Objectives

Cortical and cancellous bone healing processes appear to be histologically different. They also respond differently to anti-inflammatory agents. We investigated whether the leucocyte composition on days 3 and 5 after cortical and cancellous injuries to bone was different, and compared changes over time using day 3 as the baseline.

Aims

Post-operative migration of cemented acetabular components as measured by radiostereometric analysis (RSA) has a strong predictive power for late, aseptic loosening. Also, radiolucent lines predict late loosening. Migration has been reduced by systemic bisphosphonate treatment in randomised trials of hip and knee arthroplasty. Used as a local treatment, a higher local dose of bisphosphonate can be achieved without systemic exposure. We wished to see if this principle could be applied usefully in total hip arthroplasty (THA).

Objectives

Healing in cancellous metaphyseal bone might be different from midshaft fracture healing due to different access to mesenchymal stem cells, and because metaphyseal bone often heals without a cartilaginous phase. Inflammation plays an important role in the healing of a shaft fracture, but if metaphyseal injury is different, it is important to clarify if the role of inflammation is also different. The biology of fracture healing is also influenced by the degree of mechanical stability. It is unclear if inflammation interacts with stability-related factors.

Summary

The negative impact of NSAIDs on fracture healing appears not to pertain to fractures in cancellous bone. Possibly this is because of a higher prevalence of MSCs in cancellous bone, making recruitment of distant cells via inflammatory signals less important.

Summary Statement

Atypical femoral fractures consist of a thin fracture line extending through the lateral cortex. The adjacent bone is undergoing resorption and mechanical abrasion and is often replaced with woven bone. The mechanical environment seems to inhibit healing.

Summary

These data suggest that PTH treatment for stimulation of bone healing after trauma is not much dependent on mechanical stimulation and therefore, roughly equal treatment effects might be expected in the upper and lower extremities in humans.

Introduction

Traumatized musculoskeletal tissue often exhibits prolonged time to healing, mostly due to low blood flow and innervation. Intermittent Pneumatic Compression (IPC) increases blood flow and decreases thromboembolic event after orthopedic surgery,[1] however little is known about healing effects.[2] We hypothesized that IPC could stimulate tissue repair: 1.) blood flow 2.) nerve ingrowth 3.) tissue proliferation and during immobilisation enhance 4.) biomechanical tissue properties.

Introduction: As Achilles tendon ruptures are healing slowly, many attempts are made trying to improve the healing after injury. Rat experiments have shown that injection of platelets improves tendon healing. A clinical study on patients has also shown a better outcome after injecting platelets into the rupture area. Therefore we wanted to verify the effect of platelets by measuring the mechanical properties of the healing Achilles tendon in a randomised study.

Methods: We included 30 patients with an Achilles tendon rupture. All patients left one blood unit at the hospital blood bank. From this unit of blood approximately 20 ml of platelet concentrate were gained. All patients were operated the next day using an open technique. Just before wound closure, patients were randomised into 2 groups, with one group receiving 10 ml of their own platelet concentrate. In order to measure mechanical properties, we implanted Tantalum beads on either side of the rupture giving us the chance of exactly determining the distance between the beads using RSA and thereby measuring the stiffness of the tendon. CT was used to measure the area of the rupture site. Both groups were postoperatively treated with a cast for 7 weeks. 4 weeks with the ankle in the equines position and 3 weeks in the neutral position. After cast removal, the patients started rehabilitation. The patients in both groups received exactly the same treatment after surgery. Patients were examined with CT and RSA to determine area, stiffness and modulus of elasticity. Measurements were performed at 7 weeks after operation and again at 19 weeks.

Results: 16 patients were randomised to platelet concentrate. One patient got a deep infection and another patient suffered from a rerupture of the Achilles tendon. Both patients had to be excluded after the first CT- and RSA-examination. Both patients had received platelet concentrate. There was no significant difference between both groups after 7 and 19 weeks in area, stiffness or modulus of elasticity.

Discussion: Our results indicate that platelet concentrate does not improve the mechanical properties of the healing Achilles tendon, at least when patients are treated with a cast for 7 weeks.

Studies have shown that the effect of platelet concentrate is depending on a certain loading of the Achilles tendon during the early phase of healing. Unfortunately we do not know much about loading of the Achilles tendon while having the leg in a cast, but the rigid fixation might lead to certain unloading. Further studies are needed to learn more about loading of the Achilles tendon in a cast.

