Aim

Staphylococcus aureus (SA) can cause various infections and is associated with high morbidity and mortality rates of up to 40%. Antibiotic treatment often fails to eradicate SA infections even if the causative strain has been tested susceptible in vitro. The mechanisms leading to this persistence is still largely unknown. In our work, we to reveal SA interactions with host cells that allow SA to persist at the site of infection.

Introduction

Wear debris and metal ions originating from metal on metal hip replacements have been widely shown to recruit and activate macrophages. These cells secrete chemokines and pro-inflammatory cytokines that lead to an adverse local tissue reaction (ALTR), frequently requiring early revision. The mechanism for this response is still poorly understood. It is well documented that cobalt gives rise to apoptosis, necrosis and reactive oxygen species generation. Additionally, cobalt stimulates T cell migration, although the effect on macrophage motility remains unknown. This study tests the hypothesis that cobalt ions and nanoparticles affect macrophage migration stimulating an ALTR.

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.

Implant-related infection (IRI) is closely related to the local immunity of peri-implant tissues. The generation of reactive oxygen species (ROS) in activated macrophages plays a prominent role in the innate immune response. In previous studies, we indicated that implant wear particles promote endotoxin tolerance by decreasing the release of proinflammatory cytokines. However, it is unclear whether ROS are involved in the damage of the local immunity of peri-implant tissues. In the present study, we assessed the mechanism of local immunosuppression using titanium (Ti) particles and/or lipopolysaccharide (LPS) to stimulate RAW 264.7 cells. The results indicate that the Ti particles induced the generation of a moderate amount of ROS through nicotinamide adenine dinucleotide phosphate oxidase-1 (NOX-1), but not through catalase. Pre-exposure to Ti particles inhibited ROS generation and extracellular regulated protein kinase (ERK) activation in LPS-stimulated macrophages. These findings indicate that chronic stimulation by Ti particles may lead to a state of oxidative stress and persistent inflammation, which may result in the attenuation of the immune response of macrophages to bacterial components such as LPS. Eventually, immunosuppression develops in peri-implant tissues, which may be a risk factor for IRI.

Aim. In the current study we aim to characterize the use of cationic host defense peptides (HDPs) as alternative antibacterial agents to include into novel antibacterial coatings for orthopedic implants. Staphyloccous aureus represent one the most challenging cause of infections to treat by traditional antibacterial therapies. Thanks to their lack of microbial resistance described so far, HDPs represent an attractive therapeutic alternative to antibiotics. Furthermore, HDPs have been showed to control infections via a dual function: direct antimicrobial activity and regulation of immune response. However, HDPs functions characterization and comparison is controversial, as changing test conditions or cell type used might yield different effects from the same peptide. Therefore, before moving towards the development of HDP-based coatings, we need to characterize and compare the immunomodulatory and antibacterial functions under the same conditions in vitro of 3 well-known cathelicidins: human LL-37, chicken CATH-2, and bovine-derived IDR-1018. Method. S. aureus, strain SH1000, was incubated with different concentrations of each HDP and bacterial growth was monitored overnight. Primary human monocytes were isolated from buffy coats using Ficoll-Paque density and CD14 microbeads, and differentiated for 7 days to macrophages. After 24h incubation in presence of LPS and HDPs, macrophages cytokines production was measured by ELISA. Macrophages cultured for 24h in presence of HDPs were infected with serum-opsonized S. aureus. 30 min and 24h after infection, bacterial phagocytosis and intracellular killing by macrophages were measured by flow cytometry and colony forming units (CFU) count respectively. Results. All HDPs efficiently inhibit macrophages LPS-mediated activation, as observed by a reduced production of TNF-α and IL-10. Despite a comparable anti-inflammatory action, only CATH-2 shows direct antibacterial properties at concentrations 10-times lower than those needed to stimulate immune cells. Although stimulation with HDPs fails to improve macrophages ability to kill intracellular S. aureus, IDR-1018 decreases the proportion of cells phagocytosing bacteria. Conclusions. In addition to a strong anti-inflammatory effect provided by all HDPs tested, CATH-2 has direct antibacterial effects while IDR-1018 reduces the proportion of macrophages infected by S. aureus. Use of these HDPs in combination with each other or with other conventional antibacterial agents could lead the way to the design of novel antibacterial coatings for orthopedic implants

