Summary

Our statistical shape analysis showed that size is the primary geometrical variation factor in the medial meniscus. Shape variations are primarily focused in the posterior horn, suggesting that these variations could influence cartilage contact pressures.

INTRODUCTION

Electron beam melting is a promising technique to produce surface structures for cementless implants. Biomimetic apatite coatings can be used to enhance bone ingrowth. The goal of this study was to evaluate bone ingrowth of an E-beam produced structure with biomimetic coating and compare this to an uncoated structure and a conventionally made implant surface.

Introduction

The different types of treatment for osteonecrosis of the femoral head have not led to a consensus about which treatment is best for the different stages. Particularly in the later stages of osteonecrosis, the disease still progresses to destruction of the femoral dome. The purpose of our study was to check the outcome of bone impaction grafting used for the head-preserving treatment of severe femoral head osteonecrosis. In order to preserve the femoral head, the sphericity and mechanical properties of the femoral dome must be contained and further collapse prevented.

Bone impaction grafting (BIG) is a surgical technique for the restoration of bone stock loss with impaction of autograft or allograft bone particles (BoP). The goal of a series in-vitro and in-vivo experiments was to assess the suitability of deformable pure Ti (titanium) particles (TiP, FONDEL MEDICAL BV, Rotterdam, The Netherlands) for application as a full bone graft substitute in cemented revision total hip arthroplasty. TiP are highly porous (interconnective porosity before impaction 85 to 90%). In-vitro acetabular reconstructions were made in Sawbones (SAWBONES EUROPE, Malmö, Sweden) to evaluate migration by roentgen stereo photogrammetric analysis and shear force resistance by a lever out experiment. In-vitro femoral TiP reconstructions (SAWBONES, Malmö, Sweden) were used to evaluate micro-particle release and subsidence. Mature Dutch milk goats were used for two in-vivo experiments.

A non-loaded femoral defect model was used to compare osteoconduction of bioceramic coated TiP with BoP and ceramic particles (CeP).

Blood samples were taken for toxicological analysis.

In-vitro: TiP were as deformable as BoP and created an entangled graft layer (porosity after impaction 70 to 75%). Acetabular TiP reconstructions were more stable and resistant to subsidence and shear force than BoP reconstructions (lever-out moment 56 ± 12 Nm respectively 12 ± 4 Nm, p < 0.001). After initial setting, femoral subsidence rates were smaller than seen in femoral bone impaction grafting (0.45 ± 0.04 mm after 300 000 loading cycles). Impaction generated 1.3 mg particles/g TiP (particle Ø 0.7–2 000 μm, tri-modal size distribution). In-vivo: Bioceramic coated (10 −40 μm) TiP showed bone ingrowth rates comparable to BoP and CeP. Reconstructed acetabular defects showed rapid bone ingrowth into the layer of TiP. Serum titanium concentrations slowly increased from 0.60 ± 0.28 parts per billion (ppb) preoperatively to 1.06 ± 0.70 ppb at fifteen weeks postoperatively (p = 0.04).

Mechanical studies showed very good initial mechanical properties of TiP reconstructed defects. The in-vitro study showed micro-particle generation, but in the short-term goat studies, histology showed very few particles and no negative biological effects were found. The in-vivo acetabular study showed very favorable bone ingrowth characteristics into the TiP layer and a much thinner interface with the cement layer compared to similar defects reconstructed with BoP or mixtures of BoP with CeP. Further analysis in a human pilot study should proof that TiP is an attractive and safe alternative for allograft bone in impaction grafting revision arthroplasty.

Introduction: The effect of different temperatures to antibiotics is unknown. What is the dose-response curve of bone chips impregnated with different kinds of antibiotics?

Material and Methods: Five different antibiotic pills and solutions (cefazolin, clindamycin, linezolid, oxacillin, vancomycin) were stored at −80°C, −20°C, 4°C, 20°C and 37°C. Also, bone chips were impregnated with cefazolin and vancomycin solution and were stored at −80°C and −20°C. After 1 month, 6 months and 1 year, reaction of the antibiotics to Staphylococcus epidermidis was measured using an inoculated iso-agar. Activity of the antibiotics was measured as the diameter of the Staphylococcus epidermidis-free zone.

