Włodzimierz Jarmundowicz

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. Patients 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.

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 patients 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.

The olfactory bulb and olfactory mucosa obtained from human cadaver donors as a source of olfactory ensheathing cells.

During the last decade, olfactory ensheathing cells (OECs) have been successfully applied in multiple experimental approaches aimed to repair damaged mammalian spinal cord. Some of these experiments have consequently been translated into clinical trials. Finding a reliable source of human OECs that is easily accessible and can ensure a sufficient number of cells is a major prerequisite for conducting studies on OEC‐mediated spinal cord regeneration. Here, we present a procedure for obtaining olfactory bulbs (OBs) and olfactory mucosa (OM) simultaneously from adult cadaver heart‐beating donors for OEC isolation and analyze some of the factors that may condition successful OEC culture. We show that the results of OEC culture from OBs (10 cases) correlated significantly with warm ischemia time (WIT) as well as the initial viability of the isolated cells. Efficient OEC culture was possible when the WIT for the OB was up to 20 min. Brain damage, assessed by determination of S100B serum level, was not related to the success of OEC culture from the OB. Cadaver OM (7 cases) was shown to be a more reliable source of human OECs than the OB. In most of the examined cases the efficacy of culturing OECs from cadaver OM obtained even 180 min after cardiac arrest was comparable to that of living patients. The method of obtaining OBs and OM from cadavers enables the use of an alternative source of primary adult human OECs for further preclinical and clinical studies on their neurotrophic properties.

Prospective study on the efficacy of low-field intraoperative magnetic resonance imaging in neurosurgical operations.

BACKGROUND AND PURPOSE The application of intraoperative magnetic resonance imaging (iMRI) is related to a series of challenges of both a technical and an organizational nature. We present our experience in the application of low-field iMRI in everyday neurosurgical practice. MATERIAL AND METHODS A group of 58 patients operated on using low-field iMRI was subject to prospective controlled observation. The significance of differences in the range of preparation time, duration and direct operation results between the iMRI group and controls was analysed. The influence of epidemiological and demographic factors and technical aspects related to iMRI application on direct outcome of the surgery was assessed. RESULTS Twenty-eight tumour resections using craniotomy, 17 transsphenoidal resections of pituitary adenomas and 13 stereotactic procedures were conducted in the group of 24 men and 34 women operated on using iMRI. The control group was not significantly different in terms of epidemiological and demographic factors. The preparation and operation times were significantly longer in the iMRI group (p < 0.001 and p = 0.002, respectively). Longer duration of the surgery was not related to an increased frequency of complications. A higher percentage of postoperative improvement in neurological status (31% vs. 14%, p = 0.045), lower complication percentage (10% vs. 28%, p = 0.03) and a similar time of hospitalization (13 ± 7 vs. 12 ± 4 days, p = 0.33) were noted in the iMRI group. CONCLUSIONS The application of low-field iMRI prolongs the duration of neurosurgical procedures but does not negatively influence their safety. It is associated with above-average functional results and a lower percentage of total complications.

Usefulness of perfusion weighted magnetic resonance imaging with signal-intensity curves analysis in the differential diagnosis of sellar and parasellar tumors: Preliminary report

PURPOSE The most common pituitary tumors are adenomas, which however may be mimicked by other tumors that can show a very similar appearance in plain MRI. The aim of our study was to evaluate the usefulness of perfusion weighted MR imaging (PWI), including signal-intensity curves analysis in the differential diagnosis of sellar/parasellar tumors. METHODS Forty-one patients with sellar/parasellar tumors (23 macroadenomas, 10 meningiomas, 5 craniopharyngiomas, 1 intrasellar hemangioblastoma, 1 intrasellar prostate cancer metastasis, 1 suprasellar glioma), underwent plain MRI followed by PWI using a 1.5T unit. In each tumor, the mean and maximum values of relative cerebral blood volume (rCBV), as well as the relative peak height (rPH) and the relative percentage of signal intensity recovery (rPSR) were calculated. RESULTS The high perfusion tumors were: macroadenomas, meningiomas, squamous-papillary type of craniopharyngiomas, hemangioblastoma, glioma and metastasis. The low perfusion neoplasms included adamantinomatous type of craniopharyngiomas. By comparing adenomas and meningiomas, we found statistically significant differences in the mean and maximum rCBV values (p=0.026 and p=0.019, respectively), but not in rPH and rPSR. The maximum rCBV values >7.14 and the mean rCBV values >5.74 with the typical perfusion curve were very suggestive of the diagnosis of meningioma. There were differences between adenomas and other high perfusion tumors in rPH and rPSR values. CONCLUSIONS PWI can provide additional information helpful in differential diagnosis of sellar/parasellar tumors. In our opinion PWI, as an easy to perform and fast technique should be incorporated into the MR protocol of all intracranial neoplasms including sellar/parasellar tumors.

