In the present work, we test four thin coatings for titanium implants, namely, bioglass, GB14, β-Tricalciumphosphate (TCP) and hydroxyapatite (HA) with and without incorporated copper ions for their osteointegrative capacity. Rabbit drill hole model for time intervals up to 24 weeks was used in this study. Implant fixation was evaluated by measuring shear strength of implant/bone interface. Quantitative histological analysis was performed for the measurements of bone contact area. Implants with and without copper ions were compared after 24 weeks. Thin coatings of GB14, HA or TCP on titanium implants demonstrated high shear strength during entire test up to 24 weeks. Results confirmed osteointegrative properties of the coatings and did not reveal any negative effect of copper ions on osteointegration. The integration of copper in degradable osteoconductive coatings with a thickness of approx. 20 µm represents a promising method of achieving antibiotic shielding during the entire period of bone healing while at the same time improving the osteointegration of the implants.

In the present work, we test four thin coatings for titanium implants, namely, bioglass, GB14, Beta-Tricalciumphosphate (β-TCP) and hydroxyapatite (HA) with and without incorporated copper ions for their osteointegrative capacity. A rabbit drill hole model for time intervals up to 24 weeks was used in this study. Implant fixation was evaluated by measuring shear strength of the implant/bone interface. Quantitative histological analysis was performed for the measurements of bone contact area. Implants with and without copper ions were compared after 24 weeks. Thin coatings of GB14, HA or TCP on titanium implants demonstrated high shear strength during the entire test period of up to 24 weeks. Results confirmed osteointegrative properties of the coatings and did not reveal any negative effect of copper ions on osteointegration. The integration of copper in degradable osteoconductive coatings with a thickness of approx. 20 μm represents a promising method of achieving antibacterial shielding during the entire period of bone healing while at the same time improving osteointegration of the implants.

Introduction The use of scaffolds in tissue engineering is becoming increasingly important as solutions need to be found to preserve human tissues such as bone or cartilage. Various factors, including cells, biomaterials, cell and tissue culture conditions, play a crucial role in tissue engineering. The in vivo environment of the cells exerts complex stimuli on the cells, thereby directly influencing cell behavior, including proliferation and differentiation. Therefore, to create suitable replacement or regeneration procedures for human tissues, the conditions of the cells’ natural environment should be well mimicked. Therefore, current research is trying to develop 3-dimensional scaffolds (scaffolds) that can elicit appropriate cellular responses and thus help the body regenerate or replace tissues. In this work, scaffolds were printed from the biomaterial polycaprolactone (PCL) on a 3D bioplotter. Biocompatibility testing was used to determine whether the printed scaffolds were suitable for use in tissue engineering. Material and Methods An Envisiontec 3D bioplotter was used to fabricate the scaffolds. For better cell-scaffold interaction, the printed polycaprolactone scaffolds were coated with type-I collagen. Three different cell types were then cultured on the scaffolds and various tests were used to investigate the biocompatibility of the scaffolds. Results Reproducible scaffolds could be printed from polycaprolactone. In addition, a coating process with collagen was developed, which significantly improved the cell-scaffold interaction. Biocompatibility tests showed that the PCL-collagen scaffolds are suitable for use with cells. The cells adhered to the surface of the scaffolds and as a result extensive cell growth was observed on the scaffolds. The inner part of the scaffolds, however, remained largely uninhabited. In the cytotoxicity studies, it was found that toxicity below 20% was present in some experimental runs. The determination of the compressive strength by means of the universal testing machine Z005 by ZWICK according to DIN EN ISO 604 of the scaffolds resulted in a value of 68.49 ± 0.47 MPa.

In the present work, an ex vivo organ model using human bone (explant) was developed for the evaluation of the initial osseointegration behavior of implant materials. The model was tested with additive manufactured Ti6Al4V test substrates with different 3D geometries. Explants were obtained from patients who underwent total knee replacement surgery. The tibial plateaus were used within 24 h after surgery to harvest bone cylinders (BC) from the anterior side using hollow burrs. The BCs were brought into contact with the test substrate and inserted into an agarose mold, then covered with cell culture media and subjected to the external load of 500 g. Incubation was performed for 28 days. After 28d the test substrate was removed for further analysis. Cells grown out BC onto substrate were immunostained with DAPI and with an antibody against Collagen-I and alkaline phosphatase (ALP) for visualization and cell counting. We show that cells stayed alive for up to 28d in our organ model. The geometry of test substrates influences the number of cells grown onto substrate from BCs. The model presented here can be used for testing implant materials as an alternative for in vitro tests and animal models.

