Transplantation of cultured olfactory mucosal cells rescues optic nerve axons in a rat glaucoma model.

PURPOSE: To determine whether transplantation of olfactory mucosal cells (OMCs) is able to rescue the loss of optic nerve axons after the intraocular pressure (IOP) is elevated in rats. METHODS: The IOP was raised by injection of magnetic microspheres into the anterior chamber of the eye. OMCs cultured from the adult olfactory mucosa were transplanted into the region of the optic disc. RESULTS: We demonstrated that although the raised IOP returned to its normal level at six weeks, there was an irreversible 58% loss of optic nerve axons in the control group. However, the loss of the axons was reduced to 23% in the group with the transplanted OMCs. The Pattern Electroretinograms (pERG) showed that the decrement of the voltage amplitudes in association with the raised IOP was significantly alleviated in the group with transplantation of OMC. CONCLUSIONS: Transplantation of OMCs is able to rescue loss of optic nerve axons induced by raised IOP in the rats. The pERG recording suggested that the functional activities of the axons are also protected. TRANSLATIONAL RELEVANCE: The results demonstrated the ability of the transplanted OMCs to protect against the loss of the optic nerve axons and the loss of function caused by raised IOPs. The findings provide a basis for future human clinical trials by autografting OMCs from autologous nasal epithelial biopsies to treat or delay glaucoma diseases.

Partial Recovery of Proprioception in Rats with Dorsal Root Injury after Human Olfactory Bulb Cell Transplantation.

Transplanted human olfactory ensheathing cells (hOECs) were mixed with collagen into a unilateral transection of four dorsal roots (C6-T1) in a rat model. By mixing with collagen, the limited numbers of hOEC were maximized from an olfactory bulb biopsy and optimize cavity filling. Cyclosporine was administered daily to prevent immune rejection. Forelimb proprioception was assessed weekly in a vertical climb task. Half of the rats receiving hOEC transplants showed some functional improvement ("responders") over six weeks of the study while the other half did not ("nonresponders") and performed similarly to "injured only" rats. Transplanted cells were seen at both one week and six weeks after the surgical procedure; many were concentrated within the lesion cavity, but others were found with elongated processes in the overlying connective tissue. There were some fibers in the injury area associated with transplanted cells that were immunostained for neurofilament and TUJ1. Responder and nonresponder rats were compared with regard to microglial activation within the deep dorsal horn of cervical levels C7, C8 and also axon loss within the cuneate fasciculus at cervical level C3. Little difference was seen in microglial activation or axonal loss that could account for the improved proprioception in the responders group. This preliminary study is the first to transplant human olfactory bulb cells into a rat model of dorsal root injury; by refining each component part of the procedure, the repair potential of OECs can be maximized in a clinical setting.

Transplantation of Cultured Olfactory Bulb Cells Prevents Abnormal Sensory Responses During Recovery From Dorsal Root Avulsion in the Rat.

The central branches of the C7 and C8 dorsal roots were avulsed close to their entry point into the spinal cord in adult rats. The forepaw responses to heat and cold stimuli were tested at 1, 2, and 3 weeks after injury. Over this period, the paws were sensitive to both stimuli at 1-2 weeks and returned toward normal at 3 weeks. Immunohistology showed no evidence of axonal regeneration into the spinal cord in a control group of rats with avulsion only, implying that adjacent dorsal roots and their corresponding dermatomes were involved in the recovery. In a further group of rats, a mixture of bulbar olfactory ensheathing cells and olfactory nerve fibroblasts were transplanted into the gap between the avulsed roots and the spinal cord at the time of avulsion. These rats showed no evidence of either loss of sensation or exaggerated responses to stimuli at any of the time points from 1 to 3 weeks. Immunohistology showed that the transplanted cells formed a complete bridge, and the central branches of the dorsal root fibers had regenerated into the dorsal horn of the spinal cord. These regenerating axons, including Tuj1 and CGRP immunoreactive fibers, were ensheathed by the olfactory ensheathing cells. This confirms our previous demonstration of central regeneration by these transplants and suggests that such transplants may provide a useful means to prevent the development of abnormal sensations such as allodynia after spinal root lesions.

Astrocytic role in synapse formation after injury.

