It is reported that more than 10 million Japanese suffer from arthrosis. To cure these cartilage defects, total joint replacements, which are the most popular treatment methods for severe disease situation, have been operated as about two hundred thousand cases a year in Japan. Although the implants made of either ceramics, metals or plastics have high wear resistance quality, it becomes apparent that the endurance life of the artificial joints in considerable cases is limited by aseptic loosening to between 10–15 years. Here we focused on a poly(vinyl alcohol) (PVA) hydrogel as an artificial cartilage tissue to make an improvement of friction surface of the artificial joints. In this paper, we observed morphology of wear particles and assessed immune responses of wear particles from the hydrogel for confirming the validity of the gel as a biotribological material. We prepared 20 w/w% of PVA hydrogel by repeated freezing-thawing method. The number of the freezing-thawing cycles was five times. Polymerization degree and saponification degree of PVA (Kishida Chemical Co. Ltd., Japan) were 2000 and 98.4–99.8 mol%, respectively. To collect the wear particles of PVA hydrogel, we processed wear testing by using a purpose-build wear test machine of reciprocating pin-on-plate tribometer as shown in figure 1. We installed a Co-Cr-Mo ball of 26 mm in diameter as a stationary upper specimen and a PVA hydrogel plate of 2 mm in thickness as reciprocating lower specimen in a water bath. The lubricant was a distilled water containing eluted PVA which PVA-FT gel had been soaked in, filtered by 0.22 μm and autoclaved, subsequently. Siding speed was 50 mm/s and the total sliding distance was 3 km. We observed the wear particles which had been dried in a desiccator, by scanning electron microscope (SEM; SU8000, Hitachi High-Technologies). Additionally, to investigate the effect of the wear particles on response of phagocytosis of macrophages, here we used THP-1 cell line from Human acute monocytic leukemia as a macrophage, which was purchased from JCRB Cell Bank, and attached the macrophages on a dish after stimulating THP-1 by phorbol 12-myristate 13-acetate (PMA; Wako). After the wear test, we harvested the lubricant in aseptic clean hood and applied the particles to the macrophages to clarify the effect of wear particles of PVA hydrogel on immune response of the cell. To assess cytokine biosynthesis as immune responses, we assayed IL-1β and TNF-α biosynthesis in culture medium by ELISA (Thermo scientific), respectively. Figure 2 shows an SEM image of PVA hydrogel after wear test under 4.9 N loading. We observed the wear particles of varied sizes. When applying the wear particles to the macrophages in RPMI-1640 supplement with 10 v/v% fetal bovine serum, it seemed that there were not enough change on cytokine synthesis in culture medium between with/without the particles

Objectives. This study aimed to characterise and qualitatively grade the severity of the corrosion particles released into the hip joint following taper corrosion. Methods. The 26 cases examined were CoC/ABG Modular (n = 13) and ASR/SROM (n = 13). Blood serum metal ion levels were collected before and after revision surgery. The haematoxylin and eosin tissue sections were graded on the presence of fibrin exudates, necrosis, inflammatory cells and corrosion products. The corrosion products were identified based on visible observation and graded on abundance. Two independent observers blinded to the clinical patient findings scored all cases. Elemental analysis was performed on corrosion products within tissue sections. X-Ray diffraction was used to identify crystalline structures present in taper debris. Results. The CoC/ABG Modular patients had a mean age of 64.6 years (49.4 to 76.5) and ASR/SROM patients had a mean age of 58.2 years (33.3 to 85.6). The mean time in situ for CoC/ABG was 4.9 years (2 to 6.4) and ASR/SROM was 6.1 years (2.5 to 8.1). The blood serum metal ion concentrations reduced following revision surgery with the exception of Cr levels within CoC/ABG. The grading of tissue sections showed that the macrophage response and metal debris were significantly higher for the ASR/SROM patients (p < 0.001). The brown/red particles were significantly higher for ASR/SROM (p < 0.001). The taper debris contained traces of titanium oxide, chromium oxide and aluminium nitride. Conclusion. This study characterised and qualitatively graded the severity of the corrosion particles released into the hip joint from tapers that had corrosion damage. Cite this article: S. Munir, R. A. Oliver, B. Zicat, W. L. Walter, W. K. Walter, W. R. Walsh. The histological and elemental characterisation of corrosion particles from taper junctions. Bone Joint Res 2016;5:370–378. DOI: 10.1302/2046-3758.59.2000507