The authors report on the manufacturing of mechanically stable β-tricalcium phosphate (β-TCP) structural hybrid scaffolds via the combination of additive manufacturing (CerAM VPP) and Freeze Foaming for engineering a potential bone replacement. In the first step, load bearing support structures were designed via FE simulation and 3D printed by CerAM VPP. In the second step, structures were foamed-in with a porous and degradable calcium phosphate (CaP) ceramic that mimics porous spongiosa. For this purpose, Fraunhofer IKTS used a process known as Freeze Foaming, which allows the foaming of any powdery material and the foaming-in into near-net-shape structures. Using a joint heat treatment, both structural components fused to form a structural hybrid. This bone construct had a 25-fold increased compressive strength compared to the pure CaP Freeze Foam and excellent biocompatibility with human osteoblastic MG-63 cells when compared to a bone grafting Curasan material for benchmark.

Just like menisci, articular cartilage is exposed to constant and varying stresses. Injuries to the meniscus are associated with the development of gonarthritis. Both the articular cartilage and the menisci are subject to structural changes under gonarthritis. The aim of this study was to investigate biomechanical alterations in articular cartilage and the menisci under gonarthritis by applying an indentation method. The study assessed 11 menisci from body donors as controls and 21 menisci from patients with severe gonarthritis. For the simultaneous examination of the articular cartilage and the menisci, we only tested the joint surfaces of the tibial plateau covered by the corresponding menisci. Over the posterior horn of the meniscus, the maximum applied load—the highest load registered by the load cell—of the arthritic samples of 0.02 ± 0.02 N was significantly greater (p = 0.04) than the maximum applied load of the arthritis-free samples of 0.01 ± 0.01 N. The instantaneous modulus (IM) at the center of the arthritic cartilage covered by the meniscus with 3.5 ± 2.02 MPa was significantly smaller than the IM of the arthritis-free samples with 5.17 ± 1.88 MPa (p = 0.04). No significant difference was found in the thickness of the meniscus-covered articular cartilage between the arthritic and arthritis-free samples. Significant correlations between the articular cartilage and the corresponding menisci were not observed at any point. In this study, the biomechanical changes associated with gonarthritis affected the posterior horn of the meniscus and the mid region of the meniscus-covered articular cartilage. The assessment of cartilage thickness as a structural characteristic of osteoarthritis may be misleading with regard to the interpretation of articular cartilage’s biomechanical properties.

Studies on osteoarthritis of the knee have examined isolated aspects of the meniscus biomechanically and histologically, but not the difference between instantaneous modulus (IM) in healthy and diseased samples. The aim of this study is to investigate the difference in the biomechanical behavior and proteoglycan content between arthritic and arthritis-free menisci. In addition, the relationship between the biomechanical behavior and proteoglycan content should be investigated. A novel indentation-based method was used, which allows the mapping of the entire meniscus, without damaging it for histological examinations. A total of 26 arthritic and 14 arthritis-free samples were examined in the present study. A Mach-1 Model V500css test machine was used for biomechanical testing. A position grid was placed over each sample allowing all measurements (indentation test and thickness measurements) to be taken at the same position. All sections were then graded for Safranin O staining intensity for proteoglycan content. The maximum applied load of our arthritic samples above the posterior horn was statistically significantly higher (p = 0.01) at 0.02 ± 0.02 N than the maximum applied load of the arthritis-free samples at 0.01 ± 0.01 N. The proteoglycan content of the meniscus, evaluated by the Safranin O score, correlated statistically significantly with the maximum applied load over the entire meniscus (p = 0.04, 95% CI: 0.06–0.71). The present study showed that in the final stage of gonarthritis, the attenuation behavior of the meniscus was significantly lower compared to the arthritis-free knee. The mapping of IM and histological examination of the meniscus showed a direct correlation between changes in proteoglycan content and altered mechanical properties of the meniscus in gonarthritis.

Background Arthrofibrosis is a painful and restraining complication that occurs after about 10% of total knee arthroplasty and cruciate ligament surgery. The pathogenesis of arthrofibrosis has not yet been fully understood. Stress signals stimulate immune cells, and fibroblast differentiates into myofibroblast, which produce a large amount of collagen. Xylosyltransferases also appear to be involved in these pathways. They catalyze proteoglycan biosynthesis, which is involved in tissue remodeling and myofibroblast differentiation. The aim of this study was to investigate the relationship between the disease arthrofibrosis and the expression of the two isoforms of xylosyltransferases I and II. Methods Tissue samples from 14 patients with arthrofibrosis were compared with tissue samples from seven healthy controls. The xylosyltransferases were detected by immunohistochemistry. The tissues were divided into four different areas of interest: vessels, synovialis, cell-poor and cell-rich fibrosis, or cell-poor and cell-rich areas in the control group. A quantification of the results was performed by modification of the immunoreactive score according to Remmele and Stegner. Results Xylosyltransferase I was expressed in the various tissue types at varying rates. Xylosyltransferase I expression was considerably and significantly stronger than that of xylosyltransferase II. The following sequences of xylosyltransferase I and xylosyltransferase II expression were determined as follows: vessels >> cell-rich fibrosis > cell-poor fibrosis > synovialis. A positive correlation between the number of positive fibroblasts and the immunoreactive scoring system (IRS) was documented. Conclusions The significant positive correlation of xylosyltransferase -I expression with increasing number of fibroblasts demonstrates a high myofibroblast differentiation rate, which implies a gradual event as the pathogenesis of arthrofibrosis.