In 1969 a paper entitled Neuronal plasticity in the septal nuclei of the adult rat proposed that new synapses are formed in the adult brain after injury (Raisman, 1969). The quantitative electron microscopic study of the timed responses to selective partial denervation of the neuropil of the adult rat septal nuclei after distant transection of the hippocampal efferent axons in the fimbria showed that the new synapses arise by sprouting of surviving adjacent synapses which selectively take over the previously denervated sites and thus restore the number of synapses to normal. This article presents the evidence for the role of perisynaptic astrocytic processes in the removal and formation of synapses and considers its significance as one of the three major divisions of the astrocytic surface in terms of the axonal responses to injury and regeneration. This article is part of a Special Issue entitled SI:50th Anniversary Issue.

Functional Repair of Rat Corticospinal Tract Lesions Does Not Require Permanent Survival of an Immunoincompatible Transplant.

Cell transplantation is one of the most promising strategies for repair of human spinal cord injuries. Animal studies from a number of laboratories have shown that transplantation of olfactory ensheathing cells cultured from biopsies of the olfactory bulb mediate axonal regeneration and remyelination and restore lost functions in spinal cord injuries. For translation from small laboratory experimental injuries to the large spinal cord injuries encountered in human patients the numbers of cells that can be obtained from a patient's own olfactory bulb becomes a serious limiting factor. Furthermore, removal of an olfactory bulb requires invasive surgery and risks unilateral anosmia. We here report that xenografted mouse bulbar olfactory ensheathing cells immunoprotected by daily cyclosporine restore directed forepaw reaching function in rats with chronic C1/2 unilateral corticospinal tract lesions. Once function had been established for 10 days, cyclosporine was withdrawn. Thirteen out of 13 rats continued to increase directed forepaw reaching. Immunohistochemistry shows that in all cases neurofilament-positive axons were present in the lesion, but that the grafted cells had been totally rejected. This implies that once grafted cells have acted as bridges for axon regeneration across the lesion site their continued presence is no longer necessary for maintaining the restored function. This raises the possibility that in the future a protocol of temporary immunoprotection might allow for the use of the larger available numbers of immunoincompatible allografted cells or cell lines, which would avoid the need for removing a patient's olfactory bulb.

60 YEARS OF NEUROENDOCRINOLOGY: MEMOIR: Geoffrey Harris and my brush with his unit.

Geoffrey Harris is chiefly known for his demonstration of the control of the pituitary gland by the portal vessels coming from the hypothalamus. This does not do justice to his extraordinary contribution to biology. Harris' life's work was central in demonstrating the brain/body interactions by which animals and humans adapt to their environment, and above all the control of that most crucial and proximate of all evolutionary events - reproduction. In this brief review, I have tried to put Geoffrey Harris' work in the context of the scientific thinking at the time when he began his work, and above all, the contribution of his mentor, FHA Marshall, on whose towering shoulders Harris rose. But this is mainly my personal story, in which I have tried to show the debt that my work owed to Harris and especially to my dear friend, the late Keith Brown-Grant in Harris' team. I myself was never an endocrinologist, but over a short period in the early 1970s, under the influence of such inspirational mentors, and using purely anatomical methods, I was able to demonstrate sexual dimorphism and hormone-dependent sexual differentiation in the connections of the preoptic area, regeneration of the median eminence, the ultrastructure of apoptosis, the requirement for the suprachiasmatic nuclei in reproductive rhythms, the existence of non-rod or cone photoreceptors in the albino rat retina and, later, the expression of vasopressin by solitary (one in 600) magnocellular neurons in the polydipsic di/di Brattleboro mutant rat; this phenomenon was subsequently shown to be due to a+1 reading frameshift. I end this brief overview by mentioning some of the abiding and fascinating mysteries of the endocrine memory of the brain that arise from Harris' work on the control of the endocrines, and by pointing out how the current interest in chronobiology emphasises what a Cinderella the endocrine mechanisms have become in current brain imaging studies.

An energy theory of glaucoma.