Background. Polyethylene (PE) as a bearing material for total joint replacements (TJR) represents the golden standard for the past forty years. However, over the past decade it becomes apparent that PE wear and the biological response to wear products are the limiting factor for the longevity of TJRs. For this reason research has focused onto PE wear particle analysis. A particle analysis highly depends on the methodological work and results often show discrepancies between different research groups. From there, our hypothesis was, that an often unattended influencing factor is the optical magnification which has been used for particle analyses. Material and Methods. In the present study samples of a previous conducted knee wear simulator test were used. Wear particles were isolated from the bovine serum using an established method. 1. Briefly the serum was digested with hydrochloric acid and a continuous stirring and heating. Particles were filtered onto 20nm alumina filters and analyzed using high resolution field emission gun scanning electron microscopy (FEG-SEM). Filters were analyzed on the same points using three different magnifications: 5000, 15000 and 30000. To describe the size and morphology of the particles the equivalent circle diameter (ECD), aspect ratio (AR), roundness (R) and form factor (FF) were specified according to ASTM F 1877-05. The estimated total number (ETN) of particles was calculated based on the number of particles recovered on the filter, the analyzed area, the dilution, evaporation and the total serum volume. Results. The results showed significant differences between the different magnifications. Examples of the analyzed pictures are depicted in Fig. 1. The results are summarized in Tab. 1. In particular the size of the particles highly depends on the choosen optical magnification which becomes apparent in a more than twofold increase when comparing wear particles at magnification of 5k or 30k (p≤0.001). The mean particle diameter distribution (Fig. 2) also shows a shift in the distribution of wear particles: A higher magnification results in a higher fraction of smallest particles (e.g. over 50 percent between 0–0.2µm with magnification 30.000 compared to only 3 percent with a magnification of 5.000) and nearly no particles above 1µm could be verified. The results regarding the particle morphology show smaller but significant differences. The ETN of particles quadruplicates when comparing results with magnifications of 5.000 and 30.000. Conclusion. This study shows great differences in particle size, which can be directly attributed to the differences in optical magnification. According to ASTM F1877-05 a magnification of 10.000 for the analysis of wear particles between 0.1–1µm is recommended. However, results show that this magnification is not sufficient especially for particle sizes below 0.2µm, which account for the greatest number of particles. To the authors opinion a more detailed recommendation concerning the magnification is needed. Additional, a standardized evaluation system which takes the magnification into account is necessary to allow comparison of different research groups

Summary Statement. Antioxidant containing UHMWPE particles induced similar levels of in vitro macrophage proliferation and in vivo inflammation in the mouse air pouch model as UHMWPE particles alone. Benefit of antioxidant in reducing wear particle induced inflammation requires further investigation. Introduction. Wear particles derived from UHMWPE implants can provoke inflammatory reaction and cause osteolysis in the bone, leading to aseptic implant loosening. Antioxidants have been incorporated into UHMWPE implants to improve their long term oxidative stability. However it is unclear if the anti-inflammatory property of the antioxidant could reduce UHMWPE particle induced inflammation. This study evaluated the effect of cyanidin and vitamin E on UHMWPE induced macrophage activation and mouse air pouch inflammation. Methods. Four types of UHMWPE were used: (1) compression molded (CM) conventional GUR1020 (PE); (2) CM GUR1020 blended with 300 ppm cyanidin (C-PE); (3) CM GUR1020 blended with 1000 ppm α-tocopherol (BE-PE); and (4) CM GUR1020, gamma irradiated at 100kGy, diffused with α-tocopherol, and sterilised at 30kGy (DE-PE). Particles were generated by cryomilling. Particle count, size, and aspect ratio were determined using SEM and Image Pro. Each particle group was cultured with RAW264.7 macrophage cells at four concentrations (0.625, 1.25, 2.5, and 5 μg/mL) in a standard medium for 4 days. Cell numbers were quantified using MTT assay. Cytokine expression (IL-1β, TNFα, and IL-6) was measured using RT-PCR and ELISA. Particles were also suspended in PBS at 2 concentrations (0.2 or 1 mg) and injected into subcutaneous air pouches in BALB/c mice. Control animals were injected with PBS alone. Six days post-injection air pouches were harvested, half of which were fixed for histology to measure membrane thickness and inflammatory cell quantity. Remaining air pouches were frozen and analyzed by ELISA for cytokine production. Data were analyzed using one-way ANOVA with post hoc testing. P<0.05 was considered significant. Results. All 4 materials showed similar particle characteristics after cryomilling. Particle size ranged from 1 to 19 μm with 33% of particle population smaller than 2 μm. All particle groups supported macrophage proliferation, showing an inverse correlation between proliferation rate and particle dose. Gene expression of IL-1β and TNFα also showed an inverse correlation with particle dose. Expression of IL-1β, TNFα, and IL-6 appeared lower in cells cultured with C-PE than the other 3 materials. The accumulative protein productions of IL-1β and TNFα were significantly lower while IL-6 production was moderately lower in C-PE, BE-PE and DE-PE when compared to PE. Injection of polyethylene particles increased the air pouch membrane thickness significantly compared to the PBS control in all particle types and doses. Higher particle dose induced thicker membrane in all 4 materials. A similar trend was also observed in the percentage of inflammatory cell infiltration in the pouch membrane. C-PE and DE-PE particles at low dose and C-PE particles at high dose induced lower levels of IL-1β and TNFα than PE. IL-6 production was similar between PE and other 3 groups. Discussion/Conclusion. Antioxidant incorporated in UHMWPE did not alter the level of macrophage proliferation and air pouch inflammation induced by UHMWPE particles, although it reduced cytokine gene expression. Future investigation in a synovial joint environment is desired to evaluate the chronic inflammation response to antioxidant containing UHMWPE wear particles and to verify the effect of antioxidant in UHMWPE properties

