Microscopic Evidences That Bone Marrow Mononuclear Cell Treatment Improves Sciatic Nerve Regeneration After Neurorrhaphy

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of low-level laser therapy (LLLT) on peripheral nerve regeneration using fibrin glue derived from snake venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mesenchymal stem cells engrafted in a fibrin scaffold stimulate Schwann cell reactivity and axonal regeneration following sciatic nerve tubulization

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Muscle reinnervation in one or two stages?: experimental study in rats with end-to-side nerve graft

OBJETIVO: Comparar a reinervação muscular com enxerto de nervo em um e dois tempos operatórios, utilizando a neurorrafia término-lateral (NTL) sem lesão do nervo doador. MÉTODOS: Vinte ratos foram distribuídos em quatro grupos. O grupo 1 (G1), um estágio, recebeu o enxerto que foi suturado ao nervo tibial (NT), por meio de NTL, e seu coto livre foi suturado por NTL ao coto distal do nervo peroneal (NP), seccionado a um centímetro do NT, na mesma cirurgia. O grupo 2 (G2), dois estágios, recebeu o enxerto de nervo na primeira cirurgia, como já descrito. Dois meses depois, na segunda cirurgia, o NP foi seccionado e seu coto distal ligado ao coto distal do enxerto como em G1. O grupo controle de normalidade (Gn) recebeu o enxerto da mesma forma, apenas. E o grupo controle de denervação (Gd), além de receber o enxerto, teve o NP seccionado e seus cotos sepultados na musculatura adjacente, com a finalidade de denervar o músculo tibial cranial (MTC), alvo deste estudo. Os parâmetros utilizados para avaliar a reinervação do MTC foram massa muscular, diâmetro mínimo da fibra muscular e área. RESULTADOS: O grupo G2 apresentou superioridade (p<0,0001) em relação ao G1 na massa do MTC, no diâmetro mínimo e na área das fibras musculares. Na comparação entre os quatro grupos, estes mesmos parâmetros tiveram sua expressão máxima em Gn e mínima em Gd, como era esperado. CONCLUSÃO: A reinervação muscular em dois estágios apresenta melhor resultado quando comparada à técnica em um tempo.

Muscle graft as a substitute for peripheral nerve graft in rats

OBJETIVO: Avaliar a aplicabilidade do uso de músculo autógeno, tratado de diversas maneiras, em substituição aos enxertos de nervo. MÉTODOS: Os ratos foram separados em sete grupos que receberam, como tratamento a uma lesão nervosa padronizada, os seguintes tipos de enxertos: músculo fresco, músculo fixado com formol 10%, músculo congelado em freezer, músculo congelado em refrigerador, músculo denervado, nervo periférico e um grupo ficou sem qualquer tratamento. Foi avaliado o aspecto histológico das fibras nervosas no segmento reparado. RESULTADOS: A avaliação do segmento nervoso reparado mostrou que existiam axônios em quase todos os grupos, mas a metodologia empregada não possibilitou caracterizar adequadamente as diferenças entre os grupos. CONCLUSÃO: Este estudo mostrou a migração de axônios por meio de todos os enxertos utilizados.

Inside-out vein graft and inside-out artery graft in rat sciatic nerve repair

Although veins and arteries present similar wall structures, there are differences which may be relevant in peripheral nerve reconstruction. Inside-out vein grafts (IOVG) have been satisfactorily used to repair both motor and sensitive nerves. However, the inside-out artery graft (IOAG) is a new technique and not fully investigated. Our study presents comparative morphological data on nerve regeneration achieved with IOVG and IOAG in the repair of Wistar rat sciatic nerves. Jugular veins and aorta arteries were harvested from donor animals and used inside-out to bridge a 10-mm gap. Animals were sacrificed at 10 weeks to evaluate nerve regeneration. Both techniques presented great variability in nervous tissue, though some animals showed satisfactory results. Different intensities of scarring processes might have interfered with nerve regeneration. Although IOVG and IOAG techniques showed similar morphometric results, in general, IOVG presented a closer-to-normal nerve organization than IOAG.