Furthermore we have also done a clinical examination of all patients, as we know from a previous study that there is a correlation between early mechanical properties and clinical outcome but we have not evaluated the clinical results of this study yet.

Healing of tendons is sensitive to mechanical loading, and the callus strength is reduced by ¾ after 14 days, if loading is prevented. Exogenous GDFs stimulate tendon healing. This response is influenced by loading: without loading, cartilage and bone formation is initiated. This suggests that BMP signalling is crucial during tendon healing, and that it is influenced by mechanical loading. We investigated if mechanical loading influences BMP signalling in intact and healing tendons, and how BMP gene expression changes during healing.

The Achilles tendon was transected in rats and left to heal. Half of the rats had one Achilles tendon unloaded by injection of Botox in the calf muscles. Ten tendons were analyzed before transection and for each of four time points. Gene expression for OP-1, GDF-5, -6, -7, Follistatin, Noggin, BMP-receptor 1b and BMP-receptor 2 were analysed with real-time PCR.

Loading had no detectable effects on intact tendons. During repair, loading decreased follistatin by more than half (p=0.0001), and increased GDF-5 (p=0.02). All genes showed changes during repair (p=0.0001), but the time sequences differed. GDF-5 and GDF-7 were generally more expressed than OP-1 and GDF-6. GDF-5 and GDF-7 were more expressed in normal tendons than during repair. Noggin was never detected.

Our results suggest that GDF-5 is specific for the mature tendon, and not much involved in repair. This contrasts to GDF-7, which is involved in both. OP-1 and GDF-6 seem to be involved in early healing. There was less expression of follistatin in loaded tendons during healing. The mechanosensitivity is likely of most importance at day 14 and 21 since the difference in strength between loaded and unloaded tendons is huge. An Anova with only these time points reveals effects of loading on GDF-5 and follistatin (p=0.0001 for both) and significant differences between the days for most variables.

Local dysregulation of the proteolytic matrix metalloproteinases (MMPs) and their tissue inhibitors of metalloproteinases (TIMPs) is a feature of tendon degeneration and rupture.^1,2 To assess the role of systemic MMPs and TIMPs in tendon rupture we compared serum MMPs and TIMPs between patients who have previously suffered Achilles tendon rupture and healthy controls. We also followed serum MMPs and TIMPs prospectively in patients with acute tendon rupture.

At three years after injury, we measured serum MMP-1, -2, -3, -7, -8, -9 and -13 and TIMP-1 and -2 in eight patients who had suffered Achilles tendon rupture. Serum was also obtained from 12 blood donors with similar age and sex distribution. In another eight patients, MMPs and TIMPs were followed over time, with samples taken at the time of Achilles tendon injury, and after 4, 8 and 24 weeks. MMPs were determined using Fluorokine Multi Analyte Profiling kits while TIMPs were analysed using ELISA (R& D systems). The study was approved by the ethics committee and written informed consent was obtained from all patients.

Patients who had previously suffered tendon rupture had increased levels of MMP-2 (median difference (m.d.) 10 %; p = 0.01), MMP-7 (m.d. 15 %; p = 0.02) and TIMP-2 (m.d. 36%; p = 0.02), as compared to controls. In patients with acute tendon rupture, MMP-2 was the only MMP or TIMP to change significantly over time (p = 0.009). MMP-7 appeared to be higher than control values already at the time of rupture. MMP-13 could not be detected in any sample.

In conclusion, patients with a history of tendon rupture had elevated serum levels of MMP-2, MMP-7 and TIMP-2. Changes in MMP-7 might be present already at the time of rupture. This suggests that disturbances in proteolytic control might render tendons prone to rupture.

Introduction: Many claim that an inflammatory reaction to wear debris particles is the main cause for prosthetic loosening. We have rat model in which bone resorption can be induced both by fluid pressure and particles. We compared the bone resorptive potency of particles and fluid pressure.

