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Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_II | Pages 88 - 88
1 Feb 2012
Shyamsundar S Morgan R Birch M Campbell P McCaskie A Fenwick S
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Clinical proteomics is an exciting new sub-discipline of proteomics that involves the application of proteomic technologies at the bedside to identify new biomarkers, associated with specific diseases. In this study to compare serum protein profiles between identical age-matched groups of fracture and non-fracture controls, we looked at the initial proteomic profile of 10 patients who had fractures and compared them to age-matched controls to see if there was any specific difference indicative of fracture.

Materials and Methods

10 patients with single fractures of the long bones, wrist or ankle gave a blood sample upon presentation at the fracture clinic. 10 healthy, age-matched, non-fracture volunteers also donated blood. Plasma was isolated and the albumin and IgG fractions removed before loading equal amounts of each sample onto 2 dimensional polyacrylamide gels for analysis by isoelectric point in the first dimension and molecular mass in the second dimension. Protein profiles between fracture patients and non-fracture controls were contrasted using Phoretix 2D analysis software.

Data analysis differentiated between the average gel of the patient group and the average gel of the control group. More than 300 protein spots were observed in both the control and patient group. Seven protein spots were identified which showed a statistically significant (p<0.05) difference between the control and patient samples. Of these, three spots (X, Y, Z) were clear, distinct and present in at least 80% of these gels. All the three spots were up regulated in the patient group as opposed to the control group. These proteins are currently being investigated further by MALDI-TOF TOF for specific protein identification.

Discussion

Proteomic analysis is already a powerful tool in the identification of disease markers. We aim to show here that there are differences seen in blood plasma profiles in fracture patients compared to non-fracture healthy controls. The differences seen may help us to understand the fracture repair process better.


Orthopaedic Proceedings
Vol. 90-B, Issue SUPP_II | Pages 391 - 391
1 Jul 2008
Mills L Noble B Fenwick S Simpson H
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Introduction: Atrophic nonunion is a well recognised complication of long bone fractures. Clinical trials show that BMP-2 accelerates healing and reduces nonunion in open tibial fractures. We are interested in a natural small molecule that has been previously demonstrated to stimulate angiogenesis in vivo. Our aim is to assess the two treatments in the prevention of nonunion. The small animal model we used is a non-critical size defect of the tibia deprived it of its blood supply by surgical stripping of the periosteum and curetting of the local endosteum thus closely reflecting the clinical situation. The outcomes were measured by radiographic assessment and histology.

Methods: Wistar rats were treated with either the angiogenic molecule (0.1% or 0.003%), BMP-2 or vehicle alone (PBS) soaked in a type I collagen sponge. All animals underwent a 2mm osteotomy, stripping of the periosteum and endosteum proximally and distally for the length of the diameter of the tibia. Fluorescent markers were injected at 2 weekly intervals. The rats were sacrificed at 8 weeks. Both tibiae were disarticulated; fixator and soft tissues were removed and AP and lateral X-rays were taken. Subjective assessment of the healing on X-ray was carried out in two ways; using a radiographic scoring system and by grey scale analysis. The samples were embedded, sectioned and stained for new bone formation.

Results: Bridging or potential to bridge was seen in a number of animals on x-ray. Bridging or potential to bridge was judged to be present in 72.22% of the BMP-2 group and 66.67% of the high dose group compared to 22.22% of the control group. Histological analysis is being performed to confirm these findings.

Discussion: Atrophic nonunion is a serious clinical complication, unfortunately BMP-2 is a highly costly treatment option and therefore alternative molecular therapies are much sought after. We describe here an angiogenic molecule has some potential in preventing formation of nonunion.