Also, five cefazolin and vancomycin solutions were used to impregnate bone chips and to make dose-response curves. Furthermore, 1 gram bone chips was impregnated with 5ml cefazolin or 5ml vancomycin solution.

Results: A decrease of the diameter free zone Staphylococcus epidermidis was seen when oxacillin and cefazolin solutions were stored at 37°C for 1 month and when vancomycin was stored for 6 months (37°C). Also, when cefazolin and oxacillin solutions were stored at 20°C, a decrease was noticed. The storage of other antibiotic solutions, pills and bone/antibiotics composite showed no differences in reaction. Dose-response curves show that with increasing antibiotic quantity, diameter free zone increases according to a logarithmic function.

Conclusion and discussion: With the dose-response curve the optimal concentration(s) for local application can be determined. It gives the opportunity to determine the amount of antibiotics present in the patient locally. Freezing of antibiotics does not affect the activity of the investigated antibiotics.

Meniscectomy, induces osteoarthritis. Options for repair of a damaged meniscus are an allograft meniscus, an implant made of natural scaffold materials (the collagen meniscus implant; CMI) or an implant made of polymers.

Allograft menisci and the CMI are already clinically used for a considerably number of years. In this educational lecture the focus is on a comparison between the three implant types and the status of a tissue-engineered meniscus.

The allograft meniscus is already used for at least ten years. It is intended for the younger patient with a previous total meniscectomy, with moderate cartilage degeneration and with a good alignment of the knee. The clinical outcome is based on function and pain scores. In this lecture the functional scores, the survival rate and the histology of allograft menisci will be highlighted.

The CMI meniscus implant is intended for a different patient group. To enable implantation of the CMI the rim of the native meniscus should be intact. Patient series that should demonstrate the efficacy of this type of implant are still small and are mainly of the inventors of the implant. In general patients tolerated the implant well. Tissue ingrowth and remodelling into a fibro-cartilaginous tissue was found in animals and patients.

Polymers may be a good alternative for the allograft and CMI implant. Previously they were used to guide vascularized new repair tissue through an ingrowth channel to the avascular lesion. We developed a porous polymer meniscus scaffold with properties to allow tissue infiltration and regeneration of a neomeniscus. It was implanted in dog knees and compared with total meniscectomy. The tissue infiltration and redifferentiation in the scaffold, the stiffness of the scaffold, and the articular cartilage degeneration were evaluated.

Three months after implantation, the implant was completely filled with fibrovascular tissue. After 6 months, the central areas of the implant contained cartilage-like tissue with abundant collagen type II and proteoglycans in their matrix. The foreign-body reaction remained limited to a few giant cells in the implant. The compression modulus of the implant-tissue construct still differed significantly from that of the native meniscus, even at 6 months. Cartilage degeneration was observed both in the meniscectomy group and in the implant group.

The improved properties of these polymer implants resulted in a faster tissue infiltration and in phenotypical differentiation into tissue resembling that of the native meniscus. However, the material characteristics of the implant need to be improved to prevent degeneration of the articular cartilage.

We investigated the feasibility of using porous titanium particles (TiP) to reconstruct femoral bone defects in revision hip replacement surgery in stead of using morzelised bone grafts. Questions regarding handling, initial stability and titanium particle release were addressed.

Seven composite femurs (Sawbones) were reamed and filled, stepwise, with 32 grams of large (Ø 3.15 – 4 mm) and 9 grams of smaller (Ø 2.8 – 3.15 mm) pure, 85% porous TiP. Subsequently an Exeter stem was cemented into the graft layer. All reconstructions were loaded axially (0–3000 N) for 300,000 loading cycles at 2 Hz. Subsidence of the stem was measured with radio stereometric analysis (RSA) and possible titanium particle release was measured using the laser diffraction technique.

The TiP were impacted into a > 3 mm (SD 1.43 mm) thick, highly entangled, graft layer. An average cement mantle of > 2 mm (SD 0.86 mm) was measured and little cement penetration was observed. The average subsidence of only 0.45 mm (SD 0.04 mm) was measured after 300 000 loading cycles. Most titanium particles were found directly after impaction. Most of these particles (87%) were smaller than 10 μm and could therefore be potentially harmful since they can induce osteolysis.

We can conclude that:

A graft layer of impacted TiP can be constructed,

These data warrant further animal tests to assess the biological response to these released impaction particles. Also, animal tests should clarify possible particle release upon loading and its effects.

