Aheed Osman

The cell culture expansion of bone marrow stromal cells from humans with spinal cord injury: implications for future cell transplantation therapy

Study design:Previous studies have shown that transplantation of bone marrow stromal cells (MSCs) in animal models of spinal cord injury (SCI) encourages functional recovery. Here, we have examined the growth in cell culture of MSCs isolated from individuals with SCI, compared with non-SCI donors.Setting:Centre for Spinal Studies, Midland Centre for Spinal Injuries, RJAH Orthopaedic Hospital, Oswestry, UK.Methods:Bone marrow was harvested from the iliac crest of donors with long-term SCI (>3 months, n=9) or from non-SCI donors (n=7). Mononuclear cells were plated out into tissue culture flasks and the adherent MSC population subsequently expanded in monolayer culture. MSC were passaged by trypsinization at 70% confluence and routinely seeded into new flasks at a density of 5 × 103 cells per cm2. Expanded cell cultures were phenotypically characterized by CD-immunoprofiling and by their differentiation potential along chondrocyte, osteoblast and adipocyte lineages. The influence of cell-seeding density on the rate of cell culture expansion and degree of cell senescence was examined in separate experiments.Results:In SCI, but not in non-SCI donors the number of adherent cells harvested at passage I was age-related. The proliferation rate (culture doubling times) between passages I and II was significantly greater in cultures from SCI donors with cervical lesions than in those with thoracic lesions. There was no significant difference, however, in either the overall cell harvests at passages I or II or in the culture doubling times between SCI and non-SCI donors. At passage II, more than 95% of cells were CD34−ve, CD45−ve and CD105+ve, which is characteristic of human MSC cultures. Furthermore, passage II cells differentiated along all three mesenchymal lineages tested. Seeding passage I–III cells at cell densities lower than 5 × 103 cells per cm2 significantly reduced culture doubling times and significantly increased overall cell harvests while having no effect on cell senescence.Conclusion:MSCs from individuals with SCI can be successfully isolated and expanded in culture; this is encouraging for the future development of MSC transplantation therapies to treat SCI. Age, level of spinal injury and cell-seeding density were all found to relate to the growth kinetics of MSC cultures in vitro, albeit in a small sample group. Therefore, these factors should be considered if either the overall number or the timing of MSC transplantations post-injury is found to relate to functional recovery.

The developing landscape of diagnostic and prognostic biomarkers for spinal cord injury in cerebrospinal fluid and blood.

Study design:Review study.Objectives:The identification of prognostic biomarkers of spinal cord injury (SCI) will help to assign SCI patients to the correct treatment and rehabilitation regimes. Further, the detection of biomarkers that predict permanent neurological outcome would aid in appropriate recruitment of patients into clinical trials. The objective of this review is to evaluate the current state-of-play in this developing field.Setting:Studies from multiple countries were included.Methods:We have completed a comprehensive review of studies that have investigated prognostic biomarkers in either the blood or cerebrospinal fluid (CSF) of animals and humans following SCI.Results:Targeted and unbiased approaches have identified several prognostic biomarkers in CSF and blood. These proteins associate with cellular damage following SCI and include components from neurons, oligodendrocytes and reactive astrocytes, that is, neurofilament proteins, glial fibrillary acidic protein, Tau and S100 calcium-binding protein β. Unbiased approaches have also identified microRNAs that are specific to SCI, as well as other cell damage-associated proteins.Conclusions:The discovery and validation of stable, specific, sensitive and reproducible biomarkers of SCI is a rapidly expanding field of research. So far, few studies have utilised unbiased approaches aimed at the discovery of biomarkers within the CSF or blood in this field; however, some targeted approaches have been successfully used. Several studies using various animal models and some with small human patient cohorts have begun to pinpoint biomarkers in the CSF and blood with putative prognostic value. An increased sample size will be required to validate these biomarkers in the heterogeneous clinical setting.

The evolution of national care pathways in spinal cord injury management

Spinal cord injury service delivery in the UK has improved over the years. The modern concept of Spinal Cord Injury care commenced after the Second World War. Patients started to live longer after the improvement in the medical and nursing care which was delivered in Spinal Cord Injury Centres across the country. In the last five years, spinal cord injury care has seen major advances. The Spinal Cord Injury Strategy Board was established in 2010 which led to the development of the Spinal Injury Pathway, the Spinal Injury National database and the creation of the clinical reference group for Spinal Cord Injury. This led to improvement in the care of evidence-based management.

