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P Tos

Publications and source records attributed to P Tos.

15 recordsLinked to original sources
PubMed2007

Origin and history of end-to-side neurorrhaphy.

This historical note offers a perspective concerning the origin of the employment of end-to-side (termino-lateral) anastomosis for nerve repair and summarizes the works that have been published on this surgical technique through the first part of the 20th Century. While the origin of end-to-side neurorrhaphy is usually dated to the beginning of the 20th Century, some works referring to this technique were published earlier, the first of which dates as far back as 1873. A number of interesting clinical and experimental studies have been carried out on end-to-side nerve anastomosis during the first years of the twentieth century. However, this literature is not easily detectable through current online scientific databases. In this paper we will give an overview of these early works. This history contributes interesting information to the debate surrounding this surgical concept and adds perspective to the use of a technique that has attracted a great deal of attention over the last 15 years.

Anastomosis, Surgical↗
PubMed2007

Comparison of fresh and predegenerated muscle-vein-combined guides for the repair of rat median nerve.

Over the last 10 years, we have investigated a particular type of bioengineered nerve guide, the muscle-vein-combined tube, which is made by filling a vein with skeletal muscle. In our previous studies we have always used fresh skeletal muscle to fill vein conduits. In the present study we compared the use of fresh and predegenerated (freeze-thawed) skeletal muscle for muscle-vein-combined nerve guides. In this study, a 10-mm-long rat median nerve defect was repaired using either type of nerve guide. The samples were analyzed 5 and 30 days after surgery by light and electron microscopy. In addition, reverse transcription polymerase chain reaction (RT-PCR) was carried out to investigate the expression of mRNAs coding for glial markers, as well as glial growth factor (NRG1) and its receptors (erbB2 and erbB3). Results showed differences between the two types of nerve guides at postoperative day 5; however, no difference was detected at day 30 suggesting that both types of tissue-engineered conduit are effective for repairing peripheral nerve defects in this experimental model.

Animals↗
PubMed2006

Lack of topographic specificity in nerve fiber regeneration of rat forelimb mixed nerves.

Multiple nerve repair by means of a Y-shaped nerve guide represents a good model for studying the specificity of peripheral nerve fiber regeneration. Here we have used it for investigating the specificity of axonal regeneration in mixed nerves of the rat forelimb model. The left median and ulnar nerves, in adult female rats, were transected and repaired with a 14-mm Y-shaped conduit. The proximal end of the Y-shaped conduit was sutured to the proximal stump of either the median nerve or the ulnar nerve. Ten months after surgery, rats were tested for functional recovery of each median and ulnar nerve. Quantitative morphology of regenerated myelinated nerve fibers was then carried out by the two-dimensional disector technique. Results showed that partial recovery of both median and ulnar nerve motor function was regained in all experimental groups. Performance in the grasping test was significantly lower when the ulnar nerve was used as the proximal stump. Ulnar test assessment showed no significant difference between the two Y-shaped repair groups. The number of regenerated nerve fibers was significantly higher in the median nerve irrespectively of the donor nerve, maintaining the same proportion of myelinated fibers between the two nerves (about 60% median and 40% ulnar). On the other hand, nerve fiber size and myelin thickness were significantly larger in both distal nerves when the median nerve was used as the proximal donor nerve stump. G-ratio and myelin thickness/axon diameter ratio returned to normal values in all experimental groups. These results demonstrate that combined Y-shaped-tubulization repair of median and ulnar nerves permits the functional recovery of both nerves, independently from the proximal donor nerve employed, and that tissue, and not topographic, specificity guides nerve fiber regeneration in major forelimb mixed nerves of rats.

Animals↗
PubMed2004

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.

Animals↗
PubMed2003

Schwann-cell proliferation in muscle-vein combined conduits for bridging rat sciatic nerve defects.

Among the various grafting procedures that have been studied as alternatives to traditional fresh nerve autografts for the repair of severed peripheral nerves, muscle-vein-combined graft conduits have recently been devised and successfully employed. In the present study, the early presence, origin, and proliferation activity of Schwann cells (SCs) along this particular type of biological graft conduit have been investigated, using antibodies directed against glial fibrillar acid protein (GFAP), a protein that is specifically expressed in glial cells, and proliferating cell nuclear antigen (PCNA), a protein that is expressed by cells during DNA synthesis. Results showed that the muscle-vein-combined graft was progressively invaded by a number of GFAP-immunopositive SCs, many of which were also found to be immunopositive for PCNA, thus demonstrating that their proliferation continues to occur inside the graft. Among the molecules that could be involved in the stimulation of Schwann-cell proliferation is neuregulin-1 (NRG-1) that mediates its effects by binding to the ErbB receptor tyrosine kinase family. In the present study, the authors report on the RT-PCR analysis for NRG-1 and ErbB3 mRNAs, showing an overall increase in the content of these transcripts inside the muscle-vein-combined graft. These results suggest that the muscle-vein-combined graft conduit constitutes an environment favorable to potentiate Schwann-cell proliferation during the early regeneration phases.