Effect of electrical stimulation of the cranial tibial muscle after end-to-side neurorrhaphy of the peroneal nerve in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Localization and irregular distribution of Na,K-ATPase in myelin sheath from rat sciatic nerve

Sodium, potassium adenosine triphosphatase (Na,K-ATPase) is a membrane-bound enzyme that maintains the Na+ and K+ gradients used in the nervous system for generation and transmission of bioelectricity. Recently, its activity has also been demonstrated during nerve regeneration. The present study was undertaken to investigate the ultrastructural localization and distribution of Na,K-ATPase in peripheral nerve fibers. Small blocks of the sciatic nerves of male Wistar rats weighing 250-300g were excised, divided into two groups, and incubated with and without substrate, the para-nitrophenyl phosphate (pNPP). The material was processed for transmission electron microscopy, and the ultra-thin sections were examined in a Philips CNI 100 (TM) electron microscope. The deposits of reaction product were localized mainly on the axolemma, on axoplasmic profiles, and irregularly dispersed on the myelin sheath, but not in the unmyelinated axons. In the axonal membrane, the precipitates were regularly distributed on the cytoplasmic side. These results together with published data warrant further studies for the diagnosis and treatment of neuropathies with compromised Na,K-ATPase activity. (c) 2007 Elsevier Ltd. All rights reserved.

Experimental surgery of the facial nerve. Assessment of the intraperitoneal use of exogenous gangliosides

Compression and section of the facial nerve were performed in 48 rats in order to study the anatomopathological alterations occurring after daily intraperitoneal injections of 100 mg of exogenous gangliosides (Sinaxial®) for 45, 90, 180 days. In groups submitted to nerve compression, the histopathological changes were discrete and in the 180-day subgroups the nerve was practically normal. In animals submitted to section and neurorrhaphy there was formation of an amputation neuroma, a granuloma around the suture, axonal unstructuration and inter and perineural fibrosis. No significant differences were observed between the groups submitted or not to injection of exogenous gangliosides, indicating that the major factors involved in the quality of nerve regeneration were the technique and the formation of fibrosis and of an amputation neuroma.

Inside-out vs. standard vein graft to repair a sensory nerve in rats

Nerve regeneration in a sensory nerve was obtained by the application of different techniques: inside-out vein graft (IOVG group) and standard vein graft (SVG group). These techniques provide a good microenvironment for axon regeneration in motor nerves, but their efficiency for regeneration of sensory nerves is controversial. The saphenous nerve was sectioned and repaired by the inside-out and standard vein graft techniques in rats. After 4, 12, and 20 weeks the graft and the distal stump were observed under electron microscopy. In each studied period, the pattern, diameters, and thickness of the myelin sheaths of the regenerated axons were measured in the graft and distal stump. A comparative study about the regenerated nerve fibers by these two different techniques was performed. Regenerated nerve fibers were prominent in both vein grafts 4 weeks after the surgical procedures. On the other hand, in the distal stump, regenerated nerve fibers were observed only from 12 weeks. In both inside-out vein graft and standard vein graft statistical difference was not observed about the diameters and thickness of the myelinated fibers after 20 weeks. On the other hand, the inside-out group had greater regenerated axon number when compared to the standard group. There is a capillary invasion in both graft and distal stump, especially in the IOVG group. The regenerated axons follow these capillaries all the time like satellite microfascicles. After 20 weeks, the diameters of regenerated fibers repaired by the standard vein graft technique were closer to the normal fibers compared to the inside-out vein graft. On the other hand, the pattern of these regenerated axons was better in the IOVG group.

Implante de tubo de silicone com e sem colágeno na regeneração de nervos em eqüinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

ESTUDO DO EFEITO DE GANGLIOSIDEOS EXOGENOS NA REGENERACAO NERVOSA APOS COMPRESSAO E NEURORRAFIA NO NERVO CIATICO DO RATO

The authors studied the possible inductive effect of exogenous gangliosides administrated intra-peritoneal and intra-muscular, in 33 rats, after compression and sutures of sciatic nerve. In the animals suffering compression of the sciatic nerve only, the myelin sheath was preserved with good regeneration of the nerve. There was no statistics difference between the two groups with or without the administration of gangliosides. In the group of rats where section of the sciatic nerve and suture were performed, neuroma of amputation as well as dense cicatricial tissue with inflammatory reaction of foreign body type was noted around the sutures. In conclusion, the formation of neuroma of amputation associated with severe inflammatory reaction had an important role in determining the regeneration of the nerve.