The aim of this study was to predefine the pore structure of β-tricalcium phosphate (β-TCP) scaffolds with different macro pore sizes (500, 750, and 1000 µm), to characterize β-TCP scaffolds, and to investigate the growth behavior of cells within these scaffolds. The lead structures for directional bone growth (sacrificial structures) were produced from polylactide (PLA) using the fused deposition modeling techniques. The molds were then filled with β-TCP slurry and sintered at 1250 °C, whereby the lead structures (voids) were burnt out. The scaffolds were mechanically characterized (native and after incubation in simulated body fluid (SBF) for 28 d). In addition, biocompatibility was investigated by live/dead, cell proliferation and lactate dehydrogenase assays. The scaffolds with a strand spacing of 500 µm showed the highest compressive strength, both untreated (3.4 ± 0.2 MPa) and treated with simulated body fluid (2.8 ± 0.2 MPa). The simulated body fluid reduced the stability of the samples to 82% (500), 62% (750) and 56% (1000). The strand spacing and the powder properties of the samples were decisive factors for stability. The fact that β-TCP is a biocompatible material is confirmed by the experiments. No lactate dehydrogenase activity of the cells was measured, which means that no cytotoxicity of the material could be detected. In addition, the proliferation rate of all three sizes increased steadily over the test days until saturation. The cells were largely adhered to or within the scaffolds and did not migrate through the scaffolds to the bottom of the cell culture plate. The cells showed increased growth, not only on the outer surface (e.g., 500: 36 ± 33 vital cells/mm² after three days, 180 ± 33 cells/mm² after seven days, and 308 ± 69 cells/mm² after 10 days), but also on the inner surface of the samples (e.g., 750: 49 ± 17 vital cells/mm² after three days, 200 ± 84 cells/mm² after seven days, and 218 ± 99 living cells/mm² after 10 days). This means that the inverse 3D printing method is very suitable for the presetting of the pore structure and for the ingrowth of the cells. The experiments on which this work is based have shown that the fused deposition modeling process with subsequent slip casting and sintering is well suited for the production of scaffolds for bone replacement.

Ultra-high molecular weight polyethylene (UHMWPE) is widely used in endoprosthetics and has been the subject of countless studies. This project investigates the dependence of alendronate (AL) release on the molecular weight of the UHMWPE used (GUR1020 and GUR1050). A 0.5 wt% AL was added to the UHMWPE during the production of the moldings. In addition to the 14-day release tests, biocompatibility tests such as live dead assay, cell proliferation assay (WST) and Lactate dehydrogenase test (LDH) with MG-63 cells as well as a tensile test according to DIN EN ISO 527 were carried out. The released AL concentration was determined by HPLC. A continuous release of the AL was observed over the entire period of 2 weeks. In addition, a correlation between molar mass and AL release was demonstrated. The GUR1020 showed a release four times higher than the GUR1050. Both materials have no negative influence on the proliferation of MG-63 cells. This was also confirmed in the live/dead assay by the increase in cell count. No cytotoxicity was detected in the LDH test. The addition of 0.5 wt% AL increased the elongation at break for GUR1020 by 23% and for GUR1050 by 49%. It was demonstrated that the choice of UHMWPE has an influence on the release of AL. The particle size in particular has a strong influence on the release behavior.