Animals↗
PubMed2001

Confocal imaging of Schwann-cell migration along muscle-vein combined grafts used to bridge nerve defects in the rat.

Schwann cells guide axonal regrowth during peripheral nerve repair. In a case of a nerve lesion with substance loss, a graft conduit is necessary to enable axons to reach the distal nerve stump. If a non-nervous autograft is used, the question arises as to the presence and origin of Schwann cells along the grafted tube. We addressed this issue using a tubulization technique based on the use of an autologous vein filled with fresh skeletal muscle for the repair of sciatic nerve defects in the rat. We showed that both ends of the graft were early and progressively colonized by a number of glial fibrillar acid protein-immunopositive and S-100 immunonegative cells, an immunocytochemical pattern typical of immature Schwann cells. These cells, which were located in the interstice between grafted skeletal muscle fibers, are mainly organized into long chains oriented along the main axis of the graft and progressively colonize all the graft. Schwann cells coming from the distal nerve end are suitable for being responsible for guiding regeneration of nerve fibers along the graft toward the correct periphery (tissue specificity).

Animals↗
PubMed2001

Methodological issues in size estimation of myelinated nerve fibers in peripheral nerves.

Size estimation of myelinated nerve fibers in peripheral nerves is a very common task in neuromorphology and different dedicated morpho-quantitative procedures have been devised and used to date. Unfortunately, many reports on experimental nerve studies lack comprehensive information on the procedures that have been designed and applied for myelinated fiber size estimation. This paper addresses the issue in the light of the recent advances in quantitative morphology that have recognized the concept of unbiased estimates as the key methodological issue to be addressed in morpho-quantitative studies. The potential foundations of bias at various study levels are analysed together with indications on how to cope with them. In addition, the issue of the precision of size estimates is addressed and the various geometrical parameters that can be selected for myelinated nerve fiber size assessment are outlined. Taken together, information provided in this paper is expected to help investigators conduct an appropriate preliminary study design phase, the key step for setting up the most adequate morpho-quantitative procedure for any given research goal.

Animals↗
PubMed2000

Nerve repair by means of vein filled with muscle grafts I. Clinical results.

Peripheral nerve lesions with a long segment defect need a grafting conduit to heal. Although autogenous nerve grafting is still considered the best method for bridging nerve defects, several alternative types of conduits (biological and synthetic) have been studied. We have demonstrated in previous experimental research in rats that a graft made using a vein (providing a guide for nerve regeneration) filled with fresh skeletal muscle (to prevent vein collapse and support axon regeneration) gave similar results to traditional nerve grafts. On this basis, we decided to use the muscle-vein-combined grafts in clinical cases. From 1993 to 1997, this technique was applied for bridging both sensory and mixed nerve defects (21 cases). We report good results in 85% of our cases with a minimum follow-up of 14 months. These results, obtained on nerve defects ranging from 0.5 to 6 cm in length, seem to be superior to those reported with other kinds of artificial or biological conduits.

Adolescent↗
PubMed2000

Nerve repair by means of vein filled with muscle grafts. II. Morphological analysis of regeneration.

The morphological features of regeneration in long-distance (3 cm) muscle-vein-combined grafts were experimentally investigated in the rat sciatic nerve by means of light and electron microscopy. In the early phases of regeneration (14 days after surgery), many regenerating nerve fibers were detected along the muscle-vein-combined graft. Six months after surgery, quantitative morphometrical analysis of myelinated nerve fibers showed that both the total number and density of myelinated nerve fibers were significantly greater in regenerated nerves than in control nerves. The contrary appeared true for the mean fiber size, with fiber size significantly smaller in regenerated nerves. Ultrastructural observations allowed the description of some peculiar aspects of the relationship between muscle fibers, nerve fibers, and Schwann cells in both early and late phases of regeneration.

Animals↗
PubMed2000

Tissue specificity in rat peripheral nerve regeneration through combined skeletal muscle and vein conduit grafts.

Diffusible factors from the distal stumps of transected peripheral nerves exert a neurotropic effect on regenerating nerves in vivo (specificity). This morphological study was designed to investigate the existence of tissue specificity in peripheral nerve fiber regeneration through a graft of vein filled with fresh skeletal muscle. This tubulization technique demonstrated experimental and clinical results similar to those obtained with traditional autologous nerve grafts. Specifically, we used Y-shaped grafts to assess the orientation pattern of regenerating axons in the distal stump tissue. Animal models were divided into four experimental groups. The proximal part of the Y-shaped conduit was sutured to a severed tibial nerve in all experiments. The two distal stumps were sutured to different targets: group A to two intact nerves (tibial and peroneal), group B to an intact nerve and an unvascularized tendon, group C to an intact nerve and a vascularized tendon, and group D to a nerve graft and an unvascularized tendon. Morphological evaluation by light and electron microscopy was conducted in the distal forks of the Y-shaped tube. Data showed that almost all regenerating nerve fibers spontaneously oriented towards the nerve tissue (attached or not to the peripheral innervation field), showing a good morphological pattern of regeneration in both the early and late phases of regeneration. When the distal choice was represented by a tendon (vascularized or not), very few nerve fibers were detected in the corresponding distal fork of the Y-shaped graft. These results show that, using the muscle-vein-combined grafting technique, regenerating axons are able to correctly grow and orientate within the basement membranes of the graft guided by the neurotropic lure of the distal nerve stump.