Introduction. The complex process of inflammation and osteolysis due to wear particles still is not understood in detail. So far, Ultra-high-molecular-weight-polyethylene (UHMWPE) is the bearing material of choice in knee arthroplasty and revision knee arthroplasty, but there is a growing demand for alternative bearing materials with improved wear properties. Lately, increasing interest developed in the use of natural and carbon-fiber-reinforced-poly-ether-ether-ketones (CFR-PEEK). While there is a lack of data concerning the effects of CFR-PEEK particles on human tissue, the effects of such wear debris in vitro and in animal studies is controversially discussed. The aim of this study was to analyze human tissue containing CFR-PEEK as well as UHMWPE wear debris. The authors hypothesized no difference between the used biomaterials because of similar size parameters of the wear particles in a prior knee simulator study of this implant. Methods and Materials. Synovial tissue samples of 10 patients while knee revision surgery of a rotating hinge knee implant design (Enduro®, Aesculap, Germany) were achieved. The tibial inserts of this design were made from UHMWPE (GUR 1020), whereas the bushings and flanges are made of CFR-PEEK containing 30% polyacrylonitrile (PAN) based carbon fibers (PEEK-Optima LT1, Invibio Ltd. Thornton-Cleveleys, UK). In a prior in vitro test most of the released CFR-PEEK particles were in a size range between 0.1 and 2μm. The implant survival until revision surgery was 22 (2.5–48 min.-max.) months. As a control synovial tissue out of a patient also got knee revision surgery without any PEEK components. The tissue was fixed with 4% paraformaldehyde, embedded in paraffin, sliced into 2 µm thick sections. stained with hematoxylin and eosin in a standard process. A modified panoptical staining (preincubation in propylenglycol; >3h; 35°C) was also done which stained the UHMWPE particles turquoise. The study was approved by the ethics committee of the local university. Results. Overall, histologically a “wear-type” reaction was seen in the testing and the control group similar as described for other materials in the common literature. In all samples of the testing group the UHMWPE particles were scattered in the tissue similar to the control. Larger UHWMPE particles were incorporated in giant cells. In contrast to these findings, CFR-PEEK particles were not scattered in tissue but located only as conglomerates. In addition, these conglomerates have been found exclusively near to or in vessels. Furthermore, CFR-PEEK particles were collected in macrophages, no CFR-PEEK particles were seen in giant cells. In conclusion, the hypothesis has to be rejected. Interestingely, different behaviour of UHMWPE and PEEK particles has been found in human synovial tissue. This aspect needs further investigation concerning the cytokine expression and also the surface texture of particles. Acknowledgement. This study was supported by Aesculap, Germany

Demineralized bone matrix (DBM) is a natural, collagen-based, well-established osteoinductive biomaterial. Nevertheless, there are conflicting reports on the efficacy of this product. The purpose of this study was to evaluate whether DBM collagen structure is affected by particle size and can influence DBM osteoinductivity. Sheep cortical bone was ground and particles were divided in three fractions with different sizes, defined as large (L, 1–2 mm), medium (M, 0.5–1 mm), and small (S, < 0.5 mm). After demineralization, the three DBM samples were characterized by DTA analysis, XRD, ICP-OES, and FTIR. Data clearly showed a particle size-dependent alteration in collagen structure, with DBM-M being altered but not as much as DBM-S. The in vivo study showed that only DBM-M was able to induce new bone formation in a subcutaneous ectopic mouse model. When sheep MSC were seeded onto DBM particles before implantation, all DBM particles were able to induce new bone formation with the best incidence for DBM-M and DBM-S. Gene expression analysis performed on recovered implants supports the histological results and underlines the supportive role of MSC in DBM osteoinduction through the regulation of host cells. In conclusion, our results show a relation between DBM particle size, structural modification of the collagen and in vivo osteoinductivity. The medium particles represent a good compromise between no modification (largest particles) and excessive modification (smallest particles) of collagen structure, yielding highest osteoinduction. We believe that these results can guide researchers to use DBM particles of 0.5–1 mm size range in applications aimed at inducing new bone formation, obtaining results more comparable and reliable among different research groups. Furthermore, we suggest to carefully analyze the structure of the collagen when a collagen-based biomaterial is used alone or in association with cells to induce new bone formation

The generation of particle debris from ultra high molecular weight polyethylene (UHMWPE) against metal hip joints has been shown to cause osteolysis leading to joint loosening in the medium term. This is known as late aseptic loosening since infection is absent. 1. . In an attempt to reduce the volume of wear debris, attention has moved to metal-on-metal prostheses as the total volume of wear debris is less. However, the size, shape and number of the particles are important as well as the total volume as these affect the biological response of the body leading to aseptic loosening. The Durham Mk I Hip Joint Simulator was used to generate CoCrMo wear particles in a series of tests. Four simulator tests took place in succession, initially 50 mm Birmingham hip replacements were tested where both the head and the cup were as-cast CoCrMo alloy. A second test was conducted where the components were 38 mm and both head and cup were as-cast CoCrMo. A third test using 50 mm components was completed where both head and cup were double heat treated CoCrMo alloy and a final test took place where both components were 50 mm the head was as-cast and the cup was as-cast which had been pre-worn to 5 million cycles. Bovine serum with a concentration of 17 g/l of protein was used as a lubricant and particles were sampled every half million cycles. The volumetric wear was also obtained gravimetrically. A double enzymatic protocol was used to cleave the proteins from the particles taking great care to minimise any effect on the particles. Finally the particles were suspended in distilled, de-ionised water to enable them to be analysed using a NanoSight LM10 particle analyser. This yielded the size distribution of the particles. This was then confirmed by placing an aliquot of the suspended particles onto a carbon coated copper grid and drying them under a lamp. These particles were then imaged in the TEM. Energy Dispersive X-ray analysis was also used to obtain the chemical composition of the particles. The results indicated a strong correlation between the gravimetric wear and the number of particles. All the as-cast CoCrMo alloy tests showed a consistent particle modal average size. The double heat treated particles were shown to be smaller, with occasional large flake like particulates which were identified under the TEM. This particle data correlates extremely well with previous data from simple material testing pin on plate experiments. Previous studies have used microscopy to investigate the size and morphology of the particulate debris. 2. , however these studies are limited due to the time taken to image the particles individually. This current method allows many more particles to be analysed, thus the data accumulated is more statistically significant and may be compared with the wear volumes calculated gravimetrically