Materials and Methods: The rats received a titanium plate at the proximal tibia. A central plug was inserted. After 5 weeks of osseointegration, the central plug was changed to a piston or a hollow plug with 20mg titanium particles. Commercially pure titanium particles with 90 % of particles lesser than 3,6 microns were used. The pressure piston was subjected to a transcutanous force of 5N. Each episode of pressure comprised 20 pressure cycles at 0.17 Hz, applied twice a day. 60 rats were randomized to 6 groups for particle treatment. One group had particle implantation both at day 1 and 7. Additionally 15 rats were randomized into 3 groups with for pressure treatment. All rats were killed after 5 or 14 days. Bone resorption underneath the piston was evaluated blindedly in hematoxlyin/eosine sections and scored as 0 – 5. Differences between groups were analyzed by Kruskal Wallis and Mann-Whitney U-tests.

Results: Many specimens with titanium particles showed no visible resorption at al, and a few showed dramatic osteolysis. After 14 days, the osteolytic effect was significant. Partice refill made no difference. Titanium particles over 10 μm had minor effect. Fluid pressure always caused bone resorption, and significantly more so than particles both after 5 and 14 days.

Discussion: There was lesser variability in fluid pressure induced osteolysis, which might be due to a different signalling pathway. Titanium particles have an osteolytic effect in this model, but in spite of the massive amount of particles, the effect was less predictable than with pressure.

Background: RSA cannot discern whether a single prosthesis is fixed or migrating below the detection level. Samples of patients usually show migration values that appear to be continuously distributed. Is there a dichotomy between stable and migrating prostheses?

Methods: We analysed the migration of 147 cemented acetabular cups of 7 different designs, by use of a new set of algoritms for frequency distribution analysis called Rmix. The migration vector lengths were assumed to be a compound of log-normal distributions. The algoritm then calculated if the observed frequency distribution is best explained by one or more log-normal distributions.

Results: After 2 years there was a significant dichotomy (p=0.006) between 2 lognormal subgroups within the sample. Neither cup design, sex or operating department could explain the dichotomy into two groups, which appears to reflect the existence of two different types of behaviour. The migration along the 3 axes in space, showed a similar dichotomy. During the second year, around 80 % of the patients belonged to a distinct, normally distributed subgroup with a mean not different from 0 mm and a small variation, corresponding to the measuring error. The remainder differed significantly from this subgroup and showed migration.

Interpretation: The majority of the cups belonged to a subpopulation that appeared completely stableduring the second year. For a single type of prosthesis, the relative size of the stable subgroup might be a good index of the expected performance.

Introduction: In a rat model, fluid pressure causes more bone resorption than particles. Does pressure also cause more inflammation?

Materials and Methods: Rats received a titanium plate at the proximal tibia. A central plug was inserted. After 5 weeks of osseointegration, the central plug was changed to either a piston or a hollow plug with titanium particles. Commercially pure titanium particles with 90% of particles lesser than 3,6 microns were used. The pressure piston was subjected to a transcutanous force of 8N. Each episode of pressure comprised 20 pressure cycles at 0.17 Hz, applied twice a day. 39 rats were randomized to 3 groups: Titanium particles (n=13), fluid pressure (n=13) and controls with neither particles nor fluid pressure (n=13). The rats were killed after 3 days. 6 rats in each group were used for histology and the others for gene expression. Extraction of total RNA was performed using the TRIspin method. Primers for cat K, RANK, RANKL, OPG IL-1, IL-b, TNF-a, iNOS and COX-2 were used. Each sample was normalized to 18S rRNA. Histology was evaluated qualitatively. Differences between the groups were analyzed by Kruskal Wallis and Mann-Whitney U-test.

Results: Both particles and fluid pressure increased the expression of osteoclastic genes. Particles induced an elevated expression of IL-6 and RANK compared to both controls and fluid pressure. There was a tendency that particles induced more expression of other inflammatory genes compared to fluid pressure.

Histology: The controls showed only few osteoclasts at the bone surface. The particle group showed osteoclasts at the surface towards the particles. In contrast, the pressure group showed resorption cavities spread out inside the bone.

Discussion: Although there was more resorption in the pressure group, there was a lesser inflammatory response. This suggests that pressure-induced resorption is mediated via different pathways.

Background: In a previous randomized studiy using Röntgen Stereometric Analysis (RSA), we showed that oral bisphosphonates reduce the mean migration distance during the first 6 months. In a similar randomized study, bisphosphonates applied locally at the operation had a similar effect. These studies showed a 0.1 mm difference in mean value between groups. Does such a small difference matter? We addressed this question by use of frequency analysis.