Aneurismal subarachnoid hemorrhage: who remains for surgical treatment in the post-ISAT era?

Introduction Although there have been a number of studies on changes and trends in the management of aneurismal subarachnoid hemorrhage (aSAH) since publication of the International Subarachnoid Aneurysm Trial (ISAT), no data exist on what category of patients still remains for surgical treatment. Our goal was to investigate the changes that occurred in the characteristics of a population of aSAH patients treated surgically in the post-ISAT period in a single neurosurgical center, with limited availability of endovascular service. Material and methods The study included 402 aSAH patients treated surgically in our unit between January 2004 and December 2011. Each year, data regarding number of admissions, age, aneurysm location and size, clinical and radiological presentation, outcome and mortality rates were collected and analyzed. Results The annual number of admissions more than halved in the study period (from 69 in 2004 to 32 in 2011). There were no linear trends regarding patients’ mean age, clinical presentation and outcomes, but the number of patients in Fisher grade 4 increased and mortality slightly decreased. An unexpected, statistically significant increase occurred in the incidence of anterior communicating artery aneurysms (from 36.2% to 50%) and medium size aneurysms (from 34.7% to 56.2%) treated surgically, with a corresponding decrease in the incidence of middle cerebral artery aneurysms (from 40.5% to 34.3%) and large aneurysms (from 21.7% to 12.5%). Conclusions Unexpected trends in characteristics of aSAH patients treated surgically could be related to treatment decision modality. Trend patterns could be properly expressed in the constant availability of endovascular services.

Assessment of in vivo behavior of polymer tube nerve grafts simultaneously with the peripheral nerve regeneration process using scanning electron microscopy technique.

In this study, scanning electron microscopy (SEM) has been applied for instantaneous assessment of processes occurring at the site of regenerating nerve. The technique proved to be especially useful when an artificial implant should have been observed but have not yet been extensively investigated before for assessment of nerve tissue. For in vivo studies, evaluation of implants morphology and its neuroregenerative properties is of great importance when new prototype is developed. However, the usually applied histological techniques require separate and differently prepared samples, and therefore, the results are never a 100% comparable. In our research, we found SEM as a technique providing detailed data both on an implant behavior and the nerve regeneration process inside the implant. Observations were carried out during 12-week period on rat sciatic nerve injury model reconstructed with nerve autografts and different tube nerve grafts. Samples were analyzed with haematoxylin-eosin (HE), immunocytochemical staining for neurofillament and S-100 protein, SEM, TEM, and the results were compared. SEM studies enabled to obtain characteristic pictures of the regeneration process similarly to TEM and histological studies. Schwann cell transformation and communication as well as axonal outgrowth were identified, newly created and matured axons could be recognized. Concurrent analysis of biomaterial changes in the implant (degradation, collapsing of the tube wall, migration of alginate gel) was possible. This study provides the groundwork for further use of the described technique in the nerve regeneration studies.

Influence of calcium alginate on peripheral nerve regeneration: In vivo study

Recently, we described the influence of sodium alginate on the inflammatory infiltrate during neuroregeneration in tube nerve grafts. It was noticeable that there was the coexistence of inflammatory cells, including neutrophils, plasma cells, and macrophages with Schwann cells and axons. This may indicate a beneficial interaction between alginates and the infiltrate and the additional beneficial effect of the cells on the neuroregeneration process in the inflammatory infiltrates. In this study, we have performed in vivo evaluation of our novel tubular implant prepared by a polyurethane/polylactide blend filled with alginate fibers. The influence of filling the lumen of the tubes with sodium and calcium alginates on the regeneration process of the rat sciatic nerve was investigated. The neuroregeneration process was assessed by detailed histomorphometric studies, axon counting, and calculating the regenerative indexes. It was observed that calcium alginate had a supportive effect on nerve regeneration similar to nerve autotransplant.

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

Abstract 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 (Fi2 = 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 (Fi2 = 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 (Fi2 = 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.

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

Abstract 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.