INTRODUCTION. Loosening is concerned to be the major cause of revision in the artificial prosthesis. Wear debris of UHMWPE dispersed into the implant-bone interface are phagocytosed by macrophages releasing inflammatory cytokines such as TNF-α which leads to osteolysis and loosening eventually. It is known that the size and structure [1] as well as attached substances on particle surface such as endotoxin could affect the amount of cytokines released [2]. An in vivo study using rat femurs showed that the presence of polyethylene particles around implants could result in accumulation of lipopolysaccharide (LPS) from exogenous sources that may affect bone remodeling around implants [3]. It is also reported that LPS is transported throughout the body with lipoproteins or LPS binding proteins [4] and Circulating LPS may originate from local sites of infection or via bloodborne bacteria [5]. In this study, we evaluated the effects of LPS that attached to UHMWPE particle surface by measuring TNF-α released from macrophages. MATERIALS AND METHODS. We cultured mouse macrophage cell line RAW 264 with spherical UHMWPE particles (8.7µm and 23µm diameter in average, Mitsui chemicals Co., LTD.) and LDPE particles (3.6µm and 5.8µm diameter in average, Sumitomo Seika Chemicals Co., LTD.) using the Inverse Culture Method for 24 hours before estimating the TNF-α generation by TNF- ALPHA QUANTIKINE ELISA KIT (R&D). Spherical UHMWPE particles (10µm diameter in average, Mitsui chemicals Co., LTD.) with E.coli original LPS (Enzo Life Sciences) attached to them were incubated with cells to see the effects of LPS on the bio-reactivity tests. REAULTS AND DISCUSSIONS. Figure 1 shows the TNF-α concentration of different materials and sizes of polyethylene particles. TNF-α concentration was shown to be dose-dependent to the total surface area of particles added regardless of the materials and sizes. Figure 2 shows TNF-α concentration relative to the particle surface area inverse and non-inverse cultured. No significant difference was observed in TNF-α concentration between particles that were attached to LPS and virgin particles in non-inverse culture method. However, when cultured inversely, the effect of LPS became more significant in higher surface area range in which dose-dependent relationship was not observed. The results suggest that saturation may occur caused by size exclusion, production limitation, etc. However, LPS attached to particle surface may alter the production limitation due to increased presence of particles around macrophages. For any figures or tables, please contact the authors directly

Today's aging society is seeing an increase of patients with rheumatoid arthritis and osteoarthritis, as well as an increase in joint replacement surgery. The artificial joints used in this surgery frequently uses ultra-high molecular weight polyethylene (UHMWPE) as a bearing material. However, UHMWPE wear particles are considered to be a major factor in long-term osteolysis, and implant loosening. Many researchers have reported that the volume and size of particles are critical factors in macrophage activation, with particles in the size range of 0.1 – 1.0 μm being the most biologically active. The micro slurry-jet erosion (MSE) apparatus was introduced to minimize the amount of wear, and increase the size of UHMWPE wear particles by texturing the surfaces of Co-Cr-Mo alloy implants. The MSE apparatus uses a slurry of alumina particles (WA#8000: average diameter 1.2 μm) mixed with water. The slurry and compressed air are mixed within an injection nozzle, which is then applied to the Co-Cr-Mo alloy at high speed to achieve a desired nano-textured surface. In this study, four Co-Cr-Mo alloy surface profiles were prepared. The MSE injection nozzle was fed 40.0 mm in alternating directions across each surface with an orthogonal step of 0.5 mm. The surface M-1 was processed with an injection nozzle feed rate of 1.0 mm/s, and obtained a surface roughness of 5.7 nm. M-2 was processed with a feed rate of 2.0 mm/s, and had a surface roughness of 2.3 nm. The M-4 surface used a 40.0 mm alternating directions surface feed, but with a 1.0 mm orthogonal step, and an injection nozzle feed rate of 0.5 mm/s. It obtained a surface roughness of 4.0 nm. The G-1 surface, with a roughness of 10.0 nm, was processed with the typical lapping method, which is used in conventional artificial joints [Fig. 1]. A pin-on-disk wear tester, capable of multidirectional motion, was used to assess which surface was the most appropriate for artificial joints. The UHMWPE pins were flat ended cylinders, 12.0 mm in diameter, and were placed on the disk with a contact pressure of 6.0 MPa. Tests were carried out in 25% (v/v) fetal calf serum with sodium azide to retard bacterial growth. A sliding speed of 12.1 mm/s, and a total sliding distance of 15.0 km were applied. The wear weight of the MSE textured surface M-1 was significantly lower than the wear weight of the conventional surface. Moreover, the percentages of various wear particle sizes obtained from MSE surface texturing was significantly different from those obtained from the traditional surface