There are many studies on osteoarthritis, but only a few studies deal with human arthrosis, comparing the mechanical properties of healthy and diseased samples. In most of these studies, only isolated areas of the tibia are examined. There is currently only one study investigating the complete mapping of cartilage tissue but not the difference between instantaneous modulus (IM) in healthy and diseased samples. The aim of this study is to investigate the relationship between the biomechanical and histological changes of articular cartilage in the pathogenesis of osteoarthritis. The study compared 25 tibiae with medial gonarthrosis and 13 healthy controls. The IM was determined by automated indentation mapping using a Mach-1 V500css testing machine. A grid was projected over the sample and stored so that all measurements could be taken at the same positions (100 ± 29 positions across the tibiae). This grid was then used to perform the thickness measurement using the needle method. Samples were then taken for histological examinations using a hollow milling machine. Then Giemsa and Safranin O staining were performed. In order to determine the degree of arthrosis according to histological criteria, the assessment was made with regard to Osteoarthritis Research Society International (OARSI) and AHO scores. A significant difference (p < 0.05) could be observed in the measured IM between the controls with 3.43 ± 0.36 MPa and the samples with 2.09 ± 0.18 MPa. In addition, there was a significant difference in IM in terms of meniscus-covered and meniscus-uncovered areas. The difference in cartilage thickness between 2.25 ± 0.11 mm controls and 2.0 ± 0.07 mm samples was highly significant with p < 0.001. With regard to the OARSI and AHO scores, the samples differed significantly from the controls. The OARSI and AHO scores showed a significant difference between meniscus-covered and meniscus-uncovered areas. The controls showed significantly better viscoelastic behavior than the arthrotic samples in the measured IM. The measured biomechanical values showed a direct correlation between histological changes and altered biomechanics in gonarthrosis.

The use of both bioglass (BG) and β tricalcium phosphate (β-TCP) for bone replacement applications has been studied extensively due to the materials’ high biocompatibility and ability to resorb when implanted in the body. 3D printing has been explored as a fast and versatile technique for the fabrication of porous bone scaffolds. This project investigates the effects of using different combinations of a composite BG and β-TCP powder for 3D printing of porous bone scaffolds. Porous 3D powder printed bone scaffolds of BG, β-TCP, 50/50 BG/β-TCP and 70/30 BG/β-TCP compositions were subject to a variety of characterization and biocompatibility tests. The porosity characteristics, surface roughness, mechanical strength, viability for cell proliferation, material cytotoxicity and in vitro bioactivity were assessed. The results show that the scaffolds can support osteoblast-like MG-63 cells growth both on the surface of and within the scaffold material and do not show alarming cytotoxicity; the porosity and surface characteristics of the scaffolds are appropriate. Of the two tested composite materials, the 70/30 BG/β-TCP scaffold proved to be superior in terms of biocompatibility and mechanical strength. The mechanical strength of the scaffolds makes them unsuitable for load bearing applications. However, they can be useful for other applications such as bone fillers.

Particle-induced periprosthetic osteolysis and subsequent aseptic implant loosening are a major cause of compromising the long-term results of total joint replacements. To date, no implant has been able to mirror radically the tribological factors (friction/lubrication/wear) of in vivo tribological pairings. Carbon-Fibre Reinforced SiC-Composites (C/SiSiC), a material primarily developed for brake technology, has the opportunity to fulfil this requirement. Until now, the material itself has not been used in medicine. The aim of this investigation was to test the suitability of C/SiSiC ceramics as a new material for bearing couples in endoprosthetics. After the preparation of the composites flexural strength was determined as well as the Young's-modulus and the coefficient of friction. To investigate in vitro biological properties, MG 63 and primary human osteoblasts were cultured on C/SiSiC composites. To review the proliferation, the cytotoxicity standardized tests were used. The cell morphology was observed by light microscopy, ESEM, confocal and 3D-laserscanning microscopy. C/SiSiC possesses a high resistance to wear. Cells exhibited no significant alterations in morphology. Vitality was not impaired by contact with the ceramic composite. There was no higher cytotoxicity to observe. Regarding these results, C/SiSiC ceramics seem to be biologically and mechanically appropriate for orthopaedic applications.

Retinitis pigmentosa is a group of hereditary retinal dystrophies that normally result in photoreceptor cell death and vision loss both in animal models and in affected patients. The rd10 mouse, which carries a missense mutation in the Pde6b gene, has been used to characterize the underlying pathophysiology and develop therapies for this devastating and incurable disease. Here we show that increased photoreceptor cell death in the rd10 mouse retina is associated with calcium overload and calpain activation, both of which are observed before the appearance of signs of cell degeneration. These changes are accompanied by an increase in the activity of the lysosomal protease cathepsin B in the cytoplasm of photoreceptor cells, and a reduced colocalization of cathepsin B with lysosomal markers, suggesting that lysosomal membrane permeabilization occurs before the peak of cell death. Moreover, expression of the autophagosomal marker LC3-II (lipidated form of LC3) is reduced and autophagy flux is blocked in rd10 retinas before the onset of photoreceptor cell death. Interestingly, we found that cell death is increased by the induction of autophagy with rapamycin and inhibited by calpain and cathepsin inhibitors, both ex vivo and in vivo. Taken together, these data suggest that calpain-mediated lysosomal membrane permeabilization underlies the lysosomal dysfunction and downregulation of autophagy associated with photoreceptor cell death.