The clinical application of bone morphogenetic proteins (BMPs) offers solutions to many challenging problems in orthopaedics. However, a practical clinical problem is to obtain a controlled release of the BMPs. The attachment of heparin to biomaterials may result in an appropriate matrix for the binding, and sustained release of BMPs. Binding of growth factors to heparin stabilizes these growth factors, protects them from proteolytic degradation, and prolongs the half-life of BMPs in culture media 20-fold. We created a carrier based delivery system with a localized sustained release by loading a tricalciumphosphate/hydroxyapatite (TCP/HA) bone substitute coated with cross-linked collagen and heparin, with BMP-7.

TCP/HA granules (BoneSave^™, Stryker Orthopaedics) were coated with collagen, and subsequently the collagen was cross-linked in the presence (TCP/HA-Col-Hep) and absence (TCP/HA-Col) of heparin. BMP-7 was loaded onto the coated TCP/HA granules. Morphology of the coated collagen with and without heparin, and release kinetics of BMP-7 from the granules were analyzed. TCP/HA granules without coating were used as controls.

Analysis showed a highly porous collagen network on both TCP/HA-Col and TCP/HA-Col-Hep granules. Immersion of the granules in BMP-7 solution, resulted in the binding of 54±3% (62.9±5.4 ng BMP-7/mg granule) to the TCP/HA granules, 64±8% (69.0±9.6 ng BMP-7/mg granule) to the TCP/HA-Col granules, and 78±1% (92.9±4.8 ng BMP-7/mg granule) to the TCP/HA-Col-Hep granules. TCP/HA granules showed a burst release of BMP-7 within the first 4 h. TCP/HA-Col granules showed an initial burst release, followed by a more gradual release. In contrast, BMP-7 release from the TCP/HA-Col-Hep granules was sustained up to 21 days.

The sustained delivery system for BMP-7 developed in this study may provide a powerful tool for bone regeneration. This system could probably also be applied to deliver multiple growth factors that have affinities for heparin, which could for instance synergistically enhance osteogenesis by increasing vascularity.

Introduction : The reason why heterotopic ossification develops after total hip arthroplasty is still not known, but it is assumed that inflammatory reaction is the major driving force. In literature little is known about the cytokine levels at the site of surgery, most measurements are done in serum. This study was conducted to investigate if the levels of different pro- en anti-inflammatory cytokines are measurable in drainage fluid and, when measurable, whether we can find a difference in cytokine concentration between one and six hours postoperatively.

Materials and methods: Samples from the drainage system in 30 consecutive patients undergoing primairy total hip replacement were collected at one and six hours after closure of the wound. GM-CSF, G-CSF, IFN-γ, TNF, MCP-1, IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13 and MIP-1beta levels were measured in the drainage fluids.

Results: Measurable levels of all cytokines studied were found, except for IL-17. A significant elevation of almost all cytokines was observed between the sample after one hour and six hours postoperatively. The elevation was significant for all cytokines except IL-10 and MIP-1b. We found a strong correlation between the different pro-inflammatory cytokines. Levels are much higher than previously shown levels in serum. When computing the IL-6 to IL-10 ratio, this ratio increased from 304 (SD 256) to 12357 (SD 6788) (p< 0,000), which shows an increased predominance of the pro-inflammatory interleukines when comparing the measurements after one and six hours respectively.

Conclusion: Detectable levels of numerous cytokines can be measured in drainage fluid post-operatively. The levels of most cytokines in drainage fluid are higher in samples taken six hours after surgery as compared to samples taken after one hour. Further studies are needed to detect the relation between these cytokine concentrations and the heterotopic bone formation.

Introduction: The purpose of this study was to evaluate the long-term clinical and radiological outcome of instrumented femoral revisions after failed total hip arthroplasties using the impaction bone grafting technique with morsellized bone chips in combination with a cemented polished stem.