Methods: The 2 previous studies were combined for analysis, and designated as bisphosphonate (n=44) or control treated (n=49). We analysed the migration vector (for the center of the rigid body) by use of a set of algoritms for frequency distribution analysis called Rmix. The migration vector lengths were assumed to be a compound of log-normal distributions. The frequency analysis determined if the observed frequency distributions were best described as a single, or a sum of 2 or more lognormally distributed subgroups.

Results: After 6 months, the control patients had formed 2 subgroups, one comprising 85% of the patients. The dichotomy was significant (p=0.016).

After 2 years, the dichotomy persisted (p=0.027). In the bisphosphonate-treated patients, no dichotomies could be found. The distribution of the migration vector length appeared similar to the larger and less migrating subgroup among the controls.

Discussion: The risk of aseptic loosening for cemented knees is extremely small. However, the migrating subgroup among our control patients may be at risk of loosening, and would have run a high risk if they were young and active. This subgroup did not appear with bisphosphonate treatment

Summary: In previous comparisons we found a slight decrease in mean value with bisphosphonates. The present analysis shows that this reflects the disappearance of a small subgroup with large migration.

Soaking bone grafts in a bisphosphonate solution before implantation can prevent their resorption and increase the local bone density in rats and humans. However, recent studies suggest that pre-treatment of allografts with bisphosphonate can prevent bone ingrowth into impaction grafts. We tested the hypothesis that excessive amounts of bisphosphonate would also cause a negative response in less dense grafts. We used a model where non-impacted metaphyseal bone grafts were randomised into three groups with either no bisphosphonate, alendronate followed by rinsing, and alendronate without subsequent rinsing, and inserted into bone chambers in rats. The specimens were evaluated histologically at one week, and by histomorphometry and radiology at four weeks. At four weeks, both bisphosphonate groups showed an increase in the total bone content, increased newly formed bone, and higher radiodensity than the controls. In spite of being implanted in a chamber with a limited opportunity to diffuse, even an excessive amount of bisphosphonate improved the outcome. We suggest that the negative results seen by others could be due to the combination of densely compacted bone and a bisphosphonate.

We suggest that bisphosphonates are likely to have a negative influence where resorption is a prerequisite to create space for new bone ingrowth.

We have examined the deterioration of implant fixation after withdrawal of parathyroid hormone (PTH) in rats. First, the pull-out force for stainless-steel screws in the proximal tibia was measured at different times after withdrawal. The stimulatory effect of PTH on fixation was lost after 16 days. We then studied whether bisphosphonates could block this withdrawal effect. Mechanical and histomorphometric measurements were conducted for five weeks after implantation. Subcutaneous injections were given daily. Specimens treated with either PTH or saline during the first two weeks showed no difference in the mechanical or histological results (pull-out force 76 N vs 81 N; bone volume density 19% vs 20%). Treatment with PTH for two weeks followed by pamidronate almost doubled the pull-out force (152 N; p < 0.001) and the bone volume density (37%; ANOVA, p < 0.001). Pamidronate alone did not have this effect (89 N and 25%, respectively). Thus, the deterioration can be blocked by bisphosphonates. The clinical implications are discussed.

Thrombin has many biological properties similar to those of growth factors. In a previous study, we showed that thrombin improves healing of the rat tendo Achillis. Low molecular weight heparin (LMWH) inhibits the activity and the generation of thrombin. We therefore considered that LMWH at a thromboprophylactic dose might inhibit tendon repair.

Transection of the tendo Achillis was carried out in 86 rats and the healing tested mechanically. Low molecular weight heparin (dalateparin) was either injected a few minutes before the operation and then given continuously with an osmotic mini pump for seven days, or given as one injection before the operation. In another experiment ,we gave LMWH or a placebo by injection twice daily. The anti-factor Xa activity was analysed.

Continuous treatment with LMWH impaired tendon healing. After seven days, this treatment caused a 33% reduction in force at failure, a 20% reduction in stiffness and a 67% reduction in energy uptake. However, if injected twice daily, LMWH had no effect on tendon healing. Anti-factor Xa activity was increased by LMWH treatment, but was normal between intermittent injections.