Introduction. Macrophages phagocytes implant wear debris and produce various cytokines to evoke inflammation and periprosthetic osteolysis of aseptic loosening. It had been reported that expression of Toll-like receptor (TLR) 2 and other TLRs increased in periprosthetic tissues of aseptic loosening. Pathogen-associated molecular patterns (PAMPs) and damaged-associated molecular patterns (DAMPs) have been known as ligands of TLRs and considered to be involved in the osteolytic reactions via TLRs. Another type of immune sensors, nucleotide-binding and oligomerization domain (NOD)-like receptors (NLR) with a pyrin domain 3 (NLRP3) can also recognize PAMPs and DAMPs as their lignds, which has been presumed to participate in the local host response of macrophage cascade via phagocytosis of implant wear particles. However, the contribution of NLRP3 in periprosthetic tissues of aseptic loosening and the correlation between TLR2 and NLRP3 are still unclear. Materials and methods. TLR1, TLR2, TLR6, NLRP3, TNF-α and IL-1β of macrophages in aseptic loose periprosthetic tissues were immnohistorically evaluated and compared to osteoarthritic synovium. RAW264.7 cells, macrophagic cell line, were stimulated by titanium particles (Ti) and lipoteichoic acid (LTA)-coated Ti. The celluar reaction associated with TLR2 and NLRP3 and the correlation of them were analyzed at mRNA expression levels with small-interfering RNA of Irak2, one of adaptor molecules in TLR2 cascades. Results. Macrophages, which expressed abundant TLR2, NLRP3, TNF-α and IL-1β, were observed dominantly in foreign body granuloma of aseptic periprosthetic tissues. The features of abundant expression were quite different from osteoarthritic synovium. In vitro experiment of RAW264.7, mRNA levels of NLRP3 and TNF-α increased after stimulation of Ti. mRNA levels of TLR2, NLRP3, TNF-α and IL-1β were enhanced by LTA-coated Ti. mRNA expression level of NLRP3 were suppressed by silencing Irak2. Discussion and conclusion. This study indicated that innate immune sensors, TLR2 and NLRP3, could respond to foreign body particles in aseptic loose periprosthetic connective tissues. In this process, mRNA expression levels of TLR2 and NLRP3 in RAW264.7 were increased by phagocytosis of Ti particles, especially by LTA-coated Ti stimulation. Suppressed mRNA expression level of NLRP3 by knocked down of Irak2 indicated that TLR2 cascade could enhance activation NLRP3 cascade and/or free LTA may stimulate NLRP3 cascade directly. It may be possible that TLR2 and NLRP3 cascades in macrophages recognising PAMPs and/or DAMPs are activated each other and they play an important role of the pathogenesis of wear debris around loose hip joints

