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Stefano Geuna

Publications and source records attributed to Stefano Geuna.

At least 19 recordsLinked to original sources
PubMed2007

Ulnar nerve repair by end-to-side neurorrhaphy on the median nerve with interposition of a vein: an experimental study.

Our study used a rat animal model to verify that Schwann cell migration, collateral axonal sprouting, and regeneration were not hindered by interposition of a 10-mm vein segment between the distal stump of the transected ulnar nerve and the donor median nerve in end-to-side reconstruction. Reconstructed nerves were withdrawn at 7, 14, 30, and 60 days after surgery, then analyzed by confocal laser microscopy after immunolabeling with anti-neurofilament-200 kD (an axonal marker) and anti-S100 (a glial marker) antibodies. Results are reported at 7, 14, 30, and 60 days after surgery. Our observations indicate that chemotactic factors that stimulate end-to-side nerve regeneration can exert their action at this distance. These findings suggest a possible clinical application for this surgical technique in cases when the severed nerve stump is far from the potential donor nerve.

Animals↗
PubMed2007

Electrical stimulation based on chronaxie reduces atrogin-1 and myoD gene expressions in denervated rat muscle.

Denervation induces muscle fiber atrophy and changes in the gene expression rates of skeletal muscle. Electrical stimulation (ES) is a procedure generally used to treat denervated muscles in humans. This study evaluated the effect of ES based on chronaxie and rheobase on the expression of the myoD and atrogin-1 genes in denervated tibialis anterior (TA) muscle of Wistar rats. Five groups were examined: (1) denervated (D); (2) D+ES; (3) sham denervation; (4) normal (N); and (5) N+ES. Twenty muscle contractions were stimulated every 48 h using surface electrodes. After 28 days, ES significantly decreased the expression of myoD and atrogin-1 in D+ES compared to the D group. However, ES did not prevent muscle-fiber atrophy after denervation. Thus, ES based on chronaxie values and applied to denervated muscles using surface electrodes, as normally used in human rehabilitation, was able to reduce the myoD and atrogin-1 gene expressions, which are related to muscular growth and atrophy, respectively. The results of this study provide new information for the treatment of denervated skeletal muscle using surface ES.

Animals↗
PubMed2006

A comparison analysis of hindlimb kinematics during overground and treadmill locomotion in rats.

The convenience of the motor-driven treadmill makes it an attractive instrument for investigating rat locomotion. However, no data are available to indicate whether hindlimb treadmill kinematic findings may be compared or generalized to overground locomotion. In this investigation, we compared overground and treadmill locomotion for differences in the two-dimensional angular kinematics and temporal and spatial measurements for the hindlimb. Ten female rats were evaluated at the same speed for natural overground and treadmill walking. The walking velocity, swing duration and stride length were statistically indistinguishable between the two testing conditions. Significant differences were found between overground and treadmill locomotion for step cycle duration and stance phase duration parameters. During the stance phase of walking, the angular movement of the hip, knee and ankle joints were significantly different in the two conditions, with greater flexion occurring on the overground. Despite this, the sagittal joint movements of the hindlimb were similar between the two walking conditions, with only three parameters being significantly different in the swing. Hip height and angle-angle cyclograms were also only found to display subtle differences. This study suggests that reliable kinematic measurements can be obtained from the treadmill gait analysis in rats.

Animals↗
PubMed2006

The ulnar test: a method for the quantitative functional assessment of posttraumatic ulnar nerve recovery in the rat.

The assessment of recovery of the neuromuscular function following nerve lesion and repair is one of the main goals of peripheral nerve researchers. The forelimb model has recently seen an increase in its employment for experimental nerve repair studies especially because of the availability of the grasping test for assessing the functional recovery of one of its major nerves, the median nerve. Nerve repair studies sometimes require the use of more than one nerve to simulate severe clinical situations and, in this case, the ulnar nerve is often used together with the median nerve. However, a test for assessing ulnar nerve functional recovery is yet not available. To fill this gap, we have developed and experimentally tested a method for the functional assessment of posttraumatic ulnar nerve recovery in the rat. Animal testing using this method is simple, quick and provides the animal with minimal distress. The method proved to be effective in detecting the date on which recovery starts after ulnar nerve impairment and in following its improvement, over time. The availability of this new test is expected to further increase the employment of forelimb experimental nerve models instead of the more disabling hindlimb models.

Animals↗
PubMed2006

End-to-side (terminolateral) nerve regeneration: a challenge for neuroscientists coming from an intriguing nerve repair concept.