Methods: Thirty-three consecutive femoral reconstructions were performed between November 1991 and February 1996 using the X-Change femoral impaction system with fresh frozen morsellized bone grafts and a cemented polished Exeter stem. All patients were prospectively followed. The learning curve with this new technique is included in this report. This technique was used in twenty-four women and nine men; the average age at surgery was sixty-three years (range 33–82). Femoral bone stock defects were classified according to the Endoklinik classification as grade 1 in three hips, grade 2 in fourteen hips, grade 3 in twelve hips and grade 4 in four hips. At a minimal eight years follow-up no patient was lost to follow-up, but eight patients died (at 0.5, 3.5, 3.5, 7.0, 7.0, 7.5 and 9.0 after reconstruction). All were followed until death, none of these deaths was related to the surgery, and none had a re-revision

Results: No femoral reconstruction was re-rerevised at a mean follow-up of 10.4 years (range 8 to 13 years). However, there were three femoral fractures during follow-up (at 3, 6 and 22 months), all at the level of tip of the prostheses. All healed after plating, all femoral implants were left in situ. The average Harris hip score improved from 49 prior to surgery to 85 at review (68–100). The average migration of the stem within the cement mantle was three mm (0–14 mm), most migration was seen in the first year. Radiologically, there were no failures. With an endpoint of femoral revision for any reason, with endpoint aseptic loosening or with endpoint radiological loosening the survival rate using the Kaplan-Meier analysis was hundred per cent in all situations (one-sided 95% C.I. 100–91.3 %)

Conclusions Femoral revision using bone impaction grafting with fresh frozen bone grafts and a cemented polished stem showed an excellent survival at eight to thirteen years follow-up.

Bone impaction grafting of the femur is associated with more complications when segmental defects are present. The effect of segmental defect repair on initial stem stability was studied in an in vitro study with fresh frozen goat femora. A standardized medial segmental defect was reconstructed using a cortical strut or a metal mesh. As controls we used intact femora and femora with a non-reconstructed defect. In all four groups impacted bone grafting was performed in combination with a cemented Exeter stem. Each group contained five femora. Reconstructions were dynamically loaded up to 1500N. Migration was measured with Roentgen Stereo-photogrammetric Analysis. All cases with a non-reconstructed segmental defect failed into excessive varus rotation. None of the femora with a reconstructed defect failed. Cortical struts and metal meshes were equally effective in creating a stable stem construction (varus rotation 2.89±2.27 and 2.27±0.57, respectively). Reconstructions with a metal mesh were more reproducible, although the obtained stability was significantly lower (p< 0.01) when compared to impaction grafting in an intact femur (varus rotation 0.58±0.36).

Besides, structural grafts may negatively influence the revascularization of the underlying impacted grafts in contrast to an open wire mesh. So, an in vivo study of 12 goats was done. A standardized medial wall defect was reconstructed with a strut or a mesh in six goats per group. In all femora impaction grafting was performed in combination with a cemented Exeter stem. After six weeks the femora were harvested. A high rate of peri-prosthetic fractures was found (43% and 29% for the strut and mesh groups, respectively). Histological and micro-radiological examination showed different revascularization patterns for both reconstruction techniques. In the strut group revascularized graft was found at the edges of the defect. In the mesh group fibrous tissue and blood vessels penetrated through the mesh and a superficial zone of revascularized grafts was found. Segmental defect reconstruction with a strut reduced the amount of revascularized grafts medially behind the strut (p=0.004). This may interfere with the stability of the stem in the first period after surgery and the incorporation of the impacted grafts on the long-term.

We would recommend segmental defect reconstruction with a mesh. A regime of unloading and long-stem prostheses should be used, irrespective of the reconstruction technique

Aims: To test the outcome of acetabular revisons with impacted morsellized bone grafts and a cemented cup at a minimum follow-up of 15 years. Methods: Between 1979–1986 62 acetabular revisions (58 patients) were performed with impacted bone grafts and a cemented cup. Two cases (2 hips) were lost to FU. The average age at operation was 59.1 years. The indication was aseptic in 56 and septic loosening in 4 hips. Defects were AAOS-classiþed as ñcavitaryñ in 37 and ñcombinedñ 23 cases. Femoral-head allografts were used. Acetabular re-revisions were determined, the HHS obtained and radiographs analyzed. Results: Nineteen cases (20 hips) died within þfteen years after the operation, none had a re-revision. Eleven acetabular re-revisions were performed: 2 septic loosenings (3 and 6 years p.o.), 7 aseptic loosenings (6, 9, 11, 12, 13, 14, 14 year p.o.) and two for cup wear (12 and 17 yrs p.o.). The average HHS at follow-up was 86 (50–100). There were no additional radiographic failures. Using Kaplan-Meier analysis the survival with endpoint revision for any reason was 79% at 15 years and 71% at 20 years; for aseptic loosening of the cup 84% at 15 years and 84% at 20 years. Conclusions: This technique is attractive to reconstruct acetabular bone stock loss.

