Hans-Georg Simank

Vascular Endothelial Growth Factor Gene‐Activated Matrix (VEGF165‐GAM) Enhances Osteogenesis and Angiogenesis in Large Segmental Bone Defects

Healing of fractures is dependent on vascularization of bone, which is in turn promoted by VEGF. It was shown that 0.1 and 1 mg of pVEGF165‐GAM led to a significant increase in vascularization and bone regeneration in defects that would otherwise have led to atrophic nonunions.

Periprosthetic Mineralization Around Cementless Total Hip Endoprosthesis: Longitudinal Study and Cross-Sectional Study on Titanium Threaded Acetabular Cup and Cementless Spotorno Stem with DEXA

In a prospective longitudinal study over 2 years and a separate cross-sectional study more than 5 years after operation, we analyzed periprosthetic bone mineral density (BMD) after cementless total hip arthroplasty (THA) (press-fit cementless Spotorno stem, Mecron threaded acetabular cup) by dual-energy X-ray absorptiometry (DEXA). BMD was analyzed in a longitudinal prospective study (n = 53 patients: 29 women, 24 men) and in a separate cross-sectional study (n = 23 patients: 13 women, 10 men) with good clinical outcome (Merle d’ Aubigne score > 12). Regions of interest were defined according to Gruen (ROI 1–7) and as netto average ROI (NETAVG I) for the periprosthetic femur, and according to De Lee and Charnley (ROI I-III) and as NETAVG II for the periprosthetic acetabulum. BMD during follow-up was compared with immediate postoperative values of the affected limb. Mean precision error (CV%) was 2.6 ± 0.5% for ROI 1–7 and 1.3 ± 0.9% for ROI I–III. BMD significantly decreased in the periprosthetic femur and acetabulum during the first 3 months after operation. At the femur, BMD (NETAVG I) for women and men, respectively, was 92.4% and 87.5% at 6 months, then 89.4% and 96.2% at 2 years. ROIs around the proximal stem showed the lowest absolute values and decreased most during follow-up (to 79.9% ROI 1 and 68.2% ROI 7, respectively). Mineralization around the cup (NETAVG II), respectively, amounted to 81.1%, 82.6% at 6 months, then 80.1% and 93.8% at 2 years. The medially placed ROI II demineralized most (respectively, 72.1% and 76.7%). More than 5 years after THA, BMD in the femur showed little change, but decreased significantly to 76.4% and 79.1%, respectively, around the cup (NETAVG II). DEXA is a useful method for analyzing changes of mineralization around cup and stem of cementless THA. The results reflect the different stress on the periprosthetic bone after implantation of THA in defined ROIs, supporting earlier reported good clinical results of the Spotorno stem and increased loosening rate of threaded acetabular cups after 5 years.

Evaluation of Mineralized Collagen and α-Tricalcium Phosphate as Scaffolds for Tissue Engineering of Bone Using Human Mesenchymal Stem Cells

Owing to their plasticity and high proliferation capacity in vitro, mesenchymal stem cells (MSC) isolated from human bone marrow are promising candidates for use in tissue engineering approaches for the repair or replacement of mesenchymal tissues such as bone, cartilage or tendon. In keeping with the tissue engineering concept, these cells are cultivated on three-dimensional (3D) scaffolds to replace 3D tissue defects. Among the scaffolds tested for tissue engineering of bone, those containing phosphorus and calcium, as natural bone does, are the most promising candidates for this purpose. In this study, MSC from five patients were isolated from bone marrow. After in vitro expansion, cells were cultivated and differentiated towards the osteogenic lineage on mineralized collagen sponges and α-tricalcium phosphate (α-TCP). To analyze how appropriate these scaffolds would be for tissue engineering purposes, we established an in vitro characterization system to describe seeding efficiency, cell distribution and proliferation behavior on each scaffold. Real-time reverse transcriptase polymerase chain reaction quantification of important genes involved in osteogenic differentiation [e.g. bone sialoprotein (BSP), bone morphogenic protein 2, alkaline phosphatase and osteocalcin] was used to monitor the differentiation process of cells seeded on mineralized collagen and α-TCP. Using this in vitro characterization, we were able to demonstrate effective 3D growth of MSC on both scaffolds investigated. Improved osteogenic differentiation was observed on the scaffolds as compared to control monolayers. Of the two matrices, mineralized collagen was superior to α-TCP with regard to seeding efficacy (98 vs. 67%, p = 0.0003), increase in osteogenic marker genes (BSP expression on day 24, pcollagen vs. TCP = 0.046) and 3D cell alignment (cell infiltration up to 500 vs. 200 µm). In conclusion, our data suggest that mineralized collagen is a promising candidate for use as a scaffold in tissue engineering of bone.

