Introduction

A modified anodisation technique where a titanium surface releases bactericidal concentrations of silver was developed and called Agluna. Our hypothesis was that silver incorporation was bactericidal and had no effects on the viability of fibroblasts and osteoblasts, would have no negative effect on interfacial shear strength and bone contact in an in vivo trans-cortical implant ovine model.

Introduction: The current practice of impaction allograft to fill large defects in revision total hip replacements is sometimes useful but clinical results are inconsistent. Other studies have shown that addition of mesenchymal stem cells (MSC) in blocks of hydroxyapatite (HA) scaffold can enhance new bone formation in a critical sized defect. However, no study has been conducted on combined MSCs with morselised allograft and HA granules. It is hypothesized that impaction of allograft or HA granules seeded with MSCs or osteoprogenitors will enhance new bone formation compared with the groups without MSCs.

Materials and Methods: Six sheep were used for the study. Each sheep received 8 scaffolds which were embedded in both paraspinal muscles. Groups were: 1) 3.5g allograft, 2) 3.5g allograft with MSCs, 3) 3.5g allograft with osteoblasts; 4) 3.5g of 50:50 allograft/ HA, 5) 3.5g of 50:50 allograft/HA with MSCs, 6) 3.5g of 50:50 allograft/HA with osteoblasts; 7) a block of HA, 8) a block of HA with MSCs. The experimental scaffolds were seeded with either 10x106 MSCs/ml or 10x106 MSC-derived osteoprogenitors/ml, in 3ml autologous plasma. Grafts were impacted twenty times at 3KN. At eight weeks, samples were sectioned for histology analysis. Areas of new bone formation were measured as percentage to total available spaces. ANOVA was used for statistical analysis.

Results: Addition of MSCs increased new bone formation in allograft (4.98%), allograft/HA (5.15%) and HA block (7.09%) compared with their controls at 2.24%, 1.96% and 1.96% respectively. Statistical study showed significant increase in 50:50 allograft/HA with MSCs compared with 50:50 allograft/HA only (p=0.046) and 50:50 allograft/HA with osteoprogenitors (p=0.028). No difference was found in allograft groups. For the HA block groups, addition of MSCs showed a significant new bone increase compared to the control (p=0.028).

Conclusion: Addition of MSCs to the allograft and HA granules will enhance new bone formation after impaction which can be used for revision total hip replacements, especially when allograft and HA is mixed. However, addition of osteoprogenitors has not achieved the similar results. This study encourages a further clinical investigation of impaction tissue-engineered graft to repair bone defects in revision total joint replacements.

Introduction: Periprosthetic bone loss, brought about by wear particle induced osteolysis, presents a major challenge and compromises outcome in revision Total Hip Replacement. Poor bone stock at revision hip replacement is the main indication for impaction allografting. There are well documented limitations in the use of bone graft. Autogenous bone graft is osseoinductive, though donor site morbidity and the limited amount available restrict its use. An alternative is allogenic bone graft from cadaveric femoral heads. The drawbacks of using allograft are a limited supply and the risk of disease transmission. An alternative may be the use of bone substitute materials. Usually these are used in conjunction with allograft and therefore a number of drawbacks still apply. This study investigates the use of impaction grafting without bone graft. In this study we tested Apopore, 60% porosity, 2–5 mm hydroxyappatite (HA) granules (ApaTech Ltd) in an animal impaction model with allograft as control. Hypothesis Impaction using porous granular HA induces a similar volume of new bone compared with impaction using allograft.

Methods and Materials: Cylindrical defects of 15mm diameter were created in the medial femoral condyles of 12 sheep (6 sheep in each group) and filled with 3.5 grams of either morselised ovine allograft, washed and defatted according to North London Tissue Bank protocols, or porous HA granules impacted with a specially designed impactor, 20 times with a force of 3 KN. This force was similar to that measured during impaction grafting in clinical cases. After 6 weeks the sheep were euthanized, samples embedded in resin and the amount of bone formation measured by histomorphometric analysis.

Results: Under the impaction forces used the HA graft was more impacted than allograft. In the impacted HA graft the average pore size was smaller than for impacted allograft. After 6 weeks more new bone formation was observed at the host implant interface than the middle of the implant in both groups. At the implant host interface there was 26.64% (± 2.13%) new bone formation in the allograft and 21.13% (± 4.51%) new bone formation in the HA implant. In the middle of the implants allograft produced 11.01% (± 2.07%) new bone whilst the HA produced 7.23% (± 4.05%) new bone. Two tailed t-test showed no significance in either region, p=0.28 at the interface and p=0.40 in the middle. Allograft underwent resorption, from 39.37% at time zero to 5.66% (± 2.04%) at 6 weeks, a total reduction of 85%, where as the volume of HA granules remained the same and was 49% at time zero and 48.59% (± 1.69%) at 6 weeks. Two tailed t-test showed a significant difference (p< 0.0001) between allograft and HA at 6 weeks.

Conclusions: This study shows that granular porous HA induced a similar level of bone formation as compared with allograft. Resorption of allograft in this model allowed greater ingrowth of fibrous tissue. This makes the structural scaffold much more porous, compromising stability of the construct. The HA was not resorbed after 6 weeks and hence may be more stable. HA also has the advantage of being readily available. This study demonstrates that a bone substitute material does not need to be mixed with allograft.

Introduction: Wear particle induced osteolysis is one of the main reasons for revision total hip replacements (THRs). Loss in bone stock as a result of aseptic loosening is responsible for inferior results in revision THRs. Results from impaction grafting to fill osteolytic defects are frequently inconsistent. Our hypothesis is that the combination of autologous mesenchymal stem cells (MSCs) and allograft will enhance bone regeneration. This study asks whether: MSCs with allograft scaffolds survive at a normal impaction force during revision THRs.

Method: MSCs were isolated from a sheep iliac crest aspirate, expanded in culture and seeded onto irradiated sheep allografts (n=9). Viability of MSCs was assayed with alamar blue with absorbance measured on day 4 (before impaction). The constructs were then impacted using forces 3, 6, and 9 kN extrapolated in surgery then assayed daily for 6 days. The control was 0 kN. Samples were resin embedded after 10 days for histology and pieces of graft were taken for scanning electron microscopy (SEM).

Results: The 0KN control shows an MSC growth curve with a lag period and log phase. Compared with the control, the 3 and 6 kN showed initial reduction in cell proliferation measured by alamar blue (^p=0.015, ^p=0.002) but recovered by day 8, while 9kN showed a significant reduction (^p=0.011) over the time (Figure 1).

For cell proliferation over time, 3 and 6 kN showed no differences, but 9 kN showed a significant difference between day 4 and day 8 (^p=0.031). SEM and histological analysis showed a network of cuboidal cells on the allograft surface.

Conclusions: The results showed that MSCs recovered from impaction of 3 and 6 kN after an initial reduction in metabolism and exceeded original cell seeding densities with no significant difference in proliferation. Viability of MSCs were not effected by impaction forces up to 6 kN. This study shows that stem cells mixed with allograft are a potential method for repairing bone defects in revision total hip replacements.