Avaliação experimental dos efeitos do anestésico local ropivacaína em nervos ciáticos

This study was undertaken to investigate the effects of ropivacaine after intrafascicular injection into the sciatic nerves of albino rabbits. Twenty adult albine rabbits were used, following sedation with intramuscular ketamine (50 mg/kg) for nerve exposure by lateral incision. We considered three experimental groups: Group I:sciatic nerve control; Group II: intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine into the rigth nerves. The specimens were colected at 48 h after drugs administration; Group III. intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine in the rigth nerves. The specimens were colected at 7 days after drugs administration. The sciatic nerves were removed from these animals and fixed in Karnowisky solution for 24 hours. After partial dehydration up to 95% ethanol, they were embedded in historesin (Leica). The tissue was then sectioned at 1-2μm. Sections were stained with haematoxylin-eosin (HE); toluidine blue (TB) or picrosirius-haematoxylin (PSH). Comparing with control group the histological evidence of inflammatory reaction (migration of macrophagic cells and eosinophils-appeared soon after injection, with intense proliferation of perineurial cells. The results show that after 7 days of intrafascicular injection there was a severe fibrosis and an increase on perineurial vascularization. In group 2 the inflammatory reaction was noted near the local of the injection. Furthermore in this experiment we observed an increase on the number of epineurial lipoblasts and adipocytes. This study demonstrated that the toxic effects of ropivacaine are transient. In many cases there was an initial fascicular recover and axonal regeneration after 7 days of the injection.

Estudo comparativo experimental da injeção perineural do álcool benzílico 0,75% e do álcool etílico absoluto nos nervos palmares de eqüinos

The clinical and histopathological effects of two alcoholic neurolytics were studied in horses. Normal horses were shod with a designed shoe adaptted with 5 screws to produce solar pain. After gait and lameness score analysis, the palmar nerve of 5 horses was injected with 5 ml of 0,75% benzyl alcohol (Group A) and 5 horses were injected with 5 ml of absolute ethyl alcohol (Group B). The animals were submitted to regular lameness evaluation and solar sensibility tests during next six months. The solar sensitivity returned 5 months latter in the group injected with benzyl alcohol 0,75%, while in the group injected with absolute ethyl alcohol, the sole was still desensitized 6 months latter. The histopathological findings showed that the nerve injected with benzyl alcohol 0,75%, resulted in axonotmesis, characterized by axonal nerve degeneration, with possibilities for the nerve conduction recovery. The perineural injection of ethyl alcohol absolute, resulted in neurotmesis with difficult nerve regeneration. It was concluded that chemical neurolysis with alcohol is an option for temporary or permanent nerve blocks in horses.

Neurorrafia do ramo bucal dorsal do nervo facial em coelhos com proteção de segmento intestinal alógeno

18 rabbits, New Zealand, males, adults were used for clinical and histological evaluation of repair dorsal buccal branch of facial nerve after 15, 30 and 60 days postoperatively (PO). The animals were divided into two groups for transection and 10-0 nylon monofilament epineural suture of buccal branch. In animals in Group I, the nerve was coated with protection of jejunum allograft preserved in glycerin 98% and in group II was applied epineural suture. Both groups occurred the return of movement of the upper lip from the eighth week. There was infiltrated cellular and giant cells with fibrosis unsystematic and collagen fibers of the allograft jejunum joing to the connective tissue. At 15 and 30 days of PO, the distal nerve stumps of both groups were found with degeneration wallerian and in 60 days, regenerated fibers. The repair of the dorsal buccal branch of facial nerve with the allograft wasn't significantly different between the control rabbits as to the assessment of histological and functional recovery.