A radial array of fortified astrocytes (FASTs) is the load bearing structure of the rat optic nerve head (ONH). At the retinal end the ONH is suspended on a fluid filled extracellular space occupied by modified pigment cells which generate a glomerular-like formation of villi. We propose that regulation of fluid in and out of this space may contribute to buffering the normal fluctuations of intraocular pressure. The energy requirement for the fluid transfer process is provided by the dense vascularity of the ONH and is reflected in the giant mitochondria of the FASTs. We propose that glaucoma occurs when a maintained rise in pressure overwhelms the capacity of this regulatory system. Under these circumstances the FAST array becomes detached from its anchorage in the surrounding ONH sheath. Progressively driven backwards by the pressure, the FASTs degenerate. We propose that the degeneration of the FASTs is associated with ischemic damage caused by the backward stretching of their blood supply. Retraction of the FAST processes deprives the retinal ganglion cell axons of their energy support, resulting in axotomy. We consider that our previously observed rescue of axons and FASTs by transplantation of olfactory ensheathing cells is due to replacement of this lost energy source.

Obtaining the olfactory bulb as a source of olfactory ensheathing cells with the use of minimally invasive neuroendoscopy-assisted supraorbital keyhole approach--cadaveric feasibility study.

BACKGROUND: Obtaining the human olfactory bulb (OB) for treatment of spinal cord injuries with olfactory ensheathing cells (OECs) requires the elaboration of a surgical approach that could meet the criteria of safety and minimal invasiveness. The aim of the study was to evaluate the suitability of the keyhole supraorbital craniotomy (SOC) with an eyebrow incision for obtaining OB for therapeutic purposes. METHODS: Seventeen SOCs were performed on nine fresh adult cadavers. The procedure of obtaining OB was conducted by neuroendoscope-assisted microsurgical dissection. Technical features related to the procedure were measured and adverse events were noted. The virtual three-dimensional planning was applied in six cases to verify the authorial A-C scale published previously. RESULTS: The intact OB was obtained in 10 (59%) cases and a minor injury was discovered in another 5 (29%) cases. In 2 (12%) specimens OB was severely damaged which was correlated with the minor neural tissue injury (Fi(2) = 0.44). While no case of an evident vascular injury was noted, there were 3 (18%) incidents of unintended frontal sinus opening positively correlated with the craniotomy width (Fi(2) = 0.44). The unfavorable three-dimensional (3D) configuration of the olfactory groove area was revealed in 66% of cases and highly correlated with OB injury (Fi(2) = 1.0) but not damage. CONCLUSIONS: The SOC via an eyebrow incision may be safely and effectively applied to obtain the OB as a source of OECs in roughly 90% of cases. Virtual 3D planning is useful in preoperative qualification of potential donors.

Functional regeneration of supraspinal connections in a patient with transected spinal cord following transplantation of bulbar olfactory ensheathing cells with peripheral nerve bridging.

Treatment of patients sustaining a complete spinal cord injury remains an unsolved clinical problem because of the lack of spontaneous regeneration of injured central axons. A 38-year-old man sustained traumatic transection of the thoracic spinal cord at upper vertebral level Th9. At 21 months after injury, the patient presented symptoms of a clinically complete spinal cord injury (American Spinal Injury Association class A-ASIA A). One of the patient's olfactory bulbs was removed and used to derive a culture containing olfactory ensheathing cells and olfactory nerve fibroblasts. Following resection of the glial scar, the cultured cells were transplanted into the spinal cord stumps above and below the injury and the 8-mm gap bridged by four strips of autologous sural nerve. The patient underwent an intense pre- and postoperative neurorehabilitation program. No adverse effects were seen at 19 months postoperatively, and unexpectedly, the removal of the olfactory bulb did not lead to persistent unilateral anosmia. The patient improved from ASIA A to ASIA C. There was improved trunk stability, partial recovery of the voluntary movements of the lower extremities, and an increase of the muscle mass in the left thigh, as well as partial recovery of superficial and deep sensation. There was also some indication of improved visceral sensation and improved vascular autoregulation in the left lower limb. The pattern of recovery suggests functional regeneration of both efferent and afferent long-distance fibers. Imaging confirmed that the grafts had bridged the left side of the spinal cord, where the majority of the nerve grafts were implanted, and neurophysiological examinations confirmed the restitution of the integrity of the corticospinal tracts and the voluntary character of recorded muscle contractions. To our knowledge, this is the first clinical indication of the beneficial effects of transplanted autologous bulbar cells.

Comparison of olfactory bulbar and mucosal cultures in a rat rhizotomy model.

