局部脉冲振动刺激对兔前交叉韧带重建后本体感觉恢复的影响

doi:10.13481/j.1671-587X.20230601

摘要/Abstract

摘要：

目的探讨不同局部脉冲振动刺激治疗条件对兔前交叉韧带重建（ACLR）术后本体感觉恢复的影响，为临床实践提供参考。方法选取45只健康新西兰大白兔，15只兔制备同种异体移植肌腱，30只兔制备前交叉韧带（ACL）模型，造模后将兔随机分为对照组和9个振动刺激治疗组（振动刺激治疗1~9组），每组3只。正交实验设计振动刺激治疗参数，采集各组兔ACL样本。HE染色观察各组兔ACL本体感受器形态表现，S100免疫组织化学染色观察各组兔ACL本体感受器分型，计数各组兔ACL本体感受器细胞数，实时荧光定量PCR（RT-qPCR）法检测各组兔ACL组织中生长相关蛋白43（GAP-43） mRNA表达水平，正交分析法筛选ACLR术后康复振动刺激治疗的最优参数。结果兔移植物张力尚可，无松弛状态，前抽屉实验和Lachman实验阴性；ACL组织移植物连续性存在，与胫骨和股骨腱骨愈合。ACL本体感受器中Ruffini小体为卵圆形或树突状形态，直径25~330 μm；Pacinian小体为圆形或椭圆形感受器，直径40~220 μm；Golgi腱器官感受器呈梭形，直径140~900 μm；游离神经末梢为无髓鞘神经末梢，长度0.5~1.5 μm。移植ACL中本体感受器主要分布于ACL的胫骨和股骨附着点，主要为类Ruffini小体和类Pacinian小体，无典型的类Golgi腱器官。与对照组比较，振动刺激治疗3、振动刺激治疗5和振动刺激治疗7组兔ACL本体感受器细胞数明显增加（F=28.49，P<0.01）。各组兔ACL组织中GAP-43 mRNA表达水平比较差异无统计学意义（F=0.83，P>0.05）。振幅强度、振动频率和作用时间对局部脉冲振动刺激治疗ACLR术后本体感受器数均存在较大影响，影响程度为振幅强度>作用时间>振动频率。其中最优振幅强度为3 mm，最优振动频率为25 Hz，最佳作用时间为10 min。结论局部脉冲振动刺激治疗可促进兔ACLR术后ACL本体感受器恢复，采用低振幅、低振动频率和短时间的局部脉冲振动刺激治疗可加快ACLR术后本体感觉的恢复。

关键词: 前交叉韧带, 振动刺激, 本体感受器, 康复, 正交实验

Abstract:

Objective To discuss the effect of different local pulse vibration stimulation treatment conditions on the recovery of proprioception after anterior cruciate ligament reconstruction （ACLR） in the rabbits，and to provide the reference for clinical practice. Methods A total of 45 healthy New Zealand white rabbits were selected，15 rabbits were used to prepare the allogenic tendon grafts， and 30 rabbits were used to prepare the anterior cruciate ligament （ACL） model. After modeling， the rabbits were randomly divided into control group and nine vibration stimulation treatment groups （vibraory stimulation treatment groups 1—9）， and there were three rabbits in each group. Orthogonal experiment was used to design the vibration stimulation treatment parameters， and the ACL samples of the rabbit in various groups were acquired； HE staining was used to observe the morphology of the proprioceptors of the rabbits in various groups； S100 immunohistochemistry staining was used to observe the types of proprioceptors of the rabbits in various groups； the number of proprioceptive cells of the rabbits in various groups was counted； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of growth associated protein-43 （GAP-43） mRNA in ACL tissue of the rabbits in various groups； orthogonal analysis method was used to select the optimal parameters for the recovery vibration stimulation treatment after ACLR. Results The graft tension was satisfactory， without slack state，with negative front drawer and Lachman tests；the continuity of graft could be seen in ACL tissue of the rabbits， healing with the tibial and femoral tendons. The Ruffini corpuscles in the ACL proprioceptor were oval or dendritic， with the diameters of 25—330 μm； the pacinian corpuscles showed round or elliptical sensors， with the diameters of 40—220 μm； the Golgi tendon organs showed fusiform， with the diameters of 140—900 μm； the free nerve endings were myelin-free nerve endings， with the diameters of 0.5—1.5 μm. The proprioceptors in the transplanted ACL were mainly distributed in the tibial and femoral attachment points of the ACL， mainly Ruffini-like corpuscles and Pacinian-like corpuscles，without typical Golgi-like tendon organs. Compared with control group， the numbers of proprioceptive cells in vibratory stimulation treatment groups 3， vibratory stimulation treatment 5， and vibratory stimulation treatment 7 groups were significantly increased （F=28.49， P<0.01）. There was no significant difference in the expression of GAP-43 mRNA in the cells between various groups （F=0.83， P>0.05）. The amplitude intensity， vibration frequency，and action time had a significant effect on the number of proprioceptors in the treatment of local pulse vibration stimulation after ACLR，and the effect degree was amplitude intensity>action time>vibration frequency. The optimal amplitude intensity was 3 mm， the optimal vibration frequency was 25 Hz， and the best action time was 10 min. Conclusion The local pulse vibration stimulation treatment can promote the recovery of ACL proprioceptors in the rabbits after ACLR. Low amplitude， low vibration frequency， and short duration of local pulse vibratory treatment could be used to accelerate the recovery of proprioception after ACLR.

Key words: Anterior cruciate ligament, Vibration stimulation, Mechanoreceptor, Rehabilitation, Orthogonal experiment

中图分类号:

R684

张新强,王博,冯会成. 局部脉冲振动刺激对兔前交叉韧带重建后本体感觉恢复的影响[J]. 吉林大学学报(医学版), 2023, 49(6): 1407-1414.

Xinqiang ZHANG,Bo WANG,Huicheng FENG. Effect of local impulse vibration stimulation on proprioception recovery after anterior cruciate ligament reconstruction in rabbits[J]. Journal of Jilin University(Medicine Edition), 2023, 49(6): 1407-1414.

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参考文献 19

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Group	Number of ACL proprioceptor cells
Control	7.67±1.53
Vibratory stimulation treatment 1	9.67±1.53
Vibratory stimulation treatment 2	6.33±2.08
Vibratory stimulation treatment 3	22.33±3.51 ^*
Vibratory stimulation treatment 4	10.33±1.53
Vibratory stimulation treatment 5	24.00±4.36 ^*
Vibratory stimulation treatment 6	6.33±1.53
Vibratory stimulation treatment 7	20.33±2.52 ^*
Vibratory stimulation treatment 8	4.00±2.65
Vibratory stimulation treatment 9	5.33±1.53

Source	Class Ⅲ sum of square	Free degree	Mean square	F	P
Amplitude intensity	698.963	2	349.481	49.197	<0.01
Vibration frequency	291.630	2	145.815	20.527	<0.01
Action time	487.185	2	243.593	34.291	<0.01

Vibration stimulation parameter	Amplitude intensity	Vibration frequency	Action time
K1	10.67	15.56	16.89
K2	18.89	13.00	6.56
K3	6.67	7.67	12.78
R	12.22	7.89	10.33

Source	Class Ⅲ sum of square	Free degree	Mean square	F	P
Amplitude intensity	0.026	2	0.013	0.183	0.834
Vibration frequency	0.047	2	0.023	0.326	0.725
Action time	0.056	2	0.028	0.391	0.681

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