Processed porcine collagen tubulization versus conventional suturing in peripheral nerve reconstruction: an experimental study in rabbits.

In peripheral nerve reconstruction, various procedures are used. One of the procedures that received the most interest in the past decade is the tubulization technique for small nerve gaps. A disadvantage in the use of non-biodegradable tubes is that the material often has to be removed owing to its mechanical properties. Some investigators, in exploring the use of collagen tubes, being a natural biodegradable material, found either allogenicity or xenogenicity and immune responses that may inhibit nerve regeneration. Processed porcine collagen (PPC) is a new inert and biodegradable material that has a favorable effect on wound healing, as demonstrated by experiments on other tissues. The aim of our study was to compare the healing of nerve sutures with PPC tubes with conventional end-to-end sutures. In our experiments, we reconstructed the saphenous nerves of 27 rabbits. In series 1 (n = 12) and 2 (n = 12), PPC tubes were slid over an end-to-end nerve suture without or with a 10-mm nerve gap, respectively. In series 3 (n = 12), conventional suturing was performed in the collateral saphenous nerves of the animals of series 1. Epineurial suturing was performed. Three other non-operated saphenous nerves served as controls. The healing was studied after 3, 6, and 12 months in sections stained by monoclonal antibodies and by conventional histologic staining. Morphometric analysis of the regenerating axons was done by using confocal scanning laser microscopy (CLSM). Data analysis was carried out using a software program especially developed for this purpose. All results were evaluated statistically. Our results showed that during the healing period in the distal nerve stump, the number of axons of the PPC procedure with a 10-mm gap was significantly higher than that in the procedure without a gap. At 12 months, the mean number of axons of all procedures was significantly lower than in the non-operated nerve, and the mean axon diameter in all distal stumps did not differ significantly from that of the non-operated nerve. In the distal nerve stump, the ratio of total axon area to total fascicle area in the PPC procedure with a gap was significantly higher than that in the conventional suturing procedure. After 12 months, there was no significant difference between the percentages of axon outgrowth of the PPC procedure without a gap, the conventional suturing procedure, and the non-operated nerve (100%). The percentage of axon outgrowth in PPC with a gap was significantly higher than in the other procedures.

Silicone rubber tubulization in peripheral sensory nerve reconstruction: an experimental study in rabbits.

Silicone rubber (polydimethyl siloxane) tubes are used clinically in peripheral nerve reconstruction. A disadvantage of this procedure is that the material often has to be removed owing to its mechanical properties. The aim of our study was to investigate the healing of reconstructed sensory nerves tubulized by silicone rubber in an animal model. In our experiments, we reconstructed the saphenous nerves in 27 rabbits. In series 1 (n = 12), silicone rubber tubes were slid over a nerve suture without a gap. In series 2 (n = 12), silicone rubber tubes were slid over a 10-mm nerve gap. In series 3 (n = 12), conventional suturing was performed in the collateral saphenous nerves of the animals of the series 1. Epineurial suturing was performed. Three other collateral nonoperated saphenous nerves served as controls. The healing was studied after 3, 6, and 12 months. Morphometric analysis of the regenerating axons was performed by using our new method for quantification of nerve fibers in cross sections stained by immunohistochemistry and using confocal laser scanning microscopy. Data analysis was carried out using a software program especially developed for this purpose. Our results showed in the silicone procedures that at 12 months significantly fewer axons per fascicle area were present compared with conventional suturing. However, mean axon diameters in the distal nerve stump of the silicone procedures did not differ significantly compared with the conventional suturing procedure. The ratio of total axon area to total fascicle area in the distal nerve stumps of the silicone procedure without gap was significantly smaller compared with the conventionally sutured nerve. The percentage outgrowing axons from the proximal nerve stump into the distal one in the silicone rubber procedure without gap was 57%. This was significantly higher than in the silicone rubber procedure with 10-mm gap (48%). However, in conventional suturing, the percentage of outgrowing axons (99%) was significantly higher than in both tubulization procedures. It appeared that tubulization by silicone rubber of sutured nerves without gap did not enhance axon regeneration. Conventional suturing gave significantly better results. If a gap was present, the use of a silicone rubber tube was preferable to non-suturing.

Method for morphometric analysis of axons in experimental peripheral nerve reconstruction.

A new method for morphometric analysis of axons in experimental peripheral nerve reconstruction is presented. Twelve adult female rabbits were used. In nine animals the saphenous nerve was transected and stitched epineurially. Three animals functioned as control. After 3, 6, and 12 months, the nerves were harvested, fixed in Kryofix and embedded in Histowax. Transverse sections of 6 microm were cut, immunohistochemically stained for NF 90, and counterstained by Sirius Red. Quantification of nerve fibers in cross sections was performed by using a confocal laser scanning microscope (CLSM), and the images were stored digitally. Data analyzing was performed by the Optimas program (5.2). Calculations were done with Microsoft Excel. The total number of axons, the mean axon diameter and the percentage axon area/fascicle area were evaluated statistically. This method for morphologic analysis provides automatically complete registration of axons and so different methods of experimental nerve reconstruction can be compared in a fast and reliable way.