Evidence-based respiratory management strategies required to prevent complications and improve outcome in acute spinal cord injury patients:

Spinal injuries without neurological damage have little effects on respiratory function unless associated with injury to the chest wall. Early verticalisation or mobilisation of these patients is safe and likely to improve vital capacity. Spinal injury with cord damage has a profound effect on the mechanics of respiration and on respiratory function particularly in cervical cord injuries. Around 40% of spinal cord injuries occur in the cervical spine, a trend that is steadily increasing, with respiratory causes being responsible for death in over 20% of individuals. Loss of lung volumes and relative hypoxemia contribute to global hypoxaemia, exacerbating cord ischaemia in the acute period. Respiratory compromise results in the loss of muscle strength generation capacity and reduced lung volumes and in particular vital capacity, of up to 70%, ineffective cough and secretion clearance abilities; reductions in both lung and chest wall compliance and an additional oxygen cost of breathing due to changes in respiratory mechanics, with obstructive sleep apnoea evident in over 50% of acute tetraplegics. While some countries have specialist spinal centres to manage such catastrophic trauma with a demonstrable improvement in health outcomes attributed to their contribution, many individuals are initially admitted to local hospitals where healthcare professionals are less likely to fully appreciate the significant and continued vulnerabilities of such individuals. This article aims to provide a basic understanding of the causes and identification of the main principles of the respiratory management strategies required to maintain pulmonary health for cervical spinal cord injury patients during the initial and early post trauma phase.

Long-term outcome of paediatric spinal cord injury:

Background Spinal cord injuries are relatively uncommon in children and evidence about long-term outcomes is limited. This study was performed to determine the frequency of common long-term complications in patients sustaining spinal injury in childhood (0–18 years) and who were followed up at a single dedicated spinal injuries centre in the UK. Method A retrospective review of clinical records of all patients injured at or less than 18 years of age between 1971 and 1999. Complications studied were renal, bowel, musculoskeletal, pressure ulcers and post-traumatic syringomyelia. Long-term social outcomes of independence, employment and driving were also assessed. Results Of 69 individuals (47 males, 22 females) the median age at injury was 17 years (range 0–18 y); 68% were older than 13 years at injury and 74% had traumatic injuries. Patients had an average duration of 27 years (12–43 years) of spinal injury – half had a neurological level of T6 and above, 80% had paraplegia and 20% had quadriplegia. Discussion Patients with both complete and incomplete spinal cord injury have minimal neurological recovery. Managing medical complications is vital as only 11.5% had normal voiding and 10.6% had normal bowel function. The incidence of skin ulcers increases with duration of spinal cord injury and scoliosis is higher in the non-traumatic injury group. Spasticity is observed in 66.6% and post-traumatic syringomyelia in 11.7%. Long-term social outcomes are good with 75% patients able to do independent care, 46% could drive and 39% managed employment or higher education. Conclusions This study documents the long-term outcomes and complications of spinal cord injuries sustained in childhood. With initial active physiological conservative management of the majority of patients, patient education and ongoing support the majority of patients achieved long-term survival and led independent and productive lives.

Epidemiology, neurological and functional outcome of concomitant traumatic brain and spinal cord injury: An Oswestry experience

Background Concomitant traumatic brain injury with spinal cord injury is likely to worsen prognosis and increase hospital length of stay. This study assessed the duration of in-patient rehabilitation and outcome in patients with both traumatic brain injury and spinal cord injury. Methods Retrospective study of all patients with concomitant traumatic brain injury and spinal cord injury over a 3-year period who had 5 years of subsequent follow-up at the Midlands Centre for Spinal Injuries. Results Twenty-seven patients had concomitant injuries of which five had severe traumatic brain injury, nine had moderate traumatic brain injury and the remaining thirteen had mild traumatic brain injury with spinal cord injury of grades A–D; commonest mechanisms of injury were motor vehicle collision (55%) and falls (37%). Thirteen (48%) had tetraplegia and 14 (52%) had paraplegia. Mean functional independence measure score at admission was 52.1 and 103.4 at 5 years. Patients with mild traumatic brain injury gained a mean functional independence measure score of 67.1; the moderate and severe traumatic brain injury patients gained mean functional independence measure score of 60.1 and 69.2, respectively. The mean length of stay was 138.3, 139.4 and 153.4 days for mild, moderate and severe traumatic brain injury, respectively. Conclusion Hospital length of stay and patient’s functioning at 5 years were not affected by traumatic brain injury severity in this subgroup; however, functional independence measure on its own may not be very sensitive to cognitive deficits.