INTRODUCTION. This study is to determine the response of CRP after TKR surgery, both unilateral and simultaneous bilateral TKR. According to the previously published literature from North America and Europe CRP value peaks on the 1. st. and 2. nd. post-operative day and then gradually comes down to normal by 6–8 weeks post-operatively. AIM. To determine the trend of CRP in Indian patients undergoing TKR, both unilateral and simultaneous bilateral TKR. To see whether it follows the trend in North American and European population and to determine whether there is a difference in the CPR pattern in unilateral versus simultaneous bilateral TKR patients. MATERIAL & METHODS. Twenty six patients were included in this study. 13 patients each had unilateral TKR and simultaneous bilateral TKR. All the patients were operated by a single surgeon and assistant. Patients who had Rheumatoid arthritis and post operative adverse events like urinary tract infection were excluded from this study. All 26 patients were female patients and the mean age in the unilateral group was 67 years and in the simultaneous bilateral TKR group was 73 years. CRP levels were measured pre- operatively on 2. nd. day and 8 weeks. TKR was performed in a standard fashion. Both the groups received standard pre and post operative antibiotic prophylaxis. All patients received a posterior stabilized knee implant (Maxx Freedom Knee). RESULTS. In both the groups CRP level shot up on the 2. nd. post-operative day. Although the rise in CRP level was significantly higher in the simultaneous bilateral TKR group as against the unilateral TKR group. This difference was statistically significant. The CRP level came back to normal in about 39% of unilateral TKR patients at 8 weeks post operatively, while in majority (12 out of 13) of bilateral simultaneous TKR patient it was still elevated at 8 weeks post-op and had not come to normal. DISCUSSION. Macrophages are the important in the development of acute phase response namely CRP. The macrophages are present in the bone and bone marrow and less often in the skeletal muscle. The bone and bone marrow injury happening while performing TKR is responsible for elevation of CRP. Various North-American and European studies have shown that the CRP level increases significantly on the 1. st. postoperative day and the decreases from a peak on the 2. nd. postoperative day, attaining normal value at 6 to 8 weeks after operation. The result of our study are in variance to this published literature. Nearly 39% of our unilateral TKR patients and majority all of our simultaneous bilateral TKR patients did not achieve a normal CRP at 8 weeks after operation. These findings are significant as CRP is often used as a very sensitive indicator of post operative joint infection. Hence we conclude that the Indian TKR patients take longer time for the CRP values to become normal and the published literature regarding the normal levels of CRP in Unilateral TKR should not be extrapolated to simultaneous bilateral TKR group

Introduction. Little is known about the relationship between head-neck corrosion and its effect on periprosthetic tissues and distant organs in the majority of patients hosting apparently well-functioning devices. We studied the degree and type of taper damage and the histopathologic response in periprosthetic tissue and distant organs. Methods. A total of 50 contemporary THRs (34 primary, 16 revision) retrieved postmortem from 40 patients after 0.4–26 years were studied. Forty-three femoral stems were CoCrMo and 7 were Ti6Al4V. In every case, a CoCrMo-alloy head articulated against a cementless polyethylene cup (19 XLPE and 31 UHMWPE). H&E and IHC sections of the joint pseudocapsules and liver were graded 1–4 for the intensity of various inflammatory cell infiltrates and tissue necrosis. The nature of the tissue response in the joint capsule, liver, spleen, kidneys and lymph nodes was assessed. Wear and corrosion products in the tissues were identified using SEM and EDS. Taper surfaces were graded for corrosion damage using modified Goldberg scoring and examined by SEM to determine the acting corrosion mode. Correlations between damage scores and the histologic variables were generated using the Spearman test. Results. No correlation was seen between taper damage scores and the macrophage response in the joint pseudocapsule. The distribution of corrosion scores for heads and femoral trunnions is shown in Figure 1. Moderate or severe corrosion of the head and/or trunnion was present in 9 hips (8 CoCr/CoCr and 1 CoCr/TiAlV). One patient with bilateral hips had local ALVAL-like lymphocyte-dominated tissue reactions (Figure 2) and mild focal lymphocytic infiltrates in the liver and kidneys (Figure 3). This was associated with severe intergranular corrosion of the CoCrMo trunnion and column damage on the head taper. Particle-laden macrophages in pseudocapsules were significantly correlated with liver macrophages (r=.382, p=0.012) and liver lymphocytes (r=.367, p=0.013). Pseudocapsule macrophage responses to metallic and/or polyethylene wear particles ranged widely from minimal to marked. Focal tissue necrosis was related to high concentrations of particulate wear debris. A minimal number of metallic particle-laden macrophages were also detected in the liver and spleen; and macrophage granulomas were present in para-aortic lymph nodes, especially in revision cases. DISCUSSION. The generation of metal ions and particulates at corroded CoCrMo heads and CoCrMo or Ti6Al4V trunnions was a significant contributor to the presence of perivascular lymphocytes within the joint pseudocapsule, with 1 patient showing a histologic pattern consistent with ALVAL. Patient factors and the rate of corrosion are among variables influencing whether an ALVAL-type reaction will develop and whether or not it will become symptomatic. Macrophages in the joint pseudocapsules were positively correlated with inflammatory cells in the liver. In this study, the intensity of inflammatory infiltrates in distant organs was mild. However, several cases of organ dysfunction have been reported in association with catastrophic wear of CoCrMo components. It continues to be essential to minimize the generation of metal ions and particulates and to improve strategies for identifying and managing patients exposed to high levels of degradation products. For any figures or tables, please contact the authors directly