Low molecular weight heparin delays tendon repair if given continuously, but not if injected intermittently, probably because the anti-factor Xa activity between injections returns to normal, allowing sufficient thrombin stimulation for repair. These findings indicate the need for caution in the assessment of long-acting thrombin and factor Xa inhibitors.

During revision total hip replacement using morcellised compacted bone allograft, 16 patients were randomised to receive a graft which had been rinsed in either an ibandronate solution or in saline. Patients were assessed by dual energy x-ray absorptiometry after operation and at 3, 6, 12 and 24 months. A region of interest between the tip of the femoral stem and the distal plastic plug was chosen to measure the changes in bone density over time. The study was double-blinded. In all the control patients the bone density decreased during the first three months and then remained constant at this lower level. A large proportion of the mass of the bone graft was lost. In contrast, all patients with grafts treated with bisphosphonate showed a slight increase in bone density. The difference between the groups was highly significant at all points in time.

We conclude that rinsing the graft in a bisphosphonate solution prevents its resorption and may therefore reduce the risk of mechanical failure. The treatment is simple, inexpensive, and appears virtually free of risk.

The aim of our study was to evaluate if PTH is able to increase the trabecular density of osteoporotic bone at the site of an implant and whether the anabolic effect of PTH at this side is stronger then the effect of an osteoclast inhibitor like alendronate.

48 cement rod was inserted in the tibia of 48 female rats, of which 36 had been ovariectomized. The cement rods, which served as implants, were made of Palacos R bone cement. After implantation, the 36 ovariectomized rats were divided in 3 groups. One was injected subcutaneusly with PTH (1–34) at a dose of 60 g/kg BW. The second was injected with alendronate at a dose of 205 g/kg BW. The third with vehicle only. The remaining 12 sham operated rats were also injected with vehicle only. All injections were given three times a week and the rats were killed 2 weeks after implantation.

The tibial segments around the hole of the rods were prepared histologically. Thus the surfaces which had been in contact with the rod appeared as straight lines and could be analyzed histomorphometricly. The trabecular density of the bone closest to the implant was measured. One femur of all animals was used for measurement by DEXA.

There was a substantial increase in the trabecular density close to the rods with PTH treatment (Anova p=0.002). PTH lead to a trabecular density of 89%, where as the ovariectomized animals revealed a trabecular density of 58% and the sham operated control of 68%. No significant increase of implant related trabecular density could be found in the alendronate treated group. In this group a density of 72% was established. DEXA showed the expected differences in bone mineral content (Anova p=0.001).

In this study, intermittent PTH treatment increased implant-related trabecular density in osteoporotic bone after 2 weeks. No such positive effect could be found with alendronate treatment at such a short period of time. We think the reason for this phenomenon could be the early onset of the anabolic PTH effect on regenerating bone, whereas alendronate is thought to only inhibit bone resorption, which might lead to a later effect.

The early onset of PTH effects even in osteoporotic bone suggests that intermittent PTH treatment might lead to an increased micro-interlock between implant and bone and might therefore be considered as a possible drug to enhance incorporation of orthopedic implants.

Aims: To compare total hip arthroplasty (THA) and internal fixation in the treatment of displaced femoral neck fractures. Methods: Patients, 75 years or older, including those with mental impairment, were randomized to either internal fixation or cemented primary total THA. A total of 146 hips in 143 patients were followed for two years. After one year 23% had died, and after two years 29%. Mortality was about the same in both groups. The accumulated mortality was pronounced among the mentally impaired patients. Results: In the internal fixation group, 44% underwent further surgery. In the THA group, 18% dislocated. The dislocation rate was higher for the mentally impaired patients. The Harris hip scores were higher in the THA group. Pain was more common in the internal fixation group.

A subgroup of 100 patients was included in a study concerning nutritional status and functional capacity. The THA group fared better concerning weight change over time, locomotion and pain. The nutritional intervention did not show any measurable effects.

All patients were followed until two years postoperatively and all fracture-related hospital costs, including reoperations, were calculated. We found no difference in total costs between the treatment groups. Costs to the municipality were calculated comparing the baseline cost before surgery with the average cost per month during the first postoperative year. No difference was found between the treatment groups. Conclusions: On the basis of these results, we recommend arthroplasty for patients in this age group with normal mental function and high functional demands. When in doubt, an arthroplasty should be generously considered.