Aims: The effects of bone graft washing and bone graft particle size on initial cup stability in an acetabular model were studied. Methods: From fresh human femoral heads large (8–12 mm) and small (2–4) grafts were produced. Four different groups were tested; large washed, large unwashed, small washed and small unwashed. An AAOS type 3 combined defect was reconstructed in a synthetic acetabular model using a metal mesh and screws. Bone grafts were impacted in a standardized way using a hammer and metal impactors. Cement was pressurized and a PE cup was inserted. Cups were dynamically loaded with 1500 N and 3000 N for 15 minutes using an MTS-device, RSA was used to calculate 3-D cup migrations. Next a lever-out test of the cup was done in a displacement (16¡/minute) controlled test. After testing CT scans of the cups were made to quantify cement layer roughness, as a measurement of cement penetration. Results: Cups reconstructed with large washed bone grafts migrated signiþcantly less during mechanical compression and withstood signiþcantly higher lever-out moments. Washing improved cup stability for both small and large grafts, however only signiþcantly for the larger bone grafts. A signiþcantly higher cement penetration depth was found for all large bone grafts, washing did not inßuence cement penetration depth. Discussion: For acetabular revisions with bone impaction grafting we recommend to use large washed bone grafts.

A large series of animal experiments in goats was performed in relatevely simple bone chamber models and in very realistic loaded pre-clinical models. In this paper the focuss is on two experiments. In exp 1 we analysed the effect of rinsing of allograft bone on bone ingrowth into the bone induction chamber. We found that rinsing improves the ingrowth capacity to a level that is comparable to that of autologous bone. In experiment 2 we analysed the effect of two different reconstruction methods, e.g., a mesh or a strut graft, on the revascularization of impacted allograft bone in a femoral reconstruction. We found that new vessels can enter the impacted bone through the mesh and that this promotes an early revascularization of the bone graft.

In patients we analysed 24 biopsies of 20 patients and quantitated the amount of non-incorporated graft (remnants of original material), graft in the process of incorporation, incorporated graft (=new bone) and fibrous tissue. With increasng follow up peripods after the revision operation the amount of normal bone increased upto ca 90%. The remaining 10% consists of non-incorpated bone and fibrous tissue.

Introduction: Even in extensive osteonecrosis of the femoral head, a head-preserving method is preferable in younger patients. This study presents the results of bone impaction grafting for osteonecrosis of the femoral head.

Materials and Methods: After a core biopsy and removal of the osteonecrotic area, impacted morsellized bone allografts were used to fill the femoral head to regain sphericity, and to prevent collapse. This prospective, single-surgeon study included 28 consecutive hips in 27 patients with extensive osteonecrotic lesions (ARCO classification Stage 2 (11 hips), Stage 3 (14 hips), and Stage 4 (three hips). Fourteen hips had preoperative collapse. The mean age of the patients was 33 years (range, 15–55 years).

Results: At a mean follow-up of 42 months (range, 24–119 months), eight hips (29%) had been converted to a total hip arthroplasty. Of the 20 reconstructions that were in situ, 18 were clinically successful (90%) and 70% were radiologically successful. Patients who were younger than 30 years at surgery had a significantly better radiologic outcome, even when compared to heads with higher stages of osteonecrosis. Patients with preoperative collapse or a continuous use of corticosteroids had disappointing results.

Discussion: This method is attractive as a salvage procedure for osteonecrosis of the femoral head. It is relatively simple and quick to perform, and it does not intervene with an eventual future hip arthroplasty.

Collagen type type II destruction was studied after induction of experimental OA by ACL-transection in the rat. Damage was investigated by analysis of type II collagen neoepitope expression. Cleavage of type II collagen by collagenases (MMP’s) was detected by the Col2-3/4C-short antibody and collagen denaturation by Col2-3/4m. Rats were sacrificed after 2, 7, 14, 28 and 70 days. Immunostaining was performed using the Col2-3/4C (Collagenase-cleavage site) or the Col2-3/4m antibody (denatured type II collagen).

