经再生的效果较差,各项检测指标为各修复组最低,差异具有统计学意义。
结论在大鼠上干损伤模型中,利用膈神经及同侧颈7神经根进行端-侧缝合修复肌皮神经是有效的；采用不同的供体神经进行端-侧缝合效果存在一定差异,膈神经与同侧颈7神经根相比,是端-侧缝合相对较好的供体神经；利用神经端.侧缝合修复臂丛神经损伤的效果尚不能达到与端-端缝合相同的效果,临床使用需掌握严格的适应征。
目的应用荧光素逆行追踪法验证神经端-侧缝合修复臂丛损伤后,重建神经通路的实际功能,对再生神经进行脊髓平面的定位。
方法SD大鼠24只,随机分为4组,造成臂丛上干损伤模型,分别以膈神经、同侧颈7神经根为供体神经,按照端-侧和端-端两种缝合方式修复肌皮神经。在术后3个月进行二期手术,采用True Blue和Diamidino Yellow两种荧光素分别注入大鼠肌皮神经和相应供体神经,对再生神经纤维的来源进行逆行示踪。在二期手术后3、7、14天后对实验动物进行灌注固定,取颈段脊髓进行冰冻连续切片,以荧光显微镜进行观察,并分析不同脊髓截段荧光素标记细胞的分布特征。
结果自术后3天起,供体神经来源的背根节及脊髓前角均出现荧光标记细胞,并随时间的延长逐渐增多。以同侧颈7为供体神经组,标记细胞仅出现在该节段,而以膈神经为供体神经组,标记细胞出现在颈3-5节段。端-侧缝合组在相应脊髓前角或背根神经节出现同时具有两种荧光剂的双标细胞或在同一脊髓节段同时出现分别以两种荧光剂标记的单标细胞。
结论采用不同供体神经进行端-侧缝合联合神经移植修复臂丛可使神经再生,端-侧缝合后,供-受体神经间建立了新的神经投射联系,并具有物质转运的实际功能；荧光素逆性示踪法可以准确定位端-侧缝合后再生神经的来源。
目的比较不同时间延迟修复对神经端-侧缝合疗效的影响,对动物实验结果与临床疗效之间存在差异的原因进行初步的探讨。
方法清洁级SD成年雌性大鼠30只,随机等分为5组,将右侧肌皮神经切断作为受体神经,以同侧尺神经作为供体神经。将肌皮神经切断后旷置不同时间,分别按照：(1)损伤后即刻修复组(Od)；(2)伤后1周(7d)修复组；(3)伤后3周(21d)修复组；(4)伤后6周(42d)修复组；(5)对照组(Controlling group,CG),制作模型。饲养3月后,对实验动物进行神经电生理及病理组织学检测。
结果术后3月,延迟1周修复组在电生理、肌肉、神经组织病理学等各项指标均为各组中最佳,与其余各组相比,差异具有统计学意义；直接修复组与延迟3周修复组部分指标差异不显著；而延迟6周后进行修复组,各项指标的恢复程度较低,与其他各组具有显著性差异(p<0.05)。
结论神经损伤后间隔不同时间进行修复,会对端-侧缝合的疗效造成影响；神经预变性一周后进行端-侧缝合可以达到较好的效果,但随着预变性时间的延长,手术效果逐渐变差；失神经支配后靶肌肉萎缩程度加重,其对神经端-侧缝合后功能恢复的影响可能是造成实验结果与临床疗效之间差异的原因之一。
Objective To compare the curative effectiveness of standard end-to-side neurorrhaphy and other modifications.
Methods 126 SD rats were randomized into 7 groups. The right musculocutaneous nerves were transected in all animals. The ipsilateral ulnar nerve served as the donor nerve. In group 1 the ulnar nerve and musculocutaneous nerve were coaptated in end-to-end pattern. In group 2, the musculocutaneous nerve was coaptated with ulnar nerve in standard end-to-side pattern. Side-to-side neurorrhaphy with and without removal of the epineural sheath were undergone in group 3 and 4. In group 5 and 6, the recipient nerve was coaptated with the donor nerve in helicoid pattern, with and without removal of the Epineurium The musculocutaneous nerve in group 7 were not reconstructed, and served as controlling group. Electrophysiological and histological analyses were performed in the 1st,2nd and 3rd month postoperatively.
