Spinal Studies

 

Midland Centre for Spinal Injuries

Scientific Research Team: Sally Roberts, Sharon Owen, Sarah Turner, Karina Wright, Charlotte Hulme, Annie Kerr.

Clinical Team: Mr Birender Balain, Mr Shashank Chitgopkar, Mr Joy Chowdhury, Mr Neil Davison, Mr David C Jaffray, Mr Naveen Kumar, Mr Matthew Ockendon, Mr Aheed Osman, Mr Jayesh Trivedi, Ms Ineta Zobina and Professors Wagih El Masri and Stephen Eisenstein.

 

CAN WE MAKE CELLS OF THE INTERVERTEBRAL DISCS TURN DISCS TO BONE FOR SPINAL FUSION?

Sharon Owen (nee Brown), Sarah Turner, Birender Balain, Matthew Ockendon, Shashank Chitgopkar and Sally Roberts

Funded by the Orthopaedic Institute Ltd, Oswestry & the Garfield Weston Trust

Spinal fusion is sometimes used to treat patients with back pain thought to be caused by degenerate intervertebral discs, but it is quite an invasive surgical technique. Over the last decade or so, one of the major scientific research areas in back pain has been on the biological repair or regeneration of the degenerate intervertebral disc. We have been studying an alternative biological approach to assisting spinal fusion by encouraging the disc cells to produce bone.  If this could be achieved it could provide a much easier technique for spinal fusion and immobilising the painful section of the spine.

So far we have been able to show that cells from the centre of the disc (nucleus pulposus cells) are capable of following an osteogenic pathway when grown in monolayer with either 25-dihydroxyvitamin D3 (Vitamin D) or standard osteogenic culture medium (containing dexamethasone, ascorbic acid and β-glycerophosphate). Standard osteogenic medium was found to be the most effective at encouraging disc cells to behave like bone cells, but not to the extent of bone marrow derived MSCs which were used for comparison. 

To assess whether the disc cells in situ in the body could become osteogenic, 3D disc explants were cultured in osteogenic media with or without the additional bone-inducing factors Vitamin D and bone morphogenetic protein (BMP) 2.  Two out of eleven disc explants showed positive mineralisation and calcification, one when cultured in osteogenic media only and the other with the additional bone-inducing factors.  We are currently trying to determine if some discs have more natural ‘inhibitors’ of mineralisation than others and if that could explain why some discs appear capable of mineralising whilst others do not.

 Figure: One of the samples of surgically removed intervertebral discs which were cultured in 3-D in a medium, or ‘soup’, which encouraged disc cells to behave like bone cells and lay down a calcium phosphate structure like bone, shown here as dark brown (arrows; section stained with von Kossa).

 


 

AN IMAGING STUDY OF THE VERTEBRAL ENDPLATE IN DISC DEGENERATION

Sharon Owen (nee Brown), Victor Cassar-Pullicino, Iain McCall, Neil Broom (Auckland), Chris Sharp (Shrewsbury) and Sally Roberts

Funded by the Orthopaedic Institute Ltd, Oswestry & the Winston Churchill Memorial Trust

Low back pain remains one of the major causes of long term disability and loss of quality of life in the world. It has a two-fold impact on the economy through both a loss in productivity and an increase in the associated healthcare costs. The hard mineralised bones (vertebral bodies) in the spine are separated from each other by the soft non-mineralised intervertebral discs which enable movement within the spine.  How these two entities attach to one another is an important factor for spine stability with loss of attachment being previously highlighted as a major contributory factor in herniated (or “slipped”) discs.  Much work has been carried out in New Zealand investigating the microscopic organisation of this junction in sheep, which has been well characterised via a specialised microscopic technique using differential interference contrast optics.

Through a combination of funding from the Orthopaedic Institute Ltd and the Winston Churchill Memorial Trust (Travelling Fellowship), 20 post-mortem lumbar spines were imaged using standard X-ray and MRI scanners and scored by a radiologist. A cohort of discs showing a range of degrees of degeneration were prepared and sent over to New Zealand. A travelling scholarship awarded to Sharon Owen allowed her to follow and learn the microscopic technique under the ‘watchful eye’ of the experienced New Zealand research team.