MIDDLE-TERM RESULTS OF THREADED ACETABULAR CUPS: HIGH FAILURE RATES FIVE YEARS AFTER SURGERY

We report our results using three different threaded acetabular components (Mecring A, Mecring B and Weill) in 715 hips with a follow-up of between one and ten years (median: 99.1, 56.5, 38.3 months, respectively). All cups were implanted with one type of cementless stem. The clinical results were good or acceptable in about 70% of the hips, but signs of loosening with radiolucency and/or migration were found in 10.1%. Radiological evidence of loosening did not correlate significantly with the clinical outcome. Pain was not a reliable indicator of loosening and its absence sometimes allowed severe osteolysis to develop. Twenty-five hips were revised (3.5%) for aseptic loosening of the acetabular component. Kaplan-Meier estimates of the cumulative rate of failure showed a rapid increase five years after the initial operation, but no significant correlation with gender, age or weight. The high rate of failure indicates that further use of these acetabular components cannot be recommended. Annual radiographs are required to assess osteolysis even if the patients are free from pain.

Bone morphogenetic protein-2 and growth and differentiation factor-5 enhance the healing of necrotic bone in a sheep model.

Abstract Introduction: Osteotropic growth factors enhance bone repair, but their efficacy in an area of necrotic bone is not known. The purpose of this study was to investigate the effects and potential side effects of an intraosseous application of absorbable bone morphogenetic protein-2 (BMP-2) and growth and differentiation factor-5 (GDF-5) composites in a sheep model for partial necrosis of the femoral head. Materials and methods: The direct injection of ethanol under fluoroscopy into the superior centre of the right femoral head produced histologically documentable necrosis of the central region of the head in a previous study of ten sheep. Another 27 sheep constituted the sample to study the effects of BMP-2 and GDFJ. Necrosis was produced in the same fashion in these animals. Four weeks later nine sheep received 300 μg recombinant BMP-2 and nine sheep 300 μg recombinant GDF-5 on an absorbable carrier by surgical implantation. Nine sheep received the carrier alone (control group). The animals were sacrificed at 3, 6, and 12 weeks after implantation and both femora were harvested. Results: Bone density analysis and microscopic examination indicated that bone formation was noticeably induced as early as 3 weeks postoperatively in the growth factor treated animals. Bone regeneration was enhanced by growth factor composites. This was documented by histological scoring and histomorphometric analysis. No severe local side effects secondary to the growth factors, such as heterotopic ossification or inflammation, were observed in either group. Discussion: The application of an absorbable growth factor composite in combination with established surgical techniques is a promising approach, that may enhance the healing of devitalised bone defects. Based on these results, further studies regarding biodegradation, dosage of the protein and surgical technique are required.

A new animal model of femoral head necrosis induced by intraosseous injection of ethanol

There is no reliable animal model of the early stages of osteonecrosis of the femoral head (ONFH) for the evaluation of new therapeutic approaches. In this study, we propose a new animal model of femoral head osteonecrosis. Pure ethanol was injected into the centre of the femoral head in adult Merino sheep under fluoroscopic control. After 3, 6 and 12 weeks the animals were killed and the femoral heads were harvested. Microradiographic and histological changes were analysed and recorded. Partial necrosis was documented over a period of 12 weeks in all animals. The appearance of necrosis in combination with intact macrotexture, macrocirculation and joint cartilage is similar to the features described in early ONFH in humans. Due to its efficacy and its similarity to the early stages of ONFH in humans, this model may be suitable to evaluate new therapeutic techniques in the treatment of ONFH.

Local application of a collagen type I/hyaluronate matrix and growth and differentiation factor 5 influences the closure of osteochondral defects in a minipig model by enchondral ossification.

This pilot study evaluated the effect of growth and differentiation factor-5 (rhGDF-5) combined with a collagen type I/hyaluronate matrix (c/h) on osteochondral defect repair in a minipig model. Defects created in both medial femoral condyles of 20 minipigs were treated with c/h (n = 10), c/h + rhGDF-5 (n = 10) or were left empty. After 3 and 12 months, five animals of each group were sacrificed. Evaluation included macroscopic and histological scoring and quantitative histomorphometry of synthesized bone. C/h and c/h + rhGDF-5 treatment increased trabecular bone formation in the upper third of the defect compared to empty controls, showing significance for c/h + rhGDF-5 (p = 0.05) but not between c/h and c/h + rhGDF-5 treatment. Cartilage regeneration and macroscopic outcome were not improved by c/h or c/h + rhGDF-5 treatment. Since c/h remnants were seen even one year postoperatively in the defect, possibly inhibiting further bone and cartilage healing, other matrices in combination with rhGDF-5 may provide further improvement in osteochondral defect treatment.

