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Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVI | Pages 23 - 23
1 Aug 2012
Tayton E Purcell M Aarvold A Smith J Kalra S Briscoe A Fahmy S Shakesheff K Howdle S Dunlop D Oreffo R
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Disease transmission, availability and economic costs of allograft have resulted in significant efforts into finding an allograft alternative for use in impaction bone grafting (IBG). Biotechnology offers the combination of skeletal stem cells (SSC) with biodegradable polymers as a potential solution. Recently polymers have been identified with both structural strength and SSC compatibility that offer the potential for clinical translation.

The aim of this study was to assess whether increasing the porosity of one such polymer via super critical CO2 dissolution (SCD) enhanced the mechanical and cellular compatibility characteristics for use as an osteogenic alternative to allograft in IBG.

High molecular weight PLA scaffolds were produced via traditional (solid block) and SCD (porous) techniques, and the differences characterised using scanning electron microscopy (SEM). The polymers were milled, impacted, and mechanical comparison between traditional vs SCD created scaffolds and allograft controls was made using a custom shear testing rig, as well as a novel agitation test to assess cohesion. Cellular compatibility tests for cell number, viability and osteogenic differentiation using WST-1 assays, fluorostaining and ALP assays were determined following 14 day culture with SSCs.

SEM showed increased porosity of the SCD produced PLA scaffolds, with pores between 50-100 micrometres. Shear testing showed the SCD polymer exceeded the shear strength of allograft controls (P<0.001). Agitation testing showed greater cohesion between the particles of the SCD polymer (P<0.05). Cellular studies showed increased cell number, viability and osteogenic differentiation on the SCD polymer compared to traditional polymer (P<0.05) and allograft (P<0.001).

The use of supercritical C02 to generate PLA scaffolds significantly improves the cellular compatibility and cohesion compared to traditional non-porous PLA, without substantial loss of mechanical shear strength. The improved characteristics are critical for clinical translation as a potential osteogenic composite for use in impaction bone grafting.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVI | Pages 20 - 20
1 Aug 2012
Tayton E Fahmy S Aarvold A Smith J Kalra S Briscoe A Purcell M Shakesheff K Howdle S Dunlop D Oreffo R
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Impaction bone grafting with milled human allograft is the gold standard for replacing lost bone stock during revision hip surgery. Problems surrounding the use of allograft include cost, availability, disease transmission and stem subsidence (usually due to shear failure of the surrounding allograft).

The aim of this study was to investigate various polymers for use as substitute allograft. The ideal graft would be a composite with similar mechanical characteristics as allograft, and with the ability to form de novo bone.

High and low molecular weight (MW) forms of three different polymers (polylactic acid (PLA), poly (lactic co-glycolic) acid (PLGA) and polycaprolactone (PCL)) were milled, impacted into discs, and then tested in a custom built shear testing rig, and compared to allograft.

A second stage of the experiment involved the addition of skeletal stem cells (SSC) to each of the milled polymers, impaction, 8 days incubation, and then tests for cell viability and number, via fluorostaining and biochemical (WST-1) assays.

The shear strengths of both high/ low MW PLA, and high/low MW PLGA were significantly higher than those of milled allograft (P<0.001, P<0.001, P<0.005 and P<0.005) but high and low MW PCL was poor to impact, and had significantly lower shear strengths (P<0.005, P<0.001). Fluorostaining showed good cell survival on high MW PLA, high MW PCL and high MW PLGA. These findings were confirmed with WST-1 assays.

High MW PLA as well as high MW PLGA performed well both in mechanical testing and cell compatibility studies. These two polymers are good contenders to produce a living composite for use as substitute human allograft in impaction bone grafting, and are currently being optimised for this use via the investigation of different production techniques and in-vivo studies.


The Journal of Bone & Joint Surgery British Volume
Vol. 94-B, Issue 6 | Pages 848 - 855
1 Jun 2012
Tayton ER Smith JO Aarvold A Kalra S Dunlop DG Oreffo ROC

When transferring tissue regenerative strategies involving skeletal stem cells to human application, consideration needs to be given to factors that may affect the function of the cells that are transferred. Local anaesthetics are frequently used during surgical procedures, either administered directly into the operative site or infiltrated subcutaneously around the wound. The aim of this study was to investigate the effects of commonly used local anaesthetics on the morphology, function and survival of human adult skeletal stem cells.

Cells from three patients who were undergoing elective hip replacement were harvested and incubated for two hours with 1% lidocaine, 0.5% levobupivacaine or 0.5% bupivacaine hydrochloride solutions. Viability was quantified using WST-1 and DNA assays. Viability and morphology were further characterised using CellTracker Green/Ethidium Homodimer-1 immunocytochemistry and function was assessed by an alkaline phosphatase assay. An additional group was cultured for a further seven days to allow potential recovery of the cells after removal of the local anaesthetic.

A statistically significant and dose dependent reduction in cell viability and number was observed in the cell cultures exposed to all three local anaesthetics at concentrations of 25% and 50%, and this was maintained even following culture for a further seven days.