In an ongoing clinical trial, a spinal injured patient who received a transplant of autologous cells cultured from the olfactory bulb is showing greater functional benefit than three previous patients with transplants of mucosal origin. Previous laboratory studies of transplantation into rat spinal cord injuries show that the superior reparative benefits of bulbar over mucosal cultures are associated with regeneration of severed corticospinal tract fibers over a bridge of olfactory ensheathing cells (OECs) formed across the injury site. In a rat rhizotomy paradigm, we reported that transplantation of bulbar cell cultures also enables severed axons of the C6-T1 dorsal roots to regenerate across a bridge of OECs into the spinal cord and restore electrophysiological transmission and forepaw grasping during a climbing test. We now report a repeat of the same rhizotomy procedure in 25 rats receiving cells cultured from olfactory mucosal biopsies. In no case did the transplanted cells form a bridging pathway. No axons crossed from the severed roots to the spinal cord, and there was no restoration of forepaw grasping. This suggests that the superior clinical benefit in the patient receiving bulbar cell transplants is due to regeneration of severed fibers across the injury site, and this correlates with imaging and the pattern of functional recovery. Using present culture protocols, the yield of OECs from bulbar biopsies is around 50%, but that from mucosal biopsies is less than 5%. Improving the yield of OECs from mucosal biopsies might avoid the necessity for the intracranial approach to obtain bulbar cells.

The supraorbital keyhole approach via an eyebrow incision applied to obtain the olfactory bulb as a source of olfactory ensheathing cells--radiological feasibility study.

BACKGROUND. The purpose of the study was to test the suitability of supraorbital keyhole craniotomy with an eyebrow incision for obtaining olfactory bulb for therapeutic purposes. METHODS. Fifty three high-resolution axial head computed tomography images of patients with a mean age of 55 ± 15 years were used. The exclusion criterion was a pathology of the anterior skull base. The virtual keyhole supraorbital craniotomy was placed on each side of a three-dimensional skull model with respect to the anatomical landmarks. Trajectories of neurosurgical instrument transitions to the anterior and posterior aspects of olfactory grooves (OGs) were subsequently designed with the use of a neuronavigation planning station and measured with correction allowing the avoidance of collisions with skull base structures. Three types of anatomical configuration were divided, reflecting the extent of the correction needed to reach the bottom of OG. RESULTS. Simulation of the surgical access and consequent metrological analysis was performed on 97 skull sides - 9 (8.5%) sides were excluded due to the large frontal sinus. The mean length of the craniotomy basis was 30.71 mm, lengths of the anterior and posterior trajectories were 53.25 and 58.24 mm, respectively (p < 0.0001). In 37% of cases the value of the corrections exceeded the depth of OG. CONCLUSIONS. The supraorbital keyhole approach via an eyebrow incision may be applied to obtain the olfactory bulb as a source of olfactory ensheathing cells in over 60% of cases. Further verification and evaluation of the surgical handiness based on cadaver specimens is justifiable.

[Studies on glial isomerization of lamina cribrosa in rat].