To evaluate the mechanism by which orthopedic implant wear particles induce apoptosis in immature osteoblasts in an in-vitro setting. Stromal cells from femurs of thirty day-old Swiss Webster Mice were isolated, cultured in-vitro, and incubated with orthopedic wear particles in the micrometer size range. After incubation with wear-particles, the cells were assessed for Caspase three expression and activity in the presence or absence of specific inhibitor(s) in order to delineate potential mechanism for cellular changes previously reported. Here we report the induction of caspase three protein expression and activity with incubation of stromal cells with titanium wear particles. Caspase three activity however was not demonstrated to be up regulated in a time dependent manner or at lower concentration of particles (2 x 107 particles/ml). However, there was a significant (P< 0.05) increase in caspase three activity with titanium particle at higher concentration (4 x 107 particles/ml) that was not reversible when the extrinsic arm of the apoptotic pathway was blocked with anti-TNFƒa antibodies. Our previous studies have suggested that aseptic loosening of orthopedic implants may be independent of inflammatory processes, and may be associated with induction of programmed cell death. Our current results would strengthen this idea by demonstrating induction of expression and activity of caspase three involved in apoptosis in cells incubated with wear particles. In addition, titanium wear particles may induce apoptosis through direct cellular effects rather than through the extrinsic TNFƒa pathway. Delineating the mechanism by which wear particles induce apoptosis in immature osteoblasts will allow for the selection and/or development of inhibitors to the process of asceptic loosening by targeting a specific pathway

Introduction. Patients suffering from finger joint pain or dysfunction due to arthritis and traumatic injury may require arthroplasty and joint replacement. Single-part silicone-based implants remain the material of choice and most widely used option, although reports on their long-term clinical performance are variable. For trauma indications, patients have a high expectation of functionality necessitating the use of materials with high wear resistance and mechanical performance. A new proximal inter phalangeal (PIP) joint designed by Zrinski AG (Wurmlingen, Germany), comprising a self-mating carbon fibre reinforced polyetheretherketone (CFR-PEEK) coupling, may provide a suitable alternative. Here we describe the wear performance of the CFR-PEEK components in a PIP joint wear simulator and subsequent characterisation of the wear particles. Methods. Four proximal and distal PIP components were milled (Zrinski AG) from CFR-PEEK (Invibio Ltd, UK) and subjected to wear testing (Endo Lab ® GmbH, Germany). The test was conducted at 37°C over 5 million cycles in 25% bovine serum (refreshed every 0.5 million cycles). The load was a static force of 63N applied at a frequency of 1Hz with a flexion/extension angle of ±40°. Wear rate was determined by mass loss from each component. Pooled serum samples from the wear simulator were subjected to protein digest and the remaining particulate debris isolated by serial filtration through 10μm, 1μm and 0.1μm filters. Particle size and morphology was subsequently determined by scanning electron microscopy (SEM) (Continuum Blue, UK). Results. Both components exhibited high resistance to wear, with the proximal component resulting in a wear rate of 0.09mg/million cycles, whilst that of the distal component was 0.07mg/million cycles. Particle analysis revealed that the majority of debris generated during the wearing in phase (0.5 million cycles) was <0.5μm in diameter. During the steady state phase (0.5–3 million cycles) a large peak in particle size was observed in the 2μm diameter range, whilst in the latter stage (3–5 million cycles) peaks in particle size were seen at 0.4μm and 2μm. During each stage, both the particle count and aspect ratio remained relatively unchanged. Conclusion. Under these test conditions the CFR-PEEK coupling demonstrated a linear and consistently low wear rate over the 5 million cycle test period, with the majority of particles generated being <2μm in diameter. The low wear rate and biocompatibility demonstrated by CFR-PEEK suggests it is a suitable alternative to silicone in PIP joint prostheses. Acknowledgements. The authors would like to thank Zrinski AG, Christian Kaddick at EndoLab GmbH for the wear simulator work and Mark Yeoman at Continuum Blue Ltd. for particle analysis

We previously reported no clinical differences in short-term results in 26 patients that underwent fixed-bearing (FB) total knee arthroplasty in one knee followed by a rotating-platform (RP) version of the same implant in the contralateral knee at a later date. This study presents intermediate-term results in this unique cohort and uses optimised MRI for detection of particle disease in both knees. Patients from the original series were asked to complete questionnaires regarding both knees. In addition, both knees were evaluated with optimised MRI, which has been shown to be useful in evaluating early particle disease and osteolysis before its appearance on radiographs. Nine patients have been enrolled to date. At an average follow-up of 8.3 years for the FB side and 6.5 years for the RP side, no significant differences were found with respect to knee preference, pain, or overall satisfaction. Seven patients underwent MRI studies of both knees. Two FB knees demonstrated a massive intracapsular burden of particle disease (average 3066 mm3) with reactive synovitis, compared to no obvious particle disease in any RP knees. Osteolysis was seen around the femur in one FB knee and around the patella in two FB knees, compared to only around a single patella in the RP side. RP knees continue to demonstrate excellent patient satisfaction that is comparable to clinical results of the FB design; however, FB knees demonstrate higher rates of particle disease and early osteolysis on MRI. This is the first study to demonstrate in vivo advantages of RP over FB designs. It is unclear whether this is due to the slightly longer follow-up period for the FB knees or a decreased wear rate in the RP design; these differences may become apparent with longitudinal follow-up