The microscopic organisation of collagen fibres at the junction between the vertebral body and intervertebral disc in humans appeared to have some similarities to that previously observed in sheep by the New Zealand group, but appeared overall less organised.  Loss of connectivity at the disc-vertebra junction between the two structures appears to increase with the degree of degeneration and this may impact on the spine’s stability.  As a consequence, the biomechanical environment of the cells present at this junction and within the disc may be altered and could potentially contribute further to the deterioration of the disc.  It is hoped that this technique may be applied to other bone-cartilage interfaces to enhance the interpretation of the macroscopic images obtained in clinic.

BIOMARKER DISCOVERY AND USE IN SPINAL CORD INJURY PATIENTS: IDENTIFYING NEW TREATMENT TARGETS AND MARKERS FOR PREDICTING CLINICAL OUTCOME

 

Sharon Owen, Charlotte Hulme, Heidi Fuller, Naveen Kumar, Ineta Zobina, Joy Chowdhury, Aheed Osman, Sally Roberts, Monte Gate, Wagih El Masri and Karina Wright

Funded by the Orthopaedic Institute,Oswestry and the Midland Centre for Spinal Injuries (MCSI), Oswestry

The overall aim of this project is to investigate biomarkers for use in spinal cord injury (SCI). It involves two approaches, the first is to identify blood biomarkers which can monitor the progression of neurological deficit or recovery following spinal cord injury (SCI) in different subgroups of patients and the other being to identify new prognostic indicators and targets for the treatment of SCI. Identifying markers which can be used to help determine how a patient will recover following SCI will ensure patients can be assigned to the most appropriate treatment pathway to achieve their best potential clinical outcome. 

Since October 2014 Dr Sharon Owen (MCSI funded scientific support) and Dr Charlotte Hulme (funded by the Orthopaedic Institute) have recruited 40 acutely injured patients (within 2 weeks of their injury). Twenty-nine of these patients have also had blood samples collected at the sub-acute phase of injury (3 months post-injury) and of these patients, 15 have also had blood samples collected at 12 months after their injury (chronic injury). These blood samples will be used to monitor changes in biomarker profiles as SCI proceeds into sub-acute and chronic phases. A review of the literature has identified several biomarker candidates which will be assessed within the collected blood samples.

Initial work-up of assays to measure these biomarkers has been completed.  So far, 43 blood samples (20 acute injury; 12 sub-acute injury and 11 chronic injury) have been assessed for S 100 calcium binding protein β (S100β) and 56 samples have been assessed for neurofilament light chain (NF-L) (24 acute injury; 21 sub-acute injury and 11 chronic injury). Assessment of these markers, along with neurofilament heavy chain (NF-H) will be completed in all blood samples that are collected and then further analysis will be carried out to investigate whether any of these markers correlate with clinical outcomes of these patients.

The second approach, which aims to discover new biomarkers which have not previously been investigated for SCI, uses an ‘unbiased approach’. The field of blood biomarker identification for SCI is currently quite limited and there is a need for identifying new potential markers. Analysis of two different models of SCI has been completed and proteins which are altered longitudinally following a SCI have been identified. Several proteins which may be useful markers of SCI severity have been identified and one of these, C-reactive peptide, has been measured in 32 blood samples from human SCI patients (16 collected within 2 weeks of injury; 12 collected at 3 months after injury and 4 collected at 12 months after injury).

Specialised computer programmes have been used to assess which of these proteins may be most important in mediating the effects of SCI and therefore which might be good targets for new treatments for SCI. Similar work will be completed by analysing blood samples collected from patients at the MCSI, if they are interested in being involved. Together these studies should help identify markers which can be used to assess a patients’ potential for neurological repair following a SCI and potentially to assess how effectively SCI treatments are working. 

ASSESSING PATIENT OPINION ON TRANSLATIONAL RESEARCH FOR SPINAL CORD INJURY

Jane Parker, Jan Herman Kuiper, Clive Glass, Mohankumar Mariappan, Wagih El Masri, Joy Chowdhury, Aheed Osman, Soukaina Bahsoun, Charlotte Hulme, Karina Wright

Funded by the Orthopaedic Institute Ltd, Oswestry

The overall purpose of this project is to begin to profile the opinions and perspectives of the spinal cord injury (SCI) patient population on the emerging scientific advances and new potential treatments for them. There are no accurate published audits for the assessment of the opinion of SCI patients for the translation of scientific discoveries from the bench to the bedside in the field of SCI research, for example, using embryonic or other types of stem cells. 104 questionnaires have been returned from RJAH SCI patients in the community. Preliminary analyses of these questionnaires indicates that on-the-whole SCI patients are generally very keen to participate in research, including participating in further opinion based studies, as well as donating tissue e.g. saliva/fat/stem cells. Overall, these patients were also willing to participate in clinical trials and/or experimental treatments; however, if they were informed of every possible risk associated with these trials, they were much less willing to participate in this level of research.