OBJECTIVE: To explore the mechanism of optic nerve damage in glaucoma by study on structure of glial lamina cribrosa(LC) in rats. METHODS: Experimental study. Albino Swiss(AS) rats were divided into 3 groups. Bilateral eyes of 10 normal rats were employed to be group I (right eye ) and group II (left eye) . Group III was from the left eyes of 13 rats underwent artificially intraocular hypertension in the right eyes. All rats were perfused and fixed with electronic microscopy fixative (2% paraformaldehyde +2% glutaraldehyde). Trimmed optic nerves were embedded with resin. Serial 1.5 µm thick 'semithin' sections were cut, either (2 eyes from group III) longitudinally, through the optic nerve head (ONH) from the retinal end to the commencement of the optic nerve, or (31 eyes) transversely (cross-sections). Ultrathin sections were cut in the middle of glial LC. The morphological observation of glial LC was obtained by light microscopy and transmission electron microscopy. Bonferroni correction was used to counteract the multiple comparison of each group. RESULTS: Fortified astrocytes formed the main supportive structure of glial LC in all rats, including group I, group II and group III. Astrocytes were ranked as a fan-like radial array, firmly attached ventrally to the sheath of the LC by thick basal processes, but dividing dorsally into progressively more slender processes with only delicate attachments to the sheath. These fortified astrocytes form ventral stout basal end feet, radial array, axon free-'preterminal' layer before terminating in a complex layer of fine interdigitating delicate branches at the dorsal. LC astrocytes were highly and uniformly electron dense throughout all the cell processes. An equally striking feature of the astrocytic processes was their massive cytoskeletal 'strengthening' of longitudinal massed filaments and tubules. Especially, massive filaments accumulated as cytoskeletal cores to form 'scaffold' of fortified astrocytes. There was vulnerable area in the dorsal of glial LC. This vulnerable area was isomerisation in bilateral eyes and different rats. There was different space in the vulnerable area. These space could be divided into 3 grades, (-), (+) and (++) . The number of (-), (+) and (++)were 1, 6, 3 eyes in group I, 1, 5, 4 eyes in group II, 1, 7, 3 eyes in group III. The Kruskal-Wallis test was used for statistical evaluations. There was no statistical differences of the ratio of (-), (+) and (++) in group I, group II and group III(χ(2) = 3.35, P = 0.187>0.05;group I vs group II, Z = -1.048, P = 0.294;group I vs group III Z = -1.691, P = 0.091;group II vs group III,Z = -1.343, P = 0.179). The ratio of space (-)was significantly less than space (+) and space (++) in group I, group II and group III(χ(2) = 23.88, P < 0.05; (-) vs (+) , Z = -2.821, P = 0.005; (-) vs (++) , Z = -2.726, P = 0.006). The ratio of space (+)was much more than space (++) in group I, group II and group III(Z = -4.410, P < 0.05). CONCLUSION: Glial isomerisation in LC may play a key role in glaucomatous optic nerve damage.

Transplantation of autologous olfactory ensheathing cells in complete human spinal cord injury.

Numerous studies in animals have shown the unique property of olfactory ensheathing cells to stimulate regeneration of lesioned axons in the spinal cord. In a Phase I clinical trial, we assessed the safety and feasibility of transplantation of autologous mucosal olfactory ensheathing cells and olfactory nerve fibroblasts in patients with complete spinal cord injury. Six patients with chronic thoracic paraplegia (American Spinal Injury Association class A-ASIA A) were enrolled for the study. Three patients were operated, and three served as a control group. The trial protocol consisted of pre- and postoperative neurorehabilitation, olfactory mucosal biopsy, culture of olfactory ensheathing cells, and intraspinal cell grafting. Patient's clinical state was evaluated by clinical, neurophysiological, and radiological tests. There were no adverse findings related to olfactory mucosa biopsy or transplantation of olfactory ensheathing cells at 1 year after surgery. There was no evidence of neurological deterioration, neuropathic pain, neuroinfection, or tumorigenesis. In one cell-grafted patient, an asymptomatic syringomyelia was observed. Neurological improvement was observed only in transplant recipients. The first two operated patients improved from ASIA A to ASIA C and ASIA B. Diffusion tensor imaging showed restitution of continuity of some white matter tracts throughout the focus of spinal cord injury in these patients. The third operated patient, although remaining ASIA A, showed improved motor and sensory function of the first spinal cords segments below the level of injury. Neurophysiological examinations showed improvement in spinal cord transmission and activity of lower extremity muscles in surgically treated patients but not in patients receiving only neurorehabilitation. Observations at 1 year indicate that the obtaining, culture, and intraspinal transplantation of autologous olfactory ensheathing cells were safe and feasible. The significance of the neurological improvement in the transplant recipients and the extent to which the cell transplants contributed to it will require larger numbers of patients.

Repair involves all three surfaces of the glial cell.

We propose that severed adult CNS axons are intrinsically capable of regeneration and reestablishing lost functions and that the key to repair lies in reconfiguring the scarring response of the astrocytic network. Astrocytes are multifunctional cells with three distinct surfaces: a glia to glial surface, providing the junctions needed to incorporate the astrocytes into the network; a glia to mesodermal surface, at which astrocytes collaborate with the meningeal fibroblasts to maintain the protective covering of the CNS; and a glia to neuronal surface, which provides the routes along which axons travel. After injury, the astrocytes collaborate with the meningeal fibroblasts to form a scar, which provides the necessary defensive sealing of the opened surface of the CNS, but which also has the detrimental effect of closing off the pathways along which axons could regenerate. Incorporation of glial cells transplanted from the olfactory system into a CNS injury causes a re-arrangement of the scarred astrocyte/fibroblast complex so as to produce the alignment of the glia to neuronal surfaces needed to provide a pathway for the regeneration of severed axons. Olfactory ensheathing cells certainly have a direct stimulatory effect on axons, but without concomitant reorganization of the glial scar, this could not in itself lead to regeneration of severed axons to their targets.