This study indicates that certain local anaesthetic agents in widespread clinical use are deleterious to skeletal progenitor cells when studied in vitro; this might have relevance in clinical applications.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXIV | Pages 12 - 12
1 May 2012
Aarvold A Smith J Edwards C Tayton E Gent E Oreffo RC
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Background

Unicameral bone cysts (UBCs) are difficult to treat and have a high recurrence rate. Their pathogenesis is unknown making targeted therapies difficult. Attributed causes include venous and interstitial fluid obstruction, oxygen free radicals, lysosomal enzymes, prostaglandins and genetic factors. Skeletal stem cells (SSCs) are osteoblast precursors critical to bone formation and cyst fluid may influence their growth, however the association between SSCs and cyst fluid has never been investigated.

Aim

To investigate the effect of UBC fluid on SSC growth.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XVIII | Pages 26 - 26
1 May 2012
Jones A Aarvold A New A Dunlop D Oreffo R
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AIM

Avascular necrosis (AVN) of the femoral head is a potentially debilitating disease of the hip in young adults. Impaction bone grafting (IBG) of morcellised fresh frozen allograft is used in a number of orthopaedic conditions. This study has examined the potential of skeletal stem cells (SSC) to augment the mechanical properties of impacted bone graft and we translate these findings into clinical practice.

STUDY DESIGN

We have examined the effect of SSC density on augmentation of bone formation. An in vitro model was developed to replicate the surgical IBG process. Plain allograft was used as the control, and the SSC's seeded at a density of 5×103, 5×104 and 2×105 cells per cc of allograft for the experimental groups. All samples were cultured for 2 weeks and mechanically tested to determine shear strength using the Mohr Coulomb failure curve. The approach was translated to 3 patients with early avascular necrosis (AVN) of the femoral head. The patient's bone marrow was concentrated in theatre using a centrifugation device and the concentrated fraction of SSC's were seeded onto milled allograft. The patient's necrotic bone was drilled, curetted and replaced with impacted allograft seeded with SSC's. Osteogenic potential of concentrated and unconcentrated marrow was simultaneously compared in vitro by colony forming unit assays.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_III | Pages 99 - 99
1 Feb 2012
Aarvold A Casey A Bernard J
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Introduction

Atlanto-occipital dislocation is rare and usually fatal. Stabilisation is typically from Occiput to C2, sacrificing atlanto-axial movement. To preserve movement, screw fixation from the articular mass of C1 to the occipital condyle has been described. Amongst other structures, the hypoglossal nerve is at risk. No previous study has addressed the anatomy of the hypoglossal canal in relation to screw trajectory. We aim to identify landmarks to aid safe screw passage into the occipital condyle.

Methods

20 dry skulls provided 40 hypoglossal canals (HCs) and 40 occipital condyles (OCs). No distinction was made between sex, race or age. 9 parameters were measured for each HC, and relation to skull base was noted.


Orthopaedic Proceedings
Vol. 90-B, Issue SUPP_III | Pages 447 - 447
1 Aug 2008
Aarvold A Casey A Bernard J
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Introduction: Atlanto-Occipital dislocation is rare and usually fatal. Stabilisation is typically from Occiput to C2; sacrificing atlantoaxial movement. To preserve movement, screw fixation from the articular mass of C1 to the occipital condyle has been described. Amongst other structures, the hypoglossal nerve is at risk. No previous study has addressed the anatomy of the hypoglossal canal in relation to screw trajectory. We aim to identify landmarks to aid safe screw passage into the occipital condyle.

Methods: 20 dry skulls provided 40 hypoglossal canals (HCs) and 40 occipital condyles (OCs). No distinction was made between sex, race or age. 9 parameters were measured for each HC, and relation to skull base was noted.

Results: The mean length of the HC was 10mm (range 8 to 14). The extra-cranial foramen of the HC is located lateral to the intra-cranial foramen (30° range 19 to 45). 19 out of 20 skulls had HCs with intra-cranial foramina more caudal than their extra-cranial foramina, ie the HC angled cranially (22° range 7 to 51). 36 of 40 OCs were found to be wholly inferior to the rim of the foramen magnum, with 4 (in 2 skulls) whose bodies lay largely below, but extended above, this landmark. Every single HC studied was situated, in its entirety, superior to the rim of the foramen magnum.

Conclusions: The trajectory of the hypoglossal canal from its intra-cranial foramen is antero-supero-lateral. It is situated, in its entirety, superior to the rim of the foramen magnum. The thickest portion of the occipital condyle is antero-medial. Screw passage from posterior through the C1 articular mass ought to aim for the anterior, superior, medial quadrant of the occipital condyle, and should not pass cranial to the rim of the Foramen Magnum in order to minimise the risk to the Hypoglossal Nerve.


The Journal of Bone & Joint Surgery British Volume
Vol. 89-B, Issue 4 | Pages 542 - 544
1 Apr 2007
Aarvold A Bann S Giblin V Wotherspoon A Mudan SS

The incidence of metastatic osteosarcoma is increasing because of improved results following multi-agent chemotherapy and resection of the primary tumour. Metastases occur most commonly in the lungs, whereas bowel metastases are rare. We describe a 25-year-old female who presented with melaena six years after successful resection of an osteosarcoma of her right femur, and one year after resection of a solitary pulmonary metastasis. Imaging revealed a lesion arising within both the duodenum and the pancreas for which a Whipple’s pancreatoduodenectomy was carried out, achieving complete resection. Histological examination confirmed the diagnosis of metastatic osteosarcoma. We believe this is only the second such case reported. At 11 months post-operatively she had no detectable disease. Although rare, osteosarcoma can metastasise to the intestine. The surgeon must be aware of this complication, and that bowel metastases are potentially resectable.