Animals↗
PubMed2000

Morphological analysis of peripheral nerve regenerated by means of vein grafts filled with fresh skeletal muscle.

Clinical data have shown that a vein segment filled with fresh skeletal muscle can be considered a good autologous grafting conduit for the repair of peripheral nerve lesions. In this study, the long-term morphological organization of rat sciatic nerve fibers regenerated along a muscle-vein-combined graft conduit is further analysed by light and electron microscopy. Regenerated nerve fibers were organized into fascicles of various sizes that were clearly delimited by perineurial-like shells made by long and thin cytoplasmic processes of perineurial-like bipolar cells and by densely packed collagen fibrils. Grafted skeletal muscle fibers were still detectable among nerve fiber fascicles. However, in spite of the persistence of skeletal muscle along the graft, regenerated nerve fibers showed a good morphological pattern of regeneration, providing further evidence that the muscle-vein-combined grafting technique represents an effective surgical alternative to the classical fresh nerve autograft for the repair of peripheral nerve defects.

Animals↗
PubMed2000

Verification of the two-dimensional disector, a method for the unbiased estimation of density and number of myelinated nerve fibers in peripheral nerves.

Quantification of the number of myelinated fibers in peripheral nerves is a common requirement in quantitative morphology. This parameter provides important information on the consequences of various physiological, pathological and experimental conditions on the nerve structure and is one of the main indicators of success of peripheral nerve repair. In this paper, the theoretical rationale for the application of stereological principles to obtain unbiased estimates of the density and total number of myelinated fibers in peripheral nerves is discussed and a simple stereological method is described. The method is applied together with a systematic random sampling scheme, that was optimized for the purposes of the present study, and with sampling scheme analysis by calculating the coefficient of error (CE). The stereological method, which consists of a two-dimensional variation of the classical disector procedure (two-dimensional disector), and the sampling scheme are verified by comparing estimates with the true density and total number of myelinated fibers in peripheral nerve trunks where true values have been accurately determined by extensive counting. The verification of the 2-D disector method, both of normal and regenerated nerves, showed that estimates of density and total number of myelinated nerve fibers are unbiased. The method also proved to be efficient (time-saving): Estimation of density and total number of myelinated fibers in a single nerve takes about 2-3 hours.

Animals↗
PubMed1999

Sutureless microvascular anastomoses by a biodegradable laser-activated solid protein solder.

A new sutureless technique to successfully anastomose the abdominal aorta of rats (1.3 mm in diameter) by using a fully biodegradable, laser-activated protein solder is presented. A total of 90 rats were divided into two groups randomly. In group one, the anastomoses were performed by using conventional microsuturing technique, whereas in group two, the anastomoses were performed by using a new laser welding technique. In addition, each of the two groups were divided into five subgroups and evaluated at different follow-up periods (10 minutes, 1 hour, 1 day, 1 week, and 6 weeks). At these intervals, the anastomoses were evaluated for patency and tensile strength. Three anastomoses in each subgroup were processed for light and electron microscopy. All anastomoses were found to be patent. The mean clamp time of the anastomoses performed with conventional suturing was 20.6 minutes compared with 7.2 minutes for the laser-activated welded anastomoses (p < 0.001). The strain measurements showed a stronger mechanical bond of the sutured anastomoses in the initial phase. However, at 6 weeks the tensile strength of the laser-welded anastomoses was higher compared with the conventional suture technique. Histologic evaluations revealed a near complete resorption of the solder after 6 weeks. The junction site of the vessel ends cannot be determined on the luminal side of the artery. In conclusion, a resorbable protein used as a solder, activated by a diode laser, can provide a reliable, safe, and rapid arterial anastomosis, which could be performed by any microsurgeon faster than conventional suturing after a short learning curve.

Anastomosis, Surgical↗
PubMedDate not supplied

A stereological study of long-term regeneration of rat severed sciatic nerve repaired by means of muscle-vein-combined grafts.

This study is a stereological analysis, by the 2-D dissector method, on the long-term regeneration of myelinated nerve fibers of the rat sciatic nerve repaired by muscle-vein-combined graft, a surgical technique that has been shown to be a valid tool for the repair of peripheral nerve defects with substance loss. Quantitative analysis showed that the total number and mean density of regenerated myelinated nerve fibers was significantly higher than in control nerves. The contrary was true for fiber mean size. The morpho-quantitative parameters of regenerated fibers from nerves repaired by the muscle-vein-combined graft were similar to those observed in rats where nerve defects were repaired by direct nerve suture thus confirming the validity of this surgical technique.

Animals↗