Repair of central nervous system lesions by transplantation of olfactory ensheathing cells.

Clinical conditions affecting the central nervous system (CNS) fall into two main categories - degenerative conditions in which nerve cells are lost (Alzheimer's, Parkinson's, Huntington's disease, etc.), and traumatic insults which sever nerve fibers but leave their cell bodies and initial parts of the severed axons intact (spinal cord injury, cerebrovascular accidents, or tumors affecting fiber tracts). After injuries of this second type, the survival of the nerve cell bodies and the local sprouting at the severed ends of the proximal stumps of the axons raise the tantalizing possibility of one day learning how to induce these severed fibers to regenerate to their original targets and restore lost functions. This chapter gives an overview of current research into the strategy of transplantation of olfactory ensheathing cells into axotomizing injuries.

Transplanted olfactory ensheathing cells reduce retinal degeneration in Royal College of Surgeons rats.

PURPOSE OF THE STUDY: Retinitis pigmentosa (RP) is a group of genetic disorders and a slow loss of vision that is caused by a cascade of retinal degenerative events. We examined whether these retinal degenerative events were reduced after cultured mixtures of adult olfactory ensheathing cells (OECs) and olfactory nerve fibroblasts (ONFs) were transplanted into the subretinal space of 1-month-old RCS rat, a classic model of RP. MATERIALS AND METHODS: The changes in retinal photoreceptors and Müller cells of RCS rats after cell transplantation were observed by the expression of recoverin and glial fibrillary acidic protein (GFAP), counting peanut agglutinin (PNA)-positive cone outer segments and calculating the relative apoptotic area. The retinal function was also evaluated by Flash electroretinography (ERG). To further investigate the mechanisms, by which OECs/ONFs play important roles in the transplanted retinas, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and basic fibroblast growth factor (bFGF) secretion of the cultured cells were analyzed by ELISA. The ability of OECs/ONFs to ingest porcine retinal outer segments and the amount of phagocytosis were compared with retinal pigment epithelium (RPE) cells. RESULTS: Our research showed that the transplantation of OECs/ONFs mixtures restored recoverin expression, protected retinal outer segments, increased PNA-positive cone outer segments, reduced caspase-positive apoptotic figures, downregulated GFAP, and maintained the b-wave of the ERG. Cultured OECs/ONFs expressed and secreted NGF, BDNF, and bFGF which made contributions to assist survival of the photoreceptors. An in vitro phagocytosis assay showed that OECs, but not ONFs, phagocytosed porcine retinal outer segments, and the phagocytic ability of OECs was even superior to that of RPE cells. CONCLUSIONS: These findings demonstrate that transplantation of OECs/ONFs cleaned up the accumulated debris in subretinal space, and provided an intrinsic continuous supply of neurotrophic factors. It suggested that transplantation of OECs/ONFs might be a possible future route for protection of the retina and reducing retinal degeneration in RP.

Directionality and bipolarity of olfactory ensheathing cells on electrospun nanofibers.

AIM: As a preliminary to the construction of olfactory ensheathing cells (OECs) bearing scaffold for bridging larger lesions in the spinal cord, we have investigated the response of purified cultured OECs to nanoscale fibers of varying diameter using US FDA-approved, biodegradable poly(lactic-co-glycolic-acid). MATERIALS & METHODS: Conventional electrospinning produced fibers of approximately 700 nm diameter (nano-700) while nanocomposite electrospinning with quantum dots produced significantly more uniform fibers of a reduced diameter to approximately 237 nm (nano-250). OECs from adult rat were FACS purified, cultured at low density on either a flat surface or a meshwork of randomly orientated nano-700 and nano-250 fibers, and assessed using cytomorphometric analysis of immunofluorescent confocal images and by scanning electron microscopy. RESULTS & CONCLUSION: Compared with a flat surface, culture on a nano-700 mesh increases cell attachment. Cells change from rounded to stellate forms in random orientation. Further size reduction to the nano-250 favors bipolarity in cells with unidirectional orientation as observed in the case when transplanted OECs were used to bridge areas of damage in rat spinal cords.