The last 15 years have seen a growing interest regarding a technique for nerve repair named end-to-side (terminolateral) neurorrhaphy. This technique is based on the concept that nerve fiber regeneration along the distal stump of a transected nerve, the proximal stump of which was lost, can be obtained by just suturing the proximal end of its distal stump to the epinerium of a neighbor healthy and undamaged donor nerve. A large body of experimental studies have shown that end-to-side neurorrhaphy, in fact, is able to induce collateral sprouting from donor nerve's axons which is at the basis of the massive repopulation of the distal nerve stump. The regenerating nerve fibers eventually reinnervate the periphery of the severed nerve leading to a recovery of the lost function the degree of which varies depending on factors that still have to be elucidated. Surprisingly, this puzzling concept of nerve regeneration has attracted very little attention from basic neuroscientists so far and, thus, the present paper is intended to call for more biological research on it by overviewing the relevant literature and indicating the several unanswered questions that this concept asks to the neuroscience community.

Animals↗
PubMed2006

Morphological characterization of GFP stably transfected adult mesenchymal bone marrow stem cells.

Increasing attention is being given to the use of adult rather than embryonic stem cells, both for research and for the development of transplantation treatments for human disease. In particular, mesenchymal bone marrow stem cells have been studied extensively because of their ability to self-renew and to give rise to various differentiated cell types, and because of the relative ease with which they can be obtained and cultured. In addition, the possibility of labelling stem cells with green fluorescent protein before transplantation has opened new and promising perspectives for their use in basic research. Because no structural or ultrastructural description of adult mesenchymal stem cells is available in the literature, this paper describes their morphology as revealed by light, confocal and electron microscopy, focusing on cells that are particularly suitable for transplantation studies, i.e. those derived from rat bone marrow transfected with green fluorescent protein. The results provide a basis for experimental studies of the differentiation of these cells in normal and pathological tissues.

Animals↗
PubMed2005

Schwann cell behavior after nerve repair by means of tissue-engineered muscle-vein combined guides.

Schwann cells play a critical role in peripheral nerve regeneration. When a non-nervous conduit is used to bridge a nerve defect, the conduit is soon colonized by a number of Schwann cells that make a pathway for regrowing axons. By using electron microscopy, immunohistochemistry, and reverse transcriptase-polymerase chain reaction analysis, we have investigated the behavior of migratory glial cells along a particular type of autologous tissue-engineered conduit made of a vein filled with fresh skeletal muscle, using the rat sciatic nerve model. With this particular type of autograft, our data show that many Schwann cells soon take up a close relationship with grafted muscle fibers, and especially with their basal lamina, which appears to serve as a migration pathway for them. The early and massive colonization of the conduit is sustained by both Schwann cell migration and proliferation, as demonstrated by PCNA immunostaining. Later, as they meet regenerating axons, Schwann cells become closely associated with them and eventually lose their connections with grafted muscle fibers because of the formation of perineurial envelopes. Because previous studies showed that alpha(2a-2b) NRG1 is overexpressed at early stages along the muscle-vein combined tubes, we have also investigated mRNA expression of its two receptors, erbB2 and erbB3. Both messengers are overexpressed, although with different time courses. Overall, our results provide some morphological and biochemical bases for explaining the effectiveness of fresh muscle-vein combined nerve guides and throw an interesting light on the possible role of alpha(2a-2b) NRG1 through the erbB2/erbB3 heterodimer receptor for nerve regeneration inside non-nervous conduits.

Animals↗
PubMed2005

The revolution of counting "tops": two decades of the disector principle in morphological research.

Twenty years have passed since the publication of the seminal paper enunciating the disector principle by an author using the pseudonym D.C. Sterio. During this time, methods based on the revolutionary principle of counting "tops" have become progressively better known and have been included in several commercially available systems for quantitative morphology. Analysis of the number of published studies citing Sterio's paper on the ISI Web of Knowledge database showed that its scientific "impact factor" has almost continuously risen since its publication, indicating the growing knowledge about disector-based methods in the various scientific fields where morphological quantification is required. This report briefly reviews the first two decades of disector use, pointing to its advantages as well as to shortcomings that have recently been addressed in critical papers and have given rise to a lively debate on the role of counting tops in quantitative morphology today.

Animals↗
PubMed2005

Nerve repair by means of tubulization: literature review and personal clinical experience comparing biological and synthetic conduits for sensory nerve repair.