The first changes after the ACL-transsection were chondrocyte death at the margins of the articular cartilage of both tibia and femur. At day seven a pannus-like tissue protruded from the synovial tissue over the dead cartilage. Underneath the pannus-like tissue a marked staining for the collagenase-cleavage site was observed. The dead cartilage was replaced by fibrocartilage within 4 weeks after which the staining for the collagenase cleavage neoepitope had completely disappeared.

In contrast with the peripheral cartilage, in the central part of the medial tibia and femur dead chondrocytes were found on week 2 until the last time point examined, which was not replaced by fibrocartilage in this timespan. In these areas, loss of proteoglycans, fibrillation of superficial cartilage and staining for denatured type II collagen was found. Both cartilage damage and staining for denatured collagen increased with time. Only light collagenase cleavage site staining was observed on all time points in this central location.

OA in rats after ACL-transsection can be divided in two stages. An early phase lasting about 4 weeks, in which chondrocyte remodelling of the dead cartilage follows death at the cartilage margins. In this phase marked degradation of type II collagen by collagenases occurs. The second phase, characterised by cartilage damage in the central tibia and femur, shows increased staining for denatured type II collagen but little staining for the collagenase cleavage neoepitope.

It has been generally accepted that dynamic mechanical load is important for normal bone physiology, remodeling and fracture healing. Impacted morsellized grafts can be seen as healing of many small fractured bone parts, involving bone remodelling, apposition and formation of new bone. Therefore load may be stimulative for the incorporation of this type of graft.

In a pilot study we observed a positive effect of load on the density of incorporated bone after 12 weeks. Based on these results we hypothesised that physiological loading has a stimulatory effect on the early stage of bone graft incorporation. To test this idea we implanted fresh frozen allograft bone chips in 12 goats and loaded these grafts with the newly developed subcutaneous pressure implant ( Lamerigts et al., Biomaterials2000; 21: 741–7). The goats were divided in two groups: non-loaded and loaded. The loaded group was subjected to a loading regime of 3 MPa for 5 days/week (1 Hz, one hour/day). After 5 weeks the bone mineral density was measured with quantitative CT scanning, followed by routine histology and histomorphometry.

Bone mineral density was not affected by load. Histology revealed microscopic evidence of normal bone graft incorporation as seen in previous studies. The amount of active incorporating bone was higher under load (p< 0.05).

The formation of a new bony structure was not affected by load in this early stage of bone graft incorporation. However, load resulted in a more active graft incorporation after 5 weeks. The difference between the loaded and non-loaded group might be partially obscured by a low level of physiological loading in the non-loaded group induced by the daily activity of the animals.

Type I and II collagen-based scaffolds, with and without attached chondroitine sulphate (CS), were implanted without additional chondrocytes into full-thickness defects in the trochlea of young adult rabbits. We hypothesise that the chemical composition of the matrix will have a direct effect on the speed of repopulation and the phenotypic expression of the subchondral repair cells.

Evaluation of the repair process was performed with routine histology and with two quantitative histological grading systems, four and twelve weeks after implantation.

Four weeks after implantation, type I collagenous scaffolds were completely filled with a cartilage-like repair tissue. By contrast, type II collagenous scaffolds showed a superficial zone of cartilaginous tissue, and in many defects chondrocyte-like cells at the interface of the implant material with the subchondral bone. In collagen type II filled lesions larger areas of the scaffolds were completely devoid of repair tissue. Control defects showed a repair reaction that was very similar to that observed in defects filled with a type I scaffold.

After 12 weeks, the subchondral defect was largely replaced by bone and the differences between the scaffolds were less pronounced. The quantitative blind score of the sections confirmed that the scores of the control defect and of the collagen type I based scaffolds were slightly higher as compared to the type II based scaffolds. Irrespective of the type of scaffold, there was a trend that the scaffolds with CS scored slightly higher than those without CS.

We conclude that different types of scaffold induce different repair reactions. Collagen types I based scaffolds seem superior to guide progenitor cells from a subchondral origin into the defect. Repair cells in collagen type II based scaffolds seem to assume a chondrocyte-like phenotype, which could have a negative effect on the mobility of the repair cells.