Effects of local application of growth and differentiation factor-5 (GDF-5) in a full-thickness cartilage defect model.

Our purpose in this study was to investigate the effects of growth and differentiation factor-5 (GDF-5) on cartilage and subchondral bone in a rabbit model harboring an osteochondral defect for a period of 6 months. Absorbable composites were implanted in adult rabbits (18 controls, 18 animals with collagen-I matrix, and 18 animals with matrix plus GDF-5). After 4, 8, or 24 weeks the specimens were studied by histology, microcomputed tomography (microCT) and flow-cytometric analysis (FACS). Implantation of GDF-5 resulted in an improved histological appearance. This was the result of significantly improved defect filling at 4 and 8 weeks. At 24 weeks, however, there was no difference between the groups. The histological examination disclosed a predominance of fibrocartilage, and remodeled subchondral bone was also observed. MicroCT documented normal bone density in all groups, excluding subchondral sclerosis. At 24 weeks, FACS analysis revealed high apoptotic activity in the GDF-5-treated group. As far as we are aware, this is the first report of the effects of GDF-5 in a full-thickness cartilage defect model. We assume that recombinant GDF-5 contained on the carrier is probably able to accelerate the regeneration of the osteochondral defect owing to its availability. However, control of the protein delivery may require further investigation.

Improving the diagnosis of septic arthritis by use of a pediatric blood culture system

BACKGROUND Prognosis and outcome of bacterial joint infections are dependent on the fast and reliable identification of pathogens in the synovial fluid. Previous studies have suggested the possible advantage of using a blood culture system in contrast to conventional culture methods. PATIENTS AND METHODS A total of 101 synovial specimens from patients presenting with symptoms suggesting septic arthritis were taken by aspiration with a sterile syringe. We compared the diagnostic results of automated analysis in a blood culture system against conventional culture on solid agar plates. RESULTS Some 67 specimens (66.3%) were found to be negative in both preparations, while samples from 21 patients (20.8%) yielded the same microorganisms. In 13 cases (12.9%), the isolation of a pathogen was possible only with the blood culture method, whereas the conventional method never yielded a positive result when the blood culture was negative. Thus, the diagnostic yield was significantly improved by use of the blood culture system (P <0.001). CONCLUSION The use of a commonly available blood culture system offers a fast, reliable and cost-effective approach for the diagnosis of septic arthritis and should therefore be considered as an useful alternative to conventional culture methods.

Implant stability in the treatment of MRSA bone implant infections with linezolid versus vancomycin in a rabbit model

The increasing prevalence of methicillin‐resistant Staphylococcus aureus (MRSA) infections represents a significant healthcare burden. Vancomycin and linezolid exhibit potent clinical and microbiological activity in MRSA infections. Our purpose was to investigate the efficacy of linezolid versus vancomycin in experimental implant infections and the influence on implant stability in a rabbit model. Thirty‐six female New Zealand White rabbits received surgical insertion of titanium implants into their distal femurs and were randomly assigned to six groups (A: infected, no treatment; B: infected, vancomycin; C: infected, linezolid; D: no infection, no treatment; E/F: no infection, vancomycin or linezolid, respectively). Antibiotics were administered, and plasma levels determined. Bone‐implant specimens were tested for mechanical stability of fixation. Quantitative histomorphometry of bone and soft tissue was performed using computerized image analysis. Plasma levels of linezolid and vancomycin were within the respective therapeutic ranges. Microbiological analysis of specimens from infected rabbits showed MRSA tissue colonization in all untreated animals, in two of six vancomycin‐treated animals, and in none of the linezolid‐treated animals. Antibiotic treatment improved mechanical stability significantly (p = 0.004) with both vancomycin and linezolid. Mechanical testing correlated with histomorphometry results. A significant negative correlation was found between displacement of the implant and the percentage of calcified tissue around the implant, and a significant positive correlation was found between displacement of the implant and the amount of noncalcified tissue. Our data indicate that both treatment regimens improved implant stability.