Hans-Arne Hansson

Nerve regeneration across an extended gap: A neurobiological view of nerve repair and the possible involvement of neuronotrophic factors

We have compared the anatomic and functional regeneration of a transected sciatic nerve following regrowth from its proximal stump through either preformed empty mesothelial chambers or autologous nerve grafts bridging a 10 mm gap. Within the mesothelial chambers an organized multifascicular nerve trunk forms between the proximal and distal stumps. After 3 months, distal segment cross sections from the mesothelial chamber and nerve graft groups did not differ with respect to axonal density or distribution of axonal diameters. Mean conduction velocities across the gaps were also similar, although the nerve graft group had a wider distribution of velocities. Little or no regeneration was evident when the gap between the nerve stumps was left empty. These results suggest that if the regrowing proximal stump is in an appropriate environment, it can form a well organized and oriented nerve trunk. In the mesothelial chambers, the regenerating nerve is surrounded by a loose cellular stroma and a small amount of interstitial fluid, which was found to contain trophic activity for cultured rodent sensory neurons. Such factors may also support nerve regeneration in vivo.

Superficial repair of severed flexor tendons in synovial environment: An experimental, ultrastructural study on cellular mechanisms

Severed and subsequently sutured rabbit flexor tendons were kept free and isolated in the synovial cavity of the knee joint. In one series the tendon specimens were surrounded by a dialyzing membrane to avoid cell seeding from the synovial fluid. At different intervals of time over a period of 3 weeks, the tendons were studied morphologically with special reference to scanning electron microscopy. Adhesions were not observed and, with synovial fluid as the nutrient medium, the tendons showed an intrinsic ability to repair in the superficial layers, also bridging the suture gap. Moreover, cell seeding, mainly of macrophages, from the synovial fluid could be demonstrated on the very surface of the tendon. When this cell seeding was prevented, the fibroplasia in the superficial layer of the tendon did decrease slightly, but tendon cell morphology was that of active fibroblasts. The results support the concept that flexor tendons may show intrinsic fibroplasia when nourished by synovial fluid, while macrophages, mainly, of extrinsic origin, contribute to restoration of the tendon surface.

Insulin-Like Growth Factor I Promotes Nerve Regeneration: An Experimental Study on Rat Sciatic Nerve

Insulin-like growth factor I (IGF-I; somatomedin C) has previously been demonstrated, with immunohistochemical methods, to accumulate locally at the site of trauma of an injured peripheral nerve. In the experiments reported here a Y-shaped silicone-chamber system was used to test if local infusion of IGF-I had supportive effects on nerve regeneration. The proximal end of a cut sciatic nerve was inserted into one channel of the Y-shaped chamber and the length and growth direction of the regenerating myelinated axons were evaluated after 1 month. When IGF-I (250 micrograms/ml 0.5 microliters/h) was infused into one channel by an osmotic pump, the length of the regenerating axons increased significantly compared to the control groups with no IGF-I added. In some instances the regenerating axons grew towards the osmotic pump. It is concluded that local infusion of IGF-I at appropriate concentration promotes regeneration of a peripheral nerve. It exerts a neuronotrophic but not a clear chemotactic effect.

Lamellar bodies in Purkinje nerve cells experimentally induced by electric field

Rabbits reared from mating to the age of almost two months outdoors in a substation in an electric (E)-field of 14 kV/m (undisturbed field, 50 Hz AC) only gained about half the weight of their controls, either protected by a Faradays cage or being kept outside measurable E-field. The Purkinje nerve cells of cerebellum showed important alterations of the endoplasmic reticulum with disintegration of Nissl bodies, disappearance of hypolemmal cisterns and formation of numerous lamellar bodies. Concomitantly, there was a reduction in number of microtubules and increase in neurofilaments. These structural neuronal changes were reflected by changes in the behaviour of the rabbits which become slow in movements.

Detection of β-endorphin in the cerebrospinal fluid after intrastriatal microinjection into the rat brain

We have investigated to what extent microinjected beta-endorphin could migrate from the rat brain parenchyma into the CSF compartment. Exogenous rat beta-endorphin (0.1 nmol) was microinjected into the left striatum 1 mm from the lateral ventricle in anesthetized male rats. CSF samples were collected at different time points up to 2 h post-injection from a catheter affixed to the atlanto-occipital membrane of the cisterna magna. Radioimmunoassay and mass spectrometry were performed on the CSF samples, and brain sections were immunostained for beta-endorphin and mu-opioid receptors. The beta-endorphin injected rats showed a marked increase in beta-endorphin immunoreactive (IR) material in the CSF, with a peak at 30-45 min post-injection, and this beta-endorphin-IR material existed mainly as the intact beta-endorphin peptide. The immunohistochemistry results revealed the appearance of distinct beta-endorphin-IR cell bodies in the globus pallidus and the bed nucleus of stria terminalis supracapsular part, regions distant from the injection site, at 2 h post-injection of exogenous beta-endorphin. The beta-endorphin-IR in several of the globus pallidus cell bodies colocalized with the mu-opioid receptor-IR at the cell surface. These findings show that upon delivery of synthetic beta-endorphin, there is a significant intracerebral spread of the injected peptide, reaching regions far from the site of injection via diffusion in the extracellular space and flow in the cerebrospinal fluid. This may be of relevance when interpreting studies based on intracerebral injections of peptides, and advances our knowledge regarding the migration of compounds within the brain.

Dorsal root ganglion nerve cells transiently express increased immunoreactivity of the calcium-binding protein S-100β after sciatic nerve transection

Transiently increased immunoreactivity of the calcium binding protein S-100beta was demonstrated in spinal ganglion nerve cells after sciatic nerve transection. Neuropeptide Y (NPY), normally not seen in these nerve cells, appeared concomitantly. The transiently elevated co-expression of S-100beta and NPY is proposed to reflect an increased demand of neurotrophic and neuroprotective compounds in reactive neurons, tentatively regulating calcium ions.

Restoration of Superficially Damaged Flexor Tendons in Synovial Environment

Flexor tendon from the synovial sheath region of rabbits was resected and placed as a free body in a synovial knee joint recess. Structural changes of the surface of the graft caused by the instruments used to handle the tendon during the surgery (jewellers microforceps), was analysed by electron microscopy at various time intervals up to 6 weeks. The superficial coat of amorphous material, i.e. ground substance, on the surface of the tendon was dishevelled by the instruments. The injured areas were crowded with monocytes and macrophages during the first days. An increasing number of fibroblasts and mesothelial-like cells covered the damaged area in less than two weeks. The superficial coat of ground substance was not completely restored until after 3 weeks or more. The results obtained are discussed in relation to functional aspects.