Summary Statement. Using the latest Next Generation Sequencing technologies, we have investigated miRNA expression profiles in human trabecular bone from total hip replacement (THR) revision surgery where wear particle associated osteolysis was evident. Introduction. A major problem in orthopaedic surgery is aseptic loosening of prosthetic implants caused by wear particle associated osteolysis. Wear debris is known to impact on a variety of cellular responses and genes in multiple pathways associated with the development of the periprosthetic osteolysis. MicroRNAs (miRNAs) act as negative regulators of gene expression and the importance of miRNAs in joint pathologies has only recently been addressed. However, miRNA profiles in osteolytic bone are largely unknown. Using the latest Next Generation Sequencing technologies, we have investigated miRNA expression profiles in human trabecular bone sourced from bone discarded during total hip replacement (THR) revision surgery where wear particle associated osteolysis was evident. Patients and Methods. Three groups of gender and age-matched patients (n=9 per group) were recruited for this study including patients undergoing revision surgery, primary THR patients and healthy subjects. Total RNAs were prepared from trabecular bone specimens. The cDNA libraries were constructed using a TruSeq Small RNA Sample Preparation kit, and then sequenced on an Illumina HiSeq2000 sequencer. All good quality tags were aligned against the reference sequences containing human chromosomal sequences and 18s and 28s rRNA sequences were analysed using Bowtie software. We used miRBase v19 to identify the start positions of all mature miRNA and the edgeR package to analyse differential expression. Osteogenesis pathway-related gene expression was also investigated using RT-qPCR Array assay. Results. We observed a significant difference in expressed miRNAs between revision and primary THR groups, including upexpressed miR127, miR-409, miR-211 and miR-146a. Importantly, the miR-127 (3.1 fold, p=0.005) and miR-146a (3.5 fold, p=0.001) were not only upexpressed in the revision group vs primary group, but also upexpressed in the revision group vs the healthy group. Thus, miR-127 and miR-146a may have potential as both biomarkers to predict osteolysis and as therapeutic targets. The miR-127 and miR-146a are critical in bone diseases because some of their target genes play an important role in osteogenesis. We have thus studied osteogenic genes and confirmed that SMAD4, RUNX2, FGFR1, TGFβ1, COL1A1 and WNT4 were downregulated. Our data also revealed that miR-93 and miR-204a were downexpressed (−3.7 fold, p=0.023; −2.5, p=0.003 respectively) and t IL-6 and IL-6R, which had been reported as miR-204 target genes, were upexpressed. Discussion and Conclusion. Our results showed that upexpressed miR-127, miR-146a, miR-204a and miR-93 in trabecular bone from revision THR may be the key negative regulators in either osteogenic genes involved in osteogenic differentiation of bone formation or inflammatory genes involved in osteoclastogenesis. Aberrant miRNA expressions identified in the revision THR group may also suggest the existence of genetic risk factors favouring the development of osteolysis in certain specific subgroups of patients. An in-depth understanding of the roles of these regulatory miRNAs in the skeleton warrants further investigation

The ability of optimised MRI to detect periarticular bony and soft tissue pathology in the post-arthroplasty hip is well documented; specifically it is able to detect early stages of particle disease well before osteolysis is apparent on radiographs. This is a prospective study designed to utilise MRI for the detection of early particle disease in asymptomatic patients after total hip arthroplasty. Patients who underwent routine non-cemented THA were recruited from three different groups: metal-on-polyethylene, ceramic-on-ceramic, and ceramic-on-polyethylene bearing surfaces. All patients enrolled underwent optimised MRI one to three years (mean 1.7) after the index procedure. Images were analyzed for the presence of synovial proliferation, fibrous membrane formation or osteolysis. Particle disease was correlated with type of bearing surface, pain, activity level, patient satisfaction, and clinical outcome scales. Thirty-two hips have been enrolled in the study to date. Early particle disease was seen in two of seven metal-on-polyethylene hips (29%), four of twelve ceramic-on-ceramic hips (33%), and six of thirteen ceramic-on-polyethylene hips (46%). Focal osteolysis was seen in one patient with a ceramic-on-polyethylene hip. These values were not statistically significant among the groups. The presence of early particle disease did not correlate with pain, activity level, patient satisfaction, or other clinical outcome scales. This study allows patients with a well functioning total hip arthroplasty to be prospectively followed with MRI. It is the first to document the natural history of particle disease in vivo and considerably enhances our knowledge of periarticular pathology in the post-operative hip. These results demonstrate early particle disease is relatively common yet asymptomatic; they do not demonstrate advantages of any bearing couple over another for protection against particle disease at short-term follow-up

Endoprosthetic reconstruction for pathologic acetabular fractures is associated with a high risk of periprosthetic joint infection. In this setting, bone defect reconstruction utilising co-delivery of a synthetic bone substitute with an antibiotic, is an attractive treatment option from both, therapeutic and prophylactic perspective. We wished to address some concerns that remain regarding the possible presence of potentially wear inducing particles in the periprosthetic joint space subsequent to this procedure. We analysed a drain fluid sample from an endoprosthetic reconstruction of a pathologic acetabular fracture with implantation of a gentamicin eluting, biphasic bone graft substitute, consisting of 40% hydroxyapatite (HA) and 60% calcium sulphate (CERAMENT G), into the residual peri-acetabular bone defect. This sample was divided into two 1.5ml subsamples, to one of which 100mg HA particles were added as control before burning off all organic substance at very high temperature. These heat treated samples were then examined with scanning electron microscopy (SEM) and energy dispersive x-ray analysis (EDAX) and compared to a reference sample consisting of HA particles only. On SEM, hydroxyapatite particles were readily recognisable in the control and reference samples, whereas only very few particles over 2μm were apparent in the ”pure” drain sample. EDAX revealed that very large amounts of salts were present in both drainage samples. The pure drainage sample however, contained markedly lower amounts of calcium and phosphate compared to reference and control samples. No HA particles as such, were seen in the pure sample, however their presence cannot be excluded with absolute certainty, as some particles might have been hidden within the large salt conglomerates. We could not find clear evidence that the drain fluid really contained HA particles. More thorough investigations are needed and future analyses with prior removal of the high salt content would likely yield more conclusive results

