Ide C, Tohyama K, Tajima K, Endoh K, Sano K, Tamura M, Mizoguchi A, Kitada M, Morihara T, Shirasu M
Department of Anatomy and Neurobiology, Kyoto University Graduate School of Medicine, Japan.
Exp Neurol. 1998 Nov;154(1):99-112. doi: 10.1006/exnr.1998.6921.
Sciatic nerves were excised from 3 beagle dogs about 5 h after their sacrifice, treated three times by freezing and thawing, and stored in physiological saline for 3 months at -20 degrees C until used. Nerve segments 5 cm in length prepared from these stored nerves were transplanted to the common peroneal nerve in the right hindlimb of beagle dogs. Sixteen beagle dogs in total were used, in four treatment groups of two pairs each studied at 1 and 3 months. Five-hundred microliters basic fibroblast growth factor (bFGF) of two different concentrations (10 micrograms/300 microliters and 100 micrograms/300 microliters) which were impregnated in 0.5 ml gelatin hydrogels was applied around the sutured allografts. Autografting was also done in 4 beagle dogs, with no bFGF application. One month after the grafting, no regenerating nerves extended beyond the middle of the transplant in any of the allografts, except in the autografts in which a number of regenerated (myelinated) axons were present. Three months after the grafting, an abundance of myelinated axons was found at the middle of the graft: the numbers of axons per 10(4) micron 2 were 22.6 in the autografts and 10.6, 10.4 and 19.2 in the allografts treated with no bFGF, low-dose bFGF, and high-dose bFGF, respectively. Regenerating axons extended into the host nerve: the numbers of myelinated axons at the level 1.5 cm distal to the distal suture were 35.7, 0.9, 3.8, and 12.1 per 10(4) micron 2 in the above respective order. Although it was inferior in quality to the autograft, peripheral nerve regeneration was extensive in the distal nerve using freeze-thawed and bFGF-treated allografts at 3 months. Electromyography showed that the peroneus longus muscle responded to the electrical stimuli given at the site proximal to the transplant in all four groups. These data indicate that a 5-cm acellular nerve segment containing Schwann cell basal laminae can be used successfully as an allograft without any immunosuppressants and that exogenously applied bFGF can improve nerve regeneration by enhancing the growth of regenerating axons.
在3只比格犬处死后约5小时,取出坐骨神经,经冻融处理3次,然后在-20℃的生理盐水中保存3个月备用。从这些保存的神经中制备5厘米长的神经段,移植到比格犬右后肢的腓总神经。总共使用了16只比格犬,分为4个治疗组,每组2对,分别在1个月和3个月时进行研究。将两种不同浓度(10微克/300微升和100微克/300微升)的500微升碱性成纤维细胞生长因子(bFGF)浸渍在0.5毫升明胶水凝胶中,应用于缝合的同种异体移植物周围。4只比格犬进行了自体移植,未应用bFGF。移植后1个月,除了自体移植中有许多再生(有髓)轴突外,任何同种异体移植中均无再生神经延伸至移植中部以外。移植后3个月,在移植物中部发现大量有髓轴突:每10(4)平方微米的轴突数量在自体移植中为22.6,在未应用bFGF、低剂量bFGF和高剂量bFGF处理的同种异体移植中分别为10.6、10.4和19.2。再生轴突延伸至宿主神经:在远端缝线远端1.5厘米处,每10(4)平方微米的有髓轴突数量分别为35.7、0.9、3.8和12.1,顺序如上。虽然质量不如自体移植,但在3个月时使用冻融和bFGF处理的同种异体移植物,远端神经的周围神经再生广泛。肌电图显示,在所有4组中,腓骨长肌对在移植近端部位给予的电刺激有反应。这些数据表明,含有雪旺细胞基膜的5厘米无细胞神经段可成功用作同种异体移植,无需任何免疫抑制剂,并且外源性应用bFGF可通过促进再生轴突的生长来改善神经再生。