A new model of upper cervical spinal contusion inducing a persistent unilateral diaphragmatic deficit in the adult rat.

Research on spinal cord trauma requires models reflecting the contusion mechanisms encountered in clinical situation. The aim of this study was to develop in the adult rat a reproducible model of upper cervical spinal cord contusion inducing persistent unilateral diaphragm deficit. After dura and pia matter removal, weight drop and compression were targeted at the ventro-lateral funiculi which contain the bulbospinal descending respiratory pathways that command the phrenic motoneurons innervating the diaphragm. At 7 days post-injury, the left diaphragm activity recorded in contused rats (27.4 +/- 5.1% of the contralateral activity) was significantly lower than in the sham group (97.6 +/- 1.2%). This respiratory deficit still persisted 1 month later. Histology showed a reproducible left C2-lateralized lesion that involved both white and gray matter including the ventro-lateral funiculi. This C2 contusion model provides a basis for testing both regenerative and neuroprotective strategies aimed at improving functional respiratory recovery after spinal cord trauma.

[Transplantation of olfactory glial cells after spinal injury. I--From experimental data to repair strategy after central injury]. / Transplantation de cellules gliales olfactives après traumatisme médullaire. I--Des données expérimentales aux visées réparatrices après lésion centrale.

Ensheathing olfactory glial cells (OEC) can be considered, with stem cells, as the other most important cell type for developing therapeutic cellular transplantation strategies following lesion of the central nervous system (CNS) and particularly in the case of spinal cord injury. OECs are macroglial cells whose precursors are located in the olfactory mucosa. OEC ensheath the axons of the sensory olfactory neurons, from the peripheral mucosa to the central olfactory bulbs. These glial cells constitute one of the rare macroglial cells which, after removal in the adult mammal, can survive in culture and multiply. After post-traumatic transplantation in the CNS, these cells have induced several instances of functional recovery after injury of different neural systems. The "OEC transplantation effect" consists in modifying the central inhibitory environment to make it more propitious for axonal regrowth and cell survival (reduction of the glial scar; releasing of numerous survival and neurotrophic factors, and of surface, extracellular matrix and adhesion molecules). In addition to the fact that OEC can ensheath and/or myelinate central axons, migrate in the CNS and accompany the growing axons over a relatively long distance, they also can be obtained from olfactory mucosa. OEC thus constitute a preferential candidate for autologous transplantation for the purposes of repair.

[Transplantation of olfactory ensheathing cells after spinal injury. II--Experimental variability and clinical perspectives]. / Transplantation de cellules gliales olfactives après traumatisme médullaire. II--Variabilités expérimentales et perspectives cliniques.

Over recent years, a certain number of experimental investigations have studied the effect of the transplantation of olfactory ensheathing glial cells (OEC) after spinal traumatism in animal, the rat in particular. Some of these studies have reported improvements in motor (mainly locomotor, postural and respiratory) and sensory function. While these new data provide additional support for the interest of the strategy of EOC transplantation to minimise the incapacitating effects of spinal pathologies in clinical therapy, it nonetheless remains necessary to continue experiments on animal models in order to better understand and master certain important points: beneficial effects according to the nature and composition of the transplants; therapeutic impact according to the type of pathology and the nature of the traumatism; influence of the dose effect; migration of the transplanted OECs (distance, pathways); active principles of the transplants; beneficial effect on various functions, in particular at the level of the vesico-sphincteric area; long-term innocuousness; long-term posttraumatic efficacy. Although therapeutic trials are in progress in certain countries (Australia, China, Portugal), it would nonetheless appear essential that these somewhat obscure points should be better understood before any clinical application might be seriously envisaged, in order to respect the principles of precaution, maximum efficacy and observance of the prevailing ethical rules.

Phrenic rehabilitation and diaphragm recovery after cervical injury and transplantation of olfactory ensheathing cells.

Functional respiratory recovery was evaluated by recording diaphragm and phrenic nerve activity several months after cervical cord hemisection followed by olfactory ensheathing cell (OEC) transplantation. The intact side was taken as a control in each rat. Sham-transplanted rats did not recover respiratory activity from the ipsilateral lesioned side. By contrast, ipsilateral phrenic and diaphragmatic activities recovered in transplanted rats amounted to 80.7% and 73% of their controls, respectively. After contralateral acute C1 section eliminating any contralateral influence from crossed compensatory pathways, the ipsilateral phrenic activity remained at 57.5% of the control, indicating that the phrenic recovery originated from the ipsilateral side. Supralesional stimulation in these rats elicited sublesional ipsilateral postsynaptic phrenic responses showing that transplantation helped ipsilateral fibers to again transmit nervous messages to the phrenic target, leading to substantial functional recovery. The origin of mechanisms involved in respiratory recovery (regeneration, resurrection, sprouting, sparing, demasking of latent pathways) is discussed.

Functional reconnections established by central respiratory neurons regenerating axons into a nerve graft bridging the respiratory centers to the cervical spinal cord.

The present work investigated, in adult rats, the long-term functional properties and terminal reconnections of central respiratory neurons regenerating axons within a peripheral nerve autograft bridging two separated central structures. A nerve graft was first inserted into the left medulla oblongata, in which the respiratory centers are located. Three months later, a C3 left hemisection was performed, and the distal tip of the graft was implanted into the C4 left spinal cord at the level of the phrenic nucleus, a natural central inspiratory target. Six to eight months after medullary implantation, the animals (n = 12) were electrophysiologically investigated to test 1) the phrenic target reinnervation by analyzing the phrenic responses elicited by bridge electrical stimulation and 2) the bridge innervation by unitary recordings of the spontaneous activity of regenerated axons within the nerve bridge. In the control group (n = 6), the medullary site of implantation corresponded to the dorsolateral medulla, a region known to be an unsuitable site for inducing respiratory axonal regrowth after nerve grafting. Stimulation of the nerve bridge never elicited phrenic nerve response, and no respiratory units were found within the nerve bridge. In the experimental group (n = 6), the proximal tip of the nerve bridge was implanted within the ventrolateral medulla at the level of the respiratory centers. Electrical stimulation of the nerve bridge induced phrenic nerve responses that reflected a postsynaptic activation of the phrenic target. Subsequent unitary recordings from teased fibers within the bridge revealed the presence of regenerated inspiratory fibers exhibiting discharge patterns typical of medullary inspiratory neurons, which normally make synaptic contacts with the inspiratory phrenic target. These results indicate that, when provided with an appropriate denervated target, central respiratory neurons with regenerated axons along a nerve bridge can remain functional for a long period and can make precise and specific functional reconnections with central homotypic target neurons.