Particulate debris has been demonstrated to have a major role in the aseptic loosening of artificial joints. Biological activity is stimulated by particles of size 0.1 to 10 microns with maximum affect being seen in particles of approximately 0.5 microns. This study investigated the particle size of wear debris produced at the stem/cement interface in cemented artificial hips comparing different materials and surface finishes of the stem. Materials and Methods: The stem surfaces of three cemented femoral prostheses with different surfaces were used as templates, one smooth (Exeter, How-medica), one slightly rough (Elite, DePuy) and one very rough (Capital, 3M). Three commonly used implant materials were compared, cobalt chrome, stainless steel and titanium alloy. Test plates were manufactured in each material and with each of the three surface finishes. The plates were opposed to cement pins in a sliding wear tester. The debris produced was collected, filtered and examined under an electron microscope. Representative samples of particles produced by each test series were measured and aspect ratios calculated. Plates were also measured pre and post test to ensure no significant change in surface roughness during testing. Results: Each of the materials demonstrated an increase in mean particle size with increasing roughness of the test surfaces. This was more marked for titanium alloy than for cobalt chrome. For any given surface roughness the size of particles produced was smallest for the hardest material (cobalt chrome) and largest for the softest material (titanium alloy). Conclusion: The majority of particles produced were within the biologically active range. Particle size of debris was related to surface roughness with rougher surfaces producing larger particles. Harder materials produced smaller particles than softer materials and more particles in the biologically active range

Summary. Particulate wear debris with different chemical composition induced similar periprosthetic tissue reactions in patients with loosened uncemented and cemented titanium hip implants, which suggests that osteolysis can develop independent of particle composition. Introduction. Periprosthetic osteolysis is a serious long-term complication in total hip replacements (THR). Wear debris-induced inflammation is thought to be the main cause for periprosthetic bone loss and implant loosening. The aim of the present study was to compare the tissue reactions and wear debris characteristics in periprosthetic tissues from patients with failed uncemented (UC) and cemented (C) titanium alloy hip prostheses. We hypothesised that implant wear products around two different hip designs induced periprosthetic inflammation leading to osteolysis. Patients & Methods. Thirty THR-patients undergoing revision surgery were included: Fifteen patients had loose cemented titanium stems (Titan. ®. , DePuy) and 15 had well-fixed uncemented titanium stems (Profile, DePuy), but loose or worn uncemented metal-backed cups with conventional UHMWPE liners (Gemini, Tropic and Tri-Lock Plus, DePuy; Harris/Galante and Trilogy, Zimmer). A semi-quantitative histological evaluation was performed in 59 sections of periprosthetic tissues using light microscopy. Wear particles were counted by polarised light and high resolution dark-field microscopy. Additionally, particle composition was determined by SEM-EDXA following particle isolation using an enzymatic digestion method. Blood metal ions were determined with high resolution-ICP-MS. Results. The implants in the uncemented group were revised after a median of 15.7 years (range: 7.25–19.3) due to osteolysis and high wear of the polyethylene liner and metal backing resulting in gross metallosis, and/or cup loosening. The average lifetime of implants in the cemented group was only 6.5 years (range: 1.5–11.75) due to early stem loosening with large osteolysis pockets in the femur close to the cement mantle. Tissue examination revealed similar results for both groups: numerous mononuclear histiocytes and chronic inflammatory cells, a few neutrophils and multinucleated giant cells, and to some extent necrosis. The amount of metal particles per histiocyte positively correlated with the tissue reactions in the cemented, but not in the uncemented group. A higher particle load (medians: C: 14727 vs. UC: 1382 particles/mm. 2. , p<0.0001) was found in tissues adjacent to cemented stems, which contained mainly submicron ZrO. 2. particles. Particles containing pure Ti or Ti alloy elements (size range: 0.21 to 6.46 µm) were most abundant in tissues from the uncemented group. Here, also PE was more frequent, but accounted only for a small portion of total particles (2.8 PE/mm. 2. ). The blood concentrations of titanium (range: 3.8–138.5 microgram/L) and zirconium (cemented cases, range: 0.6–3.5 microgram/L) were highly elevated in cases with high abrasive wear and metallosis. Discussion/Conclusion. Phagocytosis of different wear particles by histiocytes induced a similar chronic inflammatory reaction in the periprosthetic tissues in both groups. ZrO. 2. particles, originating from bone cement degradation, dominated in the cemented group, while in the uncemented group the high abundance of pure Ti and Ti alloy particles of various sizes indicates wear of the metal-backed cups. The low density of polyethylene particles in the tissues suggests that they are not solely responsible for the tissue reactions and accompanying osteolysis. Our findings suggest that the chemical composition of wear particles plays a minor role in the mechanism of osteolysis. Particle size, load and ionic exposure might be more important