Nerve repair is usually accomplished by direct suture when the two stumps can be approximated without tension. In the presence of a nerve defect, the placement of an autologous nerve graft is the current gold standard for nerve restoration. However, over the last 20 years, an increasing number of research articles reported on the use of non-nervous tubes (tubulization) for repairing nerve defects. The clinical employment of tubes (both biological and synthetic) as an alternative to autogenous nerve grafts is mainly justified by the limited availability of donor tissue for nerve autografts and the related morbidity. In addition, tubulization was proposed as an alternative to direct nerve sutures in order to create optimal conditions for nerve regeneration over the short empty space intentionally left between two nerve stumps. This paper outlines recent important advances in this field. Different tubulization techniques proposed so far are described, focusing in particular on studies that reported on the employment of tubes with patients. Our personal clinical experience on tubulization repair of sensory nerve lesions (digital nerves), using both biological and synthetic tubes, is presented, and the clinical results are compared. In our case series, both types of tubes led to good clinical results. Finally, we speculate about the prospects in the clinical application of tubulization for peripheral nerve repair.

Bioprosthesis↗
PubMed2005

Phototherapy for enhancing peripheral nerve repair: a review of the literature.

Posttraumatic nerve repair continues to be a major challenge of restorative medicine. Although enormous progress has been made in surgical techniques over the past three decades, functional recovery after a severe lesion of a major nerve trunk is often incomplete and sometimes unsatisfactory. It is thus particularly important to investigate clinical protocols to enhance nerve regeneration after surgical nerve repair. The present article reviews literature on one possible rehabilitation approach for enhancing nerve recovery, namely phototherapy. The number of experimental studies that have reported on the promoting action of phototherapy on peripheral nerve regeneration, together with the few known side effects related to the use of this type of physical therapy, make it possible to suggest that the time for broader clinical trials has come.

Animals↗
PubMed2004

On sampling and sampling errors in histomorphometry of peripheral nerve fibers.

Histomorphometrical assessment of regenerated peripheral nerves is a very common goal of many studies in experimental microsurgery. In this paper, the main critical issues in nerve fiber sampling for quantitative morphological assessment are addressed. The equal opportunity rule, i.e., the basic paradigm of random sampling, is described, together with an explanation of how sampling errors, in the selection of histologic fields and of the nerve fibers inside them, can produce a bias in quantitative estimates. Finally, some practical suggestions on how to cope with the most common sampling errors are provided, in order to help researchers obtain reliable histomorphometrical data on peripheral nerve fibers.

Humans↗
PubMed2004

Use of muscle-vein-combined Y-chambers for repair of multiple nerve lesions: experimental results.

Y-shaped vein conduits enriched with fresh skeletal muscle fibers were used to bridge a concomitant ulnar and median nerve transection with substance loss in rats. The proximal limb of the Y-chamber was sutured to the proximal ulnar nerve, while the two distal limbs were sutured to ulnar and median distal nerve stumps. Eight months after surgery, median nerve functional recovery was evaluated by means of the grasping test, and nerve fiber regeneration in both repaired nerves was assessed by means of design-based histomorphometry. Results showed that nerve fibers regenerated along both severed nerve trunks, and in the median nerve led to a recovery corresponding to 58% of normal nerve function. Quantitative analysis showed no significant morphological differences between myelinated nerve fibers regenerated along the two distal nerves except for the number of fibers, which was higher in the median nerve. Notably, the total number of regenerated nerve fibers in the two distal nerves was 4-fold the normal fiber number in the ulnar nerve. Besides their interest in relation to the long-lasting debate about the topographic specificity of nerve regeneration, the results of this study show an effective way to repair, in the rat experimental model, two transected nerve trunks innervating agonistic muscles in the case that the proximal stump of only one nerve is preserved.

Animals↗
PubMed2004

Functional and morphological assessment of a standardized rat sciatic nerve crush injury with a non-serrated clamp.