Results By the end of the experiment, regenerative signs were observed in all end-to-side groups. The curative effectiveness of end-to-side coaptation was not as satisfactory as that of the end-to-end neurorrhaphy. There was no significant difference in end-to-side groups (p>0.05).The most unsatisfactory result was observed in groups without removal of the epineural sheath.
Conclusion There was no significant difference in end-to-side coaptation patterns. Epineurium had negative impact to nerve regeneration after end-to-side neurorraphy. The effectiveness of end-to-side operative modification to improve the nerve regeneration was limited
Objective To evaluate the regenerate efficacy of different donor nerves in reconstructing brachial plexus injury by end-to-side coaptation
Methods 90 SD rats were subjected to brachial plexus upper thunk injury and randomized into 5 groups. The right musculocutaneous nerves of 4 groups were reconstructed by both end-to-side and end-to-end neurorraphy using phrenic nerve and ipsilateral C7 root respectively. Another group served as controlling group without treatment. Electrophysiologic and histologic examinations, accompanied with behavioral tests were performed in the 1st,2nd and 3rd month postoperatively.
Results By the end of the experiment, regenerative signs indicate that reinnervation was possible by end-to-side neurotization. But the outcomes were only passable. The application of the ipsilateral C7 root resulted unsatisfactorily.
Conclusion It is available to reconstruct the musculocutaneous nerves in rats of brachial plexus injury by end-to-side neurorraphy. The efficacy of various donor nerves is distinct. The phrenic nerve served as a better motor nerve in end-to-side coaptation, comparing with the ipsilateral C7 root. Unless there is no choice of end-to-end neurorraphy, the end-to-side coaptation would be an alternative for brachial plexus reconstruction.
Objective To explore the effectiveness of end-to-side neurorraphy for reconstruction in brachial plexus injury and the spinal localization of regenerating axons using flurorescence retrograde tracing.
Methods 24 female SD rats were subjected to brachial plexus superior trunk injurys and randomized into 4 groups:phrenic nerve to musculocutaneous nerve end-to-end transfer, phrenic nerve to musculocutaneous nerve end-to-side transfer ipsilateral cervical 7 root to musculocutaneous nerve end-to-end transfer and ipsilateral cervical 7 root to musculocutaneous nerve end-to-side transfer.3 months after surgery, the musculocutaneous nerve and the donor nerves were retrograde neurotraced with True Blue and Diamidino Yellow respectively. C2 to T2 dorsal root ganglion(DRG) and spinal cord were harvested after perfusion fixation 3,7 and 14 days later.30μm-thinkness slices were observed under the fluromicroscope.
Results Fluorescence labeled neurons were seen both in DRG and anterior horn of the spinal cord. There were more and more positively labeled neurons with pass of time. The fluorescence labeled neurons of C7 group were observed only in C7-DRG and anterior corner in correspond spinal cord.The ones of phrenic nerve were found in C3 to C5-DRG and anterior corner in. correspond spinal cord. Two kinds of single labeled neurons or double labeled neurons were observed in the same site in the groups of end-to-side neurorraphy
Conclusion Nerve regeneration is possible in an end-to-side neurorraphy brachial plexus reconstruction model. Fluorescence retrograde tracing can be used for precise neuron locating.
Objective To study the influence of different delayed-repairing time on end-to-side neurorraphy and to discuss the possible reasons of the disparity between the experimental studies and the clinical applications.
Methods 30 female SD rats were subjected to musculocutaneous nerve transsection and randomized into 5 groups. The musculocutaneous nerve were reconstructed in the end-to-side way with the ipsilateral ulnar nerve in different time. 1 week,3 weeks and 6 weeks were passed before the reconstructive surgery in delayed-reconstructed groups together with the immediately reconstructed group and the controlling group, in which the repair was abandoned.3 months after surgery, electrophysiologic and histologic examinations were taken.among the models.
Results 3 months later,1-week delayed group performed significantly better in electrophysiologic and histologic examinations than the other groups. There was no noticeable difference between immediately reconstructed group and 3-week delayed group in some tests. The outcomes were decreased in 6-week delayed group, which was significantly different with other groups (p<0.05).
Conclusion Delayed-rerairing time can influence the effectiveness of end-to-side neurorraphy after nerve injuries. The outcomes of the surgery were better after 1-week nerve pre-degeneration, but the therapeutic effect of end-to-side coaptation decreased with pass of time. Aggravation of myatrophy meight be one of the possible reasons to explain the imcompatible results between the experimental studies and the clinical applications.
引文
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