Introduction: Joint replacement surgery is one of the most common operations that take place in United Kingdom. The major problem in total hip arthroplasty is the generation of particulate wear debris and the subsequent biological responses. Wear debris induces osteolysis and a subsequent failure of the implant that lead to the liberation of greater quantities of particulate and soluble debris to bone marrow, blood, lymph nodes, liver and spleen. Recently, it has been suggested that these adverse effects depend not only on the chemical composition but also on the particulate nature of the material (size and shape). Particle size has been shown to influence the inflammatory response of macrophages to wear debris. This study evaluated whether particle size also influences the viability and mutagenic damage. Methods: Cobalt chrome alloy particles of two sizes (large 2.9±1.1μm, small 0.07±0.04 μm) were generated and characterised by Scanning Electron Microscopy. Different concentrations of particles were added to primary human fibroblasts in tissue culture. The release of cytokines in the medium was assayed by Enzyme-Linked ImunnoSorbent Assay (ELISA). Cell viability was determined by MTT conversion and the degree of DNA damage was quantitatively analysed by the Alkaline Single Cell Gel Electrophoresis (COMET) assay with image analysis. Results: Small particles initialise DNA damage at much lower volumetric concentrations (0.05 and 0.5 μm. 3. /cell) than larger particles (500 μm. 3. /cell). The difference in the doses was approximately related to the difference in surface area of the particles. DNA damage was related to a delayed decrease in cell viability, which was noted after three days of exposure. In contrast, the release of the inflammatory cytokine TNF-α and the multifunctional growth factor TGF-β-2 occurred at lower doses (0.0005 to 5 μm. 3. /cell for TNF-α and 0.5 to 50 μm. 3. /cell for TGF-β-2). No release of IL-6 was detected at any dose. Only growth factor FGF-23 was increased in similar pattern to the DNA damage. Conclusions: This study has demonstrated important differences between the mutagenicity, toxicity and inflammatory potential of small (nanometre sized) and large (micrometer sized) chrome particles

Introduction: The main problem facing the longevity of total hip replacements (THR) is wear particle induced osteolysis, particularly around the acetabular component. The articulating surfaces produce wear particles that migrate in the fibrous tissue membrane along the acetabular implant-bone interface causing osteolysis and subsequent implant loosening. The hypothesis that we investigated was that uncemented acetabular interfaces are more effective than cemented implants at resisting progressive osteolysis through bone attachment and the formation of a biological seal. Methods: THR surgery was performed in an ovine model. Implants remained in vivo for 1 year. Femoral heads were roughened in order to generate wear debris and aseptic loosening of the acetabular component. Sheep were randomly assigned to one of three experimental groups: cemented polyethylene, grit blasted or plasma sprayed porous acetabular components with a polyethylene insert. Ground Reaction Force (GRF) data was collected pre-op and at 12, 24, 36 and 52 weeks post op. Retrieved specimens were analysed radiographically, histologically and using Scanning Electron Microscopy (SEM). A mould was made of the polyethylene liner and head penetration rates quantified using a shadowgraph technique. Thin sections through the acetabuli were prepared and image analysis used to quantify fibrous tissue (FT) thickness at the bone-implant interface. Mann-Whitney U tests were used for comparative statistical analysis where p< 0.05 were classified as significant. Results: GRF demonstrated functional hips. A gradual increase was seen until week 36 followed by a decrease until retrieval suggesting the onset of aseptic loosening. 42.86% of control, 60% of grit blasted and 50% of porous coated components were deemed radiographically loose. Mean linear penetration rates demonstrated significantly less penetration within the porous cups (p=0.003, control and p=0.036, grit blasted). SEM established that wear particles generated were < 1μm in size. Light microscopy of thin sections revealed the common mechanism of loosening involving a resorption wedge at the interface with progressive bone loss. In all cases, the FT layer was greatest at the rim of the cup and gradually decreased towards the apex. The grit blasted group had the thickest FT layer adjacent to the cup. Under polarised light, wear debris was seen packed within macrophages in all sections. Discussion: GRF data demonstrated grit blasted cups to have least function. This was confirmed through histology as they had the thickest FT layer surrounding the acetabular shell suggesting increased aseptic loosening of its component due to wear particles being able to access the interface more easily. Data corroborates radiographic results. In conclusion, porous and control cups performed better than grit blasted cups. Acknowledgments: EPSRC

Deep infection occurs in 2–4% of lower limb arthroplasty resulting in increasing cost, co-morbidity and challenging revision arthroplasty surgery. Identifying the potential sources of infection helps reduce infection rates. The aim of our study is to identify the impact and potential for contamination of our hands and gowns whilst scrubbing using SSHS. A colony-forming unit (CFU) is a pathogenic particle of 0.5 micrometers to 5 micrometers. Concurrent particle counts and blood agar exposure settle plates for 3 subjects and 1 alcohol cleaned mannequin; testing a standard arthroplasty hood, a SSHS with and without the fan on for a 2 minute exposure to represent scrubbing time. Microbiological plates were incubated using a standard protocol by our local microbiology department. All SSHS were positive for gram-positive cocci with a mean colony count of 410cfu/m. 2. Comparing background counts for laminar flow (mean 0.7 particle/m. 3. ; 95% CI 0–1.4) versus scrub areas (mean 131.5 particle/m3; 95% CI 123.5–137.9; p=0.0003), however neither grew any CFU's with a 2-minute exposure. For the mannequin, the only significant result was with the fan on with a 1.5× increase in the particle count (p=0.042) and a correlating positive organism (13CFU/m. 2. ). With human subjects, however, the particle count increased by 3.75× the background count with the fan on (total p=0.004, CFU p=0.047) and all had positive cultures, mean 36 CFU/m. 2. There were no positive cultures with the standard arthroplasty hood or the SSHS with no fan on. If repeated in laminar flow, there was only a statistically significant increase with the fan on (p=0.049), but with negative cultures following a 2-minute exposure. Sterile gloves and gowns can be contaminated when scrubbing with the SSHS fan on. We recommend having the fan switched off when scrubbing until the hood and gown is in place, ideally in a laminar flow environment