It is not known if the radiation sterilisation dose (RSD) of 25 kGy affects mechanical properties and biocompability of allograft bone by alteration of collagen triple helix or cross-links. Our aim was to investigate the mechanical and biological performance, cross-links and degraded collagen content of irradiated bone allografts. Human femoral shafts were sectioned into cortical bone beams (40 × 4 × 2 mm) and irradiated at 0, 5, 10, 15, 20, and 25 kGy for three-point bending tests. Corresponding cortical bone slices were used for in vitro determination of macrophage activation, osteoblast proliferation and attachment, and osteoclast formation and fusion. Subsequently, irradiated cortical bone samples were hydrolised for determination of pyridinoline (PYD), deoxypyridinoline (DPD), and pentosidine (PEN) by high performance liquid chromatography (HPLC) and collagen degradation by the alpha chymotrypsin (ï. j. CT) method. Irradiation up to 25 kGy did not affect the elastic properties of cortical bone, but the modulus of toughness was decreased from 87% to 74% of controls when the gamma dose increased from 15 to 25 kGy. Macrophages activation, the proliferation and attachment of osteoblasts on irradiated bone was not affected. Osteoclast formation and fusion were less than 40% of controls when cultured on bone irradiated at 25 kGy, and 80% at 15 kGy. Increasing radiation dose did not significantly alter the content of PYR, DPD or PEN but increased the content of denatured collagen. Cortical allografts fragility increases at doses above 15 kGy. Decreased osteoclast viability at these doses suggests a reduction in the capacity for bone remodelling. These changes were not correlated with alterations in collagen cross-links but in degradation to the collagen secondary structure as evidenced by increased content of denatured collagen

Aim. Ultrahigh molecular weight polyethylene (UHMWPE) has been used for many years as a bearing surface in total joint replacement (TJR). However, late-state failure in TJR is predominantly caused by osteolysis mediated by wear particles. We tested our hypothesis that UHMWPE nanoparticles are important determinants in activating dendritic cells (DCs). Methods. UHMWPE wear particles generated from a knee simulator were profiled using an atomic force microscopy and fractionated into six fractions: 0.05-0.2, 0.2-0.8, 0.8-1, 1-5, 5-10, and 10-20 micrometer. Effects of each fraction, a mixture of nano-sized fractions, and a mixture of all fractions on the activation of mice spleen DCs were determined using flow cytometry with specific antibodies of anti-CD11c-APC, anti-CD80-PE, anti-CD11b-PerCp, anti-CD86-Biotin and streptavidin-FITC. Supernatant from DCs treated with wear particles were assayed for IL-1beta, IL-6, IL-12/23, TNF-alpha and IFN-gamma. Activation of human osteoclasts (OCs) by wear particles were determined using TRAP stain. Results. DCs treated with a mixture of nanoparticles showed a significant increase in CD80 expression. A similar trend was not observed when DCs were treated with solvent or media, suggesting that the increased expression of CD80 was UHMWPE nanoparticle specific. Macrophages treated with nanoparticles did not show a significant increase in the expression of CD80, suggesting that DCs may be more sensitive to activation than macrophages. These results were further supported by the increased production of cytokines, IL-1beta and IL-6. Furthermore, the mixture of nanoparticles and the mixture of all fractions directly stimulated maturation of OCs. Conclusion. This study identifies a novel mechanism where UHMWPE nanoparticles activate DCs. The high proportion of nanoparticles from prosthetic joints would suggest this mechanism is a likely pathway for cytokine production and OCs maturation, all of which involve osteolysis. The nanoparticles as mediators of periprosthetic inflammation should be considered in developing biomaterials for bearing surfaces