Our current work also indicates that there is no correlation between psychological wellbeing or physical capacity of these patients and the level of research participation they are willing to be involved with. The study is now being rolled-out to include 500 patients from the North West UK SCI Centre based in Southport. We hope the findings from this study will help to bridge this gap in the knowledge, for scientists and clinicians alike, so that the application of new potential treatments will benefit the whole SCI population in the most effective manner. The findings from this questionnaire will enable SCI clinicians to counsel patients about regenerative research and clinical trials with a better understanding of the patients’ perspective. 

  

Midland Centre for Spinal Injuries

Clinical Lead: Mr Joy Roy Chowdhury

Members of the Research Team: Dr K Wright, Dr S Owen, Dr C Hulme

Clinical Support: Mr Aheed Osman and Mr Naveen Kumar

2015-2016 year has been a very busy and productive year for the MCSI (Midland Centre for Spinal Injuries) research group, establishing new collaborations and moving research forward for Spinal Cord Injury.

We have been very fortunate to receive funding from external sponsors and the Orthopaedic Institute.

2015 has seen further integration of clinical activity in the hospital and laboratory research programmes within the Centre the majority of which are national or international collaborative projects.  

The completed projects have been presented in various national and international meetings including International Spinal Cord Society Annual Meetings. The majority of these projects have been published in peer reviewed journals. 

 

The projects MCSI is currently working on

Blood Biomarkers for Spinal cord injury

RJAH and Keele University

Researchers: K Wright(CI), H Fuller, E Murphy,  J Roy Chowdhury, AE Osman, W Elmasri and N Kumar

ECLISP – (Efficacy of consuming LCS (lactobacillus casei shirtta) in spinal cord injury patients) a UK multicentre, randomised, controlled study evaluating efficacy of LCS in prevention of antibiotic acquired diarrhoea in spinal cord injury patients

Researchers : N Kumar(PI), J Roy Chowdhury and  AE Osman

 

Translation Research for Spinal cord injury: current  and future perspective - Patient Questionnaire study- a UK multicentre study ( Southport  and Oswestry)

Researchers : K Wright(PI), H Fuller, E Murphy,  J Roy Chowdhury, AE Osman, W Elmasri and N Kumar

The MCSI team are constantly contributing to research programmes; we are keen to maintain this pattern into the future.

 

MCSI planned Research projects for 2016

Pan European Paediatric Spinal Cord (PEPSCI)

A multicentre international study on Research Priorities for Spinal Cord Injury in Children, Adolescents and Young Adults: An International Service User Survey

Researchers : K Wright(PI), N Kumar,  J Roy Chowdhury and AE Osman     

 

Transcranial magnetic field therapy for treatment spasticity

A joint collaboration with ORLAU

Researchers: Mr A Roberts, J Roy Chowdhury, N Kumar, and AE Osman     

 

Setting the SCENE: Spinal Cord Injuries Expectations, Needs and Experience- a UK multicentre study

A joint collaboration with Warwick Medical University lead by  Prof Diane Playford

Researchers: J Roy Chowdhury, N Kumar, and AE Osman     

 

FES study- safety, efficacy for bone health and quality of life for SCI

ORLAU, University of Keele and Case Western (USA)

Researchers: J Roy Chowdhury, N Kumar, and AE Osman     

 

Osteoporosis in chronic SCI – UK Multicentre Study

A joint collaboration - UK Multicentre Study Oswestry, Stanmore and Glasgow

Researchers: J Roy Chowdhury, N Kumar, and AE Osman     

 

Autonomic dysfunction and physical activity in SCI

Collaborative study with University of Liverpool.

Researchers: J Roy Chowdhury, N Kumar, and AE Osman     

 

Adult Stem Cells in Spinal Cord Injury and Repair

Researchers: Team of scientists and clinicians at the Robert Jones & Agnes Hunt Orthopaedic Hospital, University of Chester and UK SCI Centres.

 

The MCSI team are constantly contributing to research programmes; we are keen to maintain this pattern into the future.