Role of the lesion scar in the response to damage and repair of the central nervous system.

Traumatic damage to the central nervous system (CNS) destroys the blood-brain barrier (BBB) and provokes the invasion of hematogenous cells into the neural tissue. Invading leukocytes, macrophages and lymphocytes secrete various cytokines that induce an inflammatory reaction in the injured CNS and result in local neural degeneration, formation of a cystic cavity and activation of glial cells around the lesion site. As a consequence of these processes, two types of scarring tissue are formed in the lesion site. One is a glial scar that consists in reactive astrocytes, reactive microglia and glial precursor cells. The other is a fibrotic scar formed by fibroblasts, which have invaded the lesion site from adjacent meningeal and perivascular cells. At the interface, the reactive astrocytes and the fibroblasts interact to form an organized tissue, the glia limitans. The astrocytic reaction has a protective role by reconstituting the BBB, preventing neuronal degeneration and limiting the spread of damage. While much attention has been paid to the inhibitory effects of the astrocytic component of the scars on axon regeneration, this review will cover a number of recent studies in which manipulations of the fibroblastic component of the scar by reagents, such as blockers of collagen synthesis have been found to be beneficial for axon regeneration. To what extent these changes in the fibroblasts act via subsequent downstream actions on the astrocytes remains for future investigation.

Structural basis of glaucoma: the fortified astrocytes of the optic nerve head are the target of raised intraocular pressure.

Increased intraocular pressure (IOP) damages the retinal ganglion cell axons as they pass through the optic nerve head (ONH). The massive connective tissue structure of the human lamina cribrosa is generally assumed to be the pressure transducer responsible for the damage. The rat, however, with no lamina cribrosa, suffers the same glaucomatous response to raised IOP. Here, we show that the astrocytes of the rat ONH are "fortified" by extraordinarily dense cytoskeletal filaments that would make them ideal transducers of distorting mechanical forces. The ONH astrocytes are arranged as a fan-like radial array, firmly attached ventrally to the sheath of the ONH by thick basal processes, but dividing dorsally into progressively more slender processes with only delicate attachments to the sheath. At 1 week after raising the IOP by an injection of magnetic microspheres into the anterior eye chamber, the fine dorsal processes of the ONH astrocytes are torn away from the surrounding sheath. There is no indication of distortion or compression of the axons. Subsequently, despite return of the IOP toward normal levels, the damage to the ONH progresses ventrally through the astrocytic cell bodies, resulting in complete loss of the fortified astrocytes and of the majority of the axons by around 4 weeks. We propose that the dorsal attachments of the astrocytes are the site of initial damage in glaucoma, and that the damage to the axons is not mechanical, but is a consequence oflocalized loss of metabolic support from the astrocytes (Tsacopoulos and Magistretti (1996) J Neurosci 16:877-885).

Olfactory Ensheathing Cells Rescue Optic Nerve Fibers in a Rat Glaucoma Model.

PURPOSE: To determine if transplantation of olfactory ensheathing cells (OECs) can reduce loss of optic nerve axons after raised intraocular pressure (IOP) in the rat. METHODS: OECs cultured from the adult olfactory mucosa were transplanted into the region of the optic disc. The IOP was raised by injection of magnetic microspheres into the anterior chamber. RESULTS: At 4 weeks after raising the IOP, the transplanted OECs had migrated into the dorsal area of the optic nerve head (ONH) where they surrounded the optic nerve fibers with a non-myelinated ensheathment. The mean amount of damage to the ONH astrocytic area in rats was 51.0% compared with 85.8% in those without OEC transplants (P < 0.02) and the mean loss of axons in the optic nerve was 51.0% compared with 80.3% in the absence of OECs (P < 0.01). CONCLUSIONS: OECs transplanted into the region of the ONH of the rat can reduce the loss of axons and the damage to ONH astrocytes caused by raised IOP. TRANSLATIONAL RELEVANCE: Confirmation of these preliminary experimental data, further understanding of possible mechanisms of axonal protection by OECs, and the longer-term time course of protection could provide a basis for future human clinical trials of autografted OECs, which would be available from autologous nasal epithelial biopsies.