Introduction:. Wear debris from articulating joint implants is inevitable. Small debris particles are phagocytosed by macrophages. Larger particles initiate the fusion of many macrophages into multi-nucleated giant cells for particle encasement. Macrophages are recruited into inflamed tissues from the circulating monocyte population. Approximately 10% of white blood cells are monocytes which after release from the bone marrow circulate for 2–3 days, before being recruited into tissues as inflammatory macrophages or undergoing apoptosis. Circulating MRP8/14 (S100A8/A9) is a measure of monocyte recruitment, part of the monocyte-endothelial docking complex, and shed during monocyte transmigration across the endothelium. The higher the S100A8/A9 the more monocytes being recruited giving an indirect measure of debris production. Methods:. 2114 blood samples were collected from arthroplasty patients with hip or knee osteoarthritis (primary, post-traumatic and secondary), 589 before their primary arthroplasty, 1187 patients > 1 year post-arthroplasty, 101 patients before revision for aseptic loosening and 237 patients >1 year post-revision. Plasma S100A8/A9 was measured using BMA Biomedicals Elisa kit, normal levels in health adults are 0.5–3 mg/ml. Joint specific scores, WOMAC knee or Oxford Hip adjusted to percent of maximum, together with SF-12 were completed. Results:. Mean and S.D. plasma S100A8/A9 before a primary 4.89 + 3.12, >1 year post-surgery follow up 4.03 + 2.2, before revision 5.08 + 2.59 and >1 year post revision 3.89 + 1.94. Percent joint specific scores before a primary 40.33 + 15.75, >1 year post-surgery follow up 70.34 + 22.33, before revision 38.75 + 15.12 and >1 year post revision 53.88 + 22.02. SF-12 physical function scores before a primary 26.52 + 7.12, >1 year post-surgery follow up 35.76 + 11.50, before revision 24.71 + 6.47 and >1 year post revision 30.08 + 9.82. The pre-op primary to follow up S100A*/A9 fell p < 0.001 while both joint specific and SF-12 PCS improved p < 0.001. The pre-revision concentration of S100A8/A9 was significantly higher than routine follow up levels p < 0.001 while joint specific and SF-12 scores fell p < 0.001. S100A8/A9 correlates negatively with both scores p < 0. Discussion:. Osteoarthritis demonstrates enhanced monocyte recruitment before primary surgery which is significantly reduced following arthroplasty. There is a detectable and significant increase in S100A8/A9 concentrations indicative of enhanced monocyte recruitment again before revision surgery

Aims

This study aims to enhance understanding of clinical and radiological consequences and involved mechanisms that led to corrosion of the Precice Stryde (Stryde) intramedullary lengthening nail in the post market surveillance era of the device. Between 2018 and 2021 more than 2,000 Stryde nails have been implanted worldwide. However, the outcome of treatment with the Stryde system is insufficiently reported.

The most frequent cause of failure after total hip replacement in all reported arthroplasty registries is peri-prosthetic osteolysis. Osteolysis is an active biological process initiated in response to wear debris. The eventual response to this process is the activation of macrophages and loss of bone.

Activation of macrophages initiates a complex biological cascade resulting in the final common pathway of an increase in osteolytic activity. The biological initiators, mechanisms for and regulation of this process are beginning to be understood. This article explores current concepts in the causes of, and underlying biological mechanism resulting in peri-prosthetic osteolysis, reviewing the current basic science and clinical literature surrounding the topic.