Peripheral nerve researchers frequently use the rat sciatic nerve crush as a model for axonotmesis. Unfortunately, studies from various research groups report results from different crush techniques and by using a variety of evaluation tools, making comparisons between studies difficult. The purpose of this investigation was to determine the sequence of functional and morphologic changes after an acute sciatic nerve crush injury with a non-serrated clamp, giving a final standardized pressure of p = 9 MPa. Functional recovery was evaluated using the sciatic functional index (SFI), the extensor postural thrust (EPT) and the withdrawal reflex latency (WRL), before injury, and then at weekly intervals until week 8 postoperatively. The rats were also evaluated preoperatively and at weeks 2, 4, and 8 by ankle kinematics, toe out angle (TOA), and gait-stance duration. In addition, the motor nerve conduction velocity (MNCV) and the gastrocnemius-soleus weight parameters were measured just before euthanasia. Finally, structural, ultrastructural and histomorphometric analyses were carried out on regenerated nerve fibers. At 8 weeks after the crush injury, a full functional recovery was predicted by SFI, EPT, TOA, and gait-stance duration, while all the other parameters were still recovering their original values. On the other hand, only two of the histomorphometric parameters of regenerated nerve fibers, namely myelin thickness/axon diameter ratio and fiber/axon diameter ratio, returned to normal values while all other parameters were significantly different from normal values. The employment of traditional methods of functional evaluation in conjunction with the modern techniques of computerized analysis of gait and histomorphometric analysis should thus be recommended for an overall assessment of recovery in the rat sciatic nerve crush model.

Animals↗
PubMed2004

Bridging peripheral nerve defects with muscle-vein combined guides.

Various tubulization techniques can be used to bridge peripheral nerve lesions with substance loss. Among the different materials that have been used so far in alternative to traditional fresh nerve autografts, fresh muscle-vein combined conduits (made by a vein segment filled with fresh skeletal muscle) proved to be particularly effective. In this study, nerve repair of 10-mm long nerve defects by means of muscle-vein combined tubes was compared with repair by means of traditional nerve autografts in the rat sciatic nerve experimental model. Results did not reveal any significant difference between the two groups of regenerated nerves with respect to the total number, mean density and mean size of myelinated nerve fibers. In addition, we also report the results of an experimental study in the rabbit sciatic nerve model, which showed that fresh skeletal muscle enrichment of the vein segment made it possible to bridge 55-mm long nerve gaps. These results provide further evidence of the effectiveness of fresh muscle-vein combined grafts and support the view that this type of conduit can be used also for repairing long nerve gaps.

Animals↗
PubMed2004

Phototherapy promotes regeneration and functional recovery of injured peripheral nerve.

Numerous attempts have been made to enhance and/or accelerate the recovery of injured peripheral nerves. One of the methods studied is the use of phototherapy (low power laser or light irradiation) to enhance recovery of the injured peripheral nerve. A critical analysis of the literature on the employment of phototherapy for the enhancement of the regeneration process of the rat facial and sciatic nerve (after crush injury or transection followed by surgical reconstruction) is provided, together with the description of some of the most suitable basic biological mechanisms through which laser radiation exerts its action on peripheral nerve regeneration.

Animals↗
PubMed2003

Expression of alpha2a-2b neuregulin-1 is associated with early peripheral nerve repair along muscle-enriched tubes.

Using RT-PCR, we have investigated expression of isoforms beta1 (the axonal isoform) and alpha2a-2b (the mesenchymal isoform) of neuregulin-1, one of the most important known trophic factors for Schwann cells, in the rat sciatic nerve repaired by muscle-enriched non-nervous conduits (made by a vein filled with fresh skeletal muscle). Repaired nerves were harvested 2, 6 and 13 days post-operatively. Results showed that while muscle-vein combined grafts were enriched in mRNA coding for alpha2a-2b since the very early regeneration stages, isoform beta1 mRNA was not detectable inside the tubes at day 2 and 6 post-operatively while its expression at day 13 was very slight. These results suggest that Schwann cell survival and activity inside a fresh muscle-enriched non-nervous conduit graft (a key factor for successful nerve regeneration along the graft) may be supported by the mesenchymal isoform of neuregulin-1 during very early repair phases, i.e. when axons are still not present along the tube.

Animals↗
PubMed2003

On the use of the grasping test in the rat median nerve model: a re-appraisal of its efficacy for quantitative assessment of motor function recovery.

The quantitative assessment of motor function is an important requirement for studies on peripheral nerve injury and repair. So far, most studies on peripheral nerves have been performed on the sciatic nerve model using walking track analysis for assessing motor function. Alternatively, the employment of the median nerve model, which allows motor function evaluation by means of a simple behavioural test named grasping test (GT), have been more recently proposed. In this paper, the efficacy of the GT for the quantitative assessment of motor function recovery is re-appraised and a modified device for its carrying out is described. Finally, the rationale for the employment of the median nerve model as an alternative to